Optimization of immunomagnetic separation for cord blood-derived hematopoietic stem cells

BACKGROUND: There is a growing interest in cord blood as a source of primitive stem cells with the capacity for multilineage differentiation. Pure cell fractions are needed for the characterization and in vitro expansion of stem cells as well as for their use in preclinical research. However, enrichment of stem cells is challenging due to the lack of stem cell-specific markers and gentle protocols for the isolation of highly pure stem cell fractions. Protocols developed for the enrichment of peripheral blood-derived stem cells have been found to be suboptimal for cord blood. RESULTS: In this study, immunomagnetic cell sorting protocols to purify CD34+, CD133+ and Lin- cells from fresh and cryopreserved cord blood were optimized. Reproducible purities of up to 97% were reached. The selected cells were highly viable having substantial colony-forming potential. CONCLUSION: The optimized protocols enable rapid enrichment of highly pure hematopoietic stem cells from both fresh and cryopreserved cord blood

Shorter Cold Ischemia Time in Deceased Donor Kidney Transplantation Reduces the Incidence of Delayed Graft Function Especially Among Highly Sensitized Patients and Kidneys From Older Donors

Background. Long cold ischemia time (CIT) is the most important factor contributing to delayed graft function (DGF) after kidney transplant. Improvements in pretransplant procedures may reduce CIT and improve clinical outcome. Materials and Methods. Pretransplant histocompatibility tests were modernized at our laboratory in 2015, leading to significant decrease of time consumed for these enabling earlier surgery. The effects of this on kidney transplant CIT, DGF, and other clinical outcomes were studied. The study population consisted of 896 consecutive deceased donor kidney recipients, of which 442 patients received a transplant with the old crossmatch and 454 received a transplant with the new crossmatch. Results. CIT shortened from mean 20 hours 6 minutes to 15 hours 52 minutes (P <.001). The incidence of DGF was significantly reduced from 31% to 24% (P = .02). Reduction in the frequency of DGF was more pronounced among the highly sensitized patients (53% to 28%, P = .01) or in patients with pretransplant donor-specific antibodies (50% to 20%, P = .002) and among patients who received kidneys from donors older than 65 years (38% to 27%, P = .04). Conclusions. Process optimization that reduces CIT decreases occurrence of DGF, especially in highly sensitized patients and patients who receive kidneys from older donors.Peer reviewe

Febuxostat, But Not Allopurinol, Markedly Raises the Plasma Concentrations of the Breast Cancer Resistance Protein Substrate Rosuvastatin

Xanthine oxidase inhibitors febuxostat and allopurinol are commonly used in the treatment of gout. Febuxostat inhibits the breast cancer resistance protein (BCRP) in vitro. Rosuvastatin is a BCRP substrate and genetic variability in BCRP markedly affects rosuvastatin pharmacokinetics. In this study, we investigated possible effects of febuxostat and allopurinol on rosuvastatin pharmacokinetics. In a randomized crossover study with 3 phases, 10 healthy volunteers ingested once daily placebo for 7 days, 300 mg allopurinol for 7 days, or placebo for 3 days, followed by 120 mg febuxostat for 4 days, and a single 10 mg dose of rosuvastatin on day 6. Febuxostat increased the peak plasma concentration and area under the plasma concentration-time curve of rosuvastatin 2.1-fold (90% confidence interval 1.8-2.6; P = 5 x 10(-5)) and 1.9-fold (1.5-2.5; P = 0.001), but had no effect on rosuvastatin half-life or renal clearance. Allopurinol, on the other hand, did not affect rosuvastatin pharmacokinetics. In vitro, febuxostat inhibited the ATP-dependent uptake of rosuvastatin into BCRP-overexpressing membrane vesicles with a half-maximal inhibitory concentration of 0.35 mu M, whereas allopurinol showed no inhibition with concentrations up to 200 mu M. Taken together, the results suggest that febuxostat increases rosuvastatin exposure by inhibiting its BCRP-mediated efflux in the small intestine. Febuxostat may, therefore, serve as a useful index inhibitor of BCRP in drug-drug interaction studies in humans. Moreover, concomitant use of febuxostat may increase the exposure to BCRP substrate drugs and, thus, the risk of dose-dependent adverse effects.Peer reviewe

Anti-COX-2 autoantibody is a novel biomarker of immune aplastic anemia

In immune aplastic anemia (IAA), severe pancytopenia results from the immune-mediated destruction of hematopoietic stem cells. Several autoantibodies have been reported, but no clinically applicable autoantibody tests are available for IAA. We screened autoantibodies using a microarray containing >9000 proteins and validated the findings in a large international cohort of IAA patients (n = 405) and controls (n = 815). We identified a novel autoantibody that binds to the C-terminal end of cyclooxygenase 2 (COX-2, aCOX-2 Ab). In total, 37% of all adult IAA patients tested positive for aCOX-2 Ab, while only 1.7% of the controls were aCOX-2 Ab positive. Sporadic non-IAA aCOX-2 Ab positive cases were observed among patients with related bone marrow failure diseases, multiple sclerosis, and type I diabetes, whereas no aCOX-2 Ab seropositivity was detected in the healthy controls, in patients with non-autoinflammatory diseases or rheumatoid arthritis. In IAA, anti-COX-2 Ab positivity correlated with age and the HLA-DRB1*15:01 genotype. 83% of the >40 years old IAA patients with HLA-DRB1*15:01 were anti-COX-2 Ab positive, indicating an excellent sensitivity in this group. aCOX-2 Ab positive IAA patients also presented lower platelet counts. Our results suggest that aCOX-2 Ab defines a distinct subgroup of IAA and may serve as a valuable disease biomarker.Peer reviewe

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

Vauvan perushoitovideoita : kylvetys, vaipanvaihto ja vauvahieronta

Perheenlisäys on aina uusi ja jännittävä asia. Jo raskausaikana isillä on mahdollisuus tutustua syntyvään lapseen tunnustelemalla sikiön liikkeitä äidin vatsanpeitteiden päältä sekä ultraäänikuvien välityksellä. Varhaisen vuorovaikutuksen tukeminen on ensiarvoisen tärkeää vauvan kiintymyssuhteen kehittymisen kannalta. (Korhonen 2009: 311.) Vastasyntyneellä on synnynnäiset valmiudet olla kanssakäymisessä vanhempiensa kanssa (Stern 1982: 16, 45). Isien mielestä on mielenkiintoista seurata lapsen kasvua ja kehitystä, ja he toivoivat saavansa kahden keskistä aikaa luodakseen suhdetta omaan lapseen. (Premperg ym. 2008.) Vuorovaikutuksen ohella yhtä tärkeää on huolehtia vauvan päivittäisistä tarpeista. Joka neljäs isä on kokenut vanhemmuutensa taidot puutteellisiksi. Erityisesti isät kaipasivat tietoa lapsen tarpeista ja turvallisuudesta huolehtimiseen sekä yksilöllistä ohjausta vauvan hoidosta. (Puputti-Rantsi 2009: 39; Mesiäislehto-Soukka 2005: 124.) Tämä opinnäytetyö on tehty osana Urbaani vanhemmuus -hanketta. Urbaani vanhemmuus -hankkeen tavoitteena on kehittää verkkosivusto, joka tukee vanhemmuutta ja edistää perheiden hyvinvointia. Tämä opinnäytetyö on jatkoa aiemmin hankkeeseen tehdyille opin-näytetöille, joissa on selvitetty käyttäjien eli tässä tapauksessa isien toiveita sivuston sisällöstä. Tämän opinnäytetyön tarkoituksena on tuottaa kolme vauvan hoito -videota, joiden tavoitteena on antaa erityisesti ensimmäisen lapsen isille apua ja vinkkejä pienen lapsen hoitoon kotona. Videoiden tavoitteena on kuvata mahdollisimman käytännönläheisesti vauvan kylvetystä, vaipan vaihtoa ja vuorovaikutusta vauvahieronnan kautta. Nämä videot edistävät isien kasvua vanhemmuuteen, tukevat lapsivuodeosastolta saatavaa ohjausta ja antavat valmiuksia toimia vauvan kanssa kotona. Videot ovat myös hyvin sovellettavissa käytännön neuvola- sekä lapsivuodeosastotyön rinnalle ja hoitoalan opiskelijoiden opetukseen.Having a baby is always a new situation for a family. During the pregnancy the father can form a relationship with his baby by feeling the baby’s movements and participating in ultra-sound examinations. Supporting early interaction between the baby and his/her parents is one of the most important aims in early childhood. (Korhonen 2009 :311.) A newborn has a readiness to interact with his/her parents (Stern 1982: 16,45). Fathers find it fascinating to follow their baby’s growth and development, and they want to spend more time together with their babies. (Premperg ym. 2008.) It is also important to take care of baby’s basic needs such as changing diapers, bathing and interacting with the newborn. As much as one on four fathers find their baby caring skills to be inadequate. These fathers especially wish to receive individual assistance in caring for their babies. (Puputti-Rantsi 2009: 39; Mesiäislehto-Soukka 2005: 124.) This thesis is made in co-operation with “Urban parenting” -project. The aim of the “Urban parenting” -project is to create a website for families. The website will support parenthood, health and welfare of families. This thesis is a follow-up of an earlier thesis made within the same project, in which the fathers’ needs and wishes for the website have been studied. The aim of this thesis is to produce three videos of baby care. The objective of the videos is to help, guide and give practical tips in baby care for the fathers of newborns. The goal is to assist fathers in their growth to parenthood, to support the guidance given by maternity hospitals and prepare fathers and families to take care of their babies at home. The videos are also usable in neuvola, maternity hospitals, and to students of nursing or midwifery

Toiminnallinen hyvinvointipäivä : hyvinvoiva nainen

Toteutimme toiminnallisen opinnäytetyömme naisille suunnattuna hyvinvointitapahtumana, jolle annoimme nimeksi Hyvinvoiva nainen. Tapahtuman tavoitteena oli edistää Metropolia Ammattikorkeakoulun Tukholmankadun toimipisteen naisopiskelijoiden psyykkistä ja fyysistä hyvinvointia lähestymällä asiaa seksuaali- ja lisääntymisterveyden näkökulmasta. Järjestimme tapahtuman yhteistyöryhmämme kanssa Metropolia Ammattikorkeakoulun Tukholmankadun toimipisteen D-auditoriossa 15. huhtikuuta vuonna 2014. Jaoimme audi-torion neljään toiminnalliseen pisteeseen, jotka olivat rentoutumis-, voimaantumis- ja liikun-tapiste sekä yhteistyöryhmämme seksuaali- ja lisääntymisterveydenpiste. Jokaisen pisteen tarkoituksena oli edistää tapahtumaan osallistujien seksuaali- ja lisääntymisterveyttä kyseisen hyvinvoinnin osa-alueeseen liittyvän toiminnan ja tiedon kautta. Hyvinvointia sekä seksuaali- ja lisääntymisterveyttä voi edistää rentoutumisen, liikunnan ja voimaantumisen kautta. Säännöllinen liikunta ehkäisee, vähentää ja poistaa seksuaalitoiminnan häiriöitä sekä parantaa hedelmällisyyttä. Stressi heikentää seksuaalista halukkuutta, mutta rentoutusharjoitteilla voidaan lieventää stressiä. Tällöin myös seksuaalinen halukkuus lisääntyy. Seksuaalinen halukkuus on todennäköisemmin voimakkaampaa ihmisillä, joilla on hyvä itsetunto. Oman itsensä hyväksyminen sekä itsensä arvostaminen edistää seksuaalista voimantumista ja myös psyykkistä ja fyysistä hyvinvointia. Saamamme palautteen perusteella onnistuimme Hyvinvoiva nainen -tapahtumalla innostamaan, piristämään, hauskuuttamaan ja voimaannuttamaan suurinta osaa osallistujista. Lisäksi yksittäinen henkilö kertoi tapahtumaan osallistumisen tuloksesi uuden oppimisen. Palautteiden perusteella opinnäytetyöryhmämme ei kuitenkaan saa tietää, edistikö Hyvin-voiva nainen -tapahtuma osallistujien pitkäaikaista psyykkistä ja fyysistä hyvinvointia.We executed our functional thesis as a well-being event for women and named it Hyvinvoiva nainen. The aim of this event was to advance the psychic and physical well-being of female students at Metropolia University of Applied Sciences in Tukholmankatu and do this from the sexual and reproductive health´s point of view. We organized the event with our cooperation group on 15th of April 2014 in the D-auditorium of Tukholmankatu´s office of Metropolia University of Applied Sciences. The auditorium was separated in four functional spots that were relaxation, empowerment and exercise plus the sexual and reproductive health spot of our cooperation group. The aim of each spot was to advance the participants sexual and reproductive health through action and knowledge connected to each field. Well-being and sexual and reproductive health can be advanced by relaxation, exercise and empowerment. Regular exercising prevents, decreases, and removes sexual disorders and improves fertility. Stress can deteriorate sexual willingness but stress can be mitigated with relaxation exercises. Thus also sexual willingness increases. Sexual willingness is higher among people with high self-esteem. A key to healthy female sexuality is that women accept themselves and respect themselves as women. That affects also to psychological and physical well-being. Based on the feedback that we got, Hyvinvoiva nainen -event inspired, cheered up, amused and empowered many of the participants. Also one person had written that they had learned something new in the event. However based on the feedback the group will not know if the event advanced the psychic and physical well-being of the participants

Optimization of immunomagnetic separation for cord blood-derived hematopoietic stem cells

Abstract Background There is a growing interest in cord blood as a source of primitive stem cells with the capacity for multilineage differentiation. Pure cell fractions are needed for the characterization and in vitro expansion of stem cells as well as for their use in preclinical research. However, enrichment of stem cells is challenging due to the lack of stem cell-specific markers and gentle protocols for the isolation of highly pure stem cell fractions. Protocols developed for the enrichment of peripheral blood-derived stem cells have been found to be suboptimal for cord blood. Results In this study, immunomagnetic cell sorting protocols to purify CD34+, CD133+ and Lin- cells from fresh and cryopreserved cord blood were optimized. Reproducible purities of up to 97% were reached. The selected cells were highly viable having substantial colony-forming potential. Conclusion The optimized protocols enable rapid enrichment of highly pure hematopoietic stem cells from both fresh and cryopreserved cord blood.</p