Is High Resolution Melting Analysis (HRMA) Accurate for Detection of Human Disease-Associated Mutations? A Meta Analysis

BACKGROUND: High Resolution Melting Analysis (HRMA) is becoming the preferred method for mutation detection. However, its accuracy in the individual clinical diagnostic setting is variable. To assess the diagnostic accuracy of HRMA for human mutations in comparison to DNA sequencing in different routine clinical settings, we have conducted a meta-analysis of published reports. METHODOLOGY/PRINCIPAL FINDINGS: Out of 195 publications obtained from the initial search criteria, thirty-four studies assessing the accuracy of HRMA were included in the meta-analysis. We found that HRMA was a highly sensitive test for detecting disease-associated mutations in humans. Overall, the summary sensitivity was 97.5% (95% confidence interval (CI): 96.8-98.5; I(2) = 27.0%). Subgroup analysis showed even higher sensitivity for non-HR-1 instruments (sensitivity 98.7% (95%CI: 97.7-99.3; I(2) = 0.0%)) and an eligible sample size subgroup (sensitivity 99.3% (95%CI: 98.1-99.8; I(2) = 0.0%)). HRMA specificity showed considerable heterogeneity between studies. Sensitivity of the techniques was influenced by sample size and instrument type but by not sample source or dye type. CONCLUSIONS/SIGNIFICANCE: These findings show that HRMA is a highly sensitive, simple and low-cost test to detect human disease-associated mutations, especially for samples with mutations of low incidence. The burden on DNA sequencing could be significantly reduced by the implementation of HRMA, but it should be recognized that its sensitivity varies according to the number of samples with/without mutations, and positive results require DNA sequencing for confirmation

Kotkan kaupungin kiinteistöjen pelastussuunnitelmien käyttöönotto

Tämän opinnäytetyön aiheena oli kehittää Kotkan kaupungin kiinteistöjen uusien pelastussuunnitelmien käyttöönottoa. Tavoitteena oli kehittää selkeät ohjeet, joiden avulla kaupungin kiinteistöjen haltijat pystyvät laatimaan ja ylläpitämään oman kiinteistönsä pelastussuunnitelman internetpohjaisen Pelsu-pelastussuunnitelman verkko- ja mobiilityökalun avulla. Työssä käydään läpi pelastuslain määrittämiä vaatimuksia pelastussuunnitelmille. Lisäksi työssä esitellään Pelsu verkko- ja mobiilityökalun kehittänyt yritys Plan Brothers sekä tutustutaan tarkemmin myös Pelsun tarjoamiin työkaluihin ja ohjelman hyötyihin. Tämänhetkinen Kotkan kaupungin kiinteistöjen pelastussuunnitelmien tilanne käydään läpi tutkimalla nykyisiä pelastussuunnitelmia ja tuomalla esiin niissä havaitut puutteet. Ohjeiden laatiminen pelastussuunnitelmien luomista ja päivittämistä varten toteutettiin kuvakaappauksin ja tekstiosioin. Osaaminen ohjeiden tekoon saatiin osallistumalla Pelsu-perehdytykseen ja harjoittelemalla ohjelman verkko- ja mobiilityökalun käyttöä omilla tunnuksilla. Työssä valmistettiin monisivuiset mutta selkeät ohjeet, joita seuraamalla voidaan luoda kiinteistön internetpohjainen pelastussuunnitelma. Työssä annetaan myös suosituksia, joiden avulla pelastussuunnitelmat saadaan jaettua kaikille toimijoille helposti ja tehokkaasti. Lisäksi otetaan kantaa myös pelastussuunnitelmien ylläpitoon ja siihen kuinka Kotkan organisaatioiden tulisi ottaa Pelsun käyttö osaksi työrutiinejaan.The subject of the thesis was to develop the commissioning of rescue plans in the real estate services of the City of Kotka. The objective was to create explicit instructions that can be used by the tenants (of the city’s real estates) to draft and maintain the rescue plans of their premises using an internet based Pelsu web and mobile software. The study discusses the requirements that the rescue act defines for the rescue plans. The study also exhibits the company Plan Brothers that has developed the Pelsu web and mobile software. It also explores the tools and benefits that the Pelsu software provides. The current situation of the city rescue plans is gone through by studying the present rescue plans and by bringing out the flaws in them. The instructions for creating and updating the rescue plans are executed with screenshots and text parts. The know-how for creating the instructions is gathered by participating in orientation about Pelsu and by practicing the use of the web and mobile software with the author’s own user identification. The result of this thesis is a multipage but clear instructions that can be followed to create the real estate’s internet-based rescue plan. The study also gives recommendations on how the rescue plans can be distributed to all the operators easily and effectively. In addition, an opinion is given on maintenance of the rescue plans and on adoption of Pelsu by the different branches of the local authority as part of their work routine

Signal Transducers and Activators of Transcription-3 Up-Regulates Tissue Inhibitor of Metalloproteinase-1 Expression and Decreases Invasiveness of Breast Cancer

Signal transducers and activators of transcription (STAT)-3 is an oncogenic protein that is constitutively activated in many human cancers, including 30 to 60% of primary breast cancer. The biological significance of STAT3 activation in breast cancer is not fully understood. We have previously shown that STAT3 up-regulates tissue inhibitors of metalloproteinase (TIMP)-1, a cytokine known to block metalloproteinases and decrease invasiveness in certain cancer cell types. We hypothesize that STAT3 activation may modulate tumor invasiveness of breast cancer by regulating TIMP1 expression. Using MCF-7 cells transfected with tetracycline-off STAT3C (constitutively active STAT3), we generated an in vitro system in which STAT3C levels can be tightly controlled in breast cancer cells. Increasing tetracycline levels gradually decreased STAT3C and TIMP1 in a dose-dependent manner, and down-regulation of these proteins led to a reciprocal decrease in invasiveness of these cells in Matrigel. Addition of a neutralizing anti-TIMP1 antibody increased invasiveness in the same experimental system. Using immunohistochemistry and 142 primary breast tumors, we found a significant association between the expression of the phosphorylated/active form of STAT3 (pSTAT3) and that of TIMP1. Importantly, STAT3 activation correlated significantly with a lower frequency of vascular and lymphatic invasion (P = 0.015 and P = 0.0002, respectively). Our data support the concept that STAT3 activation significantly modulates the biological and clinical behavior of breast cancer