Selective targeting of activating and inhibitory Smads by distinct WWP2 ubiquitin ligase isoforms differentially modulates TGFβ signalling and EMT

Ubiquitin-dependent mechanisms have emerged as essential regulatory elements controlling cellular levels of Smads and TGFß-dependent biological outputs such as epithelial–mesenchymal transition (EMT). In this study, we identify a HECT E3 ubiquitin ligase known as WWP2 (Full-length WWP2-FL), together with two WWP2 isoforms (N-terminal, WWP2-N; C-terminal WWP2-C), as novel Smad-binding partners. We show that WWP2-FL interacts exclusively with Smad2, Smad3 and Smad7 in the TGFß pathway. Interestingly, the WWP2-N isoform interacts with Smad2 and Smad3, whereas WWP2-C interacts only with Smad7. In addition, WWP2-FL and WWP2-C have a preference for Smad7 based on protein turnover and ubiquitination studies. Unexpectedly, we also find that WWP2-N, which lacks the HECT ubiquitin ligase domain, can also interact with WWP2-FL in a TGFß-regulated manner and activate endogenous WWP2 ubiquitin ligase activity causing degradation of unstimulated Smad2 and Smad3. Consistent with our protein interaction data, overexpression and knockdown approaches reveal that WWP2 isoforms differentially modulate TGFß-dependent transcription and EMT. Finally, we show that selective disruption of WWP2 interactions with inhibitory Smad7 can stabilise Smad7 protein levels and prevent TGFß-induced EMT. Collectively, our data suggest that WWP2-N can stimulate WWP2-FL leading to increased activity against unstimulated Smad2 and Smad3, and that Smad7 is a preferred substrate for WWP2-FL and WWP2-C following prolonged TGFß stimulation. Significantly, this is the first report of an interdependent biological role for distinct HECT E3 ubiquitin ligase isoforms, and highlights an entirely novel regulatory paradigm that selectively limits the level of inhibitory and activating Smads

Human Endometrial Side Population Cells Exhibit Genotypic, Phenotypic and Functional Features of Somatic Stem Cells

During reproductive life, the human endometrium undergoes around 480 cycles of growth, breakdown and regeneration should pregnancy not be achieved. This outstanding regenerative capacity is the basis for women's cycling and its dysfunction may be involved in the etiology of pathological disorders. Therefore, the human endometrial tissue must rely on a remarkable endometrial somatic stem cells (SSC) population. Here we explore the hypothesis that human endometrial side population (SP) cells correspond to somatic stem cells. We isolated, identified and characterized the SP corresponding to the stromal and epithelial compartments using endometrial SP genes signature, immunophenotyping and characteristic telomerase pattern. We analyzed the clonogenic activity of SP cells under hypoxic conditions and the differentiation capacity in vitro to adipogenic and osteogenic lineages. Finally, we demonstrated the functional capability of endometrial SP to develop human endometrium after subcutaneous injection in NOD-SCID mice. Briefly, SP cells of human endometrium from epithelial and stromal compartments display genotypic, phenotypic and functional features of SSC

Rapid induction of pluripotency genes after exposure of human somatic cells to mouse ES cell extracts

The expression of 4 pluripotency genes (Oct4, Sox2, c-Myc and Klf4) in mouse embryonic fibroblasts can reprogramme them to a pluripotent state. We have investigated the expression of these pluripotency genes when human somatic 293T cells are permeabilised and incubated in extracts of mouse embryonic stem (ES) cells. Expression of all 4 genes was induced over 1 - 8 hours. Gene expression was associated with loss of repressive histone H3 modifications and increased recruitment of RNA polymerase II at the promoters. Lamin A/C, which is typically found only in differentiated cells, was also removed from the nuclei. When 293T cells were returned to culture after exposure to ES cell extract, the expression of the pluripotency genes continued to rise over the following 48 hours of culture, suggesting that long-term reprogramming of gene expression had been induced. This provides a methodology for studying the de-differentiation of somatic cells that can potentially lead to an efficient way of reprogramming somatic cells to a pluripotent state without genetically altering them

Control of Induction Motor in Terms of Angular Velocity Sensorless Control Using Artificial Neural Networks

On the basis of the design for a new electric power system including wind-electric and solar Power stations, coming into being at the Faculty of Electrical Engineering at Czestochowa University of Technology, the authors intend to present in some stages the possibility of using the existing renewable energy sources to the new opportunities for mini combined heat and power (CHP) plants. The resultant system will be responsible for data collection and analysis to focusing on cogeneration and implementation of the data in true applications.W niniejszym artykule rozważono zagadnienie sterowania z pośrednią orientacją wektora pola typu IFOC (Indirect Field Oriented Control), dla silnika indukcyjnego, przy pomocy sztucznych sieci neuronowych. W tym celu opracowano model symulacyjny, który umożliwia obserwację przebiegów czasowych dla wybranych wielkości, charakteryzujących pracę silnika, a w szczególności jego prędkości kątowej na wale ωm. W prezentowanej pracy podjęto również zagadnienie wykorzystania zdolności regulacyjnych sztucznych sieci neuronowych. Zaprojektowana struktura neuronowa ma za zadanie pośrednią regulację prędkości ωm dla zamodelowanej maszyny indukcyjnej. Niezbędne w tym celu było prawidłowe przeprowadzenie procesu uczenia, które w swej początkowej fazie wymagało obecności układu „nauczyciela”, zrealizowanego w postaci regulatora typu PI, o odpowiednio dobranych parametrach. Szczególnie interesujące z punktu widzenia rozważanego procesu regulacji prędkości, dla wybranej metody sterowania silnika indukcyjnego są: stabilność procesu, odporność na zakłócenia oraz zdolności adaptacyjne projektowanych struktur neuronowych

Design and implementation of three-wheeled electric vehicle

W artykule omówiono koncepcję napędu elektrycznego trójkołowego pojazdu elektrycznego, jego projekt w SolidWorks oraz poszczególne etapy montażu. Przedstawiono mikroprocesorowy system sterowania oraz elektroniczny system zarządzania napędem elektrycznym.The article discusses the concept of three-wheel electric drive electric vehicle, its design in SolidWorks and the various stages of assembly. Presented microprocessor control system and an electronic management system electric

Improvement purchasing procedure in a process of production’s preparation

Artykuł obejmuje problematykę procesu zakupów w wybranym przedsiębiorstwie branży automotive. Analizie poddano istotę zakupów zaopatrzeniowych oraz procedurę procesu zakupu. Omówiono główne zadania procesu zakupu oraz występujące w nim problemy. Przedstawiono możliwości pozyskania oszczędności przy zakupie materiałów u dostawców. Opisano także nowe rozwiązanie w dziale zakupów.The article contains of purchasing process in the production company. It provides the description of purchasing. The article covers procedure of purchasing department. It takes the main issue of purchasing process. An article is presented the possibilities for savings. Also this part of article look at the new solution in purchasing department