CUP-1 Is a Novel Protein Involved in Dietary Cholesterol Uptake in Caenorhabditis elegans

Sterols transport and distribution are essential processes in all multicellular organisms. Survival of the nematode Caenorhabditis elegans depends on dietary absorption of sterols present in the environment. However the general mechanisms associated to sterol uptake in nematodes are poorly understood. In the present work we provide evidence showing that a previously uncharacterized transmembrane protein, designated Cholesterol Uptake Protein-1 (CUP-1), is involved in dietary cholesterol uptake in C. elegans. Animals lacking CUP-1 showed hypersensitivity to cholesterol limitation and were unable to uptake cholesterol. A CUP-1-GFP fusion protein colocalized with cholesterol-rich vesicles, endosomes and lysosomes as well as the plasma membrane. Additionally, by FRET imaging, a direct interaction was found between the cholesterol analog DHE and the transmembrane “cholesterol recognition/interaction amino acid consensus” (CRAC) motif present in C. elegans CUP-1. In-silico analysis identified two mammalian homologues of CUP-1. Most interestingly, CRAC motifs are conserved in mammalian CUP-1 homologous. Our results suggest a role of CUP-1 in cholesterol uptake in C. elegans and open up the possibility for the existence of a new class of proteins involved in sterol absorption in mammals

The Roles of Cholesterol and Its Metabolites in Normal and Malignant Hematopoiesis.

Hematopoiesis is sustained throughout life by hematopoietic stem cells (HSCs) that are capable of self-renewal and differentiation into hematopoietic progenitor cells (HPCs). There is accumulating evidence that cholesterol homeostasis is an important factor in the regulation of hematopoiesis. Increased cholesterol levels are known to promote proliferation and mobilization of HSCs, while hypercholesterolemia is associated with expansion of myeloid cells in the peripheral blood and links hematopoiesis with cardiovascular disease. Cholesterol is a precursor to steroid hormones, oxysterols, and bile acids. Among steroid hormones, 17β-estradiol (E2) induces HSC division and E2-estrogen receptor α (ERα) signaling causes sexual dimorphism of HSC division rate. Oxysterols are oxygenated derivatives of cholesterol and key substrates for bile acid synthesis and are considered to be bioactive lipids, and recent studies have begun to reveal their important roles in the hematopoietic and immune systems. 27-Hydroxycholesterol (27HC) acts as an endogenous selective estrogen receptor modulator and induces ERα-dependent HSC mobilization and extramedullary hematopoiesis. 7α,25-dihydroxycholesterol (7α,25HC) acts as a ligand for Epstein-Barr virus-induced gene 2 (EBI2) and directs migration of B cells in the spleen during the adaptive immune response. Bile acids serve as chemical chaperones and alleviate endoplasmic reticulum stress in HSCs. Cholesterol metabolism is dysregulated in hematologic malignancies, and statins, which inhibit de novo cholesterol synthesis, have cytotoxic effects in malignant hematopoietic cells. In this review, recent advances in our understanding of the roles of cholesterol and its metabolites as signaling molecules in the regulation of hematopoiesis and hematologic malignancies are summarized

2015/16 seasonal vaccine effectiveness against hospitalisation with influenza a(H1N1)pdm09 and B among elderly people in Europe: Results from the I-MOVE+ project

We conducted a multicentre test-negative caseâ\u80\u93control study in 27 hospitals of 11 European countries to measure 2015/16 influenza vaccine effectiveness (IVE) against hospitalised influenza A(H1N1)pdm09 and B among people aged â\u89¥ 65 years. Patients swabbed within 7 days after onset of symptoms compatible with severe acute respiratory infection were included. Information on demographics, vaccination and underlying conditions was collected. Using logistic regression, we measured IVE adjusted for potential confounders. We included 355 influenza A(H1N1)pdm09 cases, 110 influenza B cases, and 1,274 controls. Adjusted IVE against influenza A(H1N1)pdm09 was 42% (95% confidence interval (CI): 22 to 57). It was 59% (95% CI: 23 to 78), 48% (95% CI: 5 to 71), 43% (95% CI: 8 to 65) and 39% (95% CI: 7 to 60) in patients with diabetes mellitus, cancer, lung and heart disease, respectively. Adjusted IVE against influenza B was 52% (95% CI: 24 to 70). It was 62% (95% CI: 5 to 85), 60% (95% CI: 18 to 80) and 36% (95% CI: -23 to 67) in patients with diabetes mellitus, lung and heart disease, respectively. 2015/16 IVE estimates against hospitalised influenza in elderly people was moderate against influenza A(H1N1)pdm09 and B, including among those with diabetes mellitus, cancer, lung or heart diseases

Vanhempien osallisuus varhaiskasvatuksessa

Tämän opinnäytetyön tavoitteena oli selvittää vanhempien kokemuksia osallisuudesta varhaiskasvatuksessa. Toimeksiantajanamme oli Rekulan päiväkoti Nurmeksessa. Teimme opinnäytetyön laadullisena tutkimuksena. Aineistonkeruumenetelmänä käy-timme paperista kyselylomaketta, jonka kysymyksissä hyödynsimme laadullisen tutki-muksen aineistonkeruumenetelmiä. Päiväkodin henkilökunta jakoi kyselylomakkeet lasten vanhemmille täytettäviksi ja me kävimme hakemassa täytetyt lomakkeet analysoi-tavaksi. Käytimme analyysimenetelmänä aineistolähtöistä sisällönanalyysia. Tutkimuksessa selvisi, että vanhemmat ovat pääosin todella tyytyväisiä tämänhetkisiin osallistamismenetelmiin päiväkodissa. Vanhemmat haluaisivat osallistua jatkossa enemmän päiväkodissa järjestettävään toimintaan. Vain pieni osa vanhemmista halu-aisi osallistua toiminnan suunnitteluun. Vanhempien mukaan päiväkodissa on avoin ja lämminhenkinen ilmapiiri, joka vaikuttaa yhteistyön laatuun. Vanhemmat kokevat yh-teistyön ja osallisuuden tärkeänä asiana osana varhaiskasvatusta. Tutkimustuloksia voidaan hyödyntää jatkossa osallisuuden kehittämistä varten.The aim of this thesis was to investigate the parents’ experiences in participation in ear-ly childhood education. The client was Rekula day-care center in Nurmes. The study was qualitative in nature. The data was collected through a questionnaire, which included qualitative data collection methods. The questionnaires were delivered to the parents by the staff and the filled in questionnaires were then picked up for ana-lysis by us. The method used was data-oriented content analysis. It was discovered that the parents were, in general, very satisfied with the current met-hods of participation in early childhood education used at the day-care center. The pa-rents would like to participate more in the activities of the day-care center. However, on-ly a small percentage of the parents would like to participate in the planning of activi-ties. According to the parents, the atmosphere in the day-care center is open and warm, which has a positive effect in co-operation with the educators. The parents also consi-der co-operation and participation as important factors in early childhood education. The results of this thesis can be utilized in further development of parents’ participation in early childhood education

Mammografialaitteen tekninen laadunvarmistus : Itseopiskelumateriaali osana e-Breast hanketta

Mammografiatutkimus on yksi osa rintasyövän diagnostiikassa. Kuvantamisen onnis-tumisen takaamiseksi laitteiden tulee olla teknisesti hyvässä kunnossa. Tätä seurataan päivittäin, viikoittain, kuukausittain ja vuosittain tehtävillä laadunvarmistustesteillä. Opinnäytetyössämme keskityimme Oulun yliopistollisen sairaalan Avohoitotalon GE:n mammografialaitteen laadunvarmistukseen. Perehdyimme röntgenhoitajan tekemiin testeihin. Opinnäytetyömme tuotoksena teimme mammografialaitteen laadunvarmistuksesta itseopiskelumateriaalin, jota varten kuvasimme jokaisesta laatutestistä videon. Videoilla näytetään, kuinka testit tehdään, ja mitä niistä tarkastellaan. Projektin tavoitteena oli lisätä röntgenhoitajien tietämystä mammografialaitteen laadunvarmistuksesta. Materiaalia voidaan käyttää laitteen perehdyttämiseen. Tilaaja oli eBreast –hanke. Itseopiskelumateriaali on tehty englannin kielellä. Tuotteen kehittelyssä halusimme keskittyä tuotteen käyttäjäläheisyyteen ja pyrimme kohdentamaan videon sekä röntgenhoitajaopiskelijoille että jo valmistuneille röntgen-hoitajille. Itseopiskelumateriaalin esitestasivat radiografian ja sädehoidon opiskelijat. Testaajat olivat tyytyväisiä materiaaliin, lisäsimme kommenttien perusteella vide-oihin tekstit, jotka kertovat, mitä tehdään. Jatkokehitysehdotuksemme on tehdä opinnäytetyö, jossa harjoitellaan laadunvarmistustestien tuloksien arvioimista.There are image quality tests to make sure that image quality in mammography is as good as possible. Tests must be done daily, weekly or/and monthly. Radiographers perform the tests. Our thesis was made as a product development project and was ordered by the eBreast. The aim of this project was to make a self-study material about image quality assurance in mammography for radiographers and student radiographers. The content of the material was focused on mammography image quality assurance tests and the material was directed to radiographers. The tests were done by the GE mammography device at Avohoitotalo´s mammography unit. The material was made in English. Our thesis was made as product development project. The information was checked by professionals and the material was pre-tested by the students of the degree programme in radiography and radiation therapy. The group that tested the material evaluated the layout, the content and the language of the material. The material was modified according to the feedback. The project could be developed further by making self-study material for how to eval-uate the Quality assurance tests in mammography.Itseopiskelumateriaali on osa eBreast -hanketta ja tulee hankkeen open acces -materiaaleihin

Electrophysiological characterization of the SK channel blockers methyl-laudanosine and methyl-noscapine in cell lines and rat brain slices

1. We have recently shown that the alkaloid methyl-laudanosine blocks SK channel-mediated afterhyperpolarizations (AHPs) in midbrain dopaminergic neurones. However, the relative potency of the compound on the SK channel subtypes and its ability to block AHPs of other neurones were unknown. 2. Using whole-cell patch-clamp experiments in transfected cell lines, we found that the compound blocks SK1, SK2 and SK3 currents with equal potency: its mean IC(50)s were 1.2, 0.8 and 1.8 μM, respectively. IK currents were unaffected. In rat brain slices, methyl-laudanosine blocked apamin-sensitive AHPs in serotonergic neurones of the dorsal raphe and noradrenergic neurones of the locus coeruleus with IC(50)s of 21 and 19 μM, as compared to 15 μM in dopaminergic neurones. However, at 100 μM, methyl-laudanosine elicited a constant hyperpolarization of serotonergic neurones of about 9 mV, which was inconsistently (i.e. not in a reproducible manner) antagonized by atropine and hence partly due to the activation of muscarinic receptors. 3. While exploring the pharmacology of related compounds, we found that methyl-noscapine also blocked SK channels. In cell lines, methyl-noscapine blocked SK1, SK2 and SK3 currents with mean IC(50)s of 5.9, 5.6 and 3.9 μM, respectively. It also did not block IK currents. Methyl-noscapine was slightly less potent than methyl-laudanosine in blocking AHPs in brain slices, its IC(50)s being 42, 37 and 29 μM in dopaminergic, serotonergic and noradrenergic neurones, respectively. Interestingly, no significant non-SK effects were observed with methyl-noscapine in slices. At a concentration of 300 μM, methyl-noscapine elicited the same changes in excitability in the three neuronal types than did a supramaximal concentration of apamin (300 nM). 4. Methyl-laudanosine and methyl-noscapine produced a rapidly reversible blockade of SK channels as compared with apamin. The difference between the IC(50)s of apamin (0.45 nM) and methyl-laudanosine (1.8 μM) in SK3 cells was essentially due to a major difference in their k(−1) (0.028 s(−1) for apamin and ⩾20 s(−1) for methyl-laudanosine). 5. These experiments demonstrate that both methyl-laudanosine and methyl-noscapine are medium potency, quickly dissociating, SK channel blockers with a similar potency on the three SK subtypes. Methyl-noscapine may be superior in terms of specificity for the SK channels

Mechanisms of lysosomal positioning and movement

Chemical Immunolog