Novel VHH-Based Tracers with Variable Plasma Half-Lives for Imaging of CAIX-Expressing Hypoxic Tumor Cells

Hypoxic areas are present in the majority of solid tumors, and hypoxia is associated with resistance to therapies and poor outcomes. A transmembrane protein that is upregulated by tumor cells that have adapted to hypoxic conditions is carbonic anhydrase IX (CAIX). Therefore, noninvasive imaging of CAIX could be of prognostic value, and it could steer treatment strategies. The aim of this study was to compare variants of CAIX-binding VHH B9, with and without a C-terminal albumin-binding domain with varying affinity (ABD(low) and ABD(high)), for SPECT imaging of CAIX expression. The binding affinity and internalization of the various B9-variants were analyzed using SK-RC-52 cells. Biodistribution studies were performed in mice with subcutaneous SCCNij153 human head and neck cancer xenografts. Tracer uptake was determined by ex vivo radioactivity counting and visualized by SPECT/CT imaging. Furthermore, autoradiography images of tumor sections were spatially correlated with CAIX immunohistochemistry. B9-variants demonstrated a similar moderate affinity for CAIX in vitro. Maximal tumor uptake and acceptable tumor-to-blood ratios were found in the SCCNij153 model at 4 h post injection for [(111)In]In-DTPA-B9 (0.51 ± 0.08%ID/g and 8.1 ± 0.85, respectively), 24 h post injection for [(111)In]In-DTPA-B9-ABD(low) (2.39 ± 0.44%ID/g and 3.66 ± 0.81, respectively) and at 72 h post injection for [(111)In]In-DTPA-B9-ABD(high) (8.7 ± 1.34%ID/g and 2.43 ± 0.15, respectively)(.) An excess of unlabeled monoclonal anti-CAIX antibody efficiently inhibited tumor uptake of [(111)In]In-DTPA-B9, while only a partial reduction of [(111)In]In-DTPA-B9-ABD(low) and [(111)In]In-DTPA-B9-ABD(high) uptake was found. Immunohistochemistry and autoradiography images showed colocalization of all B9-variants with CAIX expression; however, [(111)In]In-DTPA-B9-ABD(low) and [(111)In]In-DTPA-B9-ABD(high) also accumulated in non-CAIX expressing regions. Tumor uptake of [(111)In]In-DTPA-B9-ABD(low) and [(111)In]In-DTPA-B9-ABD(high), but not of [(111)In]In-DTPA-B9, could be visualized with SPECT/CT imaging. In conclusion, [(111)In]In-DTPA-B9 has a high affinity to CAIX and shows specific targeting to CAIX in head and neck cancer xenografts. The addition of ABD prolonged plasma half-life, increased tumor uptake, and enabled SPECT/CT imaging. This uptake was, however, partly CAIX- independent, precluding the ABD-tracers for use in hypoxia quantification in this tumor type

TRIM46 Controls Neuronal Polarity and Axon Specification by Driving the Formation of Parallel Microtubule Arrays

Axon formation, the initial step in establishing neuronal polarity, critically depends on local microtubule reorganization and is characterized by the formation of parallel microtubule bundles. How uniform microtubule polarity is achieved during axonal development remains an outstanding question. Here, we show that the tripartite motif containing (TRIM) protein TRIM46 plays an instructive role in the initial polarization of neuronal cells. TRIM46 is specifically localized to the newly specified axon and, at later stages, partly overlaps with the axon initial segment (AIS). TRIM46 specifically forms closely spaced parallel microtubule bundles oriented with their plus-end out. Without TRIM46, all neurites have a dendrite-like mixed microtubule organization resulting in Tau missorting and altered cargo trafficking. By forming uniform microtubule bundles in the axon, TRIM46 is required for neuronal polarity and axon specification in vitro and in vivo. Thus, TRIM46 defines a unique axonal cytoskeletal compartment for regulating microtubule organization during neuronal development

Inclusão de Alunos com Deficiência nas Aulas de Educação Física: Atitudes de Professores nas Escolas Regulares1

RESUMO a inclusão de alunos com deficiência nas escolas regulares ainda é um desafio para professores, especialmente pela insegurança gerada pela precária formação profissional e falta de estrutura de apoio. O objetivo desta investigação foi analisar as atitudes dos professores de Educação Física em relação à inclusão de alunos com deficiência no ensino regular, bem como a influência do sexo, tempo de experiência profissional e tipo de deficiência dos alunos. Para tanto, 35 professores de Educação Física de 15 escolas públicas de Londrina responderam individualmente ao Teacher Inclusion Attitudes Questionnaire (TIAQ) - Questionário de Atitudes dos Professores com relação à Inclusão. Os resultados mostraram que, de um modo geral, os professores são otimistas com relação à inclusão de alunos com deficiência em suas aulas, embora seja destacada a falta de apoio recebido pela escola como uma barreira importante no processo. O tempo de experiência e sexo do professor, bem como o tipo de deficiência do aluno, foram fatores de influência nas atitudes, reforçando que mulheres com menor tempo de experiência, especialmente com alunos com deficiência intelectual em suas turmas, apresentam atitudes mais negativas no sentido da inclusão

The attitudes of teachers towards inclusive education in Vojvodina

The aim of this study was to examine the attitudes of preschool, primary, secondary and high school teachers towards inclusive education of children with special educational needs. In addition, the study established the correlation between these attitudes and gender, education level, teaching experience, formal training in the special education field, and the duration and quality of work experience with children with special education needs. The sample comprised 322 teachers from the Serbian province of Vojvodina. The My Thinking about Inclusion Scale (Stoiber, K. C., M. Gettinger, and D. Goetz. 1998. "Exploring Factors Influencing Parents' and Early Childhood Practitioners Beliefs about Inclusion." Early Childhood Research Quarterly 13 (1): 107-131) was used. The results show that, in general, the participants held neutral attitudes towards inclusive education and more positive expectations regarding the outcomes of inclusion. This study also emphasised teaching performance in an inclusive class as a subject of great concern. The high school and preschool teachers as well as the teachers with previous positive experience with working in an inclusive environment reported more positive attitudes towards inclusive education than those from primary and secondary schools and those with negative experiences with the implementation of inclusive practices