Quantitative phosphoproteomics of cytotoxic T cells to reveal Protein Kinase D 2 regulated networks

The focus of the present study was to characterize the phosphoproteome of cytotoxic T cells and to explore the role of the serine threonine kinase PKD2 (Protein Kinase D2) in the phosphorylation networks of this key lymphocyte population. We used Stable Isotope Labeling of Amino acids in Culture (SILAC) combined with phosphopeptide enrichment and quantitative mass-spectrometry to determine the impact of PKD2 loss on the cytotoxic T cells phosphoproteome. We identified 15,871 phosphorylations on 3505 proteins in cytotoxic T cells. 450 phosphosites on 281 proteins were down-regulated and 300 phosphosites on 196 proteins were up-regulated in PKD2 null cytotoxic T cells. These data give valuable new insights about the protein phosphorylation networks operational in effector T cells and reveal that PKD2 regulates directly and indirectly about 5% of the cytotoxic T-cell phosphoproteome. PKD2 candidate substrates identified in this study include proteins involved in two distinct biological functions: regulation of protein sorting and intracellular vesicle trafficking, and control of chromatin structure, transcription, and translation. In other cell types, PKD substrates include class II histone deacetylases such as HDAC7 and actin regulatory proteins such as Slingshot. The current data show these are not PKD substrates in primary T cells revealing that the functional role of PKD isoforms is different in different cell lineages

Síntese de novos cristais líquidos discóticos derivados do 2,1,3-benzotiadiazol e 1,3,4-oxadiazol

TCC (graduação) - Universidade Federal de Santa Catarina. Centro de Ciências Físicas e Matemáticas. Curso de Química.Cristais líquidos luminescentes são materiais orgânicos que mesclam as propriedades de fluidez, auto-organização e luminescência. Por conta disso, possuem uma gama de aplicações tecnológicas como em OLEDs, sensores, semicondutores, etc. Visando o desenvolvimento de novos materiais para este tipo de aplicação, neste trabalho foram sintetizadas três novas moléculas derivadas do 2,1,3-benzotiadiazol e 1,3,4-oxadiazol, heterociclos utilizados como fluoróforos e unidades aceitadoras de elétrons para obtenção de materiais luminescentes e semicondutores, variando entre essas três moléculas, o tamanho e a quantidade de cadeias alcóxi nas periferias, obtendo-se assim materiais com diferentes estabilidades térmicas. As três moléculas sintetizadas apresentaram propriedade líquido-cristalina, sendo que duas delas (contendo quatro cadeias periféricas, uma de seis e outra de doze átomos de carbono) formaram mesofases nemáticas discóticas e a terceira (contendo oito cadeias periféricas de doze átomos de carbono) formou uma mesofase do tipo colunar hexagonal. A caracterização estrutural dessas moléculas foi feita por RMN e espectrometria de massas; as propriedades térmicas, estudadas por MOLP, DSC e TGA e as propriedades ópticas, através de espectrofotometria de absorção, emissão e fluorescência em solução. As três moléculas estudadas apresentaram emissão em 476 nm e altos rendimentos quânticos absolutos entre 0,79 e 0,85, se mostrando promissoras para posteriores estudos de aplicação em dispositivos eletrônico

Relatório De Estágio Supervisionado Desenvolvido No Laboratório Federal De Defesa Agropecuária (Lfda) - Seção Laboratorial Avançada (São José/SC)

TCC (graduação) - Universidade Federal de Santa Catarina. Centro de Ciências Físicas e Matemáticas. Curso de Química.O monitoramento e a fiscalização em alimentos são de fundamental importância para garantir a segurança alimentar e evitar que a ocorrência de fraudes traga prejuízos à saúde da população. Os frutos do mar são alimentos que estão presentes na culinária ao redor do mundo, porém podem causar sérios problemas à saúde se estiverem contaminados. Por conta disto, neste trabalho serão abordadas duas análises de rotina realizadas em diferentes amostras de frutos do mar. Uma delas é a determinação de bases voláteis totais em pescados, feita pela técnica de acidimentria e a outra, a determinação de biotoxinas marinhas em moluscos bivalves por cromatografia líquida de alta eficiência acoplada a espectrômetro de massas. Estas análises fazem parte do escopo da Seção Laboratorial Avançada de São José, que pertence ao Laboratório Federal de Defesa Agropecuária do estado do Rio Grande do Sul, sendo este, um dos laboratórios oficiais do Ministério da Agricultura, pecuária e Abastecimento (MAPA). Além disso, este trabalho também relata outras atividades realizadas durante o estágio supervisionado, como a verificação de instrumentos laboratoriais, importante para o controle de qualidade e a confiabilidade dos resultados emitidos. Com isso, foi possível aplicar os conhecimentos adquiridos durante a formação acadêmica e obter-se uma experiência profissional muito rica para conclusão do curso de Bacharelado em Química Tecnológica

Epinicion carmen in laudem… Caroli Rueberi = A bécsi Lorenz Eiseler győzelmi éneke a nemes báró, Karl Rueber úr dicséretére és annak diadalára a török felett

Laurentius Eiseler egy a sok fiatal közül, akiket jómódú szüleik külföldi iskolákba küldtek tanulni. Őt Bécsből Strasbourgba. A strasbourgi Johann Sturm alapította Academia ékesszólás-stúdiumai európai hírűek voltak a korszakban. Melchior Junius óráin több alkalommal kellett olyan szemináriumi dolgozatot írni, hogy egy-egy latin műfaj feldolgozásához valamilyen aktuális történeti témát választhatott a hallgató. Az ifjú Eiseler a Magyar Királyság hadszínterein ismert osztrák főtisztek közül egy tokaji várkapitányt választott a „győzelmi ének” műfaj gyakorlására, a neki tulajdonított törökök feletti győzelem történetének kiszínezett bemutatására. Ha a mű nem is a latinitás csúcsteljesítménye, ha a hallgató nem is volt igazán tisztában a választott történet körülményeivel, kis művével – mert megjelent nyomtatásban – tájékoztatta Elzász lakóit a tőlük távoli háború egyik eseményéről

WRITING ASSESSMENT IN MIDDLE SCHOOL STUDENTS : ANALYZING SPELLING WITHIN A MULTIDIMENSIONAL LANGUAGE FRAMEWORK

Although producing quality written expression is a vital skill, many students in the United States struggle to produce proficient written language. There are many academic and career outcomes related to the ability to produce written expression, yet many schools lack formalized writing assessment and instruction. As such, many questions remain related to individual differences in writing ability and best practices in assessment and instruction. To answer these questions, it is necessary to establish a model of written expression and what specific variables exist within the model to be used to assess written language. Modern writing assessment theory uses levels of language as a framework with commonly assessed dimensions of accuracy, complexity and productivity. This framework has yet to be firmly established in the literature, and the variables included in each level are just beginning to be explored. One salient variable in writing research, assessment and instruction is spelling ability, and how this ability may influence the production of written language. This study furthers the work by Wilson et al. (2017), Troia and colleagues (2019) and many others (e.g., Berninger et al., 2006; Flower & Hayes, 1981) with the ultimate goal of developing a model of written language to guide assessment and instruction in schools. Specifically, data were drawn from Truckenmiller and colleagues (2020) study piloting a writing assessment tool, Writing Architect, which sampled 526 students from third to eight grades; this study used sixth, seventh and eighth grades with a resulting sample size of 290 students. Results indicated spelling was a significant predictor of writing quality, in that better spelling indicated better writing quality. The same was true for text. For the sentence-level variable, a higher score indicated worse writing quality in a significant way. The word variable did not significantly predict writing quality in the model. The significant interaction between spelling and text variables suggests that the effect of text on writing quality is even higher when spelling ability is also high. Findings highlight the importance of writing and spelling instruction in school. The findings for this age group help identify how writing abilities may change over the trajectory of development and vary individually. Additionally, this analysis echoes the call for further research to establish variables for automated writing assessment.Thesis (Ph.D.)--Michigan State University. School Psychology - Doctor of Philosophy, 2022Includes bibliographical reference

Human pluripotent stem cell-derived acinar/ductal organoids generate human pancreas upon orthotopic transplantation and allow disease modelling

Objective The generation of acinar and ductal cells from human pluripotent stem cells (PSCs) is a poorly studied process, although various diseases arise from this compartment. Design We designed a straightforward approach to direct human PSCs towards pancreatic organoids resembling acinar and ductal progeny. Results Extensive phenotyping of the organoids not only shows the appropriate marker profile but also ultrastructural, global gene expression and functional hallmarks of the human pancreas in the dish. Upon orthotopic transplantation into immunodeficient mice, these organoids form normal pancreatic ducts and acinar tissue resembling fetal human pancreas without evidence of tumour formation or transformation. Finally, we implemented this unique phenotyping tool as a model to study the pancreatic facets of cystic fibrosis (CF). For the first time, we provide evidence that in vitro, but also in our xenograft transplantation assay, pancreatic commitment occurs generally unhindered in CF. Importantly, cystic fibrosis transmembrane conductance regulator (CFTR) activation in mutated pancreatic organoids not only mirrors the CF phenotype in functional assays but also at a global expression level. We also conducted a scalable proof-of-concept screen in CF pancreatic organoids using a set of CFTR correctors and activators, and established an mRNA-mediated gene therapy approach in CF organoids. Conclusions Taken together, our platform provides novel opportunities to model pancreatic disease and development, screen for disease-rescuing agents and to test therapeutic procedures.This study was funded by the Deutsche Forschungsgemeinschaft (DFG, K.L. 2544/1-1 and 1-2), the Forschungskern SyStaR to AK, BIU (Böhringer Ingelheim Ulm to AK), the Fritz-Thyssen Foundation (Az. 10.15.2.040), the German Cancer Aid (111879) and the Else-Kröner-Fresenius-Stiftung (2011_A200). AK is indebted to the Baden-Württemberg Stiftung for the financial support of this research project by the Eliteprogramme for Postdocs. AK is also an Else-Kröner-Fresenius Memorial Fellow. LP is supported by a research fellowship of the Else-Kröner-Fresenius-Stiftung. MH was supported by the International Graduate School in Molecular Medicine and the Bausteinprogramme (L.SBN. 110), Ulm University. MM is supported by a grant of Ulm University (Baustein for Senior Clinician Scientists). IGC is funded by the Interdisciplinary Center for Clinical Research (IZKF Aachen) and Start Program, RWTH Aachen University Medical School, Aachen, German

Hereditary Colorectal Tumors: A Literature Review on MUTYH-Associated Polyposis

MAP (MUTYH-associated polyposis) is a syndrome, described in 2002, which is associated with colorectal adenomas, with enhanced colorectal carcinogenesis. This review synthesizes the available literature on MAP and outlines its pathogenesis, association with colorectal tumorigenesis, screening, treatment, and the subtle differences between it and its close cousins—FAP and AFAP. The preponderance of data is collected using MAP guidelines. However, although AFAP and MAP appear similar, potentially important distinctions exist, warranting targeted diagnostic criteria and treatment approaches. We suggest that it may be prudent to screen for MAP earlier than in current clinical practice, as it has been shown that sequence variants are associated with more severe disease, presenting with an earlier onset of colorectal cancer. Finally, we issue a call-to-action for much-needed further data to establish clear clinical and diagnostic criteria

Keratin 8 phosphorylation regulates keratin reorganization and migration of epithelial tumor cells

Cell migration and invasion are largely dependent on the complex organization of the various cytoskeletal components. Whereas the role of actin filaments and microtubules in cell motility is well established, the role of intermediate filaments in this process is incompletely understood. Organization and structure of the keratin cytoskeleton, which consists of heteropolymers of at least one type 1 and one type 2 intermediate filament, are in part regulated by post-translational modifications. In particular, phosphorylation events influence the properties of the keratin network. Sphingosylphosphorylcholine (SPC) is a bioactive lipid with the exceptional ability to change the organization of the keratin cytoskeleton, leading to reorganization of keratin filaments, increased elasticity, and subsequently increased migration of epithelial tumor cells. Here we investigate the signaling pathways that mediate SPC-induced keratin reorganization and the role of keratin phosphorylation in this process. We establish that the MEK–ERK signaling cascade regulates both SPC-induced keratin phosphorylation and reorganization in human pancreatic and gastric cancer cells and identify Ser431 in keratin 8 as the crucial residue whose phosphorylation is required and sufficient to induce keratin reorganization and consequently enhanced migration of human epithelial tumor cells.</jats:p