[Association of Polymorphic Genome Variants in the 2q32.1 Locus with the Development of Vasovagal Syncope].

The vasovagal syncope (VVS) is the most common form of syncope. The mechanisms of VVS development are not entirely clear. It is known that there is a genetic predisposition to this disease, but the data on the roles of individual genes are quite contradictory. Recently, a genome-wide association study identified a locus at chromosome 2q32.1 associated with a united group of diseases, that is, syncope and collapse; among the single nucleotide polymorphisms (SNPs) of this locus, the most significant association was observed for rs12465214. In a homogeneous sample of patients diagnosed with VVS, we analyzed the association of rs12465214, rs12621296, rs17582219 and rs1344706 located on chromosome 2q32.1 with this form of syncope. In the enrolled set, only rs12621296 was associated with VVS by itself, whereas associations of other SNPs were observed only in biallelic combinations. An epistatic interaction between the components of the combination rs12621296*A + rs17582219*A was revealed. The possible involvement of individual genes on the 2q32.1 locus in the genetic architecture of the VVS is discussed.

BMP-2 and IL-1ß as Markers of Nasal Mucosa Inflammation in Rhinosinusitis with Nasal Polyps.

Rhinosinusitis with nasal polyps is characterized by chronic inflammation and hyperplasia of the nasal mucosa. The key mechanism for polyp formation is the expression of molecules that regulate proliferation and inflammation. We studied immunolocalization of bone morphogenetic protein-2 (BMP-2) and IL-1ß in the nasal mucosa in patients aged 35-70 years (n=70, mean age 57.4±1.52 years). The typology of polyps was determined depending on the distribution of inflammatory cells, subepithelial edema, the presence of fibrosis and cysts. The immunolocalization of BMP-2 and IL-1ß had the same pattern in edematous, fibrous, and eosinophilic (allergic) polyps. Goblet and connective tissue cells, microvessels and terminal sections of the glands were positively stained. BMP-2+ and IL-1ß+ cells predominated in polyps of the eosinophilic type. BMP-2/IL-1ß can be considered as a specific marker of inflammatory remodeling of the nasal mucosa in refractory rhinosinusitis with nasal polyps.

Evaluation of the Effectiveness of Bone Grafting in Alveolar Ridge Augmentation Using the Two-Stage Splitting Technique.

Stimulation of neoosteogenesis is the main mechanism of osseointegration during installation of dental implants, bone tissue recession, and alveolar process augmentation in adentia. In experiments on miniature pigs, we used the technology of two-stage splitting of the ridge of the alveolar process of the mandible in combination with a xenograft that was placed between the fragments of the split bone plate. The morphology of the reparative process and the distribution of osteogenic differentiation markers in the compact and trabecular bone of the alveolar crest were studied. Signs of reparative osteogenesis were observed in the bone regenerate that had a lamellar structure, formed osteons, and foci of woven tissue. It was found that the xenograft was replaced by newly formed trabecular bone tissue. These sites were characterized by increased expression of osteocalcin and CD44. Augmentation technology through two-stage splitting provides trophic relationship of osteoprogenitor cells and is an effective method of osteogenesis stimulation in the alveolar process.

[MiRNA miR-375 as a Multifunctional Regulator of the Cardiovascular System].

MicroRNA (miRNA) miR-375 acts as a multifunctional regulator of the activity of many physiological and pathological cellular processes by interacting with a large number of target genes. MiR-375 is involved in the regulation of the differentiation and functioning of cells of the nervous and immune systems, bone and adipose tissue, and even the life cycle of a number of viruses. Changes in miR-375 expression have been found in carcinogenesis, inflammation, and autoimmune and cardiovascular diseases. Every year, new studies appear that expand our understanding of the range of processes regulated by this miRNA. According to the latest data, miR-375 can be used as a biomarker and therapeutic target in some diseases. This review discusses the role of miR-375 in the functioning of the cardiovascular system in health and disease.

Localization of VEGF, TGF-ß1, BMP-2, and Apoptosis Factors in Hypertrophic Nonunion of Human Tubular Bones.

We studied localization of VEGF, TGF-ß1, BMP-2, caspase-3, Bcl-2, and TNFα in the callus samples obtained from 5 patients (4 women and 1 man) aged 41-53 years during planned surgery for nonunion and pseudarthrosis of the clavicle (n=1), ulna (n=1), femur (n=1), and tibia (n=2) bones. Two control groups included material of hypertrophied callus (n=3) with consolidated fractures of long bones and samples of intact bones (n=3) obtained by postmortem autopsy of subjects without pathology of the musculoskeletal system. A nonuniform distribution of the studied markers was revealed. Active expression of VEGF was observed in fibroblast-like cells of the fibrous tissue, osteoblasts of the periosteum and osteons. Osteoblasts expressing BMP-2 were localized in the periosteum and the loose connective tissue of the Haversian canals. The number of immunopositive cells expressing TGF-ß1 and TNFα in the callus exceeded that in the control and correlated with the expression of caspase-3 in fibroblast-like cells, osteoblasts, chondroblasts, and microvascular endotheliocytes. The results allow considering fracture nonunion as a result of overproduction of cytotoxic and proapoptotic factors in chronic inflammation and dysfunction of the expression of morphogenetic proteins. The morphochemical patterns of the studied markers open up prospects for the development of new methods of pharmacological correction of fracture repair.

On-Bone Fixation of Free Gingival Graft Induces an Osteoinductive Effect in Human Alveolar Bone.

We examined alveolar bone samples in the area of on-bone fixation of a free gingival graft performed during surgery in patients aged 37-55 years with a diagnosis of secondary partial adentia of the upper and lower jaws. Six months after fixation of the graft in the alveolar bone, foci of neoosteogenesis were found in the contact zone. They were characterized by the appearance of appositional lines, cords of basophilic osteoblasts, and growing osteons. An immunohistochemical study revealed an increase in the number of CD44+, CD29+, and osteocalcin+ cells in the layer of the outer circumferential lamellae, primary osteons, and the lining of the Haversian canals. TGF-ß1+ cells were located in the intertrabecular reticular tissue and wall of microvessels. The results indicate activation of mesenchymal stem cells in the area of localization of the graft and differentiating osteoblasts. The observed osteoinductive effect of free gingival graft is associated with its participation in reorganization in MSC and induction of morphogenetic molecules.

Dynamics of Renewal of Cell Populations of the Bone Tissue on the Surface of Titanium Implants with Bioactive Coating during Fracture Modeling in Rats.

Localization of PCNA, CD44, osteocalcin, Mdm2, p53, and caspase-3 on the surface of implant with calcium phosphate and hydroxyapatite coating was studied by immunocytochemical method in a model of femur fracture in rats. PCNA+, Ost+, CD44+, and Mdm2+ cells were found in the periosteum, in the layer of the outer surrounding plates, and in the connective tissue of the Haversian canals. Cell density increased on day 7 after fracture and then decreased by day 30. The number of p53+ and CASP3+ cells reached a maximum on day 14 (they were predominantly located in the periosteum and bone plates adjacent to it) and decreased by day 30. Calcium phosphate coating stimulated proliferative activity of cells at the early stages of the regeneration phase and apoptotic death at the later stages. Components of coating can be viewed as a positioning clue for differentiation of mesenchymal stromal cells. The effectiveness of reparative osteogenesis is determined by the balance of proliferative and destructive factors at the site of the fracture healing. This process can be optimized with various nanostructured materials with osteoinductive properties, in particular bioresorbable calcium phosphate coatings on titanium implants. However, the influence of these components on the state of cambial cells, their differentiation, and positioning in the repair zone is unknown.

Osteogenic and Regenerative Potential of Free Gingival Graft.

We studied immunolocalization of CD29, CD44, osteocalcin, and TGF-ß1 in the bone tissue of the mandible of miniature pigs with extra-bone fixation of a free gingival graft. Three months after surgery, neoosteogenesis foci with high expression of the studied markers were found in the contact area of the free gingival graft with the alveolar bone. The markers were localized in the layer of external circumferential lamellae, on the surface of concentric lamellae of the growing osteons, and in the connective tissue of the Haversian canals. TGF-ß1-immunopositive cells predominated in the connective tissue of the Haversian and Volkmann canals and in the adventitia and inner lining of the vascular wall. The established morphochemical patterns of osteogenous cells indicate significant reparative capabilities of a free gingival graft and allows considering it as an effective osteoinductive factor.

Brain-Derived Neurotrophic Factor (BDNF) As a Regulator of Apoptosis under Conditions of Focal Experimental Stroke.

The immunolocalization of apoptotic factors in rat neocortex was studied on the model of permanent occlusion of the middle cerebral artery with administration of exogenous BDNF. We revealed heterogeneous distribution of pro- and anti-apoptotic factors in the stroke area and in the surrounding penumbra, where caspase-3+ and p53+ cells were found. Their number was maximum on day 3 of ischemia. The number of neurons containing anti-apoptotic factors was relatively decreased. Injection of BDNF changed the distribution of apoptotic factors. In the penumbra area, BDNF enhanced the expression of Mdm2 primarily in the pyramid cells of layers V/VI and Bcl-2 in interneurons of layers II and III. Localization of p53 and caspase-3 varied at different terms of the ischemic period and showed an inverse dependence. Considering the selective neuroprotective effect of BDNF, various mechanisms of the formation of ischemic tolerance in neurons are proposed.

[Expression of pro- and anti-apoptotic molecules in the mucous membrane of the nasal cavity with polypous rhinosinusitis]. / Ekspressiya pro- i antiapoptoticheskikh molekul v slizistoi obolochke polosti nosa pri polipoznom rinosinusite.

OBJECTIVE: To study the expression of proapoptotic (p53, p21) and antiapoptotic (MDM2) factors, as well as the distribution of proliferating PCNA-immunoreactive cells in the nasal mucosa in various types of polyposis rhinosinusitis (PRS). MATERIAL AND METHODS: We studied the immunolocalization of proapoptotic (p53, p21) and antiapoptotic (MDM2) factors, as well as the distribution of proliferating PCNA immunoreactive cells in the mucous membrane of the nasal cavity for various types of polypous rhinosinusitis. RESULTS AND DISCUSSION: Comparing with the control group, increased expression of all factors is detected. The main portion of marked cells is located in the loose connective tissue of the lamina propria of mucous membrane, a relatively small number of cells are detected in the epithelial layer. The edematous, eosinophilic (allergic) type of PRS is characterized by the expression of p53 and PCNA and by the reduction of MDM2 immunoreactive cells, while the expression of p53 and p21 is reduced in fibroinflammatory (neutrophilic) type. CONCLUSION: PRS is accompanied by epithelial metaplasia, by formation of inflammatory cell infiltrates, fibrosis and edema. The results are discussed in connection with the data on the regulatory role of apoptosis in the pathomorphism of chronic productive inflammation.

Generation of four iPSC lines from two siblings with a microdeletion at the CNTN6 gene and intellectual disability.

The human induced pluripotent stem cell (iPSC) lines, ICGi009-A, ICGi009-B, ICGi013-A and ICGi013-B, were generated from skin fibroblasts of two siblings with intellectual disability. Both patients were carriers of CNTN6 gene microdeletion (Kashevarova et al., 2014). iPSC lines have normal karyotype, express pluripotency markers, are able to differentiate in vitro into derivatives of all three germ layers and represent a unique tool to study neurodevelopmental disorders.

[The Role of microRNA in the Development of Ischemic Heart Disease].

Coronary artery disease is the most clinically significant manifestation of atherosclerosis and the main cause of morbidity and mortality around the world. Atherogenesis is a complex process, involving various types of cells and regulatory molecules. MicroRNA molecules were discovered at the end of the 20th century, and nowadays are the important regulators of several pathophysiological processes of atherogenesis. The review examines data on the participation of various microRNAs in the development of atherosclerosis and its main clinical manifestations and discusses the possibility of using microRNAs as diagnostic markers for these diseases.

[The Myocardial Infarction Associated Variant in the MIR196A2 Gene and Presumable Signaling Pathways to Involve miR-196a2 in the Pathological Phenotype].

The heritable component of susceptibility to myocardial infraction (MI) remains unexplained, possibly due to the minor effects of genes, which are not obviously associated with the disease. These genes may be integrated in miRNA regulated networks associated with myocardial infarction. A systematic review of the literature led us to selecting rs2910164 (MIR146A), rs11614913 (MIR196A2), and rs3746444 (MIR499А) variants to study the association with the MI phenotype. In ethnic Russians, variant rs11614913*C (MIR196A2) was found to be associated with the risk of myocardial infraction (p = 0.023, OR = 1.74) for the first time; this association was validated in an independent cohort. The gene-gene interaction network for experimentally validated miR-196a2 target genes was built and analyzed. One of its four topological clusters contained the majority of miR-196a2 target genes associated with atherosclerosis, coronary artery disease or myocardial infarction and was enriched with the genes regulating the TGFß and class I MHC signaling pathways, platelet activation/aggregation, and the cell cycle control. This analysis points towards the role of miR-196a2 in the pathological coronary phenotypes and opens up an avenue for further investigations.

Role of Vascular Endothelial Growth Factor and Transforming Growth Factor-ß2 in Rat Bone Tissue after Bone Fracture and Placement of Titanium Implants with Bioactive Bioresorbable Coatings.

The study established enhanced expression of vascular endothelial growth factor (VEGF) in the subpopulation of osteoblasts located in the regeneration region of femoral bone fracture near the titanium implants with bioactive calcium phosphate and hydroxyapatite coatings and suppressed activity of transforming growth factor-ß2 (TGF-ß2) in chondroblasts during the two weeks after surgery. In the delayed posttraumatic period, the distribution of TGF-ß2 inversely related to its maximal activity. The data revealed the up-regulating effect of bioresorbable coatings on expression of VEGF and TGF-ß2 and their implication in the control over various stages of reparative osteogenesis.

Multilocus Analysis of Genetic Susceptibility to Myocardial Infarction in Russians: Replication Study.

In search of genetic markers of myocardial infarction (MI) risk, which have prognostic significance for Russians, we performed a replication study of MI association with genetic variants of PCSK9 (rs562556), APOE (epsilon polymorphism, rs7412 and rs429358), LPL (rs320), MTHFR (rs1801133), eNOS (rs2070744), and the 9p21 region (rs1333049) in 405 patients with MI and 198 controls. Significant MI association was observed with variants of the lipid metabolism genes (PCSK9, APOE and LPL), and of eNOS. The SNPs in the MTHFR gene and the 9p21 region were not significantly associated with MI one by one but were included in several different MI-associated allelic combinations identified by multilocus analysis. Since we have not revealed nonlinear epistatic interactions between the components of the identified combinations, we postulate that the cumulative effect of genes that form a combination arises from the summation of their small independent contributions. The prognostic significance of the additive composite model built from the PCSK9, APOE, LPL, and eNOS genes as genetic markers was assessed using ROC analysis. After we included these markers in the previously published composite model of individual genetic risk of MI, the prognostic efficacy in our sample reached AUC = 0.676. However, the results obtained in this study certainly need to be replicated in an independent sample of Russians.

[Multilocus analysis of the association of polymorphic variants of inflammation genes with ischemic stroke in Russians].

Carriage frequencies of alleles and genotypes of polymorphic loci of inflammation genes (49A>G CTLA4, 41G>A and 87C>T PDE4D, -590C>T IL4, -308A>G TNF, 252G>A LTA, 874A>T IFNG, -509С>Т, 869T>C and 915G>C TGFB1) were determined in a sample of 200 patients diagnosed with ischemic stroke and in the control group similar in gender and age (146 individuals), all ethnic Russians. The positive association of the allele PDE4D*87C (р = 0.028) and genotype TGFB1*-509Т/Т (р = 0.02) carriage with ischemic stroke was shown. The association of the disease with the carriage of the allele PDE4D*41А (р = 0.009) in individuals under the age of 60 and with carriage of the allele IFNG*874Т (р = 0.02) in individuals older than 60 was observed among the subgroups of patients stratified by age when they suffered the stroke compared to a control group of the same age. In subgroups stratified by gender, carriage of the genotype TGFB1*915G/G (р = 0.0015) was identified as a risk factor in male patients, while no significant differences between female patients and healthy women were observed. Multilocus analysis was undertaken to search for the association of several combinations of studied gene variants with ischemic stroke. The polymorphic locus-174G>C of the IL6 gene, for which an association with the disease was previously demonstrated, was also included in this analysis. The disease-predisposing biallelic combinations include the IL6*-174G, PDE4D*87C, TGFB1*-509Т and TGFB1*915G alleles. In the subgroups stratified by gender, the allelic combinations mainly include the similar risk alleles as in the total group, while between the subgroups stratified by age (patients who suffered the first stroke at the age of 18 and no older than 60 years and older than 60 years), greater differences were observed. However, a new risk allele, LTA*252G, was identified in combination with PDE4D*41А in women. These findings demonstrate the important role of inflammation in ischemic stroke. The identified single and combined markers may be used further to determine an individual risk for ischemic stroke.

Morphologic characteristics of sacra associated with assimilation of the last lumbar vertebra.

BACKGROUND: The impact of lumbosacral transitional states on biomechanics of load transmission between the spine and the legs has been sporadically reported. The aims of the study were to identify morphostructural alterations of sacra associated with assimilation of the last lumbar vertebra and to analyse them in the light of their biomechanical impact. MATERIALS AND METHODS: Linear dimensions of sacrum, its body and base and articular surfaces were measured in 31 normal and 41 transitory sacra. Nineteen sacra presented articular and 22 osseous fusion of the last lumbar vertebra. Measured parameters were compared between normal sacra and the two variations of transitory sacra. RESULTS: Sacra with articular fusion of the last lumbar vertebra showed more pronounced concavity of the sacral curvature and wider than long sacral bodies. The first sacral segment was modified, broaden, ventrally wider and elevated. Almost the whole segment bore at its sides auricular surfaces. Very small portion of the segment was non-articular with less pronounced wedging. Sacra with osseous fusion of the last lumbar vertebra showed similar concavity of the sacral curvature as normal sacra, but longer than wide sacral bodies. The ventral sloping half of the newly formed first segment bore auricular surfaces. The non-articular part was enlarged with pronounced wedging. CONCLUSIONS: The term "sacralisation" includes both types of transitory sacra with mutually different morphostructural characteristics in contrast to the normal sacra. Analysis of these morphologic variations may help in understanding the different biomechanical properties and patterns of load transmission.

Anatomy of coronary sinus ostium.

BACKGROUND: The coronary sinus is the main cardiac vein and it has become a clinically important structure especially through its role in providing access for different cardiac procedures. MATERIALS AND METHODS: The study was carried out on 100 randomly selected adult human cadaver hearts fixed in 10% formalin. The transverse and craniocaudal diameters of the coronary sinus ostium (CSO) were directly measured. The presence of the Thebesian valve was noted and the anatomical details of the valve were documented in each case in terms of the shape and extent of coverage of the CSO. RESULTS: Considerable variations in the diameter of the CSO were observed. The mean craniocaudal diameter of the CSO was 8.1 ± 1.51 mm, and the mean transverse diameter was 7.67 ± 1.72 mm. Heart specimens without Thebesian valve tended to have larger ostia. The mean craniocaudal diameter and the mean transverse diameter of the CSO were statistically larger in the specimens without Thebesian valves (p = 0.000 and p = 0.001, respectively). CONCLUSIONS: The Thebesian valves were observed in 86 hearts, and a wide variety of their morphology was seen. The majority of the Thebesian valves were semilunar in shape (74.42%). The extent to which the valve covered the ostium was variable, including remnant valves that covered < 15% of the CSO (35%), and valves that were large and covered at least 75% of the CSO (22.09%). In 3 specimens the valve completely occluded the ostium.

[Combined Effect of Genetic Factors, Age, and Smoking on the Risk of Developing Myocardial Infarction].

OBJECTIVE: to elaborate a complex model for myocardial infarction (MI) risk assessment considering the combined effect of genetic predisposition, age and smoking. MATERIALS AND METHODS: The study included two independent samples of ethnic Russians: 325 patients with MI and 185 individuals without history of cardiovascular diseases (controls) from the Moscow region, and 220 patients and 197 controls from the Republic of Bashkortostan. Genotyping of polymorphic loci of genes CRP (rs1130864), IFNG (rs2430561), TGFB1 (rs1982073), FGB (rs1800788) and PTGS1 (rs3842787) was performed. To construct the predictive models, we used logistic regression with stepwise inclusion of variables. The predictive value was evaluated by the area under the curve (AUC) in a ROC-analysis. The factor was considered as a marker at pAUC <0.05 calculated by the method of DeLong. The marker was considered effective at AUC >0.60. RESULTS: Three separate genetic variants FGB rs1800788*T, TGFB1 rs1982073*TT, CRP rs1130864*TT, and biallelic combination IFNG rs2430561*A + PTGS1 rs3842787*T whose association with MI we described earlier, were used to construct the composite genetic marker (AUC=0.66 in the training and test samples) by the logistic regression method. Adding to the obtained composite genetic marker such parameters as age and smoking allowed to create a complex MI risk marker, which was characterized by the predictive value stability (AUC=0.77 in the training sample and 0.82 in the test sample). CONCLUSION: The obtained complex model for MI risk assessment was reproduced in two independent samples of Russian ethnicity individuals from different regions of Russia with different gender identities, and allowed to have a reasonable chance (about 80%) of distinguishing patients and healthy individuals.

[Ischemic stroke as a complex polygenic disease].

In the modern world acute cerebrovascular accidents (strokes) are one of the most important sociomedical problems due to their high share in the structure of morbidity, invalidization and mortality of the population. The main part of strokes is ischemic stroke (IS). IS is a complex (multifactorial) polygenic disease, i.e. develops as a result of interactions between risk factors and genetic components that determine a joint contribution of the sets of independently acting or interacting polymorphic genes. Currently, the search for a genetic predisposition to IS uses two main methods that are based on the analysis of the association between the polymorphic regions of the genome and disease: a candidate gene approach and genome-wide association studies, followed by a meta-analysis of the received results. In this work we reviewed the literature on genetic susceptibility to IS. It showed progress in this direction, which can be the starting point for the study of the molecular mechanisms determining the pathophysiology of IS. However to a full solution of the problem of identification of the genetic risk factors applicable for the individual prognosis of predisposition to IS, still very far, mainly due to the low replication of results. The way to solve this problem lies through the study of ethnically homogeneous populations and clinically different forms of IS.