Combined Characterization of microRNA and mRNA Profiles Delineates Early Differentiation Pathways of CD133+ and CD34+ Hematopoietic Stem and Progenitor Cells

MicroRNAs (miRNAs) have been shown to play an important role in hematopoiesis. To elucidate the role of miRNAs in the early steps of hematopoiesis, we directly compared donor-matched CD133+ cells with the more differentiated CD34+CD133− and CD34−CD133− cells from bone marrow on the miRNA and mRNA level. Using quantitative whole genome miRNA microarray and sequencing-based profiling, we found that between 109 (CD133+) and 216 (CD34−CD133−) miRNAs were expressed. Quantification revealed that the 25 highest expressed miRNAs accounted for 73% of the total miRNA pool. miR-142-3p was the highest expressed miRNA with up to 2,000 copies per cell in CD34+CD133− cells. Eighteen miRNAs were significantly differentially expressed between CD133+ and CD34+CD133− cells. We analyzed their biological role by examining the coexpression of miRNAs and its bioinformatically predicted mRNA targets and luciferase-based reporter assays. We provide the first evidence for a direct regulation of CD133 by miR-142-3p as well as tropomyosin 1 and frizzled homolog 5 by miR-29a. Overexpression of miRNAs in CD133+ cells demonstrated that miR-142-3p has a negative influence on the overall colony-forming ability. In conclusion, the miRNAs expressed differentially between the CD133+ and CD34+CD133− cells are involved in inhibition of differentiation, prevention of apoptosis, and cytoskeletal remodeling. These results are highly relevant for stem cell-based therapies with CD133+ cells and delineate for the first time how the stem cell character of CD133+ cells is defined by the expression of specific miRNAs. Stem Cells 2011;29:847–85

Identification of New Hematopoietic Cell Subsets with a Polyclonal Antibody Library Specific for Neglected Proteins

The identification of new markers, the expression of which defines new phenotipically and functionally distinct cell subsets, is a main objective in cell biology. We have addressed the issue of identifying new cell specific markers with a reverse proteomic approach whereby approximately 1700 human open reading frames encoding proteins predicted to be transmembrane or secreted have been selected in silico for being poorly known, cloned and expressed in bacteria. These proteins have been purified and used to immunize mice with the aim of obtaining polyclonal antisera mostly specific for linear epitopes. Such a library, made of about 1600 different polyclonal antisera, has been obtained and screened by flow cytometry on cord blood derived CD34+CD45dim cells and on peripheral blood derived mature lymphocytes (PBLs). We identified three new proteins expressed by fractions of CD34+CD45dim cells and eight new proteins expressed by fractions of PBLs. Remarkably, we identified proteins the presence of which had not been demonstrated previously by transcriptomic analysis. From the functional point of view, looking at new proteins expressed on CD34+CD45dim cells, we identified one cell surface protein (MOSC-1) the expression of which on a minority of CD34+ progenitors marks those CD34+CD45dim cells that will go toward monocyte/granulocyte differentiation. In conclusion, we show a new way of looking at the membranome by assessing expression of generally neglected proteins with a library of polyclonal antisera, and in so doing we have identified new potential subsets of hematopoietic progenitors and of mature PBLs

The N-glycome of human embryonic stem cells

<p>Abstract</p> <p>Background</p> <p>Complex carbohydrate structures, glycans, are essential components of glycoproteins, glycolipids, and proteoglycans. While individual glycan structures including the SSEA and Tra antigens are already used to define undifferentiated human embryonic stem cells (hESC), the whole spectrum of stem cell glycans has remained unknown. We undertook a global study of the asparagine-linked glycoprotein glycans (N-glycans) of hESC and their differentiated progeny using MALDI-TOF mass spectrometric and NMR spectroscopic profiling. Structural analyses were performed by specific glycosidase enzymes and mass spectrometric fragmentation analyses.</p> <p>Results</p> <p>The data demonstrated that hESC have a characteristic N-glycome which consists of both a constant part and a variable part that changes during hESC differentiation. hESC-associated N-glycans were downregulated and new structures emerged in the differentiated cells. Previously mouse embryonic stem cells have been associated with complex fucosylation by use of SSEA-1 antibody. In the present study we found that complex fucosylation was the most characteristic glycosylation feature also in undifferentiated hESC. The most abundant complex fucosylated structures were Le^xand H type 2 antennae in sialylated complex-type N-glycans.</p> <p>Conclusion</p> <p>The N-glycan phenotype of hESC was shown to reflect their differentiation stage. During differentiation, hESC-associated N-glycan features were replaced by differentiated cell-associated structures. The results indicated that hESC differentiation stage can be determined by direct analysis of the N-glycan profile. These results provide the first overview of the N-glycan profile of hESC and form the basis for future strategies to target stem cell glycans.</p

Luonnoksesta kolmiulotteiseksi tilaksi : Virtuaalitilan 3D-mallinnus mobiilialustalle

Tämä tutkielma on osa opinnäytetyötäni. Toteutin opinnäytetyöni produktio-osan CIE:n (Center for Internet Excellence) Chiru-projektissa. Produktion tarkoituksena oli suunnitella ja toteuttaa kolmiulotteinen tila mobiililaitteelle. Kyseessä on rokkiklubi-tyylinen huone, jossa ihmiset voivat liikkua virtuaalihahmoillaan eli avatareillaan sekä esimerkiksi keskustella ja kuunnella musiikkia. Tutkielmani on prosessikuvaus siitä, miten suunnittelin ja toteutin tilan. Tutkielman tavoitteena on kerätä yhteen se oppi, jota projektissa sain, sekä antaa myös muuta tietoa mobiililaitteille mallintamisesta. Alussa kerron siitä, kuinka suunnitteluprosessi toimii ja mitä apukeinoja siinä voi hyödyntää. Loppuosassa kerron mallintamisesta ja siitä, mitä siinä tulee erityisesti ottaa huomioon kun mallinnetaan tilaa mobiilisovellukseen jossa voi olla yhtäaikaisesti useita käyttäjiä omilla avatareillaan. Raportin pohjana toimii tieto ja oppi jota kertyi produktio-osuutta tehdessä. Lisäksi olen käyttänyt kirjallisia lähteitä tukemaan raporttiani. Esille tulee kysymyksiä esimerkiksi siitä, kuinka mobiililaitteiden tehojen asettamat rajat tulevat vastaan, kuinka suuri tila voi olla, kuinka paljon siellä voi olla tavaroita ja kuinka yksityiskohtaista toteutus voi olla. Tutkielman tuloksena tein listauksen siitä, mitä kaikkea tulee ottaa huomioon suunnitellessa ja mallintaessa 3D-tilaa mobiililaitteille. Tärkeimpinä huomioina mittasuhteista sopiminen, avatarien ja käyttäjien määrä, valheellinen tilan tuntu ja sen välttäminen sekä se, kuinka säästää tiedoston koossa tekstuurien ja yhdistettyjen muotojen avulla.This study is a part of my Bachelor’s thesis. The empirical part of the thesis was made in CIE’s (Center for Internet Excellence) Chiru project. The purpose of the empirical part was to design and model a three dimensional (3D) space for mobile platform. The virtual interior is a rock club kind of a room where people can move with their avatars and, for example, chat and listen to music. The study is a description of the process of how the space was designed and modeled. The aim of the thesis was to bring together the knowledge gained through the process and also provide information of how to make a 3D model for a mobile platform. In the beginning of the thesis I will present how the design process works and what techniques can be made use of. Furthermore I will tell about the modeling and what should be taken in to consideration when modeling a space for several avatars. The knowledge and the things learned through the process of the empirical part is the base of this report. Some references were also used in order to support the study. Questions like how will the power usage of the mobile devices affect to the design of the space were discussed; how big can the space be, how many items can there be and how detailed the models can be built. As the results I made a list of the things that should be taken in to consideration when designing and modeling a 3D space for mobile platform. As most important issues, deciding about the measures, figuring out the amount of avatars and users, avoiding false sense of space and how to save in file size using textures and stitched geometry

Akuutin eteisvärikohtauksen hoitotyö : simulaatioharjoitus hoitotyön opiskelijoille

Opinnäytetyöni oli tuotekehitysprojekti, jonka toteutin yhteistyössä Oulun seudun ammattikorkeakoulun sosiaali- ja terveysalan yksikön INNOPI-hankkeen kanssa. INNOPI-hankkeeseen kuuluu simulaatio-opetus, jonka tarkoituksena on edistää opiskelijoiden teknistä ja ei-teknistä osaamista sekä potilasturvallisuutta. Projektini tulostavoitteena oli luoda skenaariosuunnitelma akuutin eteisvärinäkohtauksen hoitotyöstä ja skenaario-suunnitelman pohjalta toteutuva simulaatioharjoitus. Skenaariosuunnitelma laatutavoitteena oli tehdä siitä sisällöltään teoriatietoon perustuva ja käytännönläheinen simulaatioharjoitus, jossa opiskelija voi soveltaa oppimaansa teoriatietoa simulaatiotilanteessa ja harjaantua ei-teknisissä taidoissaan. Skenaariosuunnitelman tein simulaatio-opettajan ohjauksessa ja pohjana omalle tuotteelleni käytin jo käytössä olevaa skenaariosuunnitelmaa. Teknisten taitojen oppimistavoitteiden pohjana käytin omaa opetussuunnitelmaani ja ei-teknisten oppimistavoitteiden pohjana käytin CRM-strategiaa. Simulaatioharjoituksen potilaan esitiedot, oireet, hoidot ja hoidon vasteen suunnittelin hoitotyön ja lääketieteen kirjallisuuden avulla. Skenaariosuunnitelman esitestasin viimeisen vuoden hoitotyön opiskelijoilla ja keräsin heiltä kirjallista palautetta. Esitestauksesta saadun palautteen avulla viimeistelin tuotteeni valmiiksi. Skenaariosuunnitelma oli sitä käyttävien opettajien mielestä opetuskäyttöön soveltuva ja harjoitukseen osallistuneiden opiskelijoiden mielestä heidän teknistä ja ei-teknistä osaamistaan edistävä. Jatkossa skenaariosuunnitelmaani voidaan käyttää simulaatio-opetuksessa ja laajentaa opetustarpeiden mukaan niin, että sitä voidaan käyttää moniammatillisten tiimien taitoja ja yhteistyötä kehittämään. Asiasanat: INNOPPI, potilasturvallisuus, simulaatio-opetus, ei-tekniset taidot, mo-niammatillisuus Patient safety is one of the biggest concerns in the health care field due to ever-complicating scientific advances and technological tools used in nursing treatment. To improve patient safety, Oulu University of Applied Sciences has included new teaching methods into their curriculum. INNOPI-project – short for Innovative Learning Enviroments for Health Education 2010 – is developing new innovative learning environments where examining and treating the patients can be safely practiced. In these simulation like exercises non-technical skills such as leading, teamwork and interaction skills, can be practiced alongside technical skills. This thesis was a product development project, which produced a simulation exercise to be used in the Degree Programme of Nursing. The target group of my product is primarily teachers of nursing and its purpose is to increase the proficiency of the nursing students. I made my simulation exercise in co-operation with a teacher who had experience in simulation teaching and I used broad theoretical knowledge to build a practical simu-lation exercise. To ensure the quality of the product the simulation exercise was pre-tested with nursing students. The scenario plan was then finalized based on their feedback. Scenario plan turned out good and the teachers using it had no troubles understanding or using it as a tool of teaching. According to the students who completed the simulation the exercise had clarified the nursing treatment of acute atrium fibrillation and further advanced their proficiency. My simulation exercise will be used in simulation teaching to improve the proficiency of nursing students and to improve multi-professional co-operation. keywords: patient safety, simulation exercise, non-technical skills, multi-professional co-operatio