Pengembangan Perangkat Pembelajaran Konsep Klasifikasi Benda terhadap Keterampilan Berpikir Kritis Siswa SMP

Penelitian & pengembangan bertujuan mengevaluasi kualitas perangkat pembelajaran hasil pengembangan konsep klasifikasi benda terhadap keterampilan berpikir kritis siswa SMP. Model pengembangan menggunakan kawin silang ASSURE dan Tessmer. Subyek penelitian kevalidan tiga orang dosen pendidikan biologi, uji coba perorangan enam siswa kelas VIIc SMPN 2 Sungai Loban, uji coba kelompok kecil 15 siswa kelas VIIc SMPN 2 Sungai Loban, dan uji coba lapangan siswa kelas VII SMPN 4 Sungai Loban. Penetapan subyek berdasarkan keragaman kemampuan akademik. Penelitian dilaksanakan Januari-Juni 2015. Data kevalidan perangkat pembelajaran diperoleh dari pendapat pakar dianalisis berdasarkan modus. Pendapat siswa menggunakan lembar penilaian bahan ajar dan LKS dianalisis secara deskriptif. Data kepraktisan diperoleh dari keterlaksanaan RPP menggunakan lembar penilaian kegiatan guru dianalisis menggunakan kategori 1-4. Data keefektivan dari 1) aktivitas siswa, 2) hasil belajar, 3) sikap spiritual, 4) sikap sosial, dan 5) keterampilan berpikir kritis siswa. Data aktivitas siswa dikumpulkan menggunakan lembar pengamatan aktivitas siswa berdasarkan frekuensi aktivitas yang muncul setiap 5\u27 dinyatakan dengan %. Aktivitas siswa rendah jika < 10%. Hasil belajar menggunakan tes dan diberi skor 1 (benar) dan 0 (salah), dengan KKM ≥ 2,67. Data sikap (spiritual dan sosial) diperoleh menggunakan lembar pengamatan sikap menggunakan kategori 1-4. Data keterampilan berpikir kritis siswa diperoleh menggunakan rubric, dan dikategorikan dengan %. Hasil penelitian perangkat pembelajaran valid karena 1) silabus dinyatakan valid namun melalui perbaikan, dan 2) komponen RPP, bahan ajar, LKS, dan lembar penilaian juga valid. Praktis karena guru mitra mampu melaksanakan pembelajaran. Efektif berdasarkan 1) sebagian besar aktivitas siswa sudah baik namun mempersentasikan hasil percobaan dan menjawab dan menanggapi penyajian kelompok lain masih perlu perbaikan, 2) hasil belajar siswa telah melampaui ketuntasan minimal, 3) sikap spiritual sudah baik, 4) sikap social sebagian besar baik, 5) keterampilan berpikir kritis siswa rata-rata sudah baik. Berkenaan dengan unsur utama penelitian ini, maka dapat disimpulkan 1) merancang dan melaksanakan pengamatan dan 2) mengumpulkan data perlu ditingkatkan

Pencarian Jalur Terpendek untuk Robot Micromouse dengan Menggunakan Algoritma Backtracking

Robot Micromouse adalah robot cerdas yang dapat bergerak bebas di dalam sebuah labirin (maze) tanpa menyentuh objek sekitarnya, robot mengetahui ke arah mana harus bergerak, berapa derajat harus berputar jika menemui jalan buntu pada area labirin. Robot micromouse ini termasuk kedalam jenis Robot Mobile yaitu Autonomous Mobile Robot dimana pengendalian gerakan dari robot yang berdasarkan program kemudi yang diberikan sehingga seolah -olah robot tersebut bergerak sendiri. Arah pergerakan mobile robot ini ditentukan ketika ada respon terhadap obyek di depan, kanan dan kiri. Robot ini dibuat dengan mikrokontroller ATMEGA 8535 sebagai pengendali, sensor inframerah GP2D12 untuk mendeteksi adanya tembok atau tidak adanya tembok dan driver motor untuk menggerakkan motor sebagai aktuator. Robot ini menggunakan algoritma backtracking untuk mencari jalan terpendek dalam sebuah labirin ke tempat yang dituju

Mapping Asbestos-Cement Roofing with Hyperspectral Remote Sensing over a Large Mountain Region of the Italian Western Alps

The World Health Organization estimates that 100 thousand people in the world die every year from asbestos-related cancers and more than 300 thousand European citizens are expected to die from asbestos-related mesothelioma by 2030. Both the European and the Italian legislations have banned the manufacture, importation, processing and distribution in commerce of asbestos-containing products and have recommended action plans for the safe removal of asbestos from public and private buildings. This paper describes the quantitative mapping of asbestos-cement covers over a large mountainous region of Italian Western Alps using the Multispectral Infrared and Visible Imaging Spectrometer sensor. A very large data set made up of 61 airborne transect strips covering 3263 km2 were processed to support the identification of buildings with asbestos-cement roofing, promoted by the Valle d’Aosta Autonomous Region with the support of the Regional Environmental Protection Agency. Results showed an overall mapping accuracy of 80%, in terms of asbestos-cement surface detected. The influence of topography on the classification’s accuracy suggested that even in high relief landscapes, the spatial resolution of data is the major source of errors and the smaller asbestos-cement covers were not detected or misclassified

Clinical impact of a targeted next-generation sequencing gene panel for autoinflammation and vasculitis.

BACKGROUND: Monogenic autoinflammatory diseases (AID) are a rapidly expanding group of genetically diverse but phenotypically overlapping systemic inflammatory disorders associated with dysregulated innate immunity. They cause significant morbidity, mortality and economic burden. Here, we aimed to develop and evaluate the clinical impact of a NGS targeted gene panel, the "Vasculitis and Inflammation Panel" (VIP) for AID and vasculitis. METHODS: The Agilent SureDesign tool was used to design 2 versions of VIP; VIP1 targeting 113 genes, and a later version, VIP2, targeting 166 genes. Captured and indexed libraries (QXT Target Enrichment System) prepared for 72 patients were sequenced as a multiplex of 16 samples on an Illumina MiSeq sequencer in 150bp paired-end mode. The cohort comprised 22 positive control DNA samples from patients with previously validated mutations in a variety of the genes; and 50 prospective samples from patients with suspected AID in whom previous Sanger based genetic screening had been non-diagnostic. RESULTS: VIP was sensitive and specific at detecting all the different types of known mutations in 22 positive controls, including gene deletion, small INDELS, and somatic mosaicism with allele fraction as low as 3%. Six/50 patients (12%) with unclassified AID had at least one class 5 (clearly pathogenic) variant; and 11/50 (22%) had at least one likely pathogenic variant (class 4). Overall, testing with VIP resulted in a firm or strongly suspected molecular diagnosis in 16/50 patients (32%). CONCLUSIONS: The high diagnostic yield and accuracy of this comprehensive targeted gene panel validate the use of broad NGS-based testing for patients with suspected AID

Trehalose prodegradative role on AR aggregates in a muscle model of Spinal and Bulbar Muscular Atrophy.

Spinal and Bulbar Muscular Atrophy (SBMA) is a motor neuronal disease whose onset and progression have been recently linked also to a muscular defect. SBMA is caused by a polyglutammine tract in the exon 1 of the androgen receptor protein (ARpolyQ). When AR is activated by testosterone a fraction of the protein misfolds and become toxic to cells. Moreover if ARpolyQ is not correctly removed from cellular environment it also forms aggregates that could damage many cellular process. In this work we have studied the protein quality control system (composed of a chaperone network and two main degradative pathways: proteasome and autophagy) in a cellular muscular model of SBMA. We use C2C12 stably transfected with ARwt or ARpolyQ bearing an elongation of 100 glutammine. Initially we performed a filter trap assay (FTA) on both cell line treated with testosterone. We observed that testosterone triggers the aggregation of ARpolyQ but not of ARwt. We also observed that testosterone treatment caused mortality in C2C12 with ARpolyQ. By real time PCR we found that there was not activation of the PQC system in presence of ARpolyQ but that the expression of AchR was significantly lower than in control cell. These data suggest that ARpolyQ led to muscular atrophy. We investigate degradative systems that degrade AR and found that autophagy is highly involved in AR degradation. We facilitate autophagy towards the overexpression of HspB8. We observed that HspB8 counteracted testosterone dependent aggregation of ARpolyQ. We know that trehalose in motorneuronal model of SBMA induce the expression of HspB8. We then enhanced autophagy with trehalose and found that ARpolyQ aggregation was almost completely reverted. We co-treated cells with trehalose and bafilomycin and found that this condition abolished trehalose effect. We demonstrated that trehalose effect depend upon an efficient autophagic flux. By rtPCR we observed that trehalose enhance the expression of a wide range of genes related to autophagy. Interestingly we also found VCP overexpression in presence of trehalose. The valosin containing protein VCP is a multi-functional protein involved also in the ERAD pathway. So we inhibit VCP with DBEQ a specific inhibitor of the ATPase activity of VCP and found that testosterone dependent aggregation was significantly increased. Interestingly we found that trehalose treatment counteracted this DBEQ associated aggregation. In conclusion we characterized C2C12 as a reliable muscle model of SBMA. we also found that autophagy is highly involved in ARpolyQ degradation and consequently we demonstrated that autophagy activation rescues ARpolyQ aggregation in muscle cells. We finally observed that also the ERAD pathway plays an important role in ARpolyQ degradation. AFM-TELETHON; FONDAZIONE TELETHON; FONDAZIONE CARIPLO; FONDAZIONE ARISLA; Ministero della Sanità; Joint Programme Neurodegenerative Disease (JPND)

Site-Specific Bioconjugation of a Murine Dihydrofolate Reductase Enzyme by Copper(I)-Catalyzed Azide-Alkyne Cycloaddition with Retained Activity

Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is an efficient reaction linking an azido and an alkynyl group in the presence of copper catalyst. Incorporation of a non-natural amino acid (NAA) containing either an azido or an alkynyl group into a protein allows site-specific bioconjugation in mild conditions via CuAAC. Despite its great potential, bioconjugation of an enzyme has been hampered by several issues including low yield, poor solubility of a ligand, and protein structural/functional perturbation by CuAAC components. In the present study, we incorporated an alkyne-bearing NAA into an enzyme, murine dihydrofolate reductase (mDHFR), in high cell density cultivation of Escherichia coli, and performed CuAAC conjugation with fluorescent azide dyes to evaluate enzyme compatibility of various CuAAC conditions comprising combination of commercially available Cu(I)-chelating ligands and reductants. The condensed culture improves the protein yield 19-fold based on the same amount of non-natural amino acid, and the enzyme incubation under the optimized reaction condition did not lead to any activity loss but allowed a fast and high-yield bioconjugation. Using the established conditions, a biotin-azide spacer was efficiently conjugated to mDHFR with retained activity leading to the site-specific immobilization of the biotin-conjugated mDHFR on a streptavidin-coated plate. These results demonstrate that the combination of reactive non-natural amino acid incorporation and the optimized CuAAC can be used to bioconjugate enzymes with retained enzymatic activityope

Chronic treatment with 17-DMAG improves balance and coordination in a new mouse model of Machado-Joseph disease

Machado-Joseph disease (MJD) or spinocerebellar ataxia type 3 (SCA3) is a neurodegenerative disease currently with no treatment. We describe a novel mouse model of MJD which expresses mutant human ataxin-3 at near endogenous levels and manifests MJD-like motor symptoms that appear gradually and progress over time. CMVMJD135 mice show ataxin-3 intranuclear inclusions in the CNS and neurodegenerative changes in key disease regions, such as the pontine and dentate nuclei. Hsp90 inhibition has shown promising outcomes in some neurodegenerative diseases, but nothing is known about its effects in MJD. Chronic treatment of CMVMJD mice with Hsp90 inhibitor 17-DMAG resulted in a delay in the progression of their motor coordination deficits and, at 22 and 24 weeks of age, was able to rescue the uncoordination phenotype to wild-type levels; in parallel, a reduction in neuropathology was observed in treated animals. We observed limited induction of heat-shock proteins with treatment, but found evidence that 17-DMAG may be acting through autophagy, as LC3-II (both at mRNA and protein levels) and beclin-1 were induced in the brain of treated animals. This resulted in decreased levels of the mutant ataxin-3 and reduced intranuclear aggregation of this protein. Our data validate this novel mouse model as a relevant tool for the study of MJD pathogenesis and for pre-clinical studies, and show that Hsp90 inhibition is a promising therapeutic strategy for MJD.We would like to thank to Dr. Henry Paulson for providing the anti-ataxin-3 serum, Dr. Monica Sousa for the pCMV vector and to Eng. Lucilia Goreti Pinto, Lu s Martins, Miguel Carneiro and Celina Barros for technical assistance. This work was supported by Fundacao para a Ciencia e Tecnologia through the projects FEDER/FCT, POCI/SAU-MMO/60412/2004 and PTDC/SAU-GMG/64076/2006. This work was supported by Fundacao para a Ciencia e Tecnologia through fellowships SFRH/BPD/91562/2012 to A.S-F., SFRH/BD/78388/2011 to S. D-S., SFRH/BD/51059/2010 to A.N-C., and SFRH/BPD/79469/2011 to A.T-C.