Hearing loss (HL) is a health condition that affects more than 360 million people worldwide. The findings from previous research point at the adverse relationship between adults with hearing loss and important aspects of everyday life such as social relations, communication and work-related tasks. However, the overall picture concerning the functional and disabling aspects of adults with HL re- mains incomplete. To identify the functional and disabling aspects, a conceptual and/or theoretical framework is required. The International Classification of Functioning, Disability and Health (ICF) offer a multidimensional framework based on bio-psycho-social assumptions about health. In previous research inves- tigations in which the ICF has been used, some utility problems in the linking (relating) of data to the classification have been highlighted. The aims of the present thesis were to explore the areas of functioning and disability of relevance for adults with HL and to explore how audiological data can be linked to ICF. The aims were explored by applying the methodology of the ‘interdisciplinary evidence-based approach to functioning and disability in adults with HL’, acknowledging the merging of three perspectives designated the Researcher, the Patient and the Professional perspective. Four studies that focus on the three perspectives were conducted. All results were linked to the ICF classification. The results were merged into a model designated ‘the integrative model of functioning and disability in adults with HL’. When the three perspectives were linked, the results revealed several aspects of relevance for the target group. Bodily (individual) dimensions, such as hear- ing, auditory perception, memory, attention, energy, and emotions, were acknowledged. Aspects of everyday life such as conversations, the usage of communication strategies, family relationships and work, were highlighted. Influential environmental factors, such as noise, assistive technical devices, the design of public buildings, social support and the attitudes of people in the envi- ronment, were also identified. In conclusion, interactions seemed to be vital as almost all identified aspects highlighted or were tied to this dimension of human functioning. Further, concerning the linking of the data it was acknowledged that the ICF and the research area of adult HL do not fully comply. Suggestions for improvements in future revisions of the ICF were highlighted and discussed. / <p>Funding agency: Oticon Foundation; Hörselforskningsfonden (Swedish HearingResearch Foundation); the Foundation for Audiological Research (FAR) Forskningskommitten, Region Örebro County</p>

Master of Science / Department of Biochemistry / Michael R. Kanost / Resilin is a rubber-like cuticular protein found in many insect species. Resilin is important for jumping and flying of those insects due to the properties of high elasticity and efficient energy storage. Some recombinant proteins or peptides derived from resilin sequences have been synthesized to produce biomaterials that mimic the remarkable properties of resilin. This research focused on resilin in the red flour beetle, Tribolium castaneum. A cDNA for T. castaneum resilin was inserted into plasmid vectors for expression of resilin in Escherichia coli or Bacillus subtilus. Resilin produced in E. coli was used as antigen to produce a rabbit antiserum. Resilin synthesized by B. subtilis as a secreted protein was purified and used for biochemical studies. Resilin is highly expressed in the late pupal stage, and in hind wings, but not found in elytra of pharate adults, indicated by RT-PCR and immunoblot analysis. Recombinant resilin could be cross-linked in the presence of horseradish peroxidase and hydrogen peroxide, detected by appearance of a high molecular weight band on SDS-PAGE, which had blue fluorescence under ultraviolet light, presumably due to dityrosine linkages. RNA interference was used to knock down resilin expression in T. castaneum. Immunoblot and RT-PCR analyses indicated that resilin expression was successfully decreased by RNAi. However, the knockdown adults exhibited no apparent differences in morphology, behavior or life span from control beetles. Blue fluorescence under ultraviolet illumination has frequently been used as an indication of the presence of resilin containing dityrosine cross-links in insect tissues such as wings, wing tendons and leg joints. A similar blue fluorescence was observed in hind wings of T. castaneum. However, this fluorescence was not decreased in hind wings of beetles in which resilin expression was knocked down by RNA interference. There was a blue fluorescence in the hind wings of knockdown beetles, which was similar in distribution to that in wings of control insects. This result suggests that the observed blue fluorescence in T. castaneum hind wings is derived not only from cross-linked resilin but also from components other than resilin, perhaps other cuticular proteins that contain dityrosine cross-links.

Patients with keratoconus exhibit a biomechanically weakened cornea which loses its proper shape and thereby loses its refractive power. It is usually progressive, beginning with poor visual acuity and eventually necessitating corneal transplant. The cause is likely multifactorial, but involves the weakening of the collagen structure of the corneal stroma, resulting in characteristic thinning and conical distortion. Collagen cross-linking is the first treatment to demonstrate efficacy in halting the progression of the disease. UVA radiation is used to activate riboflavin and photochemically induce cross-linking reactions among collagen and proteoglycans within the stroma, thereby stiffening and strengthening the tissue, and preventing further loss of shape. The current standard treatment, which gained FDA approval less than one year ago, has proven to be efficacious, but has been modified very little since pioneering experiments. Optimization aims to maximize clinical effect while maintaining safety and reducing total treatment time. Major procedural modifications involve increasing light intensity over a reduced exposure duration, and varying the method of delivering riboflavin to the stroma. Theoretical modeling, informed by and scaled to experimental results, has the potential to predict clinical effect as a function of treatment parameters, enabling tailoring of individual treatments to the specific needs of each patient.

Now that the functioning of microtubules and the actin cytoskeleton has been worked out in enormous detail, the next important task is defining the structure of intermediate filaments that are far behind the other two major skeletal networks due to their inherent resistance to most structural techniques. The evolution of novel structural approaches for flexible proteins is making this possible now. In my thesis I will aim to elucidate the structure and assembly principles of lamin A nuclear intermediate filament protein. To study lamin A, I principally employed chemical cross-linking that allows the capturing of full-length protein structures in solution. I combined this with mass spectrometry approaches to identify cross-linked residues at the various stages of lamin A assembly that were additionally tracked with SILAC labelling and rotary metal shadowing TEM. Unlike previous cross-linking studies on intermediate filaments I use a zero-length self-excluding cross-linking agent EDC that is better tailored for investigation of the polar interactions between multiple unstructured or otherwise flexible charged sequences of lamins. Using this composite approach I interrogated lamin A dimeric and tetrameric assemblies. I elucidated hinge-like properties of the L12 and found indications that L1 and the region containing coil 2A and L2 and the beginning of coil 2B possess properties of linker-like flexibility and of predicted linear α-helical bundle and could act as molecular springs or compression buffers for the nuclear intermediate filaments. Further I confirm the role of the N-terminal unstructured region in lamin A assembly and for the first time show similar role for the C-terminal unstructured region flanking the rod domain of lamin A. Collected data strongly supports the model where both positively charged unstructured regions participate in extensive interaction with acidic rod termini and act as molecular bridges between these in the head-to-tail interface, confirming the uniformity of this principle between cytoplasmic and nuclear intermediate filaments. Formation of these bridges requires conformational change likely happening due to proline residues in the mitotic phosphorylation sites. Finally I suggest a mechanism of regulation of the order of assembly unique to the nuclear intermediate filament where C-terminal unstructured region blocks lateral interactions until it is tethered to the head-to-tail interface. Collected data on the dynamic behaviour of the C-terminal unstructured region and its ability to tether lamin A Ig domain may have far reaching implications for filament assembly and regulation of binding of hundreds of lamin A partner proteins presenting an important step in our understanding of relationship between lamin A structure and function and how altering the former could lead to disease.

O objetivo deste trabalho foi elucidar a interação entre a proantocianidina (PA), um agente natural e não tóxico de ligação cruzada, e o colágeno dentinário tipo I. A hipótese proposta é que o modelamento de uma dentina desmineralizada mais resistente e com melhores propriedades propiciará um substrato mais estável e íntegro para restaurações adesivas. Para isto, o estudo foi dividido em 4 fases: 1) Validação da PA como agente exógeno de ligação cruzada, testando matrizes de dentina desmineralizadas tratadas com extratos a base de PA em teste microflexural para análise do módulo de elasticidade (ME), variando o tempo de exposição (10, 30, 60, 120, 240 min), o tipo de extrato usado (semente de uva-GSE, semente de cacau-CSE, oxicoco-CRE, canela-CNE e açaí-ACE) e a longevidade (imediato, 3, 6 e 12 meses); 2) Estudar parâmetros relacionados às soluções de PA, como solubilidade (água, etanol e acetona), fonte (GSE e CSE) e concentração (0,65%, 3,25%, 6,5%, 15% e 30%), através da mensuração do ME pelo teste micro-flexural de matrizes de dentina desmineralizada; 3) Caracterização da dentina tratada com PA (GSE e CSE), sorção de água, calorimetria e degradação enzimática; 4) Analisar a resistência de união entre a dentina tratada com PA (GSE/CSE) e sistemas de adesivos comerciais (Adapter Single Bond Plus e One Step Plus), variando a concentração (6,5% e 30%), tempo de exposição (1, 10 e 60 min) e longevidade (imediato, 3, 6 e 12 meses). Foram realizadas análises de variâncias e teste de contraste de médias, quando necessário, para todos os ensaios. Os resultados de ME foram influenciados pelo tratamento com PA, dependendo do tipo de extrato usado, exibindo os melhores resultados para o maior tempo de exposição (240 min) para os extratos GSE e CSE. Embora os resultados imediatos com os extratos CRE e CNE não tenham aumentado, estes se mantiveram constantes apos 1 ano de armazenamento, o que não aconteceu para o ACE e os grupos controles. A solubilidade das soluções é influenciada diretamente pelo modo de extração e manufaturação dos extratos, sendo que GSE é solúvel em água destilada e CSE em acetona-água. A efetividade do GSE mostrou ser concentração-dependente, exibindo o maior ME para a concentração de 30%. A dentina tratada com agentes a base de PA foi mais resistente contra a colagenase, obteve uma menor sorção de água e um aumento significativo na temperatura de desnaturação quando comparada ao grupo controle. O GSE foi capaz de aumentar a resistência de união (RU) imediata para ambos os sistemas adesivos, mantendo-se constante para o Adapter Single Bond Plus ao longo do tempo. Para o One Step Plus todos os grupos mostraram diminuição da RU após 1 ano, independente do tempo de exposição. Quando a dentina foi tratada por 1 minuto, os valores de RU para o grupo 30% de GSE foram maiores do que os demais e mantiveram-se constante pelo período de armazenamento. O efeito dos agentes ricos em PA mostrou ser concentração e tempo dependente, porém uma dentina mais resistente e estável ao longo do tempo é possível de ser obtida, levando-se em conta a estrutura química, conteúdo de PA, solubilidade e outros fatores inerentes ao extrato / The aim of this study was to elucidate the interaction between proanthocyanidin based extracts (PA), a natural and non-toxic cross-linker, and type I dentin collagen. The hypothesis is that if a stronger and more stable collagen layer is chemically engineered, the resultant hybrid layer will be stronger and less prone to degradation. So, this research was divided into four phases: 1) PA validation as an exogenous cross-linking agent, testing the elastic modulus (E) of demineralized dentin treated with various PA based extracts (grape seed-GSE, cocoa seed-CSE, cranberry-CRE, cinnamon-CNE, and açai berry-ACE), exposure times (10, 30, 60, 120, 240 min), and long term effectiveness (immediate, 3, 6 and 12 months); 2) Evaluation of different parameters related to PA solutions, like solubility (water, ethanol and acetone), manufacturer (GSE and CSE) and concentration (0.65%, 3.25% 6.5%, 15% and 30%), E was also obtained by micro-flexural strength measurements of demineralized dentin matrix, 3) Characterization of PA treated dentin (GSE and CSE): water sorption, calorimetry and enzymatic degradation and 4) Analyze the bond strength ( BS) of PA treated dentin (GSE / CSE) with commercial adhesives systems (Adapter Single Bond Plus-SB and One Step Plus-OS), varying concentration (6.5% and 30%), exposure time (1, 10 and 60 min) and long-term effectiveness (immediate, 3, 6 and 12 months). Data were statistically analyzed at a 95% confidence interval. GSE and CSE extracts showed a time-dependant effect and were able to improve and stabilize the E of the organic matrix. CRE and CNE extracts were able to maintain the E of collagen matrices constant after 12 months artifical saliva storage. ACE and controls groups showed no effect over dentin organic matrix, did not prevent degradation and consequently reduced the mechanical properties. Herbal extraction process and other pharmacognostic parameters have an important influence on extract solubility as well as constitution, therefore GSE is better dissolved in water and CSE in acetone-water. GSE effect on demineralized dentin is concentration-dependant, with highest E values at 30% GSE concentration. Dentin treated with PA-based agents was more resistant against enzymatic degradation, less susceptible to water sorption and showed a higher denaturation temperature. As compared to the controls, GSE stabilized the dentin-resin bond strength values over a period of one year for both adhesives, however better results were achieved with SB. CSE showed less predictable results depending on the adhesive system used. One minute treatment with 30% GSE was capable of increasing short-term BS and stabilizing it for at least 6 months. Increased mechanical properties and stability of dentin matrix can be achieved by the use of PA-rich collagen crosslinkers regarding inherent characteristics of the natural compounds as chemical structure, solubility and PA content.

The main purpose of this study was to construct different vertical scales based on various combinations of calibration methods and proficiency estimators to investigate the impact different choices may have on these properties of the vertical scales that result: grade-to-grade growth, grade-to-grade variability, and the separation of grade distributions. Calibration methods investigated were concurrent calibration, separate calibration, and fixed a, b, and c item parameters for common items with simple prior updates (FSPU). Proficiency estimators investigated were Maximum Likelihood Estimator (MLE) with pattern scores, Expected A Posteriori (EAP) with pattern scores, pseudo-MLE with summed scores, pseudo-EAP with summed scores, and Quadrature Distribution (QD). The study used datasets from the Iowa Tests of Basic Skills (ITBS) in the Vocabulary, Reading Comprehension (RC), Math Problem Solving and Data Interpretation (MPD), and Science tests for grades 3 through 8. For each of the research questions, the following conclusions were drawn from the study. With respect to the comparisons of three calibration methods, for the RC and Science tests, concurrent calibration, compared to FSPU and separate calibration, showed less growth and more slowly decreasing growth in the lower grades, less decrease in variability over grades, and less separation in the lower grades in terms of horizontal distances. For the Vocabulary and MPD tests, differences in both grade-to-grade growth and in the separation of grade distributions were trivial. With respect to the comparisons of five proficiency estimators, for all content areas, the trend of pseudo-MLE ≥ MLE > QD > EAP ≥ pseudo-EAP was found in within-grade SDs, and the trend of pseudo-EAP ≥ EAP > QD > MLE ≥ pseudo-MLE was found in the effect sizes. However, the degree of decrease in variability over grades was similar across proficiency estimators. With respect to the comparisons of the four content areas, for the Vocabulary and MPD tests compared to the RC and Science tests, growth was less, but somewhat steady, and the decrease in variability over grades was less. For separation of grade distributions, it was found that the large growth suggested by larger mean differences for the RC and Science tests was reduced through the use of effect sizes to standardize the differences.

No description available.

No description available.

Lignocellulose can become the best opportunity for the society to reduce its dependency on the harmful petroleum based products as well as to produce clean energy. In each part of the production cycle, biomass based products have a better environmental profiles than their petroleum based counterparts. Woody biomass has a vast availability, but it suffers from recalcitrance that is mostly caused by lignin that is functioning as a matrix, surrounding and binding the carbohydrates that are currently the most valuable of the wood components. Lignin-carbohydrate (LC) bonds are believed to be a key element in this recalcitrance and research has shown that these types of bonds are common in wood. These bonds are important in an economical point of view as well, as e.g. residual lignin structures in pulp (lignins bonded to the cellulose and hemicelluloses) require expensive bleaching sequences for their removal. The LC-structures can also be exploited technically as we now have demonstrated. We developed a method that utilizes phenolic end groups that are bonded to different hemicelluloses for cross-linking. The enzyme laccase was used for the cross-linking to create a cost-efficient processing scheme to both isolate and increase the molecular weight of the hemicelluloses. Membrane filtration was used as the key separation technique, which enables the establishment of industrial scale production. The final product had improved mechanical and thermal properties and could be used e.g. as barrier film component in renewable packaging. Nanocomposite formation with nanofibrillated cellulose was also studied. This improved the film properties further. The complexes are also possible to use as model compounds for lignin-carbohydrate complexes in wood. This technique can also be seen to mimick the lignification and lignin-carbohydrate network formation phenomena in plants enabling the formation of entire networks of wood components. Our results suggests that the side chains of hemicellulose might play an important role in network formation and that hemicellulose molecules can carry more than one lignin phenolic end group to fulfill this capability. / <p>QC 20140825</p>

No description available.