Közművelődési jelenségek és jelentések. Az ÁMK

Many classroom climate studies suffer from 2 critical problems: They (a) treat climate as a student-level (L1) variable in single-level analyses instead of a classroom-level (L2) construct in multilevel analyses; and (b) rely on manifest-variable models rather than on latent-variable models that control measurement error at L1 and L2, and sampling error in the aggregation of L1 ratings to form L2 constructs. On the basis of an analysis of 2,541 students in Grades 5 or 6 from 89 classrooms, the authors demonstrate doubly latent multilevel structural equation models that overcome both of these problems. The results show that L2 classroom climate (a higher-order factor representing classroom mastery goal orientation, challenge, and teacher caring) had positive effects on self-efficacy and achievement. The authors conclude with a discussion of related issues (e.g., the meaning of L2 constructs vs. L1 residuals, the dimensionality of climate constructs at L2) and guidelines for future research

Phenotypic lentivirus screens to identify functional single domain antibodies

Manipulation of proteins is key in assessing their in vivo function. Although genetic ablation is straightforward, reversible and specific perturbation of protein function remains a challenge. Single domain antibody fragments, such as camelid-derived VHHs, can serve as inhibitors or activators of intracellular protein function, but functional testing of identified VHHs is laborious. To address this challenge, we have developed a lentiviral screening approach to identify VHHs that elicit a phenotype when expressed intracellularly. We identified 19 antiviral VHHs that protect human A549 cells from lethal infection with influenza A virus (IAV) or vesicular stomatitis virus (VSV), respectively. Both negative-sense RNA viruses are vulnerable to VHHs uniquely specific for their respective nucleoproteins. Antiviral VHHs prevented nuclear import of viral ribonucleoproteins or mRNA transcription, respectively, and may provide clues for novel antiviral reagents. In principle, the screening approach described here should be applicable to identify inhibitors of any pathogen or biological pathway. To identify the proteins essential to a biological pathway, small-molecule inhibitors or activators may be used to manipulate protein function transiently. Alternatively, screens involving mutagenesis, a reduction in levels or complete elimination of gene products are common. As applied to mammalian cells, these methods usually seek to achieve the removal of a protein from its normal biological context. Many proteins are multifunctional, or are components of multi-subunit complexes. Depletion of any single component may cause unexpected phenotypes due to the collapse of entire protein complexes. Small-molecule inhibitors often lack specificity and at best can target a fraction of all the proteins of interest. The screening of chemically diverse libraries must be paired with sophisticated methods to identify the molecular targets of any hit identified. Antibodies have been used as intracellular perturbants of protein function after microinjection or cytosolic expression of single-chain variable antibody fragments, but technical challenges have limited their application to few selected cases. In addition to conventional antibodies, the immune system of camelids generates heavy-chain-only antibodies. Their antigen binding site only consists of the variable domain of the heavy chain. This domain can be expressed on its own and is referred to as a VHH or nanobody, an entity that can retain its function in the reducing environment of the cytosol, independent of glycosylation. Many VHHs bind to their targets with affinities comparable to conventional antibodies. VHHs expressed in the cytosol can therefore act as molecular perturbants by occluding the interfaces involved in protein–protein interactions, by binding in the active sites of enzymes, or through the recognition or stabilization of distinct conformations of their targets. Both phage and yeast display, as well as mass spectrometry in combination with high-throughput sequencing, allow the identification of VHHs based on their binding properties. However, the identification of inhibitory VHHs remains a time-consuming process. VHHs obtained through biochemical screening methods must be expressed individually in the relevant cell type to test for the functional consequences of VHH expression. To address this challenge, we developed a phenotypic VHH screening method in living cells.National Institutes of Health (U.S.) (Health Pioneer Award

STRATEGI DINAS PENDIDIKAN DALAM MEMINIMALISIR ANAK PUTUS SEKOLAH DI KOTA BITUNG

AbstrakMasih tingginya angka putus sekolah di Kota Bitung menjadi suatu indicator masih lemahnya kinerja Dinas terkait yaitu Dinas Pendidikan melalui strategi yang diterapkan. Tujuan penelitian ini untuk mengetahui strategi Dinas Pendidikan dalam menekan angka anak putus sekolah di Kota Bitung. Penelitian ini menggunakan metode penelitian kualitatif deskriptif. Informan penelitian sebanyak 11 (sebelas) informan yaitu 1 orang Kepala Dinas Pendidikan Kota Bitung, 4 orang UPT Dinas Pendidikan di Kecamatan, 3 orang Kepala Sekolah tingkat SD, tingkat SMP, dan tingkat SMA, dan 3 orang masyarakat khususnya orang tua anak putus sekolah. Teknik pengumpulan data yang dipakai yaitu Teknik Observasi, Teknik Wawancara, Teknik Dokumentasi, dan Studi Pustaka, Teknik Analisa Data yang dipakai yaitu data yang diperoleh kemudian dianalisis secara bersamaan dengan proses pengumpulan data, proses analisis yang dilakukan merupakan suatu proses yang cukup panjang. Data dari hasil wawancara yang diperoleh kemudian dicatat dan dikumpulkan sehingga menjadi sebuah catatan lapangan. Berdasarkan hasil penelitian melalui hasil pengamatan, pengumpulan data dan proses wawancara yang penelti lakukan, maka penelitian mengenai strategi Dinas Pendidikan dalam menekan angka anak putus sekolah di Kota Bitung yaitu strategi sudah cukup baik dalam menekan angka anak putus sekolah, antara lain : Kebijakan Pemerintah Daerah dalam menekan angka Anak Putus Sekolah yakni berupa gerakan penuntasan wajib belajar 12 tahun sebagai wujud pemenuhan hak dasar masyarakat sesuai visi dan misi pembangunan Kota Bitung, Pemberian bantuan dana, Pemberian beasiswa pendidikan bagi masyarakat miskin, Program BSM (Bantuan Siswa Miskin), Program PIP (Program Indonesia Pintar), Program MaMa CEPAT (Cerdas Peduli Anak Tidak Sekolah), dan Sosialisasi kepada masyarakat.Kata kunci : Strategi, Dinas Pendidikan, Anak Putus Sekolah

In Vivo Detection of Succinate by Magnetic Resonance Spectroscopy as a Hallmark of SDHx Mutations in Paraganglioma

International audiencePurpose: Germline mutations in genes encoding mitochon-drial succinate dehydrogenase (SDH) are found in patients with paragangliomas, pheochromocytomas, gastrointestinal stromal tumors, and renal cancers. SDH inactivation leads to a massive accumulation of succinate, acting as an oncometabolite and which levels, assessed on surgically resected tissue are a highly specific biomarker of SDHx-mutated tumors. The aim of this study was to address the feasibility of detecting succinate in vivo by magnetic resonance spectroscopy. Experimental Design: A pulsed proton magnetic resonance spectroscopy (1 H-MRS) sequence was developed, optimized, and applied to image nude mice grafted with Sdhb À/À or wild-type chromaffin cells. The method was then applied to patients with paraganglioma carrying (n ¼ 5) or not (n ¼ 4) an SDHx gene mutation. Following surgery, succinate was measured using gas chromatography/mass spectrometry, and SDH protein expression was assessed by immunohistochemistry in resected tumors. Results: A succinate peak was observed at 2.44 ppm by 1 H-MRS in all Sdhb À/À-derived tumors in mice and in all paragangliomas of patients carrying an SDHx gene mutation, but neither in wild-type mouse tumors nor in patients exempt of SDHx mutation. In one patient, 1 H-MRS results led to the identification of an unsus-pected SDHA gene mutation. In another case, it helped define the pathogenicity of a variant of unknown significance in the SDHB gene. Conclusions: Detection of succinate by 1 H-MRS is a highly specific and sensitive hallmark of SDHx mutations. This non-invasive approach is a simple and robust method allowing in vivo detection of the major biomarker of SDHx-mutated tumors. Clin Cancer Res; 22(5); 1120–9. Ó2015 AACR