The Wnt receptor FZD1 mediates chemoresistance in neuroblastoma through activation of the Wnt/beta-catenin pathway.

The development of chemoresistance represents a major obstacle in the successful treatment of cancers such as neuroblastoma (NB), a particularly aggressive childhood solid tumour. The mechanisms underlying the chemoresistant phenotype in NB were addressed by gene expression profiling of two doxorubicin (DoxR)-resistant vs sensitive parental cell lines. Not surprisingly, the MDR1 gene was included in the identified upregulated genes, although the highest overexpressed transcript in both cell lines was the frizzled-1 Wnt receptor (FZD1) gene, an essential component of the Wnt/beta-catenin pathway. FZD1 upregulation in resistant variants was shown to mediate sustained activation of the Wnt/beta-catenin pathway as revealed by nuclear beta-catenin translocation and target genes transactivation. Interestingly, specific micro-adapted short hairpin RNA (shRNAmir)-mediated FZD1 silencing induced parallel strong decrease in the expression of MDR1, another beta-catenin target gene, revealing a complex, Wnt/beta-catenin-mediated implication of FZD1 in chemoresistance. The significant restoration of drug sensitivity in FZD1-silenced cells confirmed the FZD1-associated chemoresistance. RNA samples from 21 patient tumours (diagnosis and postchemotherapy), showed a highly significant FZD1 and/or MDR1 overexpression after treatment, underlining a role for FZD1-mediated Wnt/beta-catenin pathway in clinical chemoresistance. Our data represent the first implication of the Wnt/beta-catenin pathway in NB chemoresistance and identify potential new targets to treat aggressive and resistant NB.

The Oncogene Metadherin Modulates the Apoptotic Pathway Based on the Tumor Necrosis Factor Superfamily Member TRAIL (Tumor Necrosis Factor-related Apoptosis-inducing Ligand) in Breast Cancer.

Metadherin (MTDH), the newly discovered gene, is overexpressed in more than 40% of breast cancers. Recent studies have revealed that MTDH favors an oncogenic course and chemoresistance. With a number of breast cancer cell lines and breast tumor samples, we found that the relative expression of MTDH correlated with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) sensitivity in breast cancer. In this study, we found that knockdown of endogenous MTDH cells sensitized the MDA-MB-231 cells to TRAIL-induced apoptosis both in vitro and in vivo. Conversely, stable overexpression of MTDH in MCF-7 cells enhanced cell survival with TRAIL treatment. Mechanically, MTDH down-regulated caspase-8, decreased caspase-8 recruitment into the TRAIL death-inducing signaling complex, decreased caspase-3 and poly(ADP-ribose) polymerase-2 processing, increased Bcl-2 expression, and stimulated TRAIL-induced Akt phosphorylation, without altering death receptor status. In MDA-MB-231 breast cancer cells, sensitization to TRAIL upon MTDH down-regulation was inhibited by the caspase inhibitor Z-VAD-fmk (benzyloxycarbonyl-VAD-fluoromethyl ketone), suggesting that MTDH depletion stimulates activation of caspases. In MCF-7 breast cancer cells, resistance to TRAIL upon MTDH overexpression was abrogated by depletion of Bcl-2, suggesting that MTDH-induced Bcl-2 expression contributes to TRAIL resistance. We further confirmed that MTDH may control Bcl-2 expression partly by suppressing miR-16. Collectively, our results point to a protective function of MTDH against TRAIL-induced death, whereby it inhibits the intrinsic apoptosis pathway through miR-16-mediated Bcl-2 up-regulation and the extrinsic apoptosis pathway through caspase-8 down-regulation.

Influence of the pseudopotential on the properties of liquid rubidium at 315 K

The influence of the pseudopotential on both the structure and the self-diffusion of liquid rubidium at the melting point has been investigated by means of molecular-dynamics calculations. The model potential considered has been computed from the pseudopotential of Ashcroft, the dielectric function of Geldart and Vosko, and a Born-Mayer term. Four different values for the core radius which enters as input in the pseudopotential have been considered. In this way we have been able to observe and interpret the effect of this contribution on the properties of the liquid.

Growth promoting signaling by tenascin-C [corrected].

Tenascin-C is an adhesion-modulating extracellular matrix molecule that is highly expressed in tumor stroma and stimulates tumor cell proliferation. Adhesion of T98G glioblastoma cells to a fibronectin substratum is inhibited by tenascin-C. To address the mechanism of action, we performed a RNA expression analysis of T89G cells grown in the presence or absence of tenascin-C and found that tenascin-C down-regulates tropomyosin-1. Upon overexpression of tropomyosin-1, cell spreading on a fibronectin/tenascin-C substratum was restored, indicating that tenascin-C destabilizes actin stress fibers through down-regulation of tropomyosin-1. Tenascin-C also increased the expression of the endothelin receptor type A and stimulated the corresponding mitogen-activated protein kinase signaling pathway, which triggers extracellular signal-regulated kinase 1/2 phosphorylation and c-Fos expression. Tenascin-C additionally caused down-regulation of the Wnt inhibitor Dickkopf 1. In consequence, Wnt signaling was enhanced through stabilization of beta-catenin and stimulated the expression of the beta-catenin target Id2. Finally, our in vivo data derived from astrocytoma tissue arrays link increased tenascin-C and Id2 expression with high malignancy. Because increased endothelin and Wnt signaling, as well as reduced tropomyosin-1 expression, are closely linked to transformation and tumorigenesis, we suggest that tenascin-C specifically modulates these signaling pathways to enhance proliferation of glioma cells.

Produtividade e bienalidade da produção de cafezais adensados, sob diferentes doses de N e K

O objetivo deste trabalho foi avaliar o efeito da adubação com N e K sobre a produtividade e a bienalidade de produção de cafeeiros em plantio adensado, em estudo de longa duração. Foram avaliados dois experimentos, na região da Zona da Mata de Minas Gerais, conduzidos em blocos ao acaso, com parcelas subdivididas no tempo. No primeiro experimento, realizado no espaçamento de 1,5x0,7 m (9.523 plantas por hectare), foram avaliados os efeitos de sete doses de N + K2O na proporção 1:1 (0, 200, 400, 600, 800, 1.000 e 1.200 kg ha‑1 por ano), durante oito safras (2001/2002 a 2008/2009). Como o fertilizante utilizado tinha proporção 1:1, para obter a dose utilizada de cada elemento (N ou K2O), deve-se dividir a dose total por dois. O segundo experimento foi conduzido no espaçamento de 2,5x0,6 m (6.666 plantas por hectare), em arranjo fatorial, com cinco doses de N (0, 150, 300, 450 e 600 kg ha‑1 por ano) e cinco doses de K2O (0, 150, 300, 450 e 600 kg ha‑1 por ano), durante seis safras (2005/2006 a 2010/2011). As doses associadas à máxima produtividade de café beneficiado variaram de 424 a 560 kg ha‑1, para N, e de 21 a 338 kg ha‑1, para K2O. A adubação com N reduz os efeitos da bienalidade da produção do cafeeiro, e a adubação com K, em anos de baixa produtividade, favorece a recuperação da produtividade do cafeeiro no ano seguinte.

Role of P-glycoprotein in the uptake/efflux transport of oral vitamin K antagonists and rivaroxaban through the Caco-2 cell model.

Vitamin K antagonists (VKAs) are prescribed worldwide and remain the oral anticoagulant of choice. These drugs are characterized by a narrow therapeutic index and a large inter- and intra-individual variability. P-glycoprotein could contribute to this variability. The aim of this study was to investigate the involvement of P-gp in the transport of acenocoumarol, phenprocoumon and warfarin using an in vitro Caco-2 cell monolayer model. These results were compared with those obtained with rivaroxaban, a new oral anticoagulant known to be a P-gp substrate. The transport of these four drugs was assessed at pH conditions 6.8/7.4 in the presence or absence of the P-gp inhibitor cyclosporine A (10 μM) and the more potent and specific P-gp inhibitor valspodar (5 μM). Analytical quantification was performed by LC/MS. With an efflux ratio of 1.7 and a significant decrease in the efflux (Papp B-A), in the presence of P-gp inhibitors at a concentration of 50 μM, acenocoumarol can be considered as a weak P-gp substrate. Concerning phenprocoumon, the results suggest that this molecule is a poor P-gp substrate. The P-gp inhibitors did not affect significantly the transport of warfarin. The efflux of rivaroxaban was strongly inhibited by the two P-gp inhibitors. In conclusion, none of the three VKAs tested are strong P-gp substrates. However, acenocoumarol can be considered as a weak P-gp substrate and phenprocoumon as a poor P-gp substrate.

Aproximación al reajuste automático de centroides mediante la heurística de Lloyd para resolver el problema de las K-Medias

Peer-reviewed

The caspase-8 inhibitor FLIP promotes activation of NF-kappaB and Erk signaling pathways.

BACKGROUND: Activation of Fas (CD95) by its ligand (FasL) rapidly induces cell death through recruitment and activation of caspase-8 via the adaptor protein Fas-associated death domain protein (FADD). However, Fas signals do not always result in apoptosis but can also trigger a pathway that leads to proliferation. We investigated the level at which the two conflicting Fas signals diverge and the protein(s) that are implicated in switching the response. RESULTS: Under conditions in which proliferation of CD3-activated human T lymphocytes is increased by recombinant FasL, there was activation of the transcription factors NF-kappaB and AP-1 and recruitment of the caspase-8 inhibitor and FADD-interacting protein FLIP (FLICE-like inhibitory protein). Fas-recruited FLIP interacts with TNF-receptor associated factors 1 and 2, as well as with the kinases RIP and Raf-1, resulting in the activation of the NF-kappaB and extracellular signal regulated kinase (Erk) signaling pathways. In T cells these two signal pathways are critical for interleukin-2 production. Increased expression of FLIP in T cells resulted in increased production of interleukin-2. CONCLUSIONS: We provide evidence that FLIP is not simply an inhibitor of death-receptor-induced apoptosis but that it also mediates the activation of NF-kappaB and Erk by virtue of its capacity to recruit adaptor proteins involved in these signaling pathways.

c-Myc controls the balance between hematopoietic stem cell self-renewal and differentiation.

The activity of adult stem cells is essential to replenish mature cells constantly lost due to normal tissue turnover. By a poorly understood mechanism, stem cells are maintained through self-renewal while concomitantly producing differentiated progeny. Here, we provide genetic evidence for an unexpected function of the c-Myc protein in the homeostasis of hematopoietic stem cells (HSCs). Conditional elimination of c-Myc activity in the bone marrow (BM) results in severe cytopenia and accumulation of HSCs in situ. Mutant HSCs self-renew and accumulate due to their failure to initiate normal stem cell differentiation. Impaired differentiation of c-Myc-deficient HSCs is linked to their localization in the differentiation preventative BM niche environment, and correlates with up-regulation of N-cadherin and a number of adhesion receptors, suggesting that release of HSCs from the stem cell niche requires c-Myc activity. Accordingly, enforced c-Myc expression in HSCs represses N-cadherin and integrins leading to loss of self-renewal activity at the expense of differentiation. Endogenous c-Myc is differentially expressed and induced upon differentiation of long-term HSCs. Collectively, our data indicate that c-Myc controls the balance between stem cell self-renewal and differentiation, presumably by regulating the interaction between HSCs and their niche.

[Kepek a Keleti-Karpatokbol... Bilder aus den Ostkarpaten... Budapest, Birkholz, Vayda und Preszburg, s.d. Album de 32 phot. des Carpathes orientales par K. Divald, donateur en 1888]

Comprend : Ó-Kemenczei gereb. Unghmagye ; A Popp Ivan (1940 m), Szerban (1795 m) és Poloninka (1625 m) a Precsulka elöhavasról. Máramorosmegye ; Szaláncz várromjai az Ujhely-Kassai vasut vonalán. Albanjmegye ; Crachyt szikla a "Szinnai kö" tetején (998 méter), Zemplén és Unghmegye Határán

Jätehuoltosuunnitelma Pohjois-Karjalan keskussairaalan laajennus K:ssa

Tämän työn tarkoituksena oli suunnitella toimiva jätehuoltoPohjois-Karjalan keskussairaalan alueelle vuosina 2007-2008 rakennettavaan laajennus K:hon. Tavoitteena oli löytää kustannustehokas ja hyvin vanhan rakennuskannan logistiikan kanssa yhteen toimiva jätehuollon vaihtoehto. Suunnittelua ohjasivat lakien, asetusten ja muiden määräyksien ohella laajennuksen pitkälle edennyt suunnitteluvaihe ja vanhan rakennuskannan jätehuollon toimintatavat. Jätehuolto on yksi merkittävä osa sairaalan perustehtävää palvelevista tukitoiminnoista. Jätehuoltojärjestelmä on sairaanhoitotyötä palveleva, jos järjestelyt ovat loogisia, toimivia ja logistiset yhteydet ovat mahdollisimman lyhyitä. Nykyinen ihmistyövoiman käyttöön perustuva jätehuollon prosessi on esitetty varsin tarkasti, sillä laajennuksen jätehuoltosuunnitelman on käytännön syistä pohjauduttava nykyiseen toimintatapaan. Työssä esitetään uusia jätehuollon vaihtoehtoja kustannusarvioineen, mutta ehdotukset jäävät lähinnä visiotasolle laajennuksen pitkälle ehtineen suunnitteluvaiheen vuoksi. Konkreettiset vaihtoehdot koskevat jätejakeiden keräystä ja niiden kuljetuksia. Keskus sairaalan jätehuollon tehostaminen vaatii kokonaisvaltaisen jätehuoltosuunnitelman laatimista koko sairaalan alueelle. Sen myötä on mahdollista lisätä kustannustehokkuutta ottamalla käyttöön teknisiä ratkaisuja. Jätehuoltosuunnitelma laajennukseen on työssä tehty, mutta työn suurimmaksi saavutukseksi jää kuitenkin esitys myöhempien hakkeiden jätehuollon suunnittelun aikaistamisesta jo hankesuunnitteluvaiheeseen.

Tieliikelaitoksen t&k -projektien kannattavuuden arviointi

Työn tavoitteena oli laatia Tieliikelaitoksen tutkimus- ja kehitysprojektien kannattavuuden arvioinnin toimintamalli ja laskentamalli. Työn tavoitteena oli myös kehittääTieliikelaitoksen tutkimus- ja kehitysprojektin kannattavuuden arviointia ja tuottaa informaatiota t&k -projektien valintatilanteisiin. Työ koostuu teoriasta, haastatteluista, t&k -projektien kannattavuuden arvioinnin toimintamallista ja laskentamallista. Työssä käsitellään kirjallisuudesta ja haastatteluista esille tulleita käytäntöjä ja menetelmiä, joiden avulla tutkimus- ja kehitysprojektien kannattavuutta voidaan arvioida. Työssä kuvataan myös arviointimenetelmien käyttöä t&k -prosessin eri vaiheissa. Työn merkittävimpinä tuloksina ovat laaditut toimintamalli ja laskentamalli Tieliikelaitoksen t&k -projektien kannattavuuden arviointiin. Lisäksi työssä tuloksena oli, että arviointimenetelmiä tulee käyttää monipuolisesti ja projektien arviointi tulee olla jatkuvaa. Projektien kannattavuuden arviointi ei voi perustua pelkästään taloudellisiin menetelmiin vaan arviointiin tulee valita sekä taloudellisia että laadullisia menetelmiä. Projektien jatkuvalla arvioinnilla ja arviointimenetelmien käytön monipuolisuudella varmistetaan oikeiden ja kannattavien projektien toteuttaminen.

Phosphorylation of phosphatidylinositol-3-kinase-protein kinase B and extracellular signal-regulated kinases 1/2 mediate reoxygenation-induced cardioprotection during hypoxia.

In vivo exposure to chronic hypoxia (CH) depresses myocardial performance and tolerance to ischemia, but daily reoxyenation during CH (CHR) confers cardioprotection. To elucidate the underlying mechanism, we tested the role of phosphatidylinositol-3-kinase-protein kinase B (Akt) and p42/p44 extracellular signal-regulated kinases (ERK1/2), which are known to be associated with protection against ischemia/reperfusion (I/R). Male Sprague-Dawley rats were maintained for two weeks under CH (10% O(2)) or CHR (as CH but with one-hour daily exposure to room air). Then, hearts were either frozen for biochemical analyses or Langendorff-perfused to determine performance (intraventricular balloon) and tolerance to 30-min global ischemia and 45-min reperfusion, assessed as recovery of performance after I/R and infarct size (tetrazolium staining). Additional hearts were perfused in the presence of 15 micromol/L LY-294002 (inhibitor of Akt), 10 micromol/L UO-126 (inhibitor of ERK1/2) or 10 micromol/L PD-98059 (less-specific inhibitor of ERK1/2) given 15 min before ischemia and throughout the first 20 min of reperfusion. Whereas total Akt and ERK1/2 were unaffected by CH and CHR in vivo, in CHR hearts the phosphorylation of both proteins was higher than in CH hearts. This was accompanied by better performance after I/R (heart rate x developed pressure), lower end-diastolic pressure and reduced infarct size. Whereas the treatment with LY-294002 decreased the phosphorylation of Akt only, the treatment with UO-126 decreased ERK1/2, and that with PD-98059 decreased both Akt and ERK1/2. In all cases, the cardioprotective effect led by CHR was lost. In conclusion, in vivo daily reoxygenation during CH enhances Akt and ERK1/2 signaling. This response was accompanied by a complex phenotype consisting in improved resistance to stress, better myocardial performance and lower infarct size after I/R. Selective inhibition of Akt and ERK1/2 phosphorylation abolishes the beneficial effects of the reoxygenation. Therefore, Akt and ERK1/2 have an important role to mediate cardioprotection by reoxygenation during CH in vivo.