As habilidades manuais podem estar prejudicadas na paralisia cerebral (PC),interferindo nas atividades funcionais do cotidiano da criança, como alimentar-se, ir ao banheiro, brincar, o que ocasiona impacto negativo na vida dela e da família. Objetivo: elaborar, aplicar e analisar um protocolo de avaliação da função manual de crianças com PC, de 0 a 7 anos e 11 meses de idade. Método: estudo dividido em 2 etapas - 1) planejamento: definição de objetivo e população alvo, realização de revisão da literatura sobre instrumentos já existentes, tipos de itens e formato de instrumento, 2) construção: construção I -desenvolvimento dos itens, seleção dos instrumentos utilizados como base do protocolo, seleção dos itens relevantes por faixa etária, análise dos itens e adequação do protocolo; projeto piloto - aplicação do protocolo junto a 36 crianças de 0 a 7 anos e 11 meses, sendo 18 com PC e 18 com DT, análise dos itens e adequação do protocolo; construção II - validação de conteúdo por profissionais/pesquisadores experientes (experts) na área, elaboração e envio do questionário aos juízes especialistas, revisão dos itens (inclusão e exclusão), análise das avaliações dos especialistas e adequação do protocolo. Resultados: foram selecionados 238 itens, sendo 38 itens da Escala Lúdica Pré Escolar de Knox (ELPK-R), 126 do Inventário Portage Operacionalizado (IPO), 32 do Teste de Triagem de Desenvolvimento de Denver II (TTDDR), 38 do Inventário de Avaliação Pediátrica de Incapacidade (PEDI) e 4 do Manual de Avaliação Motora - Escala de Desenvolvimento Motor (MAM-EDM). Todos os itens foram revisados. Realizou-se o agrupamento de 161 itens recorrentes, totalizando 120 itens, os quais foram aplicados em crianças com PC e DT. Após a aplicação, em nova análise do protocolo, foram agrupados 6 itens semelhantes, divididos 2 e excluídos 7, totalizando 109 itens, os quais foram enviados em forma de questionário para os juízes especialistas. Na sequência, as avaliações dos experts foram analisadas e, então, 3 itens foram agrupados, 1 item subdividido e 7 itens excluídos, chegando-se a um protocolo final com 101 itens, divididos em 8 faixas etárias: de 0 a 5 meses, de 6 meses a 11 meses, de 1 ano a 1 ano e 11 meses e, sucessivamente, na sequência anual até de 4 anos a 4 anos e 11 meses; em seguida há uma faixa etária de 5 anos a 6 anos e 11 meses e, após, a faixa etária de 7 anos a 7 anos e 11 meses. O resultado foi então denominado de Avaliação Manual Infantil Geral e Objetiva - AMIGO. Foram observadas 2 crianças de cada grupo em cada faixa etária, sendo que as crianças com DT apresentaram, no geral, melhores pontuações quando comparadas às crianças com PC da mesma faixa etária. Considerações finais: a avaliação AMIGO foi considerada válida e alcançou seu objetivo principal. Ainda está, porém, em processo de construção e, em estudos futuros, passará por mais duas fases de construção, sendo elas a fase III - avaliação quantitativa e a fase IV - validação, para que possa ser considerada totalmente válida e segura a sua utilização na prática clínica e na pesquisa científica. / Manual skills may be affected in cerebral palsy (CP), interfering with functional activities of child\'s daily life, such as eating, toileting, playing, which causes negative impact on child´s life and family. Objective: To develop, implement and analyze an evaluation protocol to assess manual children function with CP, aged between 0-7 years and 11 months old. Method: This study was divided into two stages - 1) planning: setting goals and target population, literature review on existing instruments, types of items and instrument format, 2) Construction: Construction I -development of items, selection of instruments used as basis of the protocol, selection of relevant items by age group, item analysis and protocol adjustment; Pilot Project - protocol application with 36 children 0-7 years, 11 months, 18 CP childerb and 18 children with typical development (TD), item analysis and protocol adjustment; Construction II - Content validation by professional / experienced researchers (experts) in the area, preparation and submission of the questionnaire to expert judges, items review (item inclusion or exclusion), analysis of experts evaluations and suitability of the protocol. Results: We selected 238 items, 38 items Revised Knox Preschool Scale (RKPPS) 126 items from Operationalized Portage Inventory (IPO), 32 from Denver II Developmental Screening Test (DDST-R) 38 from Pediatric Evaluation of Disability Inventory (PEDI) and 4 from Manual of Motor Evaluation - Motor Development Scale (MAM-EDM). All items were reviewed. We grouped 161 recurring items, totalizing 120 items, which we applied both in CP children and with CP and TD children. After the implementation, we performed a new protocol analysis and 6 similar items were grouped, 2 items divided and 7 items excluded, totalizing 109 items, which were sent as questionnaire to the expert judges. Further, experts assessments were analyzed and then 3 items were grouped, 1 item subdivided, and 7 items deleted, coming to a final protocol with 101 items, divided into eight age groups: 0-5 months of 6 months to 11 months, 1 year to 1 year and 11 months and, subsequentially, the annual sequence till 4 years to 4 years and 11 months; then there is an age group between 5 years and 6 years and 11 months, and after the age of 7 years to 7 years and 11 months. The resulting product was then called Children´s Manual Assessment General and Objective - AMIGO. Two children in each group were observed at each age, and children with TD had, overall, better scores compared to children with CP in each age group. The final considerations were that AMIGO has been valid and reached its main goal. But it is still under construction, and in future studies it will go through two more stages of construction, which were phase III - quantitative assessment and phase IV - validation, in order to be considered valid and secure to be uesd in clinical practice and in scientific research.

Deep Learning is dominant in the field of computer vision, thanks to its high performance. This high performance is driven by large annotated datasets and proper evaluation benchmarks. However, two important areas in computer vision, depth-based hand segmentation, and local features, respectively lack a large well-annotated dataset and a benchmark protocol that properly demonstrates its practical performance. Therefore, in this thesis, we focus on these two problems. For hand segmentation, we create a novel systematic way to easily create automatic semantic segmentation annotations for large datasets. We achieved this with the help of traditional computer vision techniques and minimal hardware setup of one RGB-D camera and two distinctly colored skin-tight gloves. Our method allows easy creation of large-scale datasets with high annotation quality. For local features, we create a new modern benchmark, that reveals their different aspects. Specifically wide-baseline stereo matching and Multi-View Stereo (MVS), of keypoints in a more practical setup, namely Structure-from-Motion (SfM). We believe that through our new benchmark, we will be able to spur research on learned local features to a more practical direction. In this respect, the benchmark developed for the thesis will be used to host a challenge on local features. / Graduate

No description available.

Although tremor is one of the most common movement disorders, there are few effective tremor-suppressing options available to patients. One potential tremor-suppression device involves a wearable gyrostabilizer similar to those used to stabilize cameras. However, we do not currently know how to design a gyrostabilizer to suppress tremor in an optimal manner. To address this gap, we present a systematic investigation of how gyrostabilizer parameters affect tremor suppression in a single degree of freedom (DOF). A simple model of the hand with a single DOF at the wrist and a gyroscope mounted on the back of the hand was used to focus on the most basic effects. After demonstrating that a linearized version of the non-linear equations of motion provides an adequate approximation, we simulated the frequency response of the system (hand + gyroscope) to a tremorogenic input torque at the wrist. By varying system parameters one at a time, we determined the effect of individual parameters on the frequency response of the system. To minimize the bandwidth without adding significant inertia about the wrist joint, the inertia and spin speed of the flywheel should be as high as design constraints allow, whereas the distance from the wrist joint axis to the gyroscope, the precession stiffness, and the precession damping should be kept as low as possible. The results demonstrate the potential of gyroscopic tremor suppression and can serve as the foundation for further investigations of gyroscopic tremor suppression in the upper limb.

The Wnt signaling network plays critical roles in development and is implicated in human disease. Wnts comprise a complex signaling network that, upon ligand binding, activates the phosphoprotein Dishevelled (Dvl), leading to distinct outputs including polarized cell movement (known as planar cell polarity, Wnt/PCP) and stabilization of the transcription factor β-catenin (Wnt/β-catenin). The mechanisms that determine a specific output are not completely understood, especially because they share receptors and cellular effectors, such as Naked-cuticle 1 (Nkd), a Dvl-interacting protein. The Nkd protein contains a myristoylation domain and an EF-hand, a putative calcium binding domain. Genetic evidence in Drosophila demonstrates that Nkd acts as a Wnt/β-catenin antagonist, while in contrast, Nkd modulates both branches of Wnt signaling in vertebrates. We hypothesize that the specialized role of Nkd in Drosophila is due to a disrupted EF-hand that cannot not bind calcium. Indeed, this change is unique to Drosophila and is not present in closely related insects all the way up to vertebrates. To test the role of the Nkd EF-hand in Wnt signal integration, we created two different mutations in the zebrafish Nkd: one with a neutralized EF-hand, as well as a Drosophila-like EF-hand, and manipulate Nkd activity in the zebrafish. Using a combination of biochemical and functional assays, we identified a requirement for the Nkd EF-hand in Wnt/PCP but not in Wnt/β-catenin transcriptional outputs. We demonstrate that the Drosophila-like antagonizes Wnt/β-catenin more robustly than zebrafish Nkd. The EF-hand of Nkd is similar to the EF-hand of a known calcium binding protein, Recoverin, a myristoyl-swtich protein that shuttles between the membrane and the cytoplasm depending on its calcium bound state. Consistently, we observe that NkdWT, but not the two mutant forms, shows localization changes in the calcium fluxing cells that also host converging Wnt signals versus calcium quiescent cells. Our functional data suggests that the Nkd EF-hand is important for Wnt signal integration. Interestingly, Nkd only contains one EF-hand, and proteins that bind calcium tend to have multiple. Calcium binding can also be influenced by binding partners. Because of this, we investigate the role of the Nkd binding protein Dvl and their possible calcium affinity. Dvl is a pivotal point in the Wnt signaling network, leading to the output decision of a cell. EF-hand of Nkd binds to the PDZ domain of Dvl. Interestingly, the Dvl PDZ domain contains a region rich in negatively charged amino acids that could aid in binding calcium. In the same manner as Nkd, we generated a Dvl with neutralized putative EF-hand and tested its function and localization relative to wildtype Dvl. This work elucidates the elegant mechanism by which a cell receiving multiple Wnt signals integrates the information into a specific response. The Nkd EF-hand may serve to interpret the physiology of a cell receiving multiple cues and provides mechanistic insight into Wnt signal integration in vivo.

Human immunodeficiency virus type 1 (HIV-1) persists in certain CNS cell populations, despite peripheral control of the infection with modern antiretroviral therapy. Infected and/or activated cells release viral proteins, such as trans-activator of transcription (Tat) and various pro-inflammatory factors such as CCL5, creating a positive loop of neuro-inflammation. This serves as the basis for the resulting sublethal and lethal neuropathology that manifests as a spectrum of HIV-mediated CNS impairments, known as HIV-associated neurocognitive disorders (HAND). Opiates, which exist as an interlinked epidemic with HIV-1 infections, exacerbate these neurological effects through direct and indirect mechanisms that disrupt both glial and neuronal function. We hypothesize this is due to converging actions on the CCL5-CCR5 signaling axis by HIV-1 Tat and morphine co-exposure, primarily mediated at the level of the glia, whose consequent activation leads to neuronal damage. We performed repeated measure studies on mixed glia and neuron co-cultures obtained from C57Bl/6J and/or CCR5 knockout mice, treated with Tat and/or morphine for 72 hours. As established in prior studies, morphine worsened Tat-induced neurotoxicity in wild-type co-cultures; substitution of CCR5-null glia eliminated the interactive effects of Tat and morphine, but substitution of CCR5-null neurons did not. Overall, these results suggest that glial CCR5, but not neuronal CCR5, is a convergence point for the interactive effects of Tat and morphine that result in neuron loss. Additional experiments involving treatments with naloxone, a MOR antagonist, or the CCR5 antagonist maraviroc, confirmed each receptor’s role in mediating Tat + morphine toxicity. Quite surprisingly, in co-cultures of wild-type neurons and CCR5-null glia, morphine entirely protected neurons from the neurotoxic effects of Tat. We hypothesize that this effect may reflect an imbalance of neurotrophic factors, particularly BDNF and its neurotoxic precursor proBDNF, whose levels are altered in HIV+ and illicit drug-using patients and may contribute to changes in neuronal signaling and survival exhibited in HAND. Related behavioral tests of anxiety, motor and cognitive function – three areas of neurologic decline seen in HAND – were performed in inducible Tat-transgenic mice that were treated with maraviroc via oral gavage. Tat-mediated impairment was observed in the Barnes Maze, a measure of spatial memory, and was ameliorated by maraviroc. Finally, we assessed the role of CCR5 in mediating Tat and/or morphine effects on psychomotor sensitization and dendritic morphology. With both in vitro and in vivo studies, our findings support the hypothesis that CCR5 plays a central role in driving HIV-1 Tat and/or morphine-mediated neuronal damage.

The Joint Commission reported that communication failures in United States hospitals contribute to 30% of all malpractice claims, and in 2008, The Joint Commission listed the improvement of effective communication among healthcare providers as a National Patient Safety Goal. The purpose of this practice-focused project was to develop a scenario-based communication simulation for implementation approval by a panel of experts. The approved curriculum was designed to integrate a communication-based simulation scenario into the nurse residency program of a large, urban medical center to improve new graduate nurse communication skills, increase nurse communication competency and self-efficacy, and decrease communication errors. Kolb's experiential learning theory guided the simulation-based educational project. The Delphi technique was used to achieve consensus, which was achieved with 1 Delphi round. The education curriculum was presented to a 5-member expert panel that included chief nursing officers and the staff development directors. The curriculum received panel feedback and approval for implementation in the 2019 nurse residency program. Key comments from the expert panel indicated that the curriculum was approved without major changes. A simulation communication curriculum integrated into the nurse residency program may effect positive social change by decreasing errors and improving patient outcomes.

The Wnt signaling network has critical roles in development and disease. Simplified, this complex network has two distinct outputs: the Wnt/β-catenin module activates the phosphoprotein Dishevelled (Dvl) and leads to transcriptional activation while the Wnt/Planar Cell Polarity (PCP) module activates Dvl and leads to calcium release and directed cell movement. Wnt/β-catenin and Wnt/PCP share signaling components like Frizzled receptors, Dvl, and Naked (Nkd). It is an open question how converging Wnt signals diverge into separate outcomes. In this thesis, I used molecular techniques, functional studies in the zebrafish, and biochemical approaches to determine the role of Nkd in Wnt signaling. Nkd contains and EF-hand, a putative calcium binding domain, and is known to antagonize Wnt/β-catenin and disrupt Wnt/PCP signaling. We utilized a tissue that requires both Wnt/β-catenin and Wnt/PCP signaling to properly pattern the left/right axes of the embryo; the dorsal forerunner cells (DFCs). The DFCs exhibit aperiodic calcium release as they migrate to form the Kupffer's Vesicle (KV), the organ of asymmetry. Calcium inhibition in the DFCs disrupts their migration, alters KV formation, and disrupts left/right patterning. Nkd is enriched in the DFCs during migration and KV formation and endogenous Nkd knockdown in the DFCs produces the same phenotypes as calcium inhibition, making Nkd a candidate molecule for directing converging Wnt signals to distinct outcomes. To assess the role of the EF-hand in Nkd function, I created point mutations predicted to disrupt EF-hand affinity for calcium. Through functional studies in zebrafish embryos, I determined that Nkd EF-hand is necessary for Nkd function in Wnt/PCP signaling, but dispensable for Wnt/β-catenin signaling. Although Nkd has not been shown to bind calcium, our functional data with the Nkd EF-hand point mutant provides compelling evidence for a role for calcium in Nkd function in directing Wnt signaling output. EF-hand affinity for calcium is influenced by binding partners, and since Nkd binds to Dvl in the Dvl PDZ domain, we screened the domain for a region rich in amino acids that facilitate ion binding. We identified a 12-amino acid sequence in the Dvl PDZ domain with potential to create a negatively charged pocket to help coordinate calcium binding. We expressed the Nkd EF-hand (EFX) Dvl basic domain and PDZ domain (bPDZ). The purified EFX and bPDZ constructs were used to investigate the interaction between Nkd, Dvl, and calcium. I show, by circular dichroism, that the Nkd/Dvl complex undergoes a calcium-induced change in secondary structure. This reveals the mechanism by which Nkd directs Dvl from the default Wnt/β-catenin signaling module to the Wnt/PCP module in response to calcium.

In our efforts to engineer a DNA binding and cleaving protein with greater sequence discrimination, we have designed dimeric proteins derived from engrailed homeodomain and calmodulin. Previous research by our group has shown that a hydrolytically active lanthanide binding site can be incorporated into a DNA binding motif. To understand protein-DNA interaction and improve the sequence selectivity of the chimeric complex, two lanthanide-binding homodimers were designed and expressed. One of the dimers, F2, is coupled together by a flexible polypeptide linker and the other, R7C, is a disulfide cross-linked cysteine mutant at the N-terminus. Studies of fluorescence of tryptophan residues document that the overall affinity for lanthanide and calcium is similar to traditional EF-hand peptides (1-10 μM). Metal titrations monitored by circular dichroism (CD) revealed that the secondary structures of the dimers contained a lower degree of -helicity than the designed monomeric protein due to additional modifications, but because of their flexibility and their two active-site domain, hydrolytic activity was several folds faster than our previously designed proteins and peptides. Unlike earlier reports on our chimeras, F2 also demonstrated the capability to hydrolyze DNA in the presence of some biological relevant metal ions suggesting different cleavage mechanisms were carried out. Extensive DNA sequencing studies on cleavage patterns with oligonucleotide duplexes confirmed the unique sequence selectivity and kinetic properties of F2. Two engrailed homeodomain target sites, TAATTA, were favored for hydrolytic activity corresponding to one domain acting as a DNA anchor on the first target site while the other was an "opportunist" at recognizing the second site. Nonetheless, the hydrolytic behavior at the phosphodiester bond on a specific dsDNA sequence is in good agreement with the behavior of restriction endonucleases. Unlike restriction enzymes, metallated F2 has not only demonstrated the ability to cleave DNA plasmid, but it also excises the entire nucleotide on a selected sequence. This homodimer is the first example of an active and selective hydrolytic artificial nuclease based on the modular turn substitution design approach that can be a potential template for genomic modification.

The purpose and underlying motivation for this project was to examine the procedure of garment construction methods, by specifically choosing to abandon traditional rules and standards that are associated with mass production. I chose to explore domestic hand-craft made by women in New Zealand in their domestic situation, focusing on hand-stitched techniques from the past reflecting a nostalgic value which potentially contributed to the garment’s construction process. The project sought to utilize the re-using of materials by incorporating previously made hand-craft; and looked at the remaking of second-hand garments by means of deconstruction and reconstruction. Traditional hand-craft, in this context, refers to the use of craft forms not governed by principles of efficiency, mass production or technology, allowing the garments to contain unique ‘one of a kind’ hand-made qualities. My studio practice specifically focused on exploring the relationship between hand-craft and garment construction, by researching their application and integration into the garment’s structure, along with disrupting the orderly traditional production process. I was not focusing on the finished garment’s design. The project provided an opportunity to refocus my attention on the hand-made, as I perceived that the skills required to produce these were being extinguished by modern lifestyles. Exploration promoted new discoveries by exposing the construction process and revealing unpredicted combinations. The project explored these ideas, resulting in a range of women’s garments that revealed, as part of their construction, hand-work which offered a modern variation of nostalgia. This project comprised of 80% practical work and will be accompanied by an exegesis with a value of 20%.