Spatio-temporal segmentation of image sequences for non-invasive analysis of cardiovascular structure and function in Whitefish embryos.

Label-free quantitative analysis of the cardiovascular structure and function in wild fish is highly important and still unsolved problem. We developed a robust algorithm for bright field microscopy data processing, which allows reliable detection and quantification of circulatory system across the whole animal. We verified this technique by in vivo experiments on broad whitefish (Coregonus nasus) embryos. Calculated morphofunctional features of heart and vessels are quite specific due to low temperature embryonic development of this fish, and correspond well to the reference data obtained by ex vivo techniques. This research highlights the feasibility of non-invasive cardiovascular imaging and measurements valuable for behavioral, environmental, toxicological, drug discovery and many other studies.

[Pathomorphological features of the placentas and the placental sites after in vitro fertilization with a donor egg]. / Patomorfologicheskie osobennosti platsent i platsentarnykh ploshchadok posle ékstrakorporal'nogo oplodotvoreniia s primeneniem donorskoi iaitsekletki.

OBJECTIVE: To identify the structural and immunohistochemical features of the placentas and the placental sites after in vitro fertilization (IVF) with a donor egg (surrogate motherhood). SUBJECT AND METHODS: Morphological and immunohistochemical studies were performed on the placental (a placental disk) and placental bed materials obtained after caesarean delivery. The investigation enrolled 26 patients whose pregnancy occurred with IVF with a donor egg according to the surrogacy (IVF-S) program. A comparison group included 13 patients whose pregnancy occurred after IVF with their own eggs. An immunohistochemical study was conducted on paraffin sections made from biopsy material; mouse antibodies to total cytokeratin (clone AE1/AE3, 'Dako'), HLA-DR (clone TAL.1B5, 'Dako'), and CD138 (clone MI15, 'Dako') were used as primary antibodies. RESULTS: The histological examination of the placentas in the IVF-S group showed the high incidence of central ischemic heart attacks (69%), dissociated cotyledon development (61%), pathological villous immaturity mainly with the predominance of intermediate differentiated villi (46%), and massive perivillous fibrinoid deposition (73%). The obtained differences between with the study and comparison groups were significant (p<0.05). The IVF-S group was characterized by the development of lymphoplasmacytic deciduitis (1.23±0.4 and 0.5±0.3 scores). Examination of the placental site biopsy material in the IVF-S group revealed the following changes: remodeling of the spiral arteries was incomplete in more than 40% of cases, and in 30% of the spiral arteries had no gestational changes. In the comparison group, more than 90% of the spiral arteries were characterized by complete remodeling during pregnancy. There was also an increase in the count of multinucleated trophoblastic giant cells (104.56±4.21 and 65.67±14.45) and HLA-DR positive cells (41.86±5.32 and 29.00±1.87). CONCLUSION: The placentas and the placental sites of the women whose pregnancy occurred with IVF-S are characterized by the development of high lyoplasmacytic deciduitis activity and pronounced placental immune alterations manifested by the high incidence of immune responses at the sites of the closest contact between maternal and fetal tissues. The placental bed exhibited defective spiral artery remodeling, development of chronic inflammatory lesions in the perivascular areas, and an increase in the counts of HLA-DR positive cells and multinucleated trophoblastic giant cells.

[Prediction of activity and severity of endocrine ophthalmopathy by multidimensional linear regression modeling]. / Prognozirovanie aktivnosti i tiazhesti éndokrinnoi oftal'mopatii metodom mnogomernogo lineinogo regressionnogo modelirovaniia.

Endocrine ophthalmopathy (EOP) is a severe chronic autoimmune disease associated with autoimmune thyroid pathology that leads to loss of sight, cosmetic defects and quality of life decrease. EOP is difficult to give prognosis for due to various factors affecting its course and outcome. PURPOSE: To develop a reliable and precise prognosis method for EOP activity and severity based on personalized combination of risk factors in specific patients by multidimensional linear regression modeling. MATERIAL AND METHODS: A group of 139 patients (278 orbits) with newly diagnosed EOP associated with toxic diffuse goiter was observed during 1 year by an ophthalmologist and an endocrinologist; patient examination interval was 6 months. More than 250 indices were dynamically analyzed in the course of the study. Linear regression analysis was chosen as the research method; it allowed detection of linear dependencies between dependent and explanatory variables. RESULTS: More than 600 various linear regression equations were derived that enabled prediction of EOP onset risk and development timeline, estimation of activity and/or severity of the disease, duration of active period in specific patients for the immediate and long-term outcome. From the derived models, the most reliable and safest for practical application were picked out. The present study introduces nine optimized models that can be used for patient follow-up since day one. CONCLUSION: The onset risk, progression and outcome of the disease can be determined by a combination of factors revealed in the study.

Aberrant expression of selenium-containing glutathione peroxidases in clear cell renal cell carcinomas.

AIM: To find putative diagnostic markers for clear cell renal cell carcinomas (ccRCC). MATERIAL AND METHODS: Quantitative polymerase chain reaction (Q-PCR), bisulfite treatment, methylation-specific PCR, analysis on cBioPortal for Cancer Genomics. RESULTS: We have found that expression of GPX1, GPX3, and GPX4 genes was decreased in ccRCC. We have shown that the number of alanine (GCG) repeats at the amino terminus of the GPX1 protein is variable. It was reported earlier that an allele that possess 5 alanine repeats is associated with the increased cancer risk. According to the obtained data, the allele with the 5 alanine repeats was also present in a group of healthy donors. Moreover, the frequency of alleles with repeats was similar among ccRCC patients and healthy individuals. We found that decreased expression of GPXs genes was not associated with promoter methylation. To provide other explanation, an analysis on the gene copy number was performed. We have found the heterozygous deletions for GPX1 gene, amplification for GPX3 gene, and no change in gene copy number for GPX4. CONCLUSIONS: Our data support the hypothesis that GPX1, GPX3, and GPX4 genes may play a role in ccRCC cancerogenesis and therefore they might be considered as putative diagnostic markers for ccRCC.

[Performance of fragmenting operation for restoration of a correct rhythm while treating the isolated failure of a mitral valve].

Possibilities of application of the proposed procedure for intraoperative restoration of the sinus rhythm while the mitral valve prosthesis (MVP) were studied up. In 161 patients, suffering the isolated mitral valve failure stage IV, a surgical treatment was conducted in Department of surgical treatment of acquired heart failures. The left atrium plasty was conducted in 119 (73.9%) of them with the objective to reduce its size and eliminate the re-entry waves passability. Fragmentation in left atrium was conducted in lowradiofrequency regime of diathermy (25-35 W) in accordance to variant (cut and sew) of operations Maze - 3 and Maze - 4. On a hospital stage 3 patients died (hospital lethality 1.9%). MVP in combination with Maze operation in a lowradiofrequency regime have permitted to restore successfully a correct rhythm in 80.2% of patients on a hospital stage and to stabilize it during 1 year postoperatively.

[Functional tear production indices in thyroid eye disease].

Analysis of functional tear production indices in patients with Grave's disease and thyroid eye disease confirmed lacrimal gland involvement in inflammatory autoimmune process. Close direct correlation was found between tear production and main disease characteristics - severity and activity.

[Morphological changes in autoimmune dacryoadenitis associated with thyroid eye disease].

Increase of lacrimal gland (LG) volume was revealed in 1/3 of patients with thyroid eye disease (TED) using visualization methods. Authors show morphological data confirming development of autoimmune process in LG in TED, biopsy samples were studied within 6 hours after exitus. Specific histomorphological characteristics were analysed considering dystrophic and degenerative changes in LG tissue. Quantitative assessment of fibrosis and vascularization extent is determined using fibrosis and vascularization index.

Genetic and epigenetic changes of NKIRAS1 gene in human renal cell carcinomas.

UNLABELLED: Renal cell carcinoma (RCC) is the most common malignant tumor of kidney associated with the worst clinical outcome. No molecular markers for RCC diagnostics and prognosis that could be applied in clinics were described yet. Large-scale screening of 3p human chromosome genes/loci in RCC and histologically normal tissues surrounding the tumors using NotI-microarray approach demonstrated that NKIRAS1 gene contained the largest percent of genetic/epigenetic changes in RCC tumor cells. AIM: To validate the results of NotI microarray analysis and study genetic, epigenetic changes, and the expression level of NKIRAS1 gene in human RCC samples. METHODS: DNA and RNA were isolated from freshly-frozen renal tumors' samples (n = 12) and from normal tissues surrounding the tumors. Epigenetic changes (methylation status) of NKIRAS1 were detected by bisulfite sequencing. Genetic changes and expression level were analyzed by Quantitative real-time PCR (qPCR) with SYBR Green. For relative quantification 2-(DeltaDeltaCP) method was used. Nonparametric tests (Wilcoxon, Kruskal - Wallis and Mann - Whitney) were applied for statistical data analysis using the BioStat software. RESULTS: NKIRAS1 expression was downregulated in 75% of RCC samples (9 of 12) compared with surrounding normal tissue. High grade tumors (3 and 4) showed lower expression of NKIRAS1 at the mRNA level than tumors of low grade (1 and 2). No significant association was found between gene expression level and gender or age. Analysis of NKIRAS1 gene copy number was performed in 19 tumor samples. Changes in the copy number of NKIRAS1 gene were observed in 64% (9 of 14) of cRCC samples. 9 samples displayed ratio (< 0.85 and >or= 0.35), thus were considered as hemizygous deletions. 3 samples showed ratio (> 0.85) and were considered as normal copy number. Changes in NKIRAS1 gene copy number were detected in all 3 benign oncocytomas, 1 papillary cancer and 1 sarcoma, where hemizygous deletion was observed. No changes in methylation status of NKIRAS1 were found in RCC. CONCLUSIONS: We have validated the results of NotI microarray analysis of NKIRAS1 gene in RCC. It was shown the decreased expression level of NKIRAS1 in this type of tumor.

[Functional state of arterioles in patients with essential hypertension and arterial hypertension developed at the background of diabetes mellitus].

With the aim of studying linear velocity characteristics of arteriolar blood flow and reactivity of arterioles we examined 80 patients with essential arterial hypertension (AH), 83 patients with type 2 diabetes mellitus (DM) with concomitant AH, and 63 healthy people. In all patients examination included ultrasound continuous wave dopplerography of microcirculatory vascular bed at rest and after distant warming test. It was found that in patients with essential AH blood flow velocity in arterioles was maximal, increment of arteriolar blood flow in response to warming was lower than in healthy persons. In patients with type 2 DM minimal linear velocity characteristics of arteriolar blood flow and its increment in response to warming test were minimal. Results of the study evidence for principal differences of functioning and reactivity of arterioles in patients with essential AH and AH developed at the background of diabetes mellitus.

[Arteriolar circulation and arterial pressure in patients with essential hypertension].

Arteriolar circulation and its relationship with arterial pressure were studied in 127 patients with essential hypertension and 63 healthy subjects using continuous US dopplerography of the microcirculatory system at rest. Linear arteriolar blood flow rate was measured in healthy subjects and 47 patients before and after primary indapamide therapy. It was found to equal 17.7 (mean 9.1) cm/s in systole and 8.7 cm/s in diastole in the patients versus 12.2 (7.3) and 4.3 respectively in the healthy subjects (p = 0.005) giving indirect evidence of narrowed diameter of arterioles. Arteriolar circulation was shown to depend on the duration of arterial hypertension. Both AP and linear arteriolar blood flow rate decreased under effect of hypotensive therapy but there was no significant correlation between the two parameters.

[State of microcirculation and endothelial dysfunction in patients with type 2 diabetes mellitus and arterial hypertension].

In order to evaluate microcirculation and endothelial dysfunction in type 2 diabetes mellitus accompanied by arterial hypertension, 40 men aged 54.9 +/- 1.49 years suffering from these conditions were examined using ultrasound continuous-wave microcirculation dopplerography (Minimax-Doppler-K) and ischemic test. The controls, 28 practically healthy persons, were sex- and age-comparable. Compared to the controls, the patients with diabetes and concomitant arterial hypertension displayed lower linear blood flow velocity both initially and during the postischemic period, their velocity parameters recovered after ischemic test more slowly, and the parameters of blood flow were rigid within the entire study. This could result from an increased microcirculatory blood vessel (arteriolar) diameter, changes in vessel reactivity, and the presence of endothelial dysfunction in these patients.

Lung aplasia: anatomy, history, diagnosis and surgical management.

PURPOSE: The aim of the study was to define the details of the history and clarify the cause for respiratory distress, justify the need for surgical correction and suggest a rational operative technique for patients with lung aplasia. METHODS: Our experience with the management of 9 patients with lung aplasia and 1 patient with lung agenesis in the period from 1985 to 2004 is presented. All 10 patients were referred for respiratory distress. A detailed study of clinical symptoms and the data from X-ray, computed tomography (CT), tracheobronchoscopy and digital subtraction angiography (DSA) suggested that all patients had different degrees of mediastinal shift and heart rotation, tracheal kinking and compression of the aortic arch and innominate artery or emphysema of a single lung. Two of the patients required operation. RESULTS: We performed ipsilateral cephalad translocation of the diaphragm which resulted in complete recovery from respiratory distress and a significant improvement in tolerance of physical exercise in both patients. The good results in these patients were followed up for 8 and 2 years, respectively. CONCLUSIONS: We suggest that ipsilateral translocation of the diaphragm could be a useful alternative to unsuccessful symptomatic treatment in patients with lung aplasia.

[Histogenesis and structural organization of the walls of rat venae cavae and pulmonary veins]. / Gistogenez i strukturnaia organizatsiia stenki polykh i legochnykh ven krysy.

Using light and electron microscopic methods, the histogenesis and structural organization of the walls of rat venae cavae and pulmonary veins were studied in prenatal and postnatal periods of development. The special attention was paid to the appearance of the striated myocytes in the walls of these vessels during the process of ontogenesis. The time of initial divergent development of myoblastic differon was established, the stages of differentiation of striated myoblasts and the peculiarities of intercellular junctions were characterized, as well as the innervation and vascularization of the walls of venae cavae and pulmonary veins.

[Diagnosis and treatment of biatrial myxomas of the heart]. / Diagnostika i lechenie biatrial'nykh miksom serdtsa.

Of the multiple myxomas of the heart (MH) the bilateral myxomas were revealed in 57.1%. Of 4 patients before the operation the correct diagnosis was established only in one. Meticulous data on clinical signs, diagnosis and surgical treatment peculiarities of patients with bilateral MH were adduced. Morphological peculiarities of the tumor were analyzed. Special attention was paid to the necessity of clinical and surgical guard toward revelation of undiagnosed tumors, the choice of optimal approaches to the heart cavities while performing radical operation. Basing on results of histological studying of operational specimen it was established that MH is originated from transformed vessels of interatrial septum.

[Cardiomyogenesis in Rana ridibunda tadpoles during metamorphosis in normal conditions and after exposure to various concentrations of cadmium ions]. / Kardiomiogenez u ozernoi liagushki v period metamorfoza golovastikov v norme i pri vozdeistvii ionov kadmiia v razlichnykh kontsentratsiiakh.

During metamorphosis period Rana ridibunda ventricular cardiomyocytes possess strong ability to adapt to altering developmental conditions, certain peculiarities of differentiation were observed during this process, cell death and mitotic activity increases. The action of different concentrations of cadmium ions on Rana ridibunda larvae changes interrelation and intensity of basic processes of cardiac muscular histogenesis, leading to differentiation disorders, suppression of proliferative activity and intensification of cardiomyocytes death.

An extended hydrophobic interactive surface of Yersinia pestis Caf1M chaperone is essential for subunit binding and F1 capsule assembly.

A single polypeptide subunit, Caf1, polymerizes to form a dense, poorly defined structure (F1 capsule) on the surface of Yersinia pestis. The caf-encoded assembly components belong to the chaperone-usher protein family involved in the assembly of composite adhesive pili, but the Caf1M chaperone itself belongs to a distinct subfamily. One unique feature of this subfamily is the possession of a long, variable sequence between the F1 beta-strand and the G1 subunit binding beta-strand (FGL; F1 beta-strand to G1 beta-strand long). Deletion and insertion mutations confirmed that the FGL sequence was not essential for folding of the protein but was absolutely essential for function. Site-specific mutagenesis of individual residues identified Val-126, in particular, together with Val-128 as critical residues for the formation of a stable subunit-chaperone complex and the promotion of surface assembly. Differential effects on periplasmic polymerization of the subunit were also observed with different mutants. Together with the G1 strand, the FGL sequence has the potential to form an interactive surface of five alternating hydrophobic residues on Caf1M chaperone as well as in seven of the 10 other members of the FGL subfamily. Mutation of the absolutely conserved Arg-20 to Ser led to drastic reduction in Caf1 binding and surface assembled polymer. Thus, although Caf1M-Caf1 subunit binding almost certainly involves the basic principle of donor strand complementation elucidated for the PapD-PapK complex, a key feature unique to the chaperones of this subfamily would appear to be capping via high-affinity binding of an extended hydrophobic surface on the respective single subunits.

[Changes in hypophyseal-thyroid system function and in the level of atrial natriuretic factor and cyclic nucleotides in the blood of patients with ischemic heart disease exposed to ultrasonic irradiation of the precordial area]. / Izmeneniia sostoianiia gipofizarno-tireoidnoi sistemy, urovnia predserdnogo natriiureticheskogo faktora, tsiklicheskikh nukleotidov v krovi bol'nykh ishemicheskoi bolezn'iu serdtsa pod vozdeistviem ul'trazvukovogo oblucheniia prekordial'noi oblasti.

Radiation of the precordial region with ultrasound at 880 kHz, 0.4 W/cm2 for 12 minutes was examined for its impact on blood levels of thyrotropin, tri-iodothyronine, thyroxine, atrial natriuretic factor, cyclic nucleotides (cAMP and cHMP) in 70 patients with Functional Classes II-III exercise-induced angina. The radiation was found to lead to elevated plasma levels of atrial natriuretic factor, cHMP and inhibited plasma cAMP in patients with coronary heart disease as compared to the control group.