Purpose: To determine whether symptoms of dizziness and fall rates change due to routine cataract surgery and to determine the influence of visual and refractive factors on these common problems in older adults. Methods: Self-reported dizziness and falls were determined in 287 subjects (mean age of 76.5±6.3 years, 55% females) before and after routine cataract surgery for the first (81, 28%), second (109, 38%) and both eyes (97, 34%). Six-month falls rates were determined using self-reported retrospective data. Dizziness was determined using the short-form of the Dizziness Handicap Inventory. Results: The number of patients with dizziness reduced significantly after cataract surgery (52% vs. 38%; χ2 = 19.14 , p < 0.001), but the reduction in number of patients who fell in the 6-months post surgery was not significant (23% vs. 20%; χ2= 0.87, p=0.35). Multivariate logistic regression analyses found significant links between post-operative falls and change in spectacle type (increased risk if switched into multifocal spectacles). Post-operative dizziness was associated with changes in best eye visual acuity and changes in oblique astigmatic correction. Conclusions: Dizziness is significantly reduced by cataract surgery and this is linked with improvements in best eye visual acuity, although changes in oblique astigmatic correction increased dizziness. The lack of improvement in falls rate may be associated with switching into multifocal spectacle wear after surgery.

Made available in DSpace on 2014-10-09T12:35:59Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:59:37Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:08/57338-5

Made available in DSpace on 2014-10-09T12:42:17Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:00Z (GMT). No. of bitstreams: 0 / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Made available in DSpace on 2014-10-09T12:35:59Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:59:37Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / No presente trabalho estudamos a renaturação sob alta pressão hidrostática de uma forma mutante da proteína verde fluorescente (enhanced GFP, eGFP), a qual somente emite fluorescência característica quando enovelada na sua forma nativa. A abordagem do presente estudo foi focada no controle da bioatividade da proteína recombinante, a fluorescência, como alternativa à determinação de solubilidade da proteína, fator que não é um indicador ideal de enovelamento proteico adequado. A ação da alta pressão na solubilização dos corpos de inclusão (CI) de eGFP produzidos em bactérias E. coli recombinantes e no enovelamento da proteína foi estudada. A compressão dos CI de eGFP em 2,4 kbar durante 30 minutos promoveu a dissociação dos agregados. No entanto, a incubação nesta condição não favoreceu o enovelamento da eGFP. O processo de renaturação foi avaliado em diversas condições de descompressão após a dissociação em 2,4 kbar. Durante a descompressão gradual, o aumento da fluorescência foi obtido em pressões que variaram entre a pressão atmosférica e 1,38kbar. Os níveis mais elevados de fluorescência de eGFP foram obtidos por incubação durante várias horas a níveis de pressão entre 0,35 e 0,69 kbar. Esta condição de pressão se mostrou favorável à renaturação de eGFP e é possível que também possa ser utilizada para favorecer o enovelamento de outras proteínas monoméricas. Ainda utilizando a eGFP como modelo, verificamos que os CI desta proteína produzidos por bactérias cultivadas em menor temperatura (37ºC) possuem maior quantidade de proteína recombinante apresentando a fluorescência característica em 509 nm, ou seja, na sua forma nativa, do que os CI expressos em temperaturas mais elevadas (42ºC e 47ºC). A análise realizada por espectroscopia de infravermelho (FT-IR) também demonstrou que os CI produzidos em temperaturas mais brandas possuem maior grau de estruturas secundárias semelhantes às da proteína na sua forma nativa. Além disso, os CI produzidos a 37ºC também são mais facilmente solubilizados pela ação da alta pressão do que aqueles produzidos em maior temperatura. Conforme esperado, a renaturação da eGFP a partir de CI produzidos a 37ºC foi 25 vezes mais eficiente do que a obtida utilizando CI produzidos a 47ºC. No presente estudo demonstramos também que a dissociação dos agregados exercida pela ação da alta pressão (2,4 kbar) pode ser amplificada quando em associação com a incubação em baixa temperatura (-9ºC) e que a combinação destas duas propriedades físicas eleva a solubilização dos agregados em CI, com a consequente elevação dos rendimentos de renaturação de eGFP. Mostramos ainda no presente estudo que a cinética de renaturação de eGFP em 0,69 kbar é proporcional à temperatura de incubação (entre 10ºC e 50ºC). O nível mais elevado de fluorescência foi obtido quando a renaturação de eEGP foi realizada a 20ºC. A taxa de maturação do cromóforo da eGFP é mais fortemente afetada pela temperatura do que a taxa de enovelamento da proteína. Em conclusão, a temperatura de produção dos CI, a temperatura de dissociação dos agregados e a temperatura de enovelamento podem afetar muito o rendimento e a cinética da renaturação de eGFP em alta pressão. Os resultados do presente estudo podem abrir novas perspectivas para melhorias no processo de enovelamento de proteínas a partir de CI utilizando alta pressão. Também neste trabalho descrevemos a renaturação das proteínas de Xac, PilB e os produtos dos genes XAC2810 e XAC3272 nunca antes obtidas na forma solúvel. Os rendimentos de solubilização destas três proteínas foram muito altos, entre 75% e 89%. A proteína PilB renaturada em alta pressão apresentou atividade ATPasica elevada, o que nunca antes foi demonstrado para a PilB de Xac. / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:08/57338-5

Made available in DSpace on 2014-10-09T12:42:17Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:00Z (GMT). No. of bitstreams: 0 / Com a intenção de estabelecer métodos físico-químicos como uma alternativa ao bioensaio in vivo para determinação de atividade biológica, o conteúdo de hFSH, hTSH e hLH de diferentes preparações, nativas e recombinantes, foi determinado por cromatografia líquida de alta eficiência em fase reversa (RP-HPLC) e comparado ao dado obtido pelo clássico bioensaio in vivo em camundongos ou ratos (BA). Para estes hormônios foi encontrada uma relação linear entre os dois métodos: hFSH BAUI = 0,9925 RP-HPLCUI - 1,3165, r = 0,9371, p < 0,001, n = 24; hTSH BA&mu;g = 0,9790 RP-HPLC&mu;g - 0,052, r = 0,8725 , p < 0,001, n = 14; hLH BAUI = 0,8771 RP-HPLCUI + 12,41; r = 0,9786, p < 0,01, n = 5. Para outras nove preparações de hFSH e onze preparações de hTSH foi determinada a diferença média (d) entre a bioatividade predita pela RP-HPLC através destas equações e da média das bioatividades obtidas com os dois métodos. Para o hLH não foi possível determinar esta diferença em virtude das poucas amostras disponíveis. No caso do hFSH, d ± DP = -2,11 ± 3,49 % sendo a precisão de 1,16% e no caso do hTSH, d ± DP = -2,01 ± 5,56 % com precisão de 1,68%. Amostras parcialmente alteradas apresentaram diferentes graus de atividade de hFSH, hTSH e hLH que puderam ser preditas por RP-HPLC com uma aceitável concordância com os bioensaios in vivo. Estes resultados demonstraram que o emprego de um ensaio físico-químico sem o uso de animais, tal como a RP-HPLC, é uma alternativa viável ao uso do bioensaio in vivo para a determinação da potência de hFSH e hTSH, reduzindo assim o número de animais em geral utilizados para assegurar a qualidade e eficácia de um produto farmacêutico. / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Purpose: To determine whether symptoms of dizziness and fall rates change due to routine cataract surgery and to determine the influence of visual and refractive factors on these common problems in older adults. Methods: Self-reported dizziness and falls were determined in 287 subjects (mean age of 76.5±6.3 years, 55% females) before and after routine cataract surgery for the first (81, 28%), second (109, 38%) and both eyes (97, 34%). Six-month falls rates were determined using self-reported retrospective data. Dizziness was determined using the short-form of the Dizziness Handicap Inventory. Results: The number of patients with dizziness reduced significantly after cataract surgery (52% vs. 38%; χ2 = 19.14 , p<0.001), but the reduction in number of patients who fell in the 6-months post surgery was not significant (23% vs. 20%; χ2= 0.87, p=0.35). Multivariate logistic regression analyses found significant links between post-operative falls and change in spectacle type (increased risk if switched into multifocal spectacles). Post-operative dizziness was associated with changes in best eye visual acuity and changes in oblique astigmatic correction. Conclusions: Dizziness is significantly reduced by cataract surgery and this is linked with improvements in best eye visual acuity, although changes in oblique astigmatic correction increased dizziness. The lack of improvement in falls rate may be associated with switching into multifocal spectacle wear after surgery. / The Dunhill Medical Trust

Abstract Aim The aim of this study was to describe levels and patterns of three-year-old children´s physical activity. Furthermore, to investigate if there were any weight status-, gender- and parental SES differences in three-year-old children’s physical activity levels, using objective and subjective measurements from Early Stockholm Obesity Prevention Project (Early STOPP). Methods Data from 93 children, three years of age, included in the Early STOPP study was used. Children wore an actigraph GT3X+ accelerometer for at least four consecutive days including one weekend day. Average activity in counts per minute (CPM), time spent in sedentary, light PA and moderate to vigorous physical activity (MVPA) was assessed (5 s epoch) and used to examine differences between gender, weight status (ISOBMI according to Cole et al) and socioeconomic status (SES). For this reason an index measuring SES was created using subjective data; parental reported information on living conditions and background characteristics, from the Early STOPP study. Differences between weekdays and weekend days was also examined.    Results The result showed neither any differences in PA between gender nor weight status and no differences between SES-groups. There was a difference in PA levels between weekdays and weekend days and a difference in PA between housing types. The children spent more time being active on weekdays and children living in apartments were more active than children living in villas. Children spent approximately 67% of their time being sedentary and an average of 12,5 minutes in MVPA. Conclusion The study concludes that PA-levels in children three years of age are low. There was a difference between housings, suggesting that parents/guardians play an important role in young children’s PA. However more research is needed to fully understand the PA-behavior of young children and their parents. The absence of PA differences between genders implies that gender differences later in childhood is an effect of social structures rather than innate differences. Objectively measured PA on children at this age is rare and therefore this study contributes to the knowledge regarding young children’s PA-behavior. Furthermore there is also a need to establish agreed upon definitions for SES and of thresholds to use when examining PA with accelerometer. / Sammanfattning Syfte Syftet med denna studie var att beskriva tre år gamla barns fysiska aktivitetsmönster och nivåer. Samt att undersöka om det fanns några skillnader i fysisk aktivitet (FA) beroende på viktstatus, kön och socioekonomisk tillhörighet genom objektiva och subjektiva mått från Early Stockholm Obesity Prevention Project (Early STOPP). Metod Data från 93 treåriga barn inkluderade I Early STOPP studien användes. Barnen använde en Actigraph GT3X+ rörelsemätare under minst fyra sammanhängande dagar med minst en helgdag. Genomsnittlig FA i slag per minut (CPM), tid i stillasittande, lätt aktivitet samt måttlig till kraftig fysisk aktivitet (MVPA) bedömdes (5 s intervaller) och användes för att undersöka om det fanns skillnader mellan kön, viktstatus (ISOBMI enligt Cole et al) samt socioekonomisk status (SES). Av denna anledning skapades ett index för SES med hjälp av subjektiva data; föräldrarapporterad information kring bakgrund och levnadsförhållanden från Early STOPP-studien. Även skillnader i FA mellan vardagar och helger undersöktes. Resultat Resultaten visade inga skillnader i FA mellan könen, viktstatus eller socioekonomisk tillhörighet. Det fanns däremot en skillnad mellan veckodagar och helger samt mellan boendeformer. Barnen var mer aktiva under veckodagarna och barnen boende i lägenhet var mer aktiva än barnen boende i villa. Ca 67 % av tiden spenderades i stillasittande och ungefär 12,5 min per dag spenderades i MVPA. Slutsats Studien drar slutsatsen att aktivitetsnivån hos tre år gamla barn är låg. Det fanns en skillnad i FA mellan boendeformer vilket indikerar att föräldrar/vårdnadshavare spelar en viktig roll för små barns aktivitet. Dock behövs mer forskning för att till fullo förstå barn och föräldrars aktivitetsmönster och hur dessa samvarierar. Avsaknaden av skillnader i FA mellan könen indikerar att könsskillnader senare i barndomen är en effekt av sociala strukturer snarare än medfödda skillnader. Objektivt uppmätt fysisk aktivitet hos barn i den här åldern är ovanligt och därför bidrar denna studie med kunskap kring små barns FA. Avslutningsvis drar denna studie slutsatsen att det finns ett behov av internationella bestämmelser kring definitioner av SES och av tröskelvärden att använda när FA mäts med accelerometri.

The present knowledge of factors associated with older adults’ physical activity behaviour is limited. Therefore, this study trialled an innovative physical activity program for older adults, investigating effective recruitment and retention strategies, and exploring the adults’ perceptions of physical activity. A total of 573 subjects were recruited into the quasi-randomised controlled trial, located in 30 intervention and 30 control neighbourhoods in the Perth metropolitan area. The initial response rate was 74% (260/352) in the intervention group and 82% (313/382) in the control group. Self-reported questionnaires administered at three time points (baseline, 3-months, 6-months) measured physical activity levels, personal and demographic information, including perception of financial struggle, proximity to friends, and other psychosocial data. Descriptive statistics, repeated measure analysis of variance, logistic regression and generalised estimating equations were used in the analysis. Qualitative data on the participants’ perceptions of physical activity were collected through one-on-one interviews (n=16). The results showed that: 1. This cost-effective recruitment procedure facilitated the selection of a reasonably representative sample of 65 to 74 year olds from the Perth metropolitan area. Names of 7378 older adults were obtained from the Federal Electoral Roll, then 6401 potential subjects were matched to telephone numbers and phoned with subjects meeting the screening criteria invited to join the program (n = 4209). From this sample, 573 subjects were recruited. More females (63%) than males (37%) were recruited. / The study attracted a greater proportion of ‘obese’ older adults (27%) relative to state averages. 2. Over the intervention period there was a significant increase in participants’ total physical activity of 2.25 hours per week (p >.001). The General Estimating Equation analysis confirmed significant increase in physical activity from baseline to midpoint (p=.002) and to post intervention (p=.0031). Perceptions of financial struggle (p=.020) were positively correlated with physical activity time spent by participants, whereas having friends or acquaintances living nearby (p=.037) had a significant negative correlation with physical activity time. 3. At the end of the intervention, 32% of the intervention group and 25% of the control group had dropped out, resulting in an overall drop out rate of 28%. Most of the attrition occurred in the first 3 months (77%). Characteristics of individuals lost to attrition (n=86, 35%) were compared with program completers (n=162, 65%). Logistic regression analysis showed that those lost to attrition came from areas of lower socio-economic status, were overweight, were less physically active, and had a lower walking self-efficacy score and a higher loneliness score. The results suggest that to improve retention and to avoid potential bias, early assessment of these characteristics should be undertaken to identify individuals at risk of attrition. 4. Based on the finding of this research, future intervention studies should consider: the role of tertiary students as a skilled resource; the use of volunteers to contain costs; the importance of a tailored program; the appropriateness of walking as a form of physical activity for this age group; the enjoyment associated with a walking group; and the usefulness of social support. / This practical program is potentially effective and sustainable for mobilizing physically inactive older people. 5. Qualitative research highlighted the need for older adults to receive more specific information on: the benefits of physical activity; the role of pain management in physical activity; and the concept that involvement in physical activity in younger years leads to involvement when older. The older adults also expressed a desire to engage in less age appropriate activities. These results suggest that the intervention was successful in recruiting older adults into and retaining them in the intervention, documenting a need for early identification of individuals at risk of attrition. The program significantly increased the participants’ weekly mean time for physical activity and identified factors that affect their commitment to physical activity programs. This program was practical and could be used as a model for physical activity programs aimed at older adults.

Made available in DSpace on 2014-10-09T12:27:28Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:07:44Z (GMT). No. of bitstreams: 0 / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Made available in DSpace on 2014-10-09T12:41:39Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:08:52Z (GMT). No. of bitstreams: 0 / Dissertação (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP