Analysis of the potential of cancer cell lines to release tissue factor-containing microvesicles: correlation with tissue factor and PAR2 expression

BackgroundDespite the association of cancer-derived circulating tissue factor (TF)-containing microvesicles and hypercoagulable state, correlations with the incidence of thrombosis remain unclear.MethodsIn this study the upregulation of TF release upon activation of various cancer cell lines, and the correlation with TF and PAR2 expression and/or activity was examined. Microvesicle release was induced by PAR2 activation in seventeen cell lines and released microvesicle density, microvesicle-associated TF activity, and phoshpatidylserine-mediated activity were measured. The time-course for TF release was monitored over 90 min in each cell line. In addition, TF mRNA expression, cellular TF protein and cell-surface TF activities were quantified. Moreover, the relative expression of PAR2 mRNA and cellular protein were analysed. Any correlations between the above parameters were examined by determining the Pearson’s correlation coefficients.ResultsTF release as microvesicles peaked between 30–60 min post-activation in the majority of cell lines tested. The magnitude of the maximal TF release positively correlated with TF mRNA (c = 0.717; p

E-procurement in Public Organization

Tampereen kaupungin kokonaishankinnoista vain kaksi prosenttia tehtiin toiminnanohjausjärjestelmällä vuonna 2013. Järjestelmän matalasta käyttöasteesta johtuen hankintatiedot puuttuvat järjestelmästä, mikä hankaloittaa hankintojen strategista johtamista. Työn taustalla on tarve tehostaa toimintoja ja saada aikaan kustannussäästöjä sekä saada hankinnoista enemmän tietoa niiden tehokkaammaksi johtamiseksi. Tutkimuksen tarkoituksena oli tunnistaa keinoja, joiden avulla Tampereen kaupungin ostamista voidaan kehittää sähköisellä ostojärjestelmällä. Päätavoitteena oli kaupungin ostamisen nykytilan selvittäminen ja siihen soveltuvan sähköisen ratkaisun vaatimusten määrittely. Tässä työssä keskityttiin työn toimeksiantajan Tampereen Logistiikan valitsemien Tampereen kaupungin yksiköiden Infran, Kotihoidon ja Tilakeskuksen ostamisen kehittämiseen. Tutkimus toteutettiin laadullisena monimenetelmäisenä tapaustutkimuksena. Aineistona käytettiin kirjallisuutta, vanhoja Tampereen kaupungin selvityksiä sekä kohdeyksiköiden ja muiden kaupunkien hankinnoista vastaavien henkilöiden haastatteluja. Aikaisempi kirjallisuus on käsitellyt ostamisen suhdetta hankintoihin ja toimitusketjuun sekä tarkastellut sähköistä ostamista keinona tehostaa hankintoja. Tutkimuksen empiriaosuus esittelee ensin havaintoja aikaisemmista tutkimuksista sekä muiden kaupunkien benchmark-analyysistä. Toisessa osassa käsitellään ostotoimintaa ja sen kehitysmahdollisuuksia kohdeorganisaation valikoiduissa yksiköissä. Aineistoanalyysin tuloksena tunnistettiin ostamisen ongelmaksi kirjavat ostoprosessit sekä niistä aiheutuvat prosessikustannukset. Tuloksina saatiin ostamisen asettamia vaatimuksia ostojärjestelmälle, joita ovat noutojen kirjaus ja mobiilikäyttömahdollisuus. Ratkaisuehdotuksena annettiin malli uudesta prosessista, johon sitoutuu vähemmän työaikaa aikaisempaan verrattuna sekä ehdotettiin katalogien käyttöönottamista kaupungin omassa järjestelmässä

A Clinical-Genetic Risk Score for Predicting Cancer-Associated Venous Thromboembolism : A Development and Validation Study Involving Two Independent Prospective Cohorts

Venous thromboembolism (VTE) is a leading cause of death among patients with cancer. The Khorana score was developed for assessing the risk of VTE in outpatients with cancer receiving chemotherapy, but its accuracy in identifying patients at high risk has been questioned. The aim of this study was to develop and validate a clinical-genetic score that improves the assessment of VTE risk in oncology outpatients within 6 months of diagnosis. The new score was developed using the data of 364 outpatients belonging to the Spanish ONCOTHROMB 12-01 population. In this cohort, clinical data associated with the risk of VTE were collected at the time of diagnosis, including the Khorana score. These patients were also genotyped for the 51 genetic variants known to be associated with VTE. Multivariate logistic regression was performed to determine the weight of each genetic and clinical variable in relation to VTE risk, allowing a clinical-genetic risk score (the ONCOTHROMB score) to be developed. The Khorana and the ONCOTHROMB scores were then compared via the area under the receiver operating characteristic curve (AUC), calibration, and the number of patients needed to treat. The new score was then validated in a study of 263 patients in the Vienna Cancer and Thrombosis Study population.RESULTSNine genetic variants, tumor site, TNM stage, and a body mass index of > 25 kg/m2 were found to be associated with VTE and were used to build the ONCOTHROMB score, which better predicted the overall risk of VTE than did the Khorana score (AUC, 0.781 v 0.580; P <.001). Similar AUC results were recorded in the validation study the Vienna Cancer and Thrombosis Study cohort involving patients with the same type of tumor (AUC for the ONCOTHROMB score v the Khorana score: 0.686 v 0.577; P <.001) and with all type of tumors (AUC for the ONCOTHROMB score v the Khorana score: 0.720 v 0.561; P <.0001).The ONCOTHROMB score for VTE risk in outpatients with cancer, which takes into account both clinical and genetic variables, better identifies patients who might benefit from primary thromboprophylaxis than does the Khorana score

Discovery of Molecular DNA Methylation-Based Biomarkers through Genome-Wide Analysis of Response Patterns to BCG for Bladder Cancer

Background: Bacillus Calmette-Gu&eacute;rin (BCG) immunotherapy, the standard adjuvant intravesical therapy for some intermediate and most high-risk non-muscle invasive bladder cancers (NMIBCs), suffers from a heterogenous response rate. Molecular markers to help guide responses are scarce and currently not used in the clinical setting. Methods: To identify novel biomarkers and pathways involved in response to BCG immunotherapy, we performed a genome-wide DNA methylation analysis of NMIBCs before BCG therapy. Genome-wide DNA methylation profiles of DNA isolated from tumors of 26 BCG responders and 27 failures were obtained using the Infinium MethylationEPIC BeadChip. Results: Distinct DNA methylation patterns were found by genome-wide analysis in the two groups. Differentially methylated CpG sites were predominantly located in gene promoters and gene bodies associated with bacterial invasion of epithelial cells, chemokine signaling, endocytosis, and focal adhesion. In total, 40 genomic regions with a significant difference in methylation between responders and failures were detected. The differential methylation state of six of these regions, localized in the promoters of the genes GPR158, KLF8, C12orf42, WDR44, FLT1, and CHST11, were internally validated by bisulfite-sequencing. GPR158 promoter hypermethylation was the best predictor of BCG failure with an AUC of 0.809 (p-value &lt; 0.001). Conclusions: Tumors from BCG responders and BCG failures harbor distinct DNA methylation profiles. Differentially methylated DNA regions were detected in genes related to pathways involved in bacterial invasion of cells or focal adhesion. We identified candidate DNA methylation biomarkers that may help to predict patient prognosis after external validation in larger, well-designed cohorts