RNA interference screening identifies a novel role for PCTK1/CDK16 in medulloblastoma with c-Myc amplification.

Medulloblastoma (MB) is the most common malignant brain tumor in children and is associated with a poor outcome. cMYC amplification characterizes a subgroup of MB with very poor prognosis. However, there exist so far no targeted therapies for the subgroup of MB with cMYC amplification. Here we used kinome-wide RNA interference screening to identify novel kinases that may be targeted to inhibit the proliferation of c-Myc-overexpressing MB. The RNAi screen identified a set of 5 genes that could be targeted to selectively impair the proliferation of c-Myc-overexpressing MB cell lines: AKAP12 (A-kinase anchor protein), CSNK1α1 (casein kinase 1, alpha 1), EPHA7 (EPH receptor A7) and PCTK1 (PCTAIRE protein kinase 1). When using RNAi and a pharmacological inhibitor selective for PCTK1, we could show that this kinase plays a crucial role in the proliferation of MB cell lines and the activation of the mammalian target of rapamycin (mTOR) pathway. In addition, pharmacological PCTK1 inhibition reduced the expression levels of c-Myc. Finally, targeting PCTK1 selectively impaired the tumor growth of c-Myc-overexpressing MB cells in vivo. Together our data uncover a novel and crucial role for PCTK1 in the proliferation and survival of MB characterized by cMYC amplification.

Homozygosity mapping reveals novel and known mutations in Pakistani families with inherited retinal dystrophies.

Inherited retinal dystrophies are phenotypically and genetically heterogeneous. This extensive heterogeneity poses a challenge when performing molecular diagnosis of patients, especially in developing countries. In this study, we applied homozygosity mapping as a tool to reduce the complexity given by genetic heterogeneity and identify disease-causing variants in consanguineous Pakistani pedigrees. DNA samples from eight families with autosomal recessive retinal dystrophies were subjected to genome wide homozygosity mapping (seven by SNP arrays and one by STR markers) and genes comprised within the detected homozygous regions were analyzed by Sanger sequencing. All families displayed consistent autozygous genomic regions. Sequence analysis of candidate genes identified four previously-reported mutations in CNGB3, CNGA3, RHO, and PDE6A, as well as three novel mutations: c.2656C > T (p.L886F) in RPGRIP1, c.991G > C (p.G331R) in CNGA3, and c.413-1G > A (IVS6-1G > A) in CNGB1. This latter mutation impacted pre-mRNA splicing of CNGB1 by creating a -1 frameshift leading to a premature termination codon. In addition to better delineating the genetic landscape of inherited retinal dystrophies in Pakistan, our data confirm that combining homozygosity mapping and candidate gene sequencing is a powerful approach for mutation identification in populations where consanguineous unions are common.

Multidimensional Deprivation in Contemporary Switzerland Across Social Groups and Time

We have investigated the phenomenon of deprivation in contemporary Switzerland through the adoption of a multidimensional, dynamic approach. By applying Self Organizing Maps (SOM) to a set of 33 non-monetary indicators from the 2009 wave of the Swiss Household Panel (SHP), we identified 13 prototypical forms (or clusters) of well-being, financial vulnerability, psycho-physiological fragility and deprivation within a topological dimensional space. Then new data from the previous waves (2003 to 2008) were classified by the SOM model, making it possible to estimate the weight of the different clusters in time and reconstruct the dynamics of stability and mobility of individuals within the map. Looking at the transition probabilities between year t and year t+1, we observed that the paths of mobility which catalyze the largest number of observations are those connecting clusters that are adjacent on the topological space.

Review of quantitative phase-digital holographic microscopy: promising novel imaging technique to resolve neuronal network activity and identify cellular biomarkers of psychiatric disorders.

Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed.

Clic4, a novel protein that sensitizes β-cells to apoptosis.

OBJECTIVES: Chloride intracellular channel protein 4 (Clic4) is a ubiquitously expressed protein involved in multiple cellular processes including cell-cycle control, cell differentiation, and apoptosis. Here, we investigated the role of Clic4 in pancreatic β-cell apoptosis. METHODS: We used βTC-tet cells and islets from β-cell specific Clic4 knockout mice (βClic4KO) and assessed cytokine-induced apoptosis, Bcl2 family protein expression and stability, and identified Clic4-interacting proteins by co-immunoprecipitation and mass spectrometry analysis. RESULTS: We show that cytokines increased Clic4 expression in βTC-tet cells and in mouse islets and siRNA-mediated silencing of Clic4 expression in βTC-tet cells or its genetic inactivation in islets β-cells, reduced cytokine-induced apoptosis. This was associated with increased expression of Bcl-2 and increased expression and phosphorylation of Bad. Measurement of Bcl-2 and Bad half-lives in βTC-tet cells showed that Clic4 silencing increased the stability of these proteins. In primary islets β-cells, absence of Clic4 expression increased Bcl-2 and Bcl-xL expression as well as expression and phosphorylation of Bad. Mass-spectrometry analysis of proteins co-immunoprecipitated with Clic4 from βTC-tet cells showed no association of Clic4 with Bcl-2 family proteins. However, Clic4 co-purified with proteins from the proteasome suggesting a possible role for Clic4 in regulating protein degradation. CONCLUSIONS: Collectively, our data show that Clic4 is a cytokine-induced gene that sensitizes β-cells to apoptosis by reducing the steady state levels of Bcl-2, Bad and phosphorylated Bad.

A Segmentation Model for Hi-tech Business Markets

Segmentointi on strateginen työkalu, joka tehostaa yrityksen resurssien käyttöä ja siten vaikuttaa kaikkiin asiakkuuksiin liittyviin liiketoimintaprosesseihin. Työn tavoitteena oli muodostaa segmentointimalli (sisältää sekä segmentointiprosessin että kriteerit) yritysinternetmarkkinoille. Työn tuloksia voidaan kuitenkin tulkita ja soveltaa laajemmin korkean teknologian yrityspalvelumarkkinoille. Tämä tutkielma lisää tietämystämme ja tarjoaa uudenlaisen näkemyksen segmentointiin korkean teknologian yrityspalvelumarkkinoilla. Työssä kuvataan korkean teknologian ja yritys- sekä palvelumarkkinoinnin erityispiirteitä ja kuinka nämä tekijät vaikuttavat segmentointimallin. Tutkimuksessa selvitettiin kohdeyrityksen nykyiset segmentointikäytännöt henkilökohtaisin asiantuntijahaastatteluin. Haastatteluiden avulla luotiin kuva nykyisistä lähestymistavoista sekä niiden lähtökohdista, vahvuuksista ja haasteista. Haastatteluiden analysoinnin jälkeen perustettiin projekti segmentoinnin kehittämiseksi. Työ tuloksena luotiin segmentointimalli, joka tarjoaa vankan perustan segmentoinnin kehittämiselle jatkuvana prosessina. Työssä esitetään segmentoinnin integroimista yrityksen asiakkuuksiin liittyviin liiketoimintaprosesseihin, joka usein puuttuu aiemmista töistä, sekä informaationkulun tehostamista segmentoinnin hyödyntämiseksi tehokkaammin. Segmentointi on strateginen työkalu ja vaatii siksi ylemmän johdon tuen ja sitoutumisen. Oikein sovellettuna segmentointi tarjoaa liiketoiminnalle mahdollisuuden merkittäviin etuihin kuten asiakastyytyväisyyden ja kannattavuuden kehittämiseen.