Diploma thesis "Influence of regular training of spinal cord patients in Lokomat system on particular spatio-temporal parameters of their walk." is a theoretical-empirical case report. Theoretical part deals with the neuroanatomy of the spinal cord, control, analysis and possibilities of check up of physiological walk. This part is concluded by the summary of entries about the Lokomat system, its basic technical equipment, advantages / disadvantages, indications/ contra-indications of its use in practice. The empirical part deals with the examination of the influence of the training in Lokomat system on the walk of the patients with incomplete spinal cord lesion. The main subject of the research is to monitor the changes in particular spatio-temporal parameters of walk (gait speed, stride length, stride time, single support time). Powered by TCPDF (www.tcpdf.org)

O tratamento cirúrgico das neoplasias espinais intradurais primárias está indicado para conseguir ressecação completa, diminuir morbidade e aumentar a sobrevida livre de doença, em busca da cura. Há dúvidas sobre as consequências do tratamento cirúrgico influenciando a qualidade de vida. Entretanto, até o momento, não foram encontrados dados disponíveis na literatura especializada sobre a qualidade de vida dos pacientes que foram submetidos à cirurgia. Os objetivos foram avaliar a qualidade de vida e testar as propriedades psicométricas dos instrumentos utilizados, verificando a consistência interna e a confiabilidade. O estudo foi realizado no Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, no período de 2009 a 2011, com aplicação do questionário genérico The Medical Outcomes Study 36-item Short-Form Health Survey (SF-36), e das escalas específicas de McCormick e de Aminoff-Logue para a avaliação da qualidade de vida sob o ponto de vista funcional. Todos os pacientes incluídos foram avaliados uma única vez e encontravam-se no período de, no mínimo, seis meses após a data da cirurgia. O banco de dados resultante foi analisado no Departamento de Medicina Preventiva da Faculdade de Medicina da Universidade de São Paulo. A série de casos constou de 148 pacientes, dos quais 48 foram excluídos por não completarem o protocolo. Dentre os 100 casos estudados, 55 eram do sexo feminino, com média de idade de 42,3 anos e tempo de acompanhamento pós-operatório de 20 meses. A pontuação global da qualidade de vida pelo SF-36 foi de 50,5, com resultado geral do componente físico de 46,8. A consistência interna foi demonstrada segundo a validade de construto e de critério, confirmando a hipótese da relação existente entre as pontuações do SF-36 e a escala de McCormick (p = 0,003), as escalas de Aminoff-Logue componente de marcha (p = 0,025), vesical (p = 0,013) e intestinal (p = 0,004). A confiabilidade foi demonstrada em todos os oito domínios do SF-36, alcançando em cada um o alpha Cronbach, satisfazendo o critério de Nunnally > 0,85. Estes resultados sugerem associação consistente entre a qualidade de vida e as funções medulares nesta série de casos, caracterizando melhor a percepção de saúde dos pacientes submetidos ao tratamento cirúrgico. O avanço do conhecimento aplicado à prática neurocirúrgica focada no paciente aponta para a necessidade de avaliação longitudinal da qualidade de vida para auxílio no planejamento terapêutico destas doenças. A qualidade de vida pode ser avaliada pelo questionário SF36, que se correlaciona bem com as escalas específicas de McCormick e Aminoff-Logue / The surgical treatment of intradural primary spinal tumors is indicated to obtain total ressection, decrease morbidity rate and increase survival rate, aiming at the cure. There are doubts about the consequences of surgical treatment influencing the quality of life. However, until now there have been no available data in specialized literature on the quality of life of patients who have undergone this surgery. The objectives of this study were to evaluate the patient´s quality of life and to test the psychometric properties of the instruments used, verifying consistency and reliability. The study was carried out at the Clinics Hospital of the University of São Paulo Medical School, between 2009 and 2011, by means of The Medical Outcomes Study 36-item Short-Form Health Survey (SF-36) generic questionnaire and the McCormick and the Aminoff-Logue specific scales to assess the patient´s quality of life from the functional perspective. All patients included in the study were assessed only once, at least six months postoperatively. The resulting data were analyzed at the Department of Preventive Medicine of the University of São Paulo Medical School. The series of cases comprised 148 patients; 48 patients were excluded because they did not complete the protocol. The study was finally conducted with 100 patients. Fifty-five patients were women, the mean age was 42,3 years and the mean postoperative follow-up period was 20 months. The quality of life total score on the SF-36 was 50,5, and the Physical Component Summary (PCS) score was 46,8. Based upon construct validity and criterion, the internal consistency was demonstrated, confirming the hypothesized relationship between the scores of the SF-36 and the McCormick scale (p = 0.003), the Aminoff-Logue gait subscale (p = 0.025), the Aminoff-Logue micturition subscale (p = 0.013), and the AminoffLogue defecation subscale (p = 0.004). Reliability was demonstrated for all eight SF-36 domain scales and the Physical Component Summary and the Mental Component Summary of the SF-36, where in each the Cronbach alpha satisfied the Nunnally criterion of > 0,85. These results suggest a consistent relationship between the quality of life and the spinal cord functions in this series of cases, better characterizing the evaluation of the health of the patients who underwent surgical treatment. The advancement of knowledge applied to neurosurgery practice with a patient-focused approach shows the necessity of a longitudinal evaluation of the quality of life to help tailor a treatment plan for these diseases. The quality of life can be analyzed by the SF-36 questionnaire, which is well correlated with the McCormick and the Aminoff-Logue specific scales

A utilização das células progenitoras hematopoéticas (CPH) obtidas do sangue de cordão umbilical e placentário (SCUP) apresenta vários benefícios para o transplante de CPH em comparação às células provenientes de outras fontes. Dentre eles, a maior disponibilidade e a maior imaturidade imunológica das CPH, o que permite certa flexibilidade nos critérios de compatibilidade entre doador e receptor e uma menor taxa de reação do enxerto-versus-hospedeiro. A legislação brasileira e órgãos internacionais exigem a realização de vários testes para garantir a qualidade do produto hemoterápico contendo CPH a ser transplantado. O objetivo deste estudo foi confirmar que o teste para quantificação de CPH com elevada atividade da enzima ALDH1(ALDHbr) pode ser considerado um teste de adequado ou seja, é capaz de predizer quais produtos tem melhor capacidade de repopular a medula óssea do recipiente após o transplante. Para isso, foram utilizadas 40 unidades de SCUP coletadas e processadas pelo Banco de Sangue de Cordão Umbilical e Placentário do Cetebio / Fundação Hemominas. As unidades foram processadas por método automatizado e amostras do creme leucocitário (buffy coat) foram coletadas para a realização da quantificação de células ALDHbr, quantificação de células CD34+, ensaio clonogênico (CFU), hemograma e cálculo do total de células nucleadas (TCN). A citometria de fluxo foi utilizada para a quantificação das CPH ALDHbr e CD34+ e das subpopulações CD45dim e CD38+. Outras informações como idade materna, idade gestacional e sexo do recém-nascido também foram coletadas para descrição das unidades. Para verificar a viabilidade da utilização do teste de ALDH pelos BSCUP foi realizado o levantamento do seu custo. A capacidade funcional das CPH em proliferar e se diferenciar em tecido hematopoético foi avaliada por meio do ensaio clonogênico. Detectou-se correlação entre a quantidade de células ALDHbr e o número de colônias no ensaio clonogênico (p<0,001), entre o número de células ALDHbr e de células CD34+ (p=0,001) e entre o número de colônias no ensaio clonogênico e o número de células CD34+ (p<0,001). A imunofenotipagem mostrou que 46,25% das células ALDHbr eram CD45dimCD38+CD34+. Os dados sugerem que a quantificação de células ALDHbr em unidades de SCUP pode ser considerada teste adequado, de baixo custo, de execução simples, rápida e menos dependente do operador em relação ao ensaio clonogênico. / The use of the umbilical cord blood cells presents numberless benefits when compared to the cells from different sources. Among them, the ease of availability, the bigger immunological immaturity, which allows some flexibility in the compatibility between donor and receptor and less induction of reaction of graft-versus-host. The Brazilian legislation and international organizations demand the practice of various tests to guarantee the quality of the product to be transplanted. The aim of this research was to confirm that the test used to quantify ALDHbr cells can be considered a power test, meaning that it tests the ability to repopulate the bone marrow after transplant. For this study, it has been used 40 units of SCUP collected and processed by the Cetebio Umbilical Cord Blood Bank - Fundação Hemominas. The units were processed by the automatized method and the samples of the final product (buffy coat) were collected for the quantification of ALDHbr cells, quantification of CD34+ cells, clonogenic essay (CFU), hemogram and the total number of nucleated cells (TNC). It was used the flow cytometry to perform ALDH and CD34+ tests. Besides that, it was also performed the association of antibodies anti-CD34, anti-CD45 and anti-CD38 for the immunophenotyping of the units. Other information such as maternal age, fertilization age and the newborn gender were also collected for description of the units. In order to verify the viability of the use of the ALDH test by BSCUP its costs were calculated, as well as of the clonogenic essay. The results showed a significant correlation between ALDHbr cells and the clonogenic essay (p<0.001), between ALDHbr and CD34+ cells (p=0,001) and between the clonogenic essay and the quantification of CD34+ cells (p<0,001). The immunophenotyping revealed that 46,25% of ALDHbr cells were CD45dimCD38+CD34+. The data indicated that the quantification of ALDHbr cells in the SCUP units can be considered a powerful and low cost procedure, of easier and quicker execution and less operator dependent.

Este trabalho avaliou a incidência de pacientes com lesão traumática da coluna cervical causada por mergulho em águas rasas, que foram atendidos nos Hospitais da cidade de Ribeirão Preto-SP, entre janeiro de 1989 a dezembro de 1996. O estudo epidemiológico foi realizado mediante investigação feita nos prontuários médicos desses pacientes. Complementou-se as informações por meio de uma entrevista realizada com 12 deles. Com base nos números apresentados, elaborou-se um programa de prevenção decorrente desses traumatismos a implantou-se uma Campanha de prevenção das lesões cervicais por mergulho em águas rasas em Ribeirão Preto. Um Programa de Campanha foi encaminhado à Assembléia Legislativa do Estado e transformado em Projeto de Lei. Os resultados mostraram que, dos 355 casos estudados, 69 deles foram por acidentes em águas rasas, a dentre estes, 58% tiveram danos neurológicos. Os locais mais freqüentes dos acidentes foram rios, córregos, lagos, cachoeiras,com a ocorrência de 75,4% dos casos, enquanto que a incidência em piscinas foi de 24,6%. A média de idade variou entre 10 a 30 anos, havendo diferença significativa com outras faixas etárias. Quanto ao sexo, 92,8% ocorreram em homens a 7,2% com mulheres. Em relação ao estado civil, 68,2% eram solteiros e 31,9% eram casados. O período do ano em que houve um número mais elevado dos traumatismos da coluna cervical causado pelo mergulho nas chamadas águas rasas foi nos meses de estações mais quentes na região (primavera, verão a outono). Dos 17,4% dos pacientes entrevistados de um total de 40 que tiveram lesão medular, todos mostraram desconhecimento desse tipo de acidente, bem como suas conseqüências. A Campanha de prevenção teve início em setembro de 1996 e está em fase de desenvolvimento em Ribeirão Preto. O Projeto de Lei estadual nº 183 aguarda aprovação da Assembléia. Conclui-se que, a incidência de acidentes com lesão traumática da coluna cervical por mergulho em águas rasas é elevada, sendo a terceira causa de danos traumáticos. A desinformação da população no que diz respeito ao perigo de um mergulho em águas rasas é elevado. A redução deste tipo de acidente pode acontecera partir de um processo educacional da população a da atuação decisiva do poder público. / This study is an evaluation of the incidence of patients treated in the hospitals in Ribeirão Preto - SP for traumatic injury of the cervical cord caused by diving in shallow waters. The epidemiological study was held by means of an investigation of the medical charts of patients who underwent treatment from January 1989 to December 1996. The information on the charts was complemented through interviews held with 12 of these patients. Based on the results of these investigations, a campaign to prevent cervical injury caused by diving into shallow waters was elaborated and sent to the State Legislature, where it was transformed into a bill. The study revealed that in the 355 cases of cervical injury investigated, 69 were the result of accidents in shallow waters, and of these, 58% suffered neurological dysfunction. Rivers, lakes and waterfalls were most frequently cited as locations for the occurrence of these injuries (75,4% of the cases), with the incidence of accidents in swimming pools at 24,6%. The average age of patients varied between 10 to 30 years of age, with a significant difference in other age groups. In relation to sex, 92,8% of the accidents occurred in men, and only 7,2% in women. As to marital status, 68,2% were single and only 31,9% were married. The greatest number of cases of trauma of the cervical cord caused by diving in shallow waters occurred in the warmer seasons of the year (spring summer and fall). 17,4% of the patients interviewed out of a total of 40 with medullar injury, claimed not to know about this kind of accident and its consequences. The Prevention Campaign began in September 1996 and is presently being carried out in the city of Ribeirão Preto. The Bill of Law no. 183 awaits approval from the State Legislature. One may conclude that the incidence of traumatic injury of the cervical cord due to diving in shallow waters is high (third place as the cause of injury). The lack of information on the part of the population regarding the dangers of this type of accident may be one of the reasons behind these high statistics. The reduction of this type of accident may occur as the result of an educational process involving the population as well as decisive action on the part of the government.

Introdução: Várias são as propostas descritas para tratar farmacologicamente a lesão traumática da medula espinal. A metilprednisolona já foi padronizada para uso clínico. O fator estimulador de colônia de granulócitos (G-CSF) tem sido promissor em estudos experimentais e clínicos. Não há pesquisas quanto ao efeito da associação dos dois fármacos. Objetivo: Avaliar o efeito do tratamento com o fator estimulador de colônia de granulócitos associado a metilprednisolona na lesão medular aguda experimental em ratos. Material e métodos: Foram avaliados 40 ratos Wistar submetidos a lesão medular moderada com o NYU-Impactor. Os animais foram divididos em quatro grupos de 10 ratos. O Grupo Controle não recebeu tratamento; o Grupo G-CSF, foi tratado com G-CSF no momento da lesão e diariamente ao longo dos cinco dias subsequentes; o Grupo Metilprednisolona, com metilprednisolona durante 24 horas; e o Grupo G-CSF/Metilprednisolona, com metilprednisolona durante 24 horas e G-CSF no momento da lesão e ao longo de cinco dias. Os animais foram mantidos vivos durante 42 dias; a avaliação funcional foi realizada com a aplicação da escala funcional de Basso, Beattie e Bresnahan (BBB) nos dias 2, 7, 14, 21, 28, 35 e 42 subsequentes à lesão. A avaliação dos potenciais evocados motores foi realizada no dia 42 e a avaliação histológica da lesão da região da medula espinal lesada, realizada logo após a eutanásia ocorrida no dia 42. Resultados e conclusões: A associação de metilprednisolona e G-CSF no tratamento do traumatismo medular contuso experimental em ratos promoveu melhora neurológica avaliada pela escala BBB superior à melhora promovida pela metilprednisolona e G-CSF quando utilizadas isoladamente. A associação teve também efeito sinérgico que resultou em melhora nos parâmetros histológicos no local da lesão. Não houve diferença entre os grupos quanto à avaliação neurofisiológica / Introduction: There are several proposals to pharmacologically treat traumatic spinal cord injury. Methylprednisolone has already been standardized for clinical use. Granulocyte colony stimulating factor (G-CSF) has been promising in experimental and clinical studies. There is no research on the effect of the association of the two drugs. Objective: to evaluate the effect of combined treatment of the granulocyte colony-stimulating factor (G-CSF) associated with methylprednisolone in experimental acute spinal cord injury in rats. Material and methods: Forty male Wistar rats were submitted to a moderate spinal cord injury with the NYU-Impactor. The animals were divided into four groups of ten rats each. The Control Group was not treated; the G-CSF Group was treated with G-CSF at the time of injury and daily over the next five days; the Methylprednisolone Group was treated with methylprednisolone for 24 hours; the G-CSF/methylprednisolone Group, was treated with methylprednisolone for 24 hours and G-CSF at the time of injury and daily over the next five days. The animals were kept alive for 42 days; Functional evaluation was performed using the Basso, Beattie and Bresnahan (BBB) score on days 2, 7, 14, 21, 28, 35 and 42 following the spinal cord injury. Evaluation of motor evoked potentials was held and histological examination of the lesion of the spinal cord was done immediately after euthanasia on day 42. Results and conclusions: The combination of methylprednisolone and G-CSF in the treatment of experimental spinal cord injury in rats promoted neurological improvement as assessed by BBB scale with greater improvement than with methylprednisolone or G-CSF when used alone. The combination of treatment had also a synergistic effect resulting in improvement in histological parameters at the injury site. There was no difference between groups regarding neurophysiological evaluation

Human tissue banks are a potential source of cellular material for the emerging cellbased therapy industry; umbilical cord tissue (UCT) private banking is increasing in such facilities as a source of mesenchymal stem cells for future therapeutic use. However, early handling of UCT is relatively uncontrolled due to the clinical demands of the birth environment and subsequent transport logistics. It is therefore necessary to develop extraction methods that are robust to real world operating conditions,rather than idealised operation. This will be critical for all processes using primary tissue or cell sources. The research work undertaken in this PhD project was initiated by the collaboration with one of the leading private cord blood banks in the UK and later driven by the prospect of expanding the cell therapy and business potential of the bank. The investigation described in this thesis has focused on: - Developing an extraction method for human mesencymal stem cells (hMSCs) from UCT. - Understanding and minimizing the noticed variability in cell yield extracted from UCT by mapping the operating environment and assessing the risk factors before empirically determining their effect on the process. - Establishing the necessary process controls in the production of high quality hMSCs, through a series of wet experiments, targeted at narrowing down the sources of variability down to sub-process level. - Finding a novel method for assessing the cell content and viability of cords prior to processing. Therefore, helping the tissue processing facility to predict the risk of suboptimal cell yield from a given cord tissue section and processing method, given different operating ranges. - Determining the tissue storage requirements and isolation method with acceptable risk of adequate cell recovery. - Characterization of cells extracted from UCT via different extraction methods and comparison to primary cells extracted from other tissue sources. - Investigation of cryopreservation method for UCT. The result of this work provides a solid example of the type of data and analysis that will be required to inform a Quality-by-Design type approach for cell product development and manufacture. It will help tissue processing facilities and banks to predict the probability of cell yields from tissue sections given different operating ranges, and to aid and inform the experimental approach of others.

Traumatic spinal cord injury (SCI) is a devastating condition for which there is no effective clinical treatment. Neuroprotective molecules that minimize tissue loss have shown promising results; however systemic delivery may limit in vivo benefits due to short systemic half-life and minimal passage across the blood-spinal cord barrier. To overcome these limitations, an injectable intrathecal delivery vehicle comprised of hyaluronan and methylcellulose (HAMC) was developed, and previously demonstrated to be safe and biocompatible intrathecally. Here, HAMC was determined to persist in the intrathecal space for between 4-7 d in vivo, indicating it as an optimal delivery system for neuroprotective agents to reduce tissue degeneration after SCI. HAMC was then investigated as an in vivo delivery system for two neuroprotective proteins: erythropoietin (EPO) and fibroblast growth factor 2 (FGF2). Both proteins demonstrated a diffusive release profile in vitro and maintained significant bioactivity during release. When EPO was delivered intrathecally with HAMC to the injured spinal cord, reduced cavitation in the tissue and significantly improved neuron counts were observed relative to the conventional delivery strategies of intraperitoneal and intrathecal bolus. When FGF2 was delivered intrathecally from HAMC, therapeutic concentrations penetrated into the injured spinal cord tissue for up to 6 h. Poly(ethylene glycol) modification of FGF2 significantly increased the amount of protein that diffused into the tissue when delivered similarly. Because FGF2 is a known angiogenic agent, dynamic computed tomography was developed for small animal serial assessment of spinal cord hemodynamics. Following SCI and treatment with FGF2 from HAMC, moderate improvement of spinal cord blood flow and a reduction in permeability were observed up to 7 d post-injury, suggesting that early delivery of neuroprotective agents can have lasting effects on tissue recovery. Importantly, the entirety of this work demonstrates that HAMC is an effective short-term delivery system for neuroprotective agents by improving tissue outcomes following traumatic SCI.

Traumatic spinal cord injury (SCI) is a devastating condition for which there is no effective clinical treatment. Neuroprotective molecules that minimize tissue loss have shown promising results; however systemic delivery may limit in vivo benefits due to short systemic half-life and minimal passage across the blood-spinal cord barrier. To overcome these limitations, an injectable intrathecal delivery vehicle comprised of hyaluronan and methylcellulose (HAMC) was developed, and previously demonstrated to be safe and biocompatible intrathecally. Here, HAMC was determined to persist in the intrathecal space for between 4-7 d in vivo, indicating it as an optimal delivery system for neuroprotective agents to reduce tissue degeneration after SCI. HAMC was then investigated as an in vivo delivery system for two neuroprotective proteins: erythropoietin (EPO) and fibroblast growth factor 2 (FGF2). Both proteins demonstrated a diffusive release profile in vitro and maintained significant bioactivity during release. When EPO was delivered intrathecally with HAMC to the injured spinal cord, reduced cavitation in the tissue and significantly improved neuron counts were observed relative to the conventional delivery strategies of intraperitoneal and intrathecal bolus. When FGF2 was delivered intrathecally from HAMC, therapeutic concentrations penetrated into the injured spinal cord tissue for up to 6 h. Poly(ethylene glycol) modification of FGF2 significantly increased the amount of protein that diffused into the tissue when delivered similarly. Because FGF2 is a known angiogenic agent, dynamic computed tomography was developed for small animal serial assessment of spinal cord hemodynamics. Following SCI and treatment with FGF2 from HAMC, moderate improvement of spinal cord blood flow and a reduction in permeability were observed up to 7 d post-injury, suggesting that early delivery of neuroprotective agents can have lasting effects on tissue recovery. Importantly, the entirety of this work demonstrates that HAMC is an effective short-term delivery system for neuroprotective agents by improving tissue outcomes following traumatic SCI.

The aim of this thesis was to study maternal and infant effects of delayed cord clamping (≥180 seconds, DCC) compared to early (≤10 seconds, ECC) in a randomised controlled trial. Practice and guidelines regarding when to clamp the cord vary globally, and different meta-analyses have shown contradictory conclusions on benefits and disadvantages of DCC and ECC. The study population consisted of 382 term infants born after normal pregnancies and randomised to DCC or ECC after birth. The primary objective was iron stores and iron deficiency at 4 months of age, but the thesis was designed to investigate a wide range of suggested effects associated with cord clamping. Paper I showed that DCC was associated with improved iron stores at 4 months (45% higher ferritin) and that the incidence of iron deficiency was reduced from 5.7% to 0.6%. Neonatal anaemia at 2-3 days was less frequent in the DCC group, 1.2% vs. 6.3%. There were no differences between the groups in respiratory symptoms, polycythaemia, or hyperbilirubinaemia. In paper II we demonstrated that DCC versus ECC was not associated with higher risk for maternal post partum haemorrhage and rendered a comparable ratio of valid umbilical artery blood gas samples. In paper III, the Ages and Stages Questionnaire was used to assess neurodevelopment at 4 months. The total scores did not differ, but the DCC group had a higher score in the problem-solving domain and a lower score in the personal-social domain. Immunoglobulin G level was 0.7 g/L higher in the DCC group at 2–3 days, but did not differ at 4 months. Symptoms of infection up to 4 months were comparable between groups. Finally, in paper IV, iron stores and neurodevelopment were similar between groups at 12 months. Gender specific outcome on neurodevelopment at 12 months was discovered, implying positive effects from DCC on boys and negative on girls. We conclude that delaying umbilical cord clamping for 180 seconds is safe and associated with a significantly reduced risk for iron deficiency at 4 months, which may have neurodevelopmental effects at a later age.

published_or_final_version / Social Work and Social Administration / Doctoral / Doctor of Philosophy