The effect of inhibiting KPNB1-mediated nuclear import on cancer cell biology and inflammatory transcription factor signalling

Cancer remains one of the major causes of morbidity and mortality globally. Many novel and innovative approaches have been employed to develop new chemotherapeutic strategies, of which targeted therapies aim to identify a molecular lesion or dysregulated pathway that cancer cells are dependent on. Research in our laboratory and others identified the nuclear import protein, Karyopherin β1 (KPNB1), to be overexpressed in various cancers and that inhibiting its expression blocks the proliferation of cancer cells. However, little is known about the potential role of KPNB1 in other cancer cell phenotypes and inflammatory signalling pathways. The aim of this study was to investigate the anticancer and anti-inflammatory effects of inhibiting nuclear import via KPNB1 and to characterise the in vivo effect of the small molecule inhibitor of nuclear import, INI-43, on tumour formation. Using siRNA and a small molecule inhibitor, INI-43, to inhibit KPNB1 we found that cervical cancer cell migration and invasion was significantly reduced. The reduced motility of cancer cells was associated with a decrease in MMP-2 and -9 expression and an increase in TIMP-1 and -2 expression following INI-43 treatment. This corresponded with a decrease in MMP-9 gelatinase activity in KPNB1-inhibited cervical cancer cells. Extended periods of KPNB1 inhibition lead to decreased proliferation and apoptosis. These changes in cancer cell biology when KPNB1 is inhibited may in part be due to its function as a nuclear transporter of transcription factors associated with cancer cell proliferation, migration and invasion. We therefore investigated the effects of KPNB1 inhibition on the nuclear localisation and transcriptional activity of key transcription factors; NFkB and AP-1, both having been implicated in many of the hallmarks of cancer. Immunofluorescent analysis and nuclear/cytoplasmic fractionation assays showed that KPNB1 inhibition blocked the nuclear localisation of NFkB. Electromobility shift assays confirmed a reduced NFkB binding to an NFkB DNA-binding sequence in the nuclear extract of KPNB1-inhibited cells. Luciferase reporter assays containing NFkB and/or AP-1 consensus binding sites showed reduced transcriptional activity for both transcription factors following KPNB1 inhibition. Associated with these changes in NFkB and AP-1 activity was reduced inflammatory cytokines; IL-6, IL-1β, TNF-α and GM-CSF target gene expression. To further characterise the role of INI-43 as a potential chemotherapeutic, the effects on tumour growth and development were investigated in an ectopic xenograft mouse model. INI-43 treatment significantly reduced tumour growth in mice and associated with the redistribution and reduction in KPNB1 levels. INI-43 treated tumours also showed altered morphological features including; better tissue differentiation and reduced inflammatory stromal infiltration, as well as reduced Ki-67 expression. The expression of extracellular matrix components and the cytoskeletal structure of cancer cells was analysed to further investigate the role of KPNB1 inhibition in tumour development. Inhibition of KPNB1 in cancer cells caused reduced expression of both collagen type IV and MMP-9. The redistribution of B-catenin and F-actin suggested that INI-43 treatment caused a loss of mesenchymal features required for tumour progression. The nuclear transport system has been of particular interest in recent years for the development of targeted anticancer drugs. However, most studies have focused on nuclear export inhibitors with little known on the potential of nuclear import inhibitors as anticancer drugs. This study provides evidence that inhibiting the nuclear import protein, KPNB1, has anti-inflammatory and anticancer effects and shows promise as an anticancer approach requiring further investigation

PENGARUH BELANJA PEMERINTAH DAN INVESTASI TERHADAP PERTUMBUHAN EKONOMI DAN DAMPAKNYA TERHADAP PENGAGGURAN DI KABUPATEN MINAHASA SELATAN

ABSTRAKPembangunan merupakan suatu proses perbaikan atau kemajuan dengan melakukan  upaya ke arah perubahan yang lebih baik. Perubahan yang dimaksud adalah mencakup seluruh sistem yang ada dalam suatu wilayah atau negara seperti sistem politik, ekonomi, infrastruktur, pertahanan, pendidikan dan teknologi, kelembagaan serta budaya. Pembangunan ekonomi suatu daerah pada hakekatnya ialah suatu rangkaian kegiatan yang dilaksanakan secara sadar dan terus menerus untuk mewujudkan keadaan yang lebih baik secara bersama-sama dan berkesinambungan. Dalam kerangka itu, pembangunan ekonomi juga untuk memacu pemerataan pembangunan dan hasil-hasilnya dalam rangka meningkatkan kesejahteraan rakyat secara adil dan merata. Salah satu tolak ukur penting dalam menentukan keberhasilan pembangunan ekonomi adalah pertumbuhan ekonomi yang menggambarkan suatu dampak nyata dari kebijakan pembangunan yang dilaksanakan.Tujuan dari penelitian ini untuk menganalisis bagaimana pengaruh belanja pemerintah  dan investasi terhadap pengangguran melalui pertumbuhan ekonomi di Kabupaten Minahasa Selatan. Teknik analisis yang digunakan adalah analisis jalur (Path Analysis). Hasil penelitian menunjukan bahwa belanja pemerintah dan investasi berpengaruh positif terhadap pertumbuhan ekonomi di Kabupaten Minahasa Selatan akan tetapi tidak signifikan secara statistik. Belanja pemerintah berpengaruh positif terhadap pengangguran akan tetapi tidak signifikan secara statistik. Investasi dan pertumbuhan ekonomi berpengaruh negatif terhadap pengangguran di Kabupaten Minahasa Selatan. Kata Kunci: Belanja Pemerintah, Investasi, Pertumbuhan Ekonomi, PengangguranABSTRACT                    Development is a process of improvement or progress by making efforts toward a better change. The change in question covers all existing systems within a region or country such as political, economic, infrastructure, defense, education and technology, institutional and cultural systems. Economic development of a region is essentially a series of activities that are carried out consciously and continuously to realize a better state together and sustainable. Within that framework, economic development is also to stimulate equity of development and its results in order to improve the welfare of the people fairly and equitably. One important benchmark in determining the success of economic development is economic growth that illustrates a real impact of development policies implemented.                    The purpose of this study is to analyze how the influence of government spending and investment on unemployment through economic growth in South Minahasa District. Analytical technique used is path analysis (Path Analysis). The results show that government spending and investment have a positive effect on economic growth in South Minahasa regency but not statistically significant. Government spending has a positive effect on unemployment but is not statistically significant. Investment and economic growth negatively affect unemployment in South Minahasa District. Keywords: Government Expenditure, Investment, Economic Growth, Unemploymen

Moving from teaching older learners to young learners : cases of English language teachers in Turkey

This thesis explores the experiences of four English language teachers in Turkey who moved from teaching in either high schools or language schools to teaching English to young learners. The study follows these teachers in their first year of teaching in primary school, describes the changes they went through in their approach to teaching English, and identifies the influences on these changes. The background to this research was an educational reform which introduced English into the primary curriculum. After this reform, there was a shortage of English teachers at the primary level. A common solution was to recruit English teachers from other levels of the educational system. Three of the participating teachers taught in high school before they moved to teach English in the primary school. One of the participating teachers taught English to adults in a language school before moving to the primary level. The data for the study was generated through systematic interviews with the teachers, as well as regular observations of their classes, over the course of the school year. The interpretation of the data was informed by the existing literature on teaching English to young learners and the literature on teacher change. At the end of the school year, all of the teachers said they would like to continue teaching English in the primary school. In adjusting to teaching children, the teachers mainly focused on developing class management strategies. The teachers did not, however, focus on maximising opportunities for learning the target language. The main influences on the teachers' change were in-service training, textbooks, as well as interaction with and support from colleagues. The research contributes to the debate of how to best facilitate the introduction of English into primary schools, and calls for further research into teacher development in the context of such curricular reforms

Literatuurstudie over de rozeschildluis Aulacaspis rosae

Problemen met schildluis treden de laatste jaren vaker op door het wegvallen van breedwerkende bestrijdingsmiddelen. In Nederlandse kassen is schildluis traditioneel een probleem op cymbidium, maar momenteel meldt men ook vaak Aulacaspis rosae (Bouché, 1933) op roos. Chemische bestrijding van schildluis is per definitie moeilijk, doordat het insect het grootste deel van zijn leven verborgen zit onder een ondoordringbaar schildje