Generation of an induced pluripotent stem cell line, ICGi029-A, by reprogramming peripheral blood mononuclear cells of a patient suffering from hypertrophic cardiomyopathy and carrying a heterozygous p.N515del mutation in MYBPC3.

Hypertrophic cardiomyopathy (HCM) is a common cardiovascular disease. However, effective methods of its therapy have not been developed so far. To date patient-specific induced pluripotent stem cell-derived cardiomyocytes are supposed to be a useful tool for studying HCM molecular mechanisms and to help find new approaches to HCM therapy. Using non-integrating episomal vectors, we generated an iPSC line from peripheral blood mononuclear cells of an HCM patient carrying a heterozygous p.N515del mutation in MYBPC3. The iPSC line expressed pluripotency markers, gave rise to derivatives of three germ layers during spontaneous differentiation, had normal karyotype, and retained the patient-specific mutation.

Generation of induced pluripotent stem cell lines ICGi021-A and ICGi022-A from peripheral blood mononuclear cells of two healthy individuals from Siberian population.

ICGi021-A and ICGi022-A iPSC lines were obtained by reprogramming PBMCs of two healthy women of the Siberian population using episomal non-integrating vectors expressing Yamanaka factors. iPSC lines expressed pluripotency markers, had a normal karyotype and demonstrated the ability to differentiate into derivatives of the three germ layers. Clinical exome sequencing data of the original biosamples of the donors are available in the NCBI SRA database. The generated cell lines are useful as "healthy" control in biomedical studies.

Generation of induced pluripotent stem cell line ICGi018-A from peripheral blood mononuclear cells of a patient with Huntington's disease.

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by CAG repeat expansion in the HTT gene. HD patient-specific induced pluripotent stem cells (iPSCs) represent an excellent model for the disease study. We generated iPSC line from blood mononuclear cells of HD patient with 38 CAG repeats in the HTT exon 1 using integration free episomal plasmids expressing Yamanaka factors. The iPSC line retained the disease causing mutation and expressed pluripotency markers. It also displayed a normal karyotype and the ability to differentiate into derivatives of three germ layers.

[The application of the acupuncture techniques for the combined treatment of neurotic tics in the children]. / Ispol'zovanie metodov refleksoterapii v kompleksnom lechenii nevroticheskikh tikov u detei.

At present, the frequency of neurosis in the children is growing under the influence of the increasingly intensive stressful factors. The neurotic ticks are one of the manifestations of neurosis in the children characterized by involuntary movements; they are often accompanied by hyperactivity, deficit of attention, fatigue, and headaches. AIM: The objective of the present study was to evaluate the effectiveness of acupuncture used for the treatment of neurotic tics in the children. MATERIAL AND METHODS: The study was carried out at the basis of the department of children's rehabilitation of the Moscow city polyclinic #39. All the children were counselled by a neurologist, psychologist, and acupuncturist. An electroencephalogram was obtained from each patient and a questionnaire study involving the patients' parents was conducted. The patients were treated with the application of reflexotherapy and were given recommendations of a psychologist. A total of 30 children at the age from 6 to 12 years were available for the examination. The participants of the study were allocated to two groups depending on the strategy of the outpatient care. Group 1 was comprised of the patients (n=17) who were given 3 courses of reflexotherapy in addition to pharmacotherapy and psychologist's recommendations. The patients included in group 2 served as controls (n=13) and received only medications and psychotherapy without acupuncture. The effectiveness of the treatment was evaluated based on the dynamics of the patients' complaints. RESULTS AND DISCUSSION: The resullts of the study give evidence of the higher effectiveness of the treatment of the children suffering from neurotic ticks with the application of acupuncture therapy compared with the conventional approach. A few courses of reflexotherapy provided in the combination with pharmacotherapy and the compliance with the recommendations of a psychologist resulted in the more efficient elimination of the neurotic tics together with the associated hyperactivity, deficit of attention, fatigue, and headaches than the use of medication therapy and psychotherapy without acupuncture.

[Long-term survival in working-age patients after cerebrovascular accident]. / Otdalennaya vyzhivaemost' patsientov trudosposobnogo vozrasta posle tserebral'nogo insul'ta.

AIM: To assess the long-term survival in working-age patients after cerebrovascular accident (CVA) and to define basic medical and social factors determining the survival rates. MATERIAL AND METHODS: A continuous retrospective study included 756 working-age patients (<60 years) after CVA. The life-table method and Kaplan-Meier analysis were used to calculate a survival rate. RESULTS: The highest risk of death was observed in the first year of CVA. One-year survival after CVA was 81.7%, falling to 74.7% at 3 years and to 57.9% at 7 years. The five-year survival was lower among men (60%) compared to women (74.5%). The figures were higher in working population compared to non-working population (78% vs. 56%, respectively). Long-term survival rate depends on the type of CVA: the five-year survival rate after unspecified stroke was 83.0%, after ischemic stroke - 69.0%, after hemorrhagic stroke - 43.0%. The direct dependence between long-term survival and patient age has been identified: older patients have worse long-term prognosis. CONCLUSION: The seven-year survival rate in working-age patients after cerebrovascular accident was 58%. The health and social factors affecting the long-term survival rates in working-age patients are as follows: hemorrhagic stroke, unemployment, male gender and advanced age.

A POSSIBLE CYTOGENETIC ANALOGY TO GENOMIC «SPECIATION ISLANDS¼ AS REVEALED BY CHROMOSOME STUDY OF A NATURAL HYBRID VOLE.

Chromosome analysis in mammals over the last half century has largely focused on species identification. A growing number of hybrid zones of karyotypically differentiated cryptic taxa have been described. A good example is provided by two karyoforms of the 46-chromosome common vole (long known as «arvalis¼ and «obscurus¼) that make contact longitudinally from the north to the south of European Russia. The hybrid F1 karyotype displays genomic markers which distinguish arvalis and obscurus and which are cytogenetically detectable as minor variants. The apparent insignificance of the genomic region of heterozygosity associated with these markers perhaps does not prevent hybrid chromosome pair formation at meiosis but may reflect a site resistant to gene flow. A cytogenetic analogy with so-called «speciation islands¼, based, for example, on the study of the corvine hybrid zone and represented by a small number of limited genomic sites (occupying less than 1 % of the genome) (Poelstra et al., 2014), may be relevant.

Therapy of Chronic Cardiosclerosis in WAG Rats Using Cultures of Cardiovascular Cells Enriched with Cardiac Stem Cell.

We developed a protocol for preparing cardiac cell culture from rat heart enriched with regional stem cells based on clonogenic properties and proliferation in culture in a medium with low serum content. Experiments on WAG rats with experimental ischemic myocardial damage showed that implantation of autologous regional stem cells into the left ventricle reduced the volume of cicatricial tissue, promoted angiogenesis in the damaged zone, and prevented the risk of heart failure development.

Characteristics of cardiac cell cultures derived from human myocardial explants.

Primary cell cultures derived from human myocardial explants were obtained and characterized. The explant cultures contained cardiac stem cells (c-kit(+); ≈ 4%), microvascular cells (endothelial cells and pericytes), fibroblasts, and myofibroblasts. It was demonstrated that culturing of cardiac cells in cardiospheres did not promote enrichment of the cell culture with stem cells. MACS-sorted c-kit(+) cells from the explant culture were characterized by limited proliferative capacity and were capable of cardiomyogenic differentiation. The presence of microvascular cells determined general angiogenic potential of the culture.

[Molecular cytogenetic markers of cryptic species and hybrids of the common vole superspecies complex Microtus arvalis s. l].

Molecular markers of cryptic cytogenetical differentiation were shown in chromosomal polymorphic Pan-European model group of rodents Microtus arvalis s. l. by FISH analysis. The polytypy of 46-chromosomes karyotypes determined by the sites of interstitial telomeric sequences (ITS) and ribosomal DNA emphasizes the genetical isolation of M. arvalis s. s. and M. obscurus.

Cytogenetic analysis of a hybrid zone between the Moscow and Neroosa races of the common shrew (Sorex araneus) differing by a single wart-like chromosome rearrangement.

Three karyotypic variants were revealed in the Moscow-Neroosa chromosomal hybrid zone by comparative cytogenetic analysis: homozygotes of the pure parental races and complex heterozygotes (F1 hybrids). As expected, a ring-of-four configuration (RIV) was observed in diakinesis spreads of hybrids. No disturbances in the structure of the RIV were found. Distribution of telomeric repeats and rDNA on the chromosomes of an individual of the Neroosa race were studied using dual-colour FISH.

[Brain oscillations and individual variability of cardiac defense in human].

Cardiac defense response (CDR) is a specific dynamic pattern of the short- and long-latency cardiovascular reactivity (usually based on HR and/or arterial BP variables) in response an intense aversive unexpected stimulus. The CDR reflects activity of the defensive motivational system along with behavioral coping programs. The aim of the study was to estimate the putative contribution of brain oscillations into the central mechanisms of the CDR individual variability. EEG and cardiovascular concomitants of the CDR were estimated using the event-related synchroniza- tion/desynchronization (ERD/ERS) in different frequency bands as well as Finapres dependent variables obtained on the "beat-by-beat" basis. The first-ever findings evidenced significant cor- relations of the theta-2 (6-8 Hz) ERD/ERS with the sort-latency, whereas alpha-2 (10-12 Hz) ERD/ERS with the long-latency CDR variability. The hyperreactive long-latency systolic blood pressure CDR component was accompanied by the phase of deficient alpha-2 synchronization. It is suggested that upper theta oscillations are involved into central mechanisms scaling magnitude, whereas high frequency alpha oscillations may be responsible for the top-down inhibitory control of the short-and long-latency ABP stress-reactivity in the CDR.

[Racial and population variability of phenotypic (cranial) characters in the common shrew Sorex araneus L., 1758].

Variability of the cranial properties of chromosomal races Serov, Manturovo and Pechora of the common shrew were studied. A consistent increase of scull size in the Serov race with moving from the plain to highlands and a skull size decrease from low to high latitudes was detected. Interpopulation variability among different races was shown to be comparable with interracial variability or to exceed it. This suggests microevolution at the level of local populations.

Natural hybridization between extremely divergent chromosomal races of the common shrew (Sorex araneus, Soricidae, Soricomorpha): hybrid zone in European Russia.

The Moscow and Seliger chromosomal races of the common shrew differ by Robertsonian fusions and possibly whole-arm reciprocal translocations (WARTs) such that their F1 hybrids produce a chain-of-eleven configuration at meiosis I and are expected to suffer substantial infertility. Of numerous hybrid zones that have been described in the common shrew, those between the Moscow and Seliger races involve the greatest chromosomal difference. We collected 211 individuals from this zone to generate a total dataset of 298 individuals from 187 unique global positioning system (GPS) locations within the vicinity of interracial contact. We used a geographic information system (GIS) to map the location of the hybrid zone, which follows a direct route between two lakes, as would be anticipated from tension zone theory. Even within the central area of the hybrid zone, there is a much higher frequency of pure race individuals than hybrid, making this a clear example of a bimodal zone in the sense of Jiggins & Mallet (2000). The zone runs through good habitat for common shrews, but nevertheless it is very narrow (standard cline widths: 3-4 km), as would be anticipated from low hybrid fitness. There is clear potential for an interruption to gene flow and build-up of reproductive isolation. As found in some other hybrid zones, there is a high frequency of novel genetic variants, in this case, new chromosomal rearrangements. Here, we report a de novo Robertsonian fission and a de novo reciprocal translocation, both for the first time in the common shrew. There is an extraordinarily high frequency of de novo mutations recorded in F1 hybrids in the zone and we discuss how chromosomal instability may be associated with such hybrids. The occurrence of a de novo Robertsonian fission is of considerable significance because it provides missing evidence that fissions are the basis of the novel acrocentric forms found and apparently selected for in certain common shrew hybrid zones.

Presence of the 54-chromosome common vole (Mammalia) on Olkhon Island (Lake Baikal, East Siberia, Russia), and the occurrence of an unusual X-chromosome variant.

We report a new finding of the 54-chromosome sibling species of the common vole in East Siberia - the first description from Olkhon Island (Lake Baikal). The karyotype of a male specimen revealed by routine staining and C-banding demonstrates the unambiguous presence of Microtus rossiaemeridionalis Ognev, 1924 (recently often regarded as as junior synonym of Microtus levis Miller, 1908). Comparison with conspecific specimens from the European part of the species range (from the left bank of the river Volga) shows that the vole of the island population has a smaller X-chromosome due to a reduced quantity of C-positive heterochromatin. This is just the third example of this type of X-chromosome variant with previous cases on an Arctic island (Svalbard) and the West Siberian lowland (Novosibirsk) and the only one on a lake island. Although Microtus rossiaemeridionalis is largely monomorphic in its karyotype, our data show that one specific type of X-chromosome variant is remarkably widespread, though rare.

[Identification of a novel WART-like chromosome rearrangement in complex heterozygotes in an interracial hybrid zone of the common shrew Sorex araneus L].

Karyotypes uncharacteristic of pure races or hybrids were identified in the interracial hybrid zones of the common shrew Sorex araneus L. that were recently discovered in the European part of Russia. This suggests the actual existence in natural populations of WART-like rearrangements (whole-arm reciprocal translocations) along with Robertsonian fusions of acrocentrics. Demonstration of new and still rare chromosome variants is the aim of this communication.

[Meiotic inactivation of sex chromosomes in mammals].

During meiosis, heteromorphic mammalian X and Y chromosomes in males undergo transcription silencing and form a compact structure, the XY body, containing specific modifications of the chromatin. In this review, we consider the dynamics of sex chromosome inactivation and discuss the suggestion that the paternally inherited X chromosome preserve inactivated state in zygote. This state results from meiotic silencing and is prone to imprinted inactivation, which his observed in mammalian females at early embryogenetic stages.

[Cytogenetic study of the parapatric contact zone between two 46-chromosomal forms of the common vole in European Russia].

Since the first description of sympatric sibling species and allopatric chromosomal forms of the common vole (Microtus arvalis group) in 1969, the search of their range boundaries has been Continually going on up to the present time. Based on the cytogenetic material presented in the present study, the geographical distribution of two karyoforms (2n = 46: NF = 84 and NF = 72) was examined in 20 geographic localities of European Russia (within the interval between the longitude 34 degrees-56 degrees East and 51 degrees-61 degrees). The places of findings of two karyoforms, "arvalis" and "obscurus", in Russia have been supplemented and were shown to be significantly closer; whereas the well-known distribution boundaries for the "obscurus" form were specified in the northern and southern parts of Russia.A direct evidence of parapatric distribution and hybrid zone formation between the "arvalis" and "obscurus" karyoforms was obtained, which suggests the possibility of studying the evolutionary relations ships between two genomes in the Russian part of the group range under the natural experimental conditions-the hybrid zone at the boundary between the "arvalis" and "obscurus".

[Distribution of two chromosome races of the common shrew (Sorex araneus L.) in the hybrid zone: may a change of the dispersal mode maintain independent gene frequencies?].

Combination of different dispersal modes may itself, without external obstacles, lead to the appearance of subdivided populations and maintain the existence of independent population systems. The common shrew, a mammal convenient for studying different levels of intraspecific differentiation, was the object of the study. Empirical data have been used for simulation taking into account the change of dispersal modes in the population area. The obtained results agree with empirical data on the distribution of races and hybrids in the hybrid zone of chromosome races Moscow and Seliger. Change of the dispersal mode may maintain independent population dynamics and, in the case of chromosome races, prevent the migration of parental individuals into the territory of the other race.

[The structure and evolution of the MaSMC4 gene of common vole Microtus arvalis (Arvicolidae, Rodentia)].

In eukaryotes, the SMC (Structural Maintenance of Chromosomes) gene family is represented by at least six genes. Some of these genes have tissue-specific homologs. Eukaryotic SMC structural proteins are the members of biochemical complexes responsible for cohesion of sister chromatids, recombination, repair, regulation of gene expression, and formation of mitotic chromosomes. In the present study, the structure of the SMC4 sub-family gene was examined in common vole Microtus arvalis. Comparative analysis of rodent (M. arvalis, Mus musculus. and Rattus norvegicus), human, and Xenopus SMC4 orthologous genes was carried out, and the main patterns of their organization and regulation were established. The SMC4 genes contain 24 exons; open reading frame starts at exon 2. The SMC4 5' regions contain the CpG islands, extending in the region of exon-intron I and exon 2. The SMC4 genes are characterized by the presence of multiple transcription startpoints. The region of the major transcription startpoint contains the INR CCA,1TTTT element. The SMC4 5' region is characterized by the presence of putative binding site for basal transcription factor Sp and factor E2F, typical of the genes induced in the G I/S phase of the cell cycle. The divergence level of the SMC4 coding region was examined. The mean Ka/Ks ratio for the SMC4 genes examined was 0. 123. The region of exon 2 was found to be the most variable (Ka/Ks = 0.715), while the most conservative was the region coding for the C-globular domain, which contained the DA box (Ka/Ks = 0.024).