Surface plasmon resonance as a tool in the functional analisys of an immunodominant site in foot-and-mouth disease virus

A fast and direct surface plasmon resonance (SPR) method for the kinetic analysis of the interactions between peptide antigens and immobilised monoclonal antibodies (mAb) has been established. Protocols have been developed to overcome the problems posed by the small size of the analytes (< 1600 Da). The interactions were well described by a simple 1:1 bimolecular interaction and the rate constants were self-consistent and reproducible. The key features for the accuracy of the kinetic constants measured were high buffer flow rates, medium antibody surface densities and high peptide concentrations. The method was applied to an extensive analysis of over 40 peptide analogues towards two distinct anti-FMDV antibodies, providing data in total agreement with previous competition ELISA experiments. Eleven linear 15-residue synthetic peptides, reproducing all possible combinations of the four replacements found in foot-and-mouth disease virus (FMDV) field isolate C-S30, were evaluated. The direct kinetic SPR analysis of the interactions between these peptides and three anti-site A mAbs suggested additivity in all combinations of the four relevant mutations, which was confirmed by parallel ELISA analysis. The four-point mutant peptide (A15S30) reproducing site A from the C-S30 strain was the least antigenic of the set, in disagreement with previously reported studies with the virus isolate. Increasing peptide size from 15 to 21 residues did not significantly improve antigenicity. Overnight incubation of A15S30 with mAb 4C4 in solution showed a marked increase in peptide antigenicity not observed for other peptide analogues, suggesting that conformational rearrangement could lead to a stable peptide-antibody complex. In fact, peptide cyclization clearly improved antigenicity, confirming an antigenic reversion in a multiply substituted peptide. Solution NMR studies of both linear and cyclic versions of the antigenic loop of FMDV C-S30 showed that structural features previously correlated with antigenicity were more pronounced in the cyclic peptide. Twenty-six synthetic peptides, corresponding to all possible combinations of five single-point antigenicity-enhancing replacements in the GH loop of FMDV C-S8c1, were also studied. SPR kinetic screening of these peptides was not possible due to problems mainly related to the high mAb affinities displayed by these synthetic antigens. Solution affinity SPR analysis was employed and affinities displayed were generally comparable to or even higher than those corresponding to the C-S8c1 reference peptide A15. The NMR characterisation of one of these multiple mutants in solution showed that it had a conformational behaviour quite similar to that of the native sequence A15 and the X-ray diffraction crystallographic analysis of the peptide ? mAb 4C4 complex showed paratope ? epitope interactions identical to all FMDV peptide ? mAb complexes studied so far. Key residues for these interactions are those directly involved in epitope ? paratope contacts (141Arg, 143Asp, 146His) as well as residues able to stabilise a particular peptide global folding. A quasi-cyclic conformation is held up by a hydrophobic cavity defined by residues 138, 144 and 147 and by other key intrapeptide hydrogen bonds, delineating an open turn at positions 141, 142 and 143 (corresponding to the Arg-Gly-Asp motif).

Body composition, muscle strength, functional capacity, and physical disability risk in liver transplanted familial amyloidotic polyneuropathy patients

Abstract: Background: Familial amyloidotic polyneuropathy (FAP) is a neurodegenerative disease leading to sensory and motor polyneuropathies, and functional limitations. Liver transplantation is the only treatment for FAP, requiring medication that negatively affects bone and muscle metabolism. The aim of this study was to compare body composition, levels of specific strength, level of physical disability risk, and functional capacity of transplanted FAP patients (FAPTx) with a group of healthy individuals (CON). Methods: A group of patients with 48 FAPTx (28 men, 20 women) was compared with 24 CON individuals (14 men, 10 women). Body composition was assessed by dual-energy X-ray absorptiometry, and total skeletal muscle mass (TBSMM) and skeletal muscle index (SMI) were calculated. Handgrip strength was measured for both hands as was isometric strength of quadriceps. Muscle quality (MQ) was ascertained by the ratio of strength to muscle mass. Functional capacity was assessed by the six-minute walk test. Results: Patients with FAPTx had significantly lower functional capacity, weight, body mass index, total fat mass, TBSMM, SMI, lean mass, muscle strength, MQ, and bone mineral density. Conclusion: Patients with FAPTx appear to be at particularly high risk of functional disability, suggesting an important role for an early and appropriately designed rehabilitation program.

Strength, muscle quality and functional capacity in liver transplanted familial amyloidotic polineuropathy patients

Liver transplantation is the unique treatment for several end-stage diseases. Familial Amiloidotic Polineuropathy (FAP) is a neurodegenerative disease related with systemic deposition of amyloidal fiber mainly on peripheral nervous system, clinically translated by an autonomous sensitive-motor neuropathy with severe functional limitations in some cases. The unique treatment for FAP disease is a liver transplant with a very aggressive medication to muscle metabolism and force production. To our knowledge there are no quantitative characterizations of body composition, strength or functional capacity in this population.

Knee muscle strength, balance and functional independence in persons with stroke

Purpose: to investigate the relationship between knee muscle strength, balance and functional independence within the first month after stroke. Relevance: cerebral vascular disease is one of the main causes of morbidity, disability and mortality in developed countries. Problems with movement control are frequent after stroke. Lower limb weakness and impaired balance are common problems that are related with the risk of falls and are likely to interfere with the ability to perform daily life activities. Physiotherapy intervention usually starts early after stroke and addresses impairments related to movement and posture in order to improve motor recovery and restore function.

Muscle strength, balance and functional independence in persons with stroke

Cerebral vascular disease is one of the main causes of morbidity, disability and mortality in developed countries. Problems with movement control are frequent after stroke. Lower limb weakness and impaired balance are common problems that are related with the risk of falls and are likely to interfere with the ability to perform daily life activities. Physiotherapy intervention usually starts early after stroke and addresses impairments related to movement and posture in order to improve motor recovery and restore function. Purpose: to investigate the relationship between knee muscle strength, balance and functional independence within the first month after stroke.

Relationship of balance to function independence in stroke survivors

Cerebral vascular disease is the primary cause of permanent disability in Portugal. Impaired stability is considered an important feature after stroke as it is related with higher risk of falls and functional dependence. Physiotherapy intervention usually starts early after stroke in order to direct motor recovery and help patients to improve their ability to perform activities of daily living (ADL). Purpose: to investigate the relationship of balance to functionality in acute stroke patients. Methods: 16 subjects (8 women and 8 men), mean age 63,62 ± 2,16y, with unilateral ischemic stroke in the middle cerebral artery territory, who were admitted to physiotherapy department of Fernando Fonseca Hospital in Portugal, within the first month after stroke were recruited to participate in this study. All subjects have no cognitive impairment according to Mini Mental State, no history of lower extremity orthopedic problems and no other disease that could interfere with treatments. All patients gave their inform consent to participate in this study. Subjects were assessed with the Modified Barthel Index (MBI) and the Berg Balance Scale (BBS).

Análise psicométrica da escala de impacto na autonomia e participação, em pessoas com esclerose múltipla

A avaliação da Autonomia e funcionalidade das pessoas com esclerose múltipla é fundamental para a compreensão das limitações e necessidades destas pessoas. O presente estudo tem como objectivo estudar as propriedades psicométricas da Escala de Impacto Autonomia e Participação em doentes com esclerose múltipla. Participaram neste estudo 280 indivíduos com diagnóstico de esclerose múltipla, 71,4% mulheres com idade média de 39,23 anos. Procedeu-se à análise factorial exploratória, consistência interna, teste-reteste, validade convergente e discriminante. Os resultados demonstram valores psicométricos apropriados para a versão de língua portuguesa semelhantes às versões de língua alemã e de língua inglesa. Concluímos que esta escala tem boas condições psicométricas para avaliar a autonomia funcional em indivíduos com esclerose múltipla. ABSTRACT: Assessment of Autonomy and participation of patients with multiple sclerosis is essential to understand the limitations and necessities of these patients. The present study aims to examine the psychometric proprieties of the Impact on Participation and Autonomy Questionnaire (IPA) in patient with multiple sclerosis. Participants are 280 patients with a diagnostic of multiple sclerosis, 71.4% females, mean age of 39.23 years. We inspect metric properties namely, validity, construt validity and convergent and discriminant validity, and reliability, internal consistency and test-retest. The results show that IPA is a reliable and valid instrument for assessment autonomy and participation in Portuguese language, with similar sensibility than the Germany and English version. In conclusion these instruments have psychometric conditions for assessment the functional autonomy of people with multiple sclerosis.

Mortality among patients with non-affective functional psychoses in a metropolitan area of South-Eastern Brazil

High mortality rates among those suffering from schizophrenia and related psychoses have been consistently described in developed societies. However, to date there is a lack of data on this matter in Brazil. In order to examine this issue, a prospective 2-year follow-up study was carried out in S. Paulo. The sample consisted of 120 consecutive admissions to psychiatric hospitals in a defined catchment area, aged 18 to 44 years old, with clinical diagnoses of non-affective functional psychoses according to the ICD-9. After 2 years, 116 (96.7%) subjects were traced. During the study period there were 7 deaths (6.0% of those traced), 5 (4.3%) due to suicide. All but one of the suicides occurred in the first year after discharge from hospital. Age and sex Standardised Mortality Ratios (relative to rates for the population of the city of Sao Paulo) were 8.4 for overall mortality (95% confidence interval: 4.0-15.9) and 317.9 for deaths due to suicide (95% confidence interval: 125.2-668.3). These results are in agreement with previous studies, and show that in Brazil non-affective functional psychoses are life-threatening illnesses, which need adequate care, particularly when patients go back to live in the community after hospital discharge.

New insights into functional regulation in MS-based drug profiling

We present a novel data analysis strategy which combined with subcellular fractionation and liquid chromatography-mass spectrometry (LC-MS) based proteomics provides a simple and effective workflow for global drug profiling. Five subcellular fractions were obtained by differential centrifugation followed by high resolution LC-MS and complete functional regulation analysis. The methodology combines functional regulation and enrichment analysis into a single visual summary. The workflow enables improved insight into perturbations caused by drugs. We provide a statistical argument to demonstrate that even crude subcellular fractions leads to improved functional characterization. We demonstrate this data analysis strategy on data obtained in a MS-based global drug profiling study. However, this strategy can also be performed on other types of large scale biological data.

Strength, muscle quality and functional capacity in liver transplanted familial amyloidotic polineuropathy patients

Liver transplantation is the unique treatment for several end stage diseases. Familial Amiloidotic Polineuropathy (FAP) is a neurodegenerative disease related with systemic deposition of amyloidal fibre mainly on peripheral nervous system, clinically translated by an autonomous sensitive-motor neuropathy with severe functional limitations in some cases. The unique treatment for FAP disease is a liver transplant with a very aggressive medication to muscle metabolism and force production. To our knowledge there are no quantitative characterizations of body composition, strength or functional capacity in this population. The purpose of this study was to compare levels of specific strength (isometric strength adjusted by lean mass or muscle quality) and functional capacity (meters in 6 minutes walk test) between FAP patients after a liver transplant (4.1±2 months after transplant surgery) (FAPT) and a healthy group (HG).

Avaliação do perfil hemodinâmico de válvulas aórticas percutâneas: análise comparativa entre os dois dispositivos disponíveis no mercado - válvula auto-expansível e válvula expansível por balão

Mestrado em Tecnologia de Diagnóstico e Intervenção Cardiovascular. Área de especialização: Intervenção Cardiovascular.

Ligadura funcional num caso de entorse da acrómio-clavicolar, grau III: que resultados em termos de restabelecimento de força muscular?

Com o presente estudo pretendeu-se: (1) verificar se a ligadura funcional para a lesão da articulação Acrómio-Clavicular (AC), descrita por Chanussot e Danowski (2001), contribui para o restabelecimento da força muscular num paciente com lesão grau III; e, (2) observar se a ligadura funcional altera o padrão electromiográfico dos principais músculos superficiais do complexo articular do ombro: infra-espinhoso, deltóide anterior, grande peitoral, trapézio superior e trapézio inferior. Efectuou-se assim um estudo de caso do tipo observacional, descritivo transversal. Para avaliação da força muscular foi utilizado um dinamómetro isocinético e para a avaliação do nível de activação de fibras musculares foi utilizada a EMG de superfície. Os movimentos em análise foram os de rotação interna e externa, a 60º/s e 180º/s. Os resultados revelaram, na presença de ligadura, um aumento do Peak Torque dos rotadores internos, assim como da actividade do grande peitoral e trapézio inferior. Constatou-se ainda uma diminuição do Peak Torque e da percentagem do Root Mean Square do trapézio inferior, trapézio superior e grande peitoral, do ombro esquerdo, sem ligadura, quando comparado com o ombro contralateral. A ligadura funcional, numa entorse da acromio-clavicular grau III, poderá contribuir para a recuperação de força muscular e activação de fibras musculares.

Study of the influence of count’s number in myocardium in the determination of reproducible functional parameters in Gated-SPECT studies simulated with GATE

Myocardial Perfusion Gated Single Photon Emission Tomography (Gated-SPET) imaging is used for the combined evaluation of myocardial perfusion and left ventricular (LV) function. But standard protocols of the Gated-SPECT studies require long acquisition times for each study. It is therefore important to reduce as much as possible the total duration of image acquisition. However, it is known that this reduction leads to decrease on counts statistics per projection and raises doubts about the validity of the functional parameters determined by Gated-SPECT. Considering that, it’s difficult to carry out this analysis in real patients. For ethical, logistical and economical matters, simulated studies could be required for this analysis. Objective: Evaluate the influence of the total number of counts acquired from myocardium, in the calculation of myocardial functional parameters (LVEF – left ventricular ejection fraction, EDV – end-diastolic volume, ESV – end-sistolic volume) using routine software procedures.

Efetividade do Pnf-Chi® na funcionalidade do membro superior e na qualidade de vida, em mulheres submetidas a tratamento cirúrgico por cancro da mama

Mestrado em Fisioterapia.

Nintendo Wii: uso terapêutico no controlo postural em indivíduos que sofreram um acidente vascular cerebral

Mestrado em Fisioterapia.