Characterization of dermal fibroblast-derived iPSCs from a patient with low grade steatosis

AbstractPrimary fibroblasts from a low grade steatosis patient were reprogrammed by transduction of a combination of two episomal-based plasmids OCT4,SOX2, c-MYC and KLF4. iPSCs were characterized by immunocytochemistry, embryonic body-formation, DNA-fingerprint karyotype analysis and comparative transcriptome analyses with the human embryonic stem cell line H1 revealed a Pearsons correlation of 0.9251

Activation of the Renin–Angiotensin System Disrupts the Cytoskeletal Architecture of Human Urine-Derived Podocytes

High blood pressure is one of the major public health problems that causes severe disorders in several tissues including the human kidney. One of the most important signaling pathways associated with the regulation of blood pressure is the renin–angiotensin system (RAS), with its main mediator angiotensin II (ANGII). Elevated levels of circulating and intracellular ANGII and aldosterone lead to pro-fibrotic, -inflammatory, and -hypertrophic milieu that causes remodeling and dysfunction in cardiovascular and renal tissues. Furthermore, ANGII has been recognized as a major risk factor for the induction of apoptosis in podocytes, ultimately leading to chronic kidney disease (CKD). In the past, disease modeling of kidney-associated diseases was extremely difficult, as the derivation of kidney originated cells is very challenging. Here we describe a differentiation protocol for reproducible differentiation of sine oculis homeobox homolog 2 (SIX2)-positive urine-derived renal progenitor cells (UdRPCs) into podocytes bearing typical cellular processes. The UdRPCs-derived podocytes show the activation of the renin–angiotensin system by being responsive to ANGII stimulation. Our data reveal the ANGII-dependent downregulation of nephrin (NPHS1) and synaptopodin (SYNPO), resulting in the disruption of the podocyte cytoskeletal architecture, as shown by immunofluorescence-based detection of α-Actinin. Furthermore, we show that the cytoskeletal disruption is mainly mediated through angiotensin II receptor type 1 (AGTR1) signaling and can be rescued by AGTR1 inhibition with the selective, competitive angiotensin II receptor type 1 antagonist, losartan. In the present manuscript we confirm and propose UdRPCs differentiated to podocytes as a unique cell type useful for studying nephrogenesis and associated diseases. Furthermore, the responsiveness of UdRPCs-derived podocytes to ANGII implies potential applications in nephrotoxicity studies and drug screening

Derivation of the Immortalized Cell Line UM51-PrePodo-hTERT and Its Responsiveness to Angiotensin II and Activation of the RAAS Pathway

Recent demographic studies predict there will be a considerable increase in the number of elderly people within the next few decades. Aging has been recognized as one of the main risk factors for the world’s most prevalent diseases such as neurodegenerative disorders, cancer, cardiovascular disease, and metabolic diseases. During the process of aging, a gradual loss of tissue volume and organ function is observed, which is partially caused by replicative senescence. The capacity of cellular proliferation and replicative senescence is tightly regulated by their telomere length. When telomere length is critically shortened with progressive cell division, cells become proliferatively arrested, and DNA damage response and cellular senescence are triggered, whereupon the “Hayflick limit” is attained at this stage. Podocytes are a cell type found in the kidney glomerulus where they have major roles in blood filtration. Mature podocytes are terminal differentiated cells that are unable to undergo cell division in vivo. For this reason, the establishment of primary podocyte cell cultures has been very challenging. In our present study, we present the successful immortalization of a human podocyte progenitor cell line, of which the primary cells were isolated directly from the urine of a 51-year-old male. The immortalized cell line was cultured over the course of one year (~100 passages) with high proliferation capacity, endowed with contact inhibition and P53 expression. Furthermore, by immunofluorescence-based expression and quantitative real-time PCR for the podocyte markers CD2AP, LMX1B, NPHS1, SYNPO and WT1, we confirmed the differentiation capacity of the immortalized cells. Finally, we evaluated and confirmed the responsiveness of the immortalized cells on the main mediator angiotensin II (ANGII) of the renin–angiotensin system (RAAS). In conclusion, we have shown that it is possible to bypass cellular replicative senescence (Hayflick limit) by TERT-driven immortalization of human urine-derived pre-podocyte cells from a 51-year-old African male

A stem cell based in vitro model of NAFLD enables the analysis of patient specific individual metabolic adaptations in response to a high fat diet and AdipoRon interference

Non-alcoholic fatty liver disease (NAFLD) is a multifactorial disease. Its development and progression depend on genetically predisposed susceptibility of the patient towards several ‘hits’ that induce fat storage first and later inflammation and fibrosis. Here, we differentiated induced pluripotent stem cells (iPSCs) derived from four distinct donors with varying disease stages into hepatocyte like cells (HLCs) and determined fat storage as well as metabolic adaptations after stimulations with oleic acid. We could recapitulate the complex networks that control lipid and glucose metabolism and we identified distinct gene expression profiles related to the steatosis phenotype of the donor. In an attempt to reverse the steatotic phenotype, cells were treated with the small molecule AdipoRon, a synthetic analogue of adiponectin. Although the responses varied between cells lines, they suggest a general influence of AdipoRon on metabolism, transport, immune system, cell stress and signalling

Интерактивная обработка изображений, получаемых посредством лазерного монитора в режиме реального времени

В ходе исследования необходимо было создать устройство позиционирования объекта в системе лазерного монитора. В процессе исследования проводилось исследование алгоритмов определения резкости изображения. На основе выбранного метода был разработан алгоритм автоматической фокусировки изображения. Также был разработан алгоритм системы слежения за объектом на основе алгоритма нормированной кросскорреляции. В результате исследования удалось снизить искажения, вносимые оптической системой лазерного монитора и активной оптической средой, и организовать систему автоматической фокусировки изображения и систему слежения, позволяющую отслеживать перемещение объекта наблюдения с точностью до 1 пикселя.In the study, it was necessary to create a device object positioning system for the laser monitor. The study surveyed algorithms determine the sharpness of the image. Based on the selected method algorithm for automatic focus of the image was developed. There was also designeded a tracking system for the object of the algorithm based on normalized cross-correlation method. The study was able to reduce the distortions introduced by the optical system of the laser and monitor the active optical medium, and to organize a system of auto focus and image tracking system to track the movement of the object of observation up to 1 pixel

Конверсия попутных нефтяных газов С3-С4 в жидкие углеводороды на модифицированных цеолитных катализаторах

Исследованы влияние модифицирования цеолита типа MFI и активированные плазмой, на кислотные и каталитические свойства в конверсии пропан-бутановой фракции в арены.The effect of modification of MFI zeolite activated by plasma on acid and catalytic properties in the conversion of the propane-butane fraction into arenes was investigated

The MOCART (Magnetic Resonance Observation of Cartilage Repair Tissue) 2.0 Knee Score and Atlas

Objective Since the first introduction of the MOCART (Magnetic Resonance Observation of Cartilage Repair Tissue) score, significant progress has been made with regard to surgical treatment options for cartilage defects, as well as magnetic resonance imaging (MRI) of such defects. Thus, the aim of this study was to introduce the MOCART 2.0 knee score — an incremental update on the original MOCART score — that incorporates this progression. Materials and Methods The volume of cartilage defect filling is now assessed in 25% increments, with hypertrophic filling of up to 150% receiving the same scoring as complete repair. Integration now assesses only the integration to neighboring native cartilage, and the severity of surface irregularities is assessed in reference to cartilage repair length rather than depth. The signal intensity of the repair tissue differentiates normal signal, minor abnormal, or severely abnormal signal alterations. The assessment of the variables "subchondral lamina," "adhesions," and "synovitis" was removed and the points were reallocated to the new variable "bony defect or bony overgrowth." The variable "subchondral bone" was renamed to "subchondral changes" and assesses minor and severe edema-like marrow signal, as well as subchondral cysts or osteonecrosis-like signal. Overall, a MOCART 2.0 knee score ranging from 0 to 100 points may be reached. Four independent readers (two expert readers and two radiology residents with limited experience) assessed the 3 T MRI examinations of 24 patients, who had undergone cartilage repair of a femoral cartilage defect using the new MOCART 2.0 knee score. One of the expert readers and both inexperienced readers performed two readings, separated by a four-week interval. For the inexperienced readers, the first reading was based on the evaluation sheet only. For the second reading, a newly introduced atlas was used as an additional reference. Intrarater and interrater reliability was assessed using intraclass correlation coefficients (ICCs) and weighted kappa statistics. ICCs were interpreted according to Koo and Li; weighted kappa statistics were interpreted according to the criteria of Landis and Koch. Results The overall intrarater (ICC = 0.88, P < 0.001) as well as the interrater (ICC = 0.84, P < 0.001) reliability of the expert readers was almost perfect. Based on the evaluation sheet of the MOCART 2.0 knee score, the overall interrater reliability of the inexperienced readers was poor (ICC = 0.34, P < 0.019) and improved to moderate (ICC = 0.59, P = 0.001) with the use of the atlas. Conclusions The MOCART 2.0 knee score was updated to account for changes in the past decade and demonstrates almost perfect interrater and intrarater reliability in expert readers. In inexperienced readers, use of the atlas may improve interrater reliability and, thus, increase the comparability of results across studies

Анализ электрической части подстанции «Ала-Арча» и релейная защита трансформатора

Выпускная квалификационная работа 97 с.,18 рис., 25 табл., 7 источников. Выполнен анализ электрических сетей Кыргызской республики. Описано современное состояние энергетики Кыргызской Республики. Также была описана структурная схема ПС 슫Ала-Арча슻. Произведен расчет продолжительных режимов, проанализированы силовые трансформаторы.В работе выбраны и обоснованы устройства релейной защиты трансформатора.Релейная защита выполнена на основе микропроцессорного оборудования MICOM P63х. В качестве основной защиты используется дифференциальная защита с двумя участками торможения и дифференциальная отсечка, а также построена характеристика срабатывания по выбранным уставкам. Для проверки чувствительности рассчитано КЗ на шинах 10 кВ при минимальном режиме работы.Final qualifying work 97 p. , 18 fig. , 25 tab. , 7 sources. The analysis of electric networks of the Kyrgyz Republic. The present state of Energy of the Kyrgyz Republic . It is also a block diagram of the SS " Ala - Archa" has been described .The calculation of continuous operation , analyzed the power transformatory.V selected and justified relaying device transformatora.Releynaya protection is made on the basis of microprocessor equipment MICOM P63h . As the main protection differential protection with two portions of the braking and differential cut-off , as well as built on the selected tripping characteristic setting. To test the sensitivity of the calculated short-circuit on tires with a minimum of 10 kV operation

In vivo phase imaging of human epiphyseal cartilage at 7 T

PURPOSE: To assess the potential clinical utility of in vivo susceptibility-weighted imaging and quantitative susceptibility mapping of growth cartilage in the juvenile human knee at 7 T. METHODS: High-resolution gradient-echo images of the knees of six healthy children and adolescents aged 6 to 15 were acquired with a 28-channel coil at 7 T. Phase images from the coils were combined using a short echo-time reference scan method (COMPOSER). RESULTS: Veins oriented perpendicular to the static B0field appeared doubled in susceptibility-weighted imaging, but not quantitative susceptibility mapping. Veins and layers in the cartilage were visible in all children up to the age of 13. CONCLUSION: Phase imaging using susceptibility-weighted imaging and quantitative susceptibility mapping allows the in vivo visualization of veins and layers in human growth cartilage