Female and Male Phenotypes of Iron Deficiency in CHF. Additional analysis of the «The Prevalence of Iron Deficiency in Patients With Chronic Heart Failure in the Russian Federation (J-CHF-RF)¼ study.

Aim    To evaluate the incidence of iron deficiency (ID) in men and women with chronic heart failure (CHF) and to compare clinical and functional indexes in patient with and without ID depending on the gender.Material and methods    An additional analysis of the study "Prevalence of Iron Deficiency in Patients With Chronic Heart Failure in the Russian Federation (ID-CHF-RF)" was performed. The study included 498 (198 women, 300 men) patients with CHF, in whom, in addition to iron metabolism, the quality of life and exercise tolerance (ET) were studied. 97 % of patients were enrolled during their stay in a hospital. ID was defined in consistency with the European Society of Cardiology (ESC) Guidelines. Also, and additional analysis was performed according to ID criteria validated by the morphological picture of the bone marrow.Results    ID was detected in 174 (87.9 %) women and 239 (79.8 %) men (p=0.028) according to the ESC criteria, and in 154 (77.8 %) women and 217 (72.3 %) men (p=0.208) according to the criteria validated by the morphological picture of the bone marrow. Men with ID were older and had more severe CHF. They more frequently had HF functional class (FC) III and IV (63.4 % vs. 43.3 % in men without ID); higher concentrations of N-terminal pro-brain natriuretic peptide (NT-proBNP) and lower ET. HF FC III increased the probability of ID presence 3.4 times (p=0.02) and the probability of HF FC IV 13.7 times (p=0.003). This clinical picture was characteristic of men when either method of determining ID was used. In women, ID was not associated with more severe CHF.Conclusion    Based on the presented analysis, it is possible to characterize the male and female ID phenotypes. The male ID phenotype is associated with more severe CHF, low ET, and poor quality of life. In females of the study cohort, ID was not associated with either the severity of CHF or with ET.

[Iron deficiency in Russia heart failure patients. Observational cross-sectional multicenter study].

Aim    To evaluate the prevalence of iron deficiency (ID) in Russian patients with heart failure (HF).Material and methods    Iron metabolism variables were studied in 498 (198 women, 300 men) patients with HF. Data were evaluated at admission for HF (97 %) or during an outpatient visit (3 %). ID was determined according to the European Society of Cardiology Guidelines.Results    83.1 % of patients had ID; only 43.5 % of patients with ID had anemia. Patients with ID were older: 70.0 [63.0;79.0] vs. 66.0 years [57.0;75.2] (p=0.009). The number of patients with ID increased in parallel with the increase in HF functional class (FC). Among patients with ID, fewer people were past or current alcohol users (p=0.002), and a greater number of patients had atrial fibrillation (60.1 vs. 45.2 %, p=0.016). A multiple logistic regression showed that more severe HF (HF FC) was associated with a higher incidence of ID detection, whereas past alcohol use was associated with less pronounced ID. An increase in N-terminal pro-brain natriuretic peptide (NT-proBNP) by 100 pg/ml was associated with an increased likelihood of ID (odds ratio, 1.006, 95 % confidence interval: 1.002-1.011, p=0.0152).Conclusion    The incidence rate of HF patients is high in the Russian Federation (83.1 %). Only 43.5 % of these patients had anemia. The prevalence of ID in the study population increased with increases in HF FC and NT-proBNP.

[Homocysteine and markers of endothelial dysfunction in multiple sclerosis]. / Gomotsistein i markery endotelial'noi disfunktsii pri rasseyannom skleroze.

OBJECTIVE: To identify hyperhomocysteinemia and to assess its possible association with the course and other markers of endothelial damage in multiple sclerosis. MATERIAL AND METHODS: Analysis of blood serum for homocysteine, for the content of adhesion molecules sPECAM-1, matrix metalloproteinase 9, blood plasma test for von Willebrand factor antigen in patients with multiple sclerosis. The values of these indicators were analyzed depending on the course and activity of the demyelinating process, the severity of neurological disorders, as also depending on the therapy received. RESULTS: Hyperhomocysteinemia was found in more than half of patients with multiple sclerosis. A significantly higher homocysteine level was found in male patients, and hyperhomocysteinemia was associated with the activity of the process in patients with highly active multiple sclerosis. CONCLUSION: The results of the study suggest a possible association of hyperhomocysteinemia with high process activity and disease progression, as well as with mechanisms of neurodegeneration. Determination of homocysteine concentration may be one potential marker for predicting the course of the disease.

[Thrombosis of the inferior vena cava and right portions of the heart in a relapsing retroperitoneal malignant tumour]. / Tromboz nizhnei poloi veny i pravykh otdelov serdtsa pri retsidivnoi zabriushinnoi zlokachestvennoi opukholi.

Presented herein is a clinical case report regarding successful treatment of a female patient with a relapsing malignant retroperitoneal tumour complicated by disseminated thrombosis of the inferior vena cava, right atrium and right ventricle of the heart. A relapse of the malignant uterine tumour had developed 3 years after the primary operation and was represented by a large-size mass ingrowing into the infrarenal segment of the inferior vena cava, aorta, as well as jejunum. Additional examination revealed the presence of a tumorous thrombus extending from the primary tumorous node in the lumen of the inferior vena cava to the right atrium and ventricle. The procedures performed consisted in removal of the tumour with resection of the inferior vena cava, aorta, and jejunum, followed by thrombectomy from the right portions of the heart under extracorporeal circulation. The postoperative period turned out uneventful, with no complications observed. The woman was discharged on POD 15. Twelve-month postoperative follow up revealed neither relapse nor progression of the disease. Currently, the patient continues undergoing specific treatment (second-line chemotherapy). Also discussed in the article are current challenges concerning both the classification of tumour thrombosis of the inferior vena cava in retroperitoneal sarcomas, and the choice of optimal strategy and policy of treatment.

Mitochondria with Morphology Characteristic for Alzheimer's Disease Patients Are Found in the Brain of OXYS Rats.

Growing evidence suggests that mitochondrial dysfunction is closely linked to the pathogenesis of sporadic Alzheimer's disease (AD). One of the key contributors to various aspects of AD pathogenesis, along with metabolic dysfunction, is mitochondrial dynamics, involving balance between fusion and fission, which regulates mitochondrial number and morphology in response to changes in cellular energy demand. Recently, Zhang et al. ((2016) Sci. Rep., 6, 18725) described a previously unknown mitochondrial phenotype manifesting as elongated chain-linked mitochondria termed "mitochondria-on-a-string" (MOAS) in brain tissue from AD patients and mouse models of AD. The authors associated this phenotype with fission arrest, but implications of MOAS formation in AD pathogenesis remain to be understood. Here we analyze the presence and number of MOAS in the brain of OXYS rats simulating key signs of sporadic AD. Using electron microscopy, we found MOAS in OXYS prefrontal cortex neuropil in all stages of AD-like pathology, including manifestation (5-month-old rats) and progression (12-18-month-old rats). The most pronounced elevation of MOAS content (8-fold) in OXYS rats compared to Wistar controls was found at the preclinical stage (20 days) on the background of decreased numbers of non-MOAS elongated mitochondria. From the age of 20 days through 18 months, the percentage of MOAS-containing neuronal processes increased from 1.7 to 8.3% in Wistar and from 13.9 to 16% in OXYS rats. Our results support the importance of the disruption of mitochondrial dynamics in AD pathogenesis and corroborate the existence of a causal link between impaired mitochondrial dynamics and formation of the distinctive phenotype of "mitochondria-on-a-sting".

[Apoptosis as a systemic adaptive mechanism in ischemic stroke]. / Apoptoz kak sistemnyi adaptivnyi mekhanizm pri ishemicheskom insul'te.

This paper presents a literature review considering the role and mechanism of apoptosis in the pathogenesis of ischemic stroke (IS). The authors introduce a new concept: the functional request of the patient as a set of external (the nature and intensity of rehabilitation measures, characteristics of everyday life, diet, etc.) and internal (genetic factors, internal picture of the disease, availability of rental and other psychological facilities and etc.) attributes. This concept allows a new angle in understanding the pathogenesis of IS and creates fundamental and clinical potential for more successful approaches to therapy and rehabilitation after IS.

A New Concept of Action of Hemostatic Proteases on Inflammation, Neurotoxicity, and Tissue Regeneration.

Key hemostatic serine proteases such as thrombin and activated protein C (APC) are signaling molecules controlling blood coagulation and inflammation, tissue regeneration, neurodegeneration, and some other processes. By interacting with protease-activated receptors (PARs), these enzymes cleave a receptor exodomain and liberate new amino acid sequence known as a tethered ligand, which then activates the initial receptor and induces multiple signaling pathways and cell responses. Among four PAR family members, APC and thrombin mainly act via PAR1, and they trigger divergent effects. APC is an anticoagulant with antiinflammatory and cytoprotective activity, whereas thrombin is a protease with procoagulant and proinflammatory effects. Hallmark features of APC-induced effects result from acting via different pathways: limited proteolysis of PAR1 localized in membrane caveolae with coreceptor (endothelial protein C receptor) as well as its targeted proteolytic action at a receptor exodomain site differing from the canonical thrombin cleavage site. Hence, a new noncanonical tethered PAR1 agonist peptide (PAR1-AP) is formed, whose effects are poorly investigated in inflammation, tissue regeneration, and neurotoxicity. In this review, a concept about a role of biased agonism in effects exerted by APC and PAR1-AP via PAR1 on cells involved in inflammation and related processes is developed. New evidence showing a role for a biased agonism in activating PAR1 both by APC and PAR1-AP as well as induction of antiinflammatory and cytoprotective cellular responses in experimental inflammation, wound healing, and excitotoxicity is presented. It seems that synthetic PAR1 peptide-agonists may compete with APC in controlling some inflammatory and neurodegenerative diseases.

[Investigation of the surface layer of 3D-matrices for tissue engineering]. / Issledovanie poverkhnostnogo sloia 3D-matriksov dlia tkanevoi inzhenerii.

Electrospinning is a convenient and promising manufacturing method a variety of materials for tissue engineering. 3D matrices fabricated by electrospinning from solutions of polycaprolactone with human serum albumin or gelatin in 1,1,1,3,3,3-hexafluoroisopropanol were studied. The microstructure of the 3D matrices and surface of the fibers were investigated using scanning electron microscopy. Protein distribution in the surface layer was studied by modification of protein amino groups with N-(2-hydroxyethyl)phenazine and X-ray photoelectron spectroscopy. It was shown, that concentration of the proteins in the surface layer of fibers exceeded their concentration in the initial electrospun solution up to 12 times and the surface layer was enriched in the protein inversely to the concentration of the protein in solution. The minor part of the proteins was released from fibers during first 30-60 min after swelling in water. Treatment of matrices with proteinase K hydrolyzed about 1/3 of the surface exposed human serum albumin. Thus, both methods can be used to study the surface content of the materials produced by electrospinning from blends of synthetic and natural polymers, however X-ray photoelectron spectroscopy appears to be more convenient and informative.

THE FUNCTIONAL ROLE OF SMALL HEAT SHOCK PROTEIN Hsp67Bc IN DROSOPHILA MELANOGASTER.

Heat shock proteins are universal agents protecting all known organisms from environmental stresses. These proteins are classified into six families differing in their structure and function, yet having a common purpose of maintaining cellular proteins in operating condition. The small heat shock protein family has the most diversified set of effects on the organism's vital activity with a great number of its members' features studied thoroughly in a fruit fly Drosophila melanogaster, which is regarded as a convenient model object. In this review, we represent and discuss data on the role of Drosophila melanogaster Hsp67Bc protein, which was earlier identified as the closest functional ortholog of human HSPB8 small heat shock protein.

[NESPRINS--nuclear envelope proteins ensuring integrity].

This review describes the nesprins (nuclear envelope spectrin-repeat proteins), which are recently discovered family of nuclear envelope proteins. These proteins play an important role in maintaining the cellular architecture and establish the link between the nucleus and other sub-cellular compartments. Many tissue-specific diseases including lipodystrophies, hearing loss, cardiac and skeletal myopathies are associated with nesprins mutations. These proteins comprise of multiple tissue specific isoforms which contain spectrin repeats providing interaction of nesprins with other nuclear membrane proteins, cytoskeleton and intranuclear matrix. We summarize recent findings and suggestions about nesprins structural organization and function inside the cell. Human diseases caused by abnormal nesprins expression are also described.

[Peptide-agonist of protease-activated receptor (PAR 1), similar to activated protein C, promotes proliferation in keratinocytes and wound healing of epithelial layer].

Activated protein C (APC) is serine protease hemostasis, independent of its anticoagulant activity, exhibits anti-inflammatory and anti-apoptotic properties that determine the possibility of the protective effects of APC in different diseases, including sepsis and chronic wound healing. APC, binding of endothelial protein C receptor (EPCR) and specifically cleaving PAR1 receptor and releasing peptide agonist PAR1 stabilizes not only endothelial cells, but also many others, including epidermal keratinocytes of the skin. We develop the hypothesis that the cytoprotective effect of APC on the cells, involved in wound healing, seem to imitate peptide - analogous of PAR1 "tethered ligand" that activate PAR1. In our work, we synthesized a peptide (AP9) - analogue of PAR1 tethered ligand, released by APC, and firstly showed that peptide AP9 (0.1-10 мM), like to APC (0.01-100 nM), stimulates the proliferative activity of human primary keratinocytes. Using a model of the formation of epithelial wounds in vitro we found that peptide AP9, as well as protease APC, accelerates wound healing. Using specific antibodies to the receptor PAR1 and EPCR was studied the receptor mechanism of AP9 action in wound healing compared with the action of APС. The necessity of both receptors - PAR1 and EPСR, for proliferative activity of agonists was revealed. Identified in our work imitation by peptide AP9 - PAR1 ligand, APC acts on keratinocytes suggests the possibility of using a peptide AP9 to stimulate tissue repair.

[Mesenchymal stromal cells synchronize the rhythm of protein synthesis under the effect of an exogenous signal].

A comparative study was performed of dense 5-hour cultures of rat hepatocytes and equal-density cultures of mesenchymal stromal cells (MSC) isolated from human adipose tissue of rat bone marrow. The cells were grown on collagen-coated class slides in serum-free medium. Unlike in hepatocytes, no rhythm of protein synthesis was initially revealed in MSC, but such a rhythm manifested itself when the culture medium was supplemented with melatonin (2 nM, 5 min). The results of experiments with cytoplasmic calcium chelator BAPTA-AM and protein kinase inhibitor H7 indicate that the mechanism of protein synthesis synchronization in MSC consists in calcium-dependent phosphorylation of cell proteins.

[Use of mesenchymal stem cells for reparative processes activation in bone jaw tissue in experimental conditions].

In experiment on 12 Chinchilla rabbits dynamics of reparative regeneration was studied at the terms 2 and 4 months. Bone defect in mandible corner was closed by osteoplastic material Gapkol which was covered from inside by allogenic or autologic stem cells received from rabbit adipose tissue. The results of the ray tracing methods of study were verified by SEM and histological methods.

[Influence of allogenic and autologic multipotent stromal cells of adipose tissue upon bone tissue regeneration in the rabbit mandible angle defect].

Reconstructive surgery often collides with complicated task of defect recovery in the situation of deficit of maxillofacial skeleton tissues, that leads to necessary search of accessible cells source with osteogenic potentials and proper substrate for cell transplantation. In the study the regeneratory potential of stromal cells of adipose tissue was tested in the rabbit model of mandible through angle defect. Cells of allogenic and autologic nature stimulated reparative osteogenesis but their influence upon bone matrix maturation was different. When the construction with allogenic cells was transplanted reaction of its rejection was not detected on histological level.

[Evaluation of reparative regeneration of the jaw bone by microfocus roentgenography in an experiment].

In experiment on 16 grown-up chinchilla rabbits the dynamic of reparative regeneration was evaluated by digital microfocal rontgenography in the terms of 1, 2 and 4 months. Bone defect of the 8capital CHE, Cyrillic8 mm size in the region of mandible angle was caused by surgical laser Smart 2940 D+ on the right side and by physiodespenser Surgec XT on the left side. Surgical laser use let to reduce intact mother bone traumatisation and to improve remote results of bone tissue regeneration. After bone defect creation bone tissue regeneration was put into effect by all 3 callus types - endosteal, periosteal and intermediary.

[Influence of Drosophila melanogaster genotype on biological effects of endocymbiont Wolbachia (stamm wMelPop)].

Comparative analyses of symbiotic bacteria Wolbachia (stamm wMelPop reducing lifespan of flies) morphology in normal and mutant strains of Drosophila melanogaster as well as the influence of Wolbachia on the host cell ultrastructure have been done. Wolbachia infected D. melanogaster mutation strains Trithorax-like -- Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+ have been received by special flies crossing. Uninfected strain D. melanogaster white-1118 (w1118) have been obtained by antibiotic treatment of initially infected strain D. melanogaster [w]w1118. Complex of different methods and approaches let to investigate for the first time the morphology of cell structure before and after bacterial infection of insects and to value the bacterial presence effect on flies viability and reproduction of normal and mutant flies. Morphology af cytoplasmic compartments in early embryos and eggs layed by infected and uninfecyed females Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+ have been analyzed. Electron microscopy has shown that D. melanogaster embryos contain typical Wolbachia contacting with different host organelles that verify preservation of their functional activity. Atificial mitochondria and Wolbachia (wMelPop) of unusual morphology with defective bacterial membranes have been visualised in D. melanogaster [w]Trl362/TM3, Sb1 Ser y+. Wolbachia presence in ovarium cells from strains [w]Trl362/TM3, Sb1 Ser y+ and [w]Trlen82/TM3, Sb1 Ser y+ did not influence on eggs quantity layed by females. We have demonstrated for the first time that lifespan of infected and uninfected strains: D. melanogaster Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+ were similar. However the lifespan of imago from strain [w]w1118 was lower in comparison to those from strains Trl362/TM3, Sb1 Ser y+ and Trlen82/TM3, Sb1 Ser y+. It suggests that either chromosomal balancer TM3 or Trl mutation play an importance role in host-symbiotic relationship. Next experiments have revealed that lifespan of homozygotic flies decreased essentially and was close to lifespan of strain [w]w1118. Data obtained confirm that chromosomal balancer TM3 can affect on symbiont-host relationship.

[Comparison of fibroblasts-like cell differentiation capacities of human bone marrow, adipose tissue, hair papilla and dermal fibroblasts].

We compared the morphology and differentiation characteristics of the human cells from bone marrow, adipose tissue, hair papilla and skin dermis. All cell types showed fibroblastic morphology. Immunofluorescent analysis showed that adipose tissue derived stem cells (ADAS) and hair papilla cells (HPC) expressed CD105, CD49d and STRO-1, bone marrow mesenchymal stem cells (BMSC) were absent for CD49, dermal fibroblasts (DFb) expressed CD49 and STRO-1 at low level. Populations of ADAS, BMSC and HPC had similar capacities to lipid and bone differentiation. Following exposure to appropriate induction stimuli, these cells changed phenotype and expressed specific cell markers. However, the rate and extent of HPC differentiation were lower in comparison with populations of ADAS and BMSC. We propose that all investigated cell populations contain primitive progenitor cells with mesenchymal stem cell properties.

[Dynamics of the annulate lamellae in Drosophila syncytial embryos].

In eukaryotic cells, mitotic events are controlled by evolutionarily conserved cyclin-dependent kinases (cdk): these kinases phosphorylate cell proteins, which causes structural reorganization of the entire cell. Our recent studies of Drosophila syncytial embryos have demonstrated that cdk1 activity is a key factor that controls nuclear pore complex assembly/disassembly and affects the structure of cytoplasmic pores in the annulate. In this paper, we report a comparative analysis of these cytoplasmic organelles throughout the cell-cycle and throughout the development of Drosophila syncytial embryos. Based on the results obtained, it was presupposed that distribution of annulate lamellae containing cytoplasmic pores could reflect the inactivation of the mitotic kinase cdk1 in Drosophila syncytial embryos.

[Changes in Xenopus oocyte nucleus and cytoplasm organization after actin filaments depolymerization by latrunculin].

Actin-containing filaments have been visualized inside the Xenopus oocyte nuclei due to combination of fluorescence and transmission electron microscopy. It has been shown that these filaments contact with nucleoli, spherical bodies and nuclear pore complexes. The incubation of oocytes with actin-depolymerizing latrunculin causes membrane vesiculation in the cytoplasm, and disruption of the nucleoplasm and nuclear envelope integrity. We suppose that actin-containing filaments belong to crucial cell components which are involved in coordination of nuclear-cytoplasmic interactions as well as distribution and transport of intranuclear components in growing Xenopus oocytes.

[High temperature initiates changes of Wolbachia ultrastructure in the ovaries and early embryos of Drosophila melanogaster].

Electron microscopic analysis of Drosophila melanogaster (w1118) ovarian cells has demonstrated that stressful heat treatment of flies results in the appearance of electron dense granules and large lysosomes in the cytoplasm of ovarian cells, which is not related with the presence of Wolbachia, as these changes are observed in both the infected and uninfected flies. High temperature initiates essential envelope defects and other structural changes of symbiotic bacteria in the cytoplasm of ovarian cells. Some embryos developing from eggs of heat shocked flies die, however, bacteria in the survival embryos retain their typical morphology. Endosymbionts do not change their localization and their contacts with the mitochondria and endoplasmic reticulum in the ovarian cells and early embryos after heat shock treatment of the flies. The results obtained show that high temperature influences on both the host and the endosymbiont, but does not change their structural mutual interactions.