Wide-Field Multi-Parameter FLIM: Long-Term Minimal Invasive Observation of Proteins in Living Cells.

Time-domain Fluorescence Lifetime Imaging Microscopy (FLIM) is a remarkable tool to monitor the dynamics of fluorophore-tagged protein domains inside living cells. We propose a Wide-Field Multi-Parameter FLIM method (WFMP-FLIM) aimed to monitor continuously living cells under minimum light intensity at a given illumination energy dose. A powerful data analysis technique applied to the WFMP-FLIM data sets allows to optimize the estimation accuracy of physical parameters at very low fluorescence signal levels approaching the lower bound theoretical limit. We demonstrate the efficiency of WFMP-FLIM by presenting two independent and relevant long-term experiments in cell biology: 1) FRET analysis of simultaneously recorded donor and acceptor fluorescence in living HeLa cells and 2) tracking of mitochondrial transport combined with fluorescence lifetime analysis in neuronal processes

Raft-dependent endocytosis of autocrine motility factor is phosphatidylinositol-3-kinase-dependent in breast carcinoma cells

Autocrine motility factor (AMF) is internalized via a receptor-mediated, dynamin-dependent, cholesterol-sensitive raft pathway to the smooth endoplasmic reticulum that is negatively regulated by caveolin-1. Expression of AMF and its receptor (AMFR) is associated with tumor progression and malignancy; however, the extent to which the raft-dependent uptake of AMF is tumor cell-specific has yet to be addressed. By Western blot and cell surface fluorescence-activated cell sorter (FACS) analysis, AMFR expression is increased in tumorigenic MCF7 and metastatic MDA-231 and MDA-435 breast cancer cell lines relative to dysplastic MCF10A mammary epithelial cells. AMF uptake, determined by FACS measurement of protease-insensitive internalized fluorescein-conjugated AMF, was increased in MCF7 and MDA-435 cells relative to MCF-10A and caveolin-1-expressing MDA-231 cells. Uptake of fluorescein-conjugated AMF was dynamin-dependent, methyl-beta-cyclodextrin- and genistein-sensitive, reduced upon overexpression of caveolin-1 in MDA-435 cells, and increased upon short hairpin RNA reduction of caveolin-1 in MDA-231 cells. Tissue microarray analysis of invasive primary human breast carcinomas showed that AMFR expression had no impact on survival but did correlate significantly with expression of phospho-Akt. Phospho-Akt expression was increased in AMF-internalizing MCF7 and MDA-435 breast carcinoma cells. AMF uptake in these cells was reduced by phosphatidylinositol 3-kinase inhibition but not by regulators of macropinocytosis such as amiloride, phorbol ester, or actin cytoskeleton disruption by cytochalasin D. The raft-dependent endocytosis of AMF therefore follows a distinct phosphatidylinositol 3-kinase-dependent pathway that is up-regulated in more aggressive tumor cells

Surgical ulcerative gastroduodenal bleeding in patients with pyloroduodenal stenosis

Analysis of surgical treatment of 106 patients with ulcerative gastroduodenal hemorrhage arising in the presence of pyloroduodenal stenosis of varying severity. The patients were divided into 2 groups: 50 people, made a comparison group received traditional surgical treatment, and 56 people. A group of patients who received surgical treatment developed. It is shown that the application of a rational preoperative surgicaltion election tactics and postoperative therapy with the introduction of oxygen-containing solution intraenteric improve treatment outcomes

Mutational and phenotypic characterisation of hereditary hemorrhagic telangiectasia

Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant vascular dysplasia. Care delivery for HHT patients is impeded by the need for laborious, repeated phenotyping and gaps in knowledge regarding the relationships between causal DNA variants in ENG, ACVRL1, SMAD4 and GDF2, and clinical manifestations. To address this, we analyzed DNA samples from 183 previously uncharacterized, unrelated HHT and suspected HHT cases using the ThromboGenomics high-throughput sequencing platform. We identified 127 rare variants across 168 heterozygous genotypes. Applying modified American College of Medical Genetics and Genomics Guidelines, 106 variants were classified as pathogenic/likely pathogenic and 21 as nonpathogenic (variant of uncertain significance/benign). Unlike the protein products of ACVRL1 and SMAD4, the extracellular ENG amino acids are not strongly conserved. Our inferences of the functional consequences of causal variants in ENG were therefore informed by the crystal structure of endoglin. We then compared the accuracy of predictions of the causal gene blinded to the genetic data using 2 approaches: subjective clinical predictions and statistical predictions based on 8 Human Phenotype Ontology terms. Both approaches had some predictive power, but they were insufficiently accurate to be used clinically, without genetic testing. The distributions of red cell indices differed by causal gene but not sufficiently for clinical use in isolation from genetic data. We conclude that parallel sequencing of the 4 known HHT genes, multidisciplinary team review of variant calls in the context of detailed clinical information, and statistical and structural modeling improve the prognostication and treatment of HHT

Physical-Mechanical Properties of Superhard Nanocomposite Coatings on Base Zr-Ti-Si-N

Hard and super hard coatings of Zr-Ti-Si-N of from 2.8 μm to 3.5 μm thickness were fabricated using a vacuum arc source with high frequency stimulation. The samples were annealed in vacuum and in air at 1200 °C. It was found that films with a high Zr and Ti content were thermally stable up to 1180 °C. At the same time, a thin oxide layer of 180 nm to 240 nm was found on the surfaces, which protected the sample from destruction. Below 1000 °C annealing temperature in vacuum, changing of phase composition is determined by appearing of siliconitride crystallites (ß-Si3N4) with hexagonal crystalline lattice and by formation of ZrO2 oxide crystallites. Size of grains of a substitution solid solution (Zr, Ti)N varied from (10 – 12) nm to 25 nm, but Ti concentration in the solid solution increased. In the process of annealing, hardness of the best series of samples increased from (39.6 ±1.4) GPa to 53.6 GPa, which seemed to indicate that a spinodal segregation along grain interfaces was finished. When you are citing the document, use the following link http://essuir.sumdu.edu.ua/handle/123456789/3394

The association between family and community social capital and health risk behaviours in young people: an integrative review

Background: Health risk behaviours known to result in poorer outcomes in adulthood are generally established in late childhood and adolescence. These ‘risky’ behaviours include smoking, alcohol and illicit drug use and sexual risk taking. While the role of social capital in the establishment of health risk behaviours in young people has been explored, to date, no attempt has been made to consolidate the evidence in the form of a review. Thus, this integrative review was undertaken to identify and synthesise research findings on the role and impact of family and community social capital on health risk behaviours in young people and provide a consolidated evidence base to inform multi-sectorial policy and practice.<p></p> Methods: Key electronic databases were searched (i.e. ASSIA, CINAHL, Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, Database of Abstracts of Reviews of Effects, Embase, Medline, PsycINFO, Sociological Abstracts) for relevant studies and this was complemented by hand searching. Inclusion/exclusion criteria were applied and data was extracted from the included studies. Heterogeneity in study design and the outcomes assessed precluded meta-analysis/meta-synthesis; the results are therefore presented in narrative form.<p></p> Results: Thirty-four papers satisfied the review inclusion criteria; most were cross-sectional surveys. The majority of the studies were conducted in North America (n=25), with three being conducted in the UK. Sample sizes ranged from 61 to 98,340. The synthesised evidence demonstrates that social capital is an important construct for understanding the establishment of health risk behaviours in young people. The different elements of family and community social capital varied in terms of their saliency within each behavioural domain, with positive parent–child relations, parental monitoring, religiosity and school quality being particularly important in reducing risk.<p></p> Conclusions: This review is the first to systematically synthesise research findings about the association between social capital and health risk behaviours in young people. While providing evidence that may inform the development of interventions framed around social capital, the review also highlights key areas where further research is required to provide a fuller account of the nature and role of social capital in influencing the uptake of health risk behaviours.<p></p&gt

Efficiency of the "routine" external choledoch drainage method for patients with acute cholecystitis

Purpose of the study: To justify the efficiency of 'routine' choledoch drainage for patients suffering from acute cholecystitis. Materials and methods: The study was carried out at the Septic Surgery Department, Rostov-on-Don Emergency Care Hospital. The survey included 532 patients with various forms of acute cholecystitis admitted to the surgery department from 2016 through 2017. Age of the patients: 38 to 67 years. All patients were operated (laparoscopic cholecystectomy), with a 'routine' choledoch drainage performed according to an original method (RF Patent No. 2218949). Results: Results collected during the study demonstrated advisability and effectiveness of the method, which did not only prevent reoperation but also facilitated quick post-surgery organism rehabilitation. Whenever the 'routine' choledoch drainage method was used, during the postoperative care period we diagnosed choledocholithiasis in 14,8% of cases representing an indication for endoscopic papillosphincterotomy, in 82,4% of cases we noted early normalization of the total bilirubin and hemodiastase levels, improvement of the patients' condition, and shorter stay at the in-patient department. Conclusions: The range of indications for choledoch drainage in cases of acute cholecystitis should be expanded. Our method allows to reduce the number of postoperative complications and shorten the hospital stay representing both a positive factor for patients, and economic benefit for the department.Цель исследования: Обосновать эффективность использования «рутинного» дренирования холедоха у пациентов с острым холециститом. Материалы и методы: Работа выполнена на базе хирургического отделения МБУЗ ГБСМП г. Ростова-на-Дону. В исследование вошли 532 пациента с различными формами острого холецистита, находившихся на лечении в хирургическом отделении с 2016 по 2017 год. Возраст пациентов от 38 до 67 лет. Все пациенты были прооперированы (лапароскопическая холецистэктомия), выполнено «рутинное» дренирование холедоха по оригинальной методике (Федеральный патент РФ №2218949). Результаты: Полученные в ходе исследования результаты показали целесообразность и эффективность метода, которая заключается не только в предотвращении повторных операций, но и быстрой реабилитации организма после операции. При использовании «рутинного» дренирования холедоха, в послеоперационном периоде, нами был диагностирован холедохолитиазв 14,6% случаев, что явилось показанием к выполнению эндоскопической папиллосфинктеротомии, у 82,4% была отмечена ранняя нормализацию уровня общего билирубина и амилазы крови, улучшение состояния пациентов, и сокращение сроков пребывания в стационаре. Выводы: Необходимо расширить показания к выполнению дренирования холедоха при остром холецистите. Наша методика позволяет своевременно диагностировать нарушения желчеоттока, сократить количество послеоперационных осложнений, и срок пребывания пациентов в стационаре, что является не только положительным фактором для пациента, но и экономически выгодным для отделения