Title: Degenerative changes in the cervical spine with a focus on the intervertebral disc prolapse and its verification using imaging methods and 3D model Objectives: The aim of this work is to describe the problems of degenerative changes in the cervical spine with a focus on the intervertebral disc prolapse. The second part deals with the possibility of imaging methods that are able to detect pathological processes. Part of this work is the study of three cases of patients with a diagnosis of disc herniation. In conclusion, there are some possibilities of 3D modeling of the cervical spine. Methods: The theoretical part is based on the available literature. Images from MRI were obtained from medical facilities in Prague, described and assessed by a specialist. After studying the functions suitable graphical programs were created 3D models of the cervical spine. Results: Degenerative changes in the cervical spine naturally come from the aging of the organism, but a number of modifiable factors accelerating the onset. Among such factors include especially hypokinesia and unilateral overloading of cervical spine. Modern imaging techniques can in detail detect these pathological processes, but are limited by the patient's position during the investigation. The resulting series of images are used as...

Les infections à papillomavirus humains à haut-risque (HPV-HR) sont responsables de 100% des cancers cervico-utérins et de 50% des carcinomes de l'oropharynx. Les infections du col utérin sont généralement transitoires et bénignes. Cependant, en cas d'infection persistante, elles peuvent s'accompagner d'une progression vers des lésions (pré)cancéreuses du col utérin. Les facteurs viraux qui favorisent la persistance ou la clairance virale sont encore mal connus. Dans une première partie, nous avons étudié la méthylation des régions 3'L1 et LCR dans des frottis cervico-utérins HPV16+ / cytologie normale, prélevés chez 37 femmes qui présenteront soit une infection transitoire soit une infection persistante avec progression vers une lésion précancéreuse. Nous avons montré que certains îlots CpG présentaient des taux de méthylation différents en fonction de l'évolution ultérieure de l'infection HPV et pourraient être utilisés comme marqueurs prédictifs.Les cancers de l'oropharynx associés à une infection à HPV-HR présentent un pronostic plus favorable que ceux associés à une intoxication alcoolo-tabagique. Les mécanismes responsables de ce meilleur pronostic sont encore mal compris. Dans une seconde partie, nous avons caractérisé le statut HPV dans 202 cas de cancers de l'oropharynx. Au sein des 32 cancers présentant une infection HPV16 active (avec expression des ARNm E6/E7), nous avons étudié la méthylation des régions 3'L1 et LCR et l'intégration du génome viral. Nous avons montré que les niveaux de méthylation de certains sites CpG, notamment des sites de liaison à la protéine virale E2 (E2BS3 et E2BS4) sont fortement méthylés dans les formes épisomales et mixtes mais sont non méthylés dans les formes intégrées pures.Enfin dans une troisième partie, nous avons étudié des marqueurs de transition épithélio-mésenchymateuse (TEM) dans des modèles cellulaires et des cancers oropharyngés, en fonction du statut HPV. Une moindre TEM, connue pour être impliquée de façon importante dans les phénomènes d'invasion tumorale, pourrait expliquer le meilleur pronostic associé aux cancers HPV-induits. Nous avons montré que la vimentine, marqueur le plus représentatif de l'acquisition de capacités migratoires et invasives, est plus exprimée dans les cellules tumorales HPV positives que dans les cellules HPV négatives. Le pronostic différentiel observé entre les 2 étiologies des cancers oropharyngés ne serait donc pas corrélé à la TEM.Mots-clés : Cancer, HPV, col utérin, oropharynx, biomarqueurs, méthylation, intégration, TEM. / High-risk human papillomavirus (HR-HPV) infections are responsible for 100% of cervical cancers and 50% of oropharyngeal cancers. Cervical infections are usually transient and benign. Persistent infections may however progress and lead to cervical (pre)cancerous lesions. Viral factors contributing to persistence or clearance are poorly understood. In a fist part, we studied methylation of 3'L1 and LCR regions in HPV16+ / normal cytology cervical smears, taken in 37 women who will present either a transient infection, or a persistent infection with progression to a precancerous lesion. We demonstrated that some CpG sites harbored differential methylation rates in relation with later outcome of HPV infection and may be used as predictive biomarkers.HR-HPV related oropharyngeal cancers present a better prognostic that their alcohol-and-tobacco-induced counterparts. Mechanisms responsible for this better prognostic are mainly unknown. In a second part, we characterized the HPV status in 202 oropharyngeal cancers. In the 32 cases presenting with HPV16 active infections (expressing E6/E7 mRNA), we studied 3'L1 and LCR methylation and integration of viral genome. We showed that methylation rates of some CpG sites, especially in E2 binding sites (E2BS3 & 4), were strongly methylated in episomal and mixed forms but were unmethylated in purely integrated forms.In a third part, we studied epithelial-to-mesenchymal-transition (EMT) markers in cellular models and oropharyngeal cancers, according to HPV status. A lesser EMT, known as an important phenomenon implicated in tumor invasion, could explain the better prognostic associated with HPV-induced cancers. We showed that vimentin, the more representative marker for migration and invasion ability acquisition is more expressed in HPV+ tumor cells than in HPV-negative cells. So the differential prognosis observed between the 2 oropharyngeal cancer etiologies may not be linked to EMT.Key-words: Cancer, HPV, cervical cancer, oropharynx, biomarkers, methylation, integration, EMT.

Orientador: José Vicente Tagliarini / Resumo: Introdução: a doença degenerativa da coluna cervical é prevalente e incapacitante, levando a dor e sintomas neurológicos. A cirurgia de discectomia e fusão por via anterior é bem estabelecida para o seu tratamento, sendo realizada com a colocação de dispositivos interssomáticos (cages) e placas anteriores. Novos cages ancorados prometem diminuir a disfagia pós-operatória deste procedimento. Objetivo: verificar por meio de uma revisão sistemática a efetividade da utilização de cages ancorados na redução de disfagia pós-operatória sem prejuízo dos outros benefícios obtidos com a cirurgia. Métodos: foram pesquisadas as bases de dados Embase, MEDLINE, LILACS, Scopus, Web of Science e Cochrane CENTRAL. Foram reunidos estudos observacionais e experimentais que avaliaram a utilização de cages com placas anteriores e cages ancorados em pacientes com doença degenerativa da coluna cervical. Os desfechos avaliados foram disfagia, aumento do escore JOA, diminuição do escore NDI, fusão pós-operatória, aumento da lordose cervical, tempo de cirurgia, perda intraoperatória de sangue e resultados bons/excelentes pelos critérios de Odom. Para metanálise foi empregado o software RevMan 5.3 fornecido pela Colaboraçao Cochrane. Resultados: o número total de pacientes em 30 estudos selecionados foi de 2178, sendo 1089 em cada grupo. Em 48 horas, 30 dias, 90 dias e 12 meses após a cirurgia, houve menores índices de disfagia no grupo dos cages ancorados, chegando a RR 0,15 (IC95% 0,08-0,27, I2=0%) n... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: cervical degenerative disc disease is a highly prevalent and disabling disease, leading to pain and neurologic symptoms as weakness and radiculopathy. Anterior cervical diskectomy and fusion (ACDF) is a well stabilished procedure for its treatment, usually performed by placing a spacer (cage) with the addition of an anterior fixation plate. New anchored spacers have been introduced with the promise of a faster less morbid procedure. Study goals: to perform a systematic review for the analysis of postoperative dysphagia between standard and anchored anterior cervical spacers, also taking into consideration quality-of-life and radiologic parameters. Methods: a comprehensive search was performed in the MEDLINE, Scopus, Web of Science, LILACS and Cochrane CENTRAL databases. Observational and interventional studies evaluating outcomes after ACDF with conventional cage-plate construct and anchored spacers were selected. The following outcomes were evaluated: dysphagia, JOA/mJOA score, NDI score, fusion rates, cervical lordosis improvement, operative time, intraoperative blood loss and rate of good/excellent outcomes by Odom’s criteria. The RevMan software (v5.3) was used to perform meta-analysis. Results: a total of 2178 patients were enrolled in 30 studies, with 1079 patients in both control and intervention groups. At all postoperative moments (48 hours, 30 days, 90 days and 12 months), there was a significantly lower rate of dysphagia in the anchored spacer group, ... (Complete abstract click electronic access below) / Mestre

A thesis completed by published work, Submitted to the School of Public Health, Faculty of Health Sciences, University of the Witwatersrand, in fulfilment of the requirements for the degree of Doctor of Philosophy Johannesburg, South Africa December 2016. / Introduction With the largest epidemic in the world, the consequences of human immunodeficiency virus (HIV) in South Africa extend far beyond its disease burden. In fact, patterns of HIV-related infection and mortality in South Africa still reflect social cleavages and inequalities. Similarly, poverty-related issues such as poor education, unemployment and subsequent low socio-economic status, rural residence and inadequate access to health care are all implicated in human papillomavirus (HPV) associated cervical cancer-related mortality (of which South Africa also has the highest globally). Despite the knowledge of reproductive functions and sexuality being poor among adolescents in South Africa, the majority commence their sexual activity early with an estimated national average of 15 years for girls and 14 years for boys. Further, many South African adolescents engage in sexual risk-taking behaviours including concurrent partners and unprotected sexual acts that considerably increase their vulnerability to sexually transmitted infections including HIV and HPV. In recognising the unique health needs of adolescents in South Africa, the national government has already pin-pointed school health services as a strategic arm of primary health care re-engineering. The aim of this body of work is to elaborate on restructuring of adolescent health care by introducing the HIV and HPV vaccine concomitantly in South Africa via a school-based sexual and reproductive health service. Methodology Data from four studies were analysed and are presented in three published and two unpublished papers. The first study evaluated the synergism between HIV and HPV in the South African context and formed the basis of the literature review. The second study considered HIV vaccine implementation alone. The third study assessed dual HIV and HPV vaccine strategies among females and the final study compared the dual vaccination strategy against recognised biomedical HIV prevention interventions. The studies evaluated the implementation of a hypothetical HIV vaccine and the bivalent HPV vaccine both individually and in combination when administered to school-going adolescents in South Africa. The health outcomes and the cost-effectiveness of these strategies were assessed. Assumptions were made regarding the hypothetical HIV vaccine (based on HIV vaccine studies conducted to date) including a coverage rate of 60% (uncertainty range: 30-70%), vaccine efficacy of 50% (uncertainty range: 30-70%) and vaccine price per dose of US$ 12 (uncertainty range: US$ 3-24). The uncertainty ranges were tested in the sensitivity analysis. Mortality statistics, disease transition parameters (for the individual diseases and the models representing joint disease) and HPV vaccine characteristics were drawn from the South African literature. The joint effectiveness of the dual vaccine strategy was considered multiplicative. Nine year old adolescents attending South African schools in 2012 were eligible for the intervention (vaccination) that was introduced opportunistically as part of the national health initiative introducing school-based sexual and reproductive health services. The learners were targeted prior to their reported sexual debut. The HIV vaccine was considered against the comparator of HIV counselling and testing (HCT) and the national roll-out of antiretroviral therapy (ART) that constituted the standard of care in South Africa. The HPV vaccine was modelled as prevention against HPV-related cervical cancer and pre-cancerous HPV-related cervical states. The health service provider (provider) perspective was adopted and the cohort was modelled through a lifetime horizon of 70 years with annual cycles. The economic costs and health outcomes were discounted at 3% with an uncertainty range between 0% and 6% assessed. Cost valuations were for 2012 and costs were adjusted to this common year. The quality-adjusted life year (QALY) was used as the outcome measure of health related quality of life and was used to calculate the incremental cost-effectiveness ratio (ICER) of the comparator against the vaccination interventions. The core model was a semi-Markov simulation with annual cycles. The study population entered the model HIV and HPV disease free and were exposed to the risk of acquiring each disease annually. The model structure was parameterised drawing from South African data available in the literature. One-way sensitivity analyses evaluated the impact of single assumptions on cost and outcomes. Probabilistic sensitivity analysis (PSA) with a bootstrapping technique explored the uncertainty in the model and evaluated the robustness of the results. The PSA data generated determined if the intervention fell below the willingness-to-pay (WTP) threshold. As South Africa does not have a pre-defined WTP threshold, the Gross Domestic Product (GDP) per capita (for 2012) was used as a proxy in accordance with the World Health Organization’s Guide to Cost-Effective Analysis. Additionally, benchmark interventions were used in the final comparison study as a measure of cost-effectiveness. Ethical approval for the study was obtained from the Human Research Ethics Committee (Medical) of the University of the Witwatersrand. Findings The second study explored the implementation of the HIV vaccine on an individual and national, programmatic level. The simultaneous implementation of HIV vaccination services with current HIV management programmes would be cost-effective, even at relatively higher vaccine cost. At base vaccine cost of US$ 12, the ICER was US$ 43 per QALY gained, with improved ICER values yielded at lower vaccine costs. The ICER was sensitive to the duration of vaccine-mediated protection and to variations in the vaccine efficacy. Data from this work demonstrate that vaccines offering longer duration of protection and at lower cost would result in improved ICER values. Assessing this HIV vaccine model on a national programmatic level, yielded an ICER of US$ 5 per life-year gained (LYG) (95% CI US$ 3-12) compared with the comparator. This fell considerably below the national WTP threshold of cost-effectiveness. This also translated to an 11% increase in per capita costs from US$ 80 to US$ 89. National implementation of this intervention could potentially result in an estimated cumulative gain of 24 million years of life (95% CI 8–34 million years) among those adolescents aged between 10-19 years that were vaccinated. The 10 year absolute risk reduction projected by HIV vaccine implementation was 0.42% for HIV incidence and 0.41% for HIV mortality. The ICER was sensitive to the HIV vaccine efficacy, coverage and vaccine pricing in the sensitivity analysis. The third study assessed the impact of dual HIV and HPV implementation strategies. Programmes that involved the dual vaccine strategy were assessed as cost-saving. ICER values were sensitive to the HIV vaccine cost. The dual vaccine strategy resulted in 10 year absolute risk reductions in HIV incidence (5.24%), dual mortality (1.21%) and a reduction in HPV incidence (0.39%) compared with no vaccination. Importantly, the reduction in HIV incidence rate and dual mortality rate in the dual vaccine strategy exceeded the reductions noted with the use of the HIV vaccine alone. All scenarios assessed with the dual vaccine strategy were cost-effective. Lower vaccine prices and reduced discount rates were associated with improved ICER outcomes. The final study compared the biomedical interventions of oral pre-exposure prophylaxis (PrEP), voluntary medical male circumcision (VMMC) and the scaling-up of ART coverage against the vaccine strategies. When compared with other biomedical HIV prevention interventions, the dual vaccination intervention was the most cost-effective strategy (US$ 7 per QALY gained) and averted 29% of new HIV infections. VMMC (US$ 30 per QALY gained) proved more cost-effective than HIV vaccination alone (US$ 93 per QALY gained), though VMMC averted 6% more new infections than the HIV vaccine. PrEP interventions were the least cost-effective. Combined dual vaccination and VMMC strategies represent the only dominant intervention. Strategies involving oral PrEP were the least cost-effective. Conclusion The findings of this thesis have implications for school-based adolescent health care and HIV- and HPV-related disease prevention among adolescents, a highly susceptible population. The cost-effectiveness of the dual HIV and HPV vaccine strategy was demonstrated, and the improved health outcomes associated with the interventions quantified. Proposals were suggested regarding possible combinations of HIV prevention interventions that could yield the favourable health outcomes with the most efficient use of financial resources. Several important areas for future research were identified to shed light on improving adolescent health care and for optimising HIV prevention strategies. These include integrating HIV and HPV services as part of the re-engineering of primary health care in South Africa, and then formulating economic evaluations of HIV/HPV prevention strategies targeting adolescents specifically. Further, more effective methods of collecting data on socially marginalised populations such as young people need to be explored. Another vital research area is the discussion and implementation of existing school health documents with the ideals embodied in the school health programme envisaged under the National Health Insurance restructuring. Once these are integrated, the cost implication of the combined programmes need to be assessed. / MT2017

The current study uses the head and neck of giraffe (Giraffa camelopardalis) as a model for tracking the course of evolutionary change. Gould (2002) has argued that there are three main avenues of evolutionary change that result in the genesis of new morphologies. These are phylogenetic constraints, structural or allometric scaling laws of form, and specific unique adaptations. It is well known that the unique characteristic of the giraffe is its extremely long neck and yet, it only has seven cervical vertebrae. To study the neck the vertebral body lengths of different aged giraffes were measured to determine the contribution of the cervical vertebrae to the total vertebral column. The vertebrae of several extant ungulates as well as those of fossil giraffids were used as a comparison with the giraffe. CT scans were used on several giraffe skulls to study the extent of the frontal sinus in the giraffe in an attempt to explain why the giraffe evolved such a large frontal sinus. The vertebral columns and skulls of several ungulates, including the okapi (Okapia johnstoni) were also used to compare with the results obtained from the giraffe. Immunohistochemistry was used to study the medulla and spinal cord sections of the giraffe to determine if the location and size of the nuclei remained unchanged to the basic ungulate or mammalian plan in spite of the unusually long neck, or if this long neck led to changes in the nuclei found in those regions. The results of these stains were all compared to the published literature available. Although more studies need to be conducted on other ungulates to conclusively determine why giraffe have evolved a long neck, overall the results showed that the anatomy giraffe head and neck remained true to the basic mammalian plan, with very little changing in terms of it morphology. The giraffe brain and spinal cord also resembled that of a typical ungulate. This leads to the conclusion that constraints and allometric scaling laws of form play a greater role than previously thought in the evolution of extreme morphologies.

Introdução: Sabe-se que a elevada prevalência da infecção pelo HPV na população sexualmente ativa, associa-se ao desenvolvimento delesões intra-epiteliais cervicais de alto grau e de baixo grau (LIEAG e LIABG). A cirurgia dealta freqüência (CAF) veio contribuir para o tratamento da LIE, substituindo práticas invasivas e onerosas. A população contaminada pelo HPV é composta por pacientes imunocompetentes e imunodeprimidas, especialmente as infectadas pelo HIV, que poderão ter diferente resposta a esta cirurgia. Objetivos:Avaliar a efetividade da CAF no tratamento das LIEAG no colo uterino e a taxa de complicações tardias dessa modalidade terapêutica em pacientes portadoras ou não do HIV e avaliar se a infecção HIV favorece a persistência de LIEAG após a CAF. Pacientes e Métodos: Estudo observacional prospectivo longitudinal onde foram selecionadas 97 pacientes portadoras de LIEAGe tratadas com CAF divididas em dois grupos: 38 pacientes portadoras do HIV e 59 não infectadas por esse vírus, todas atendidas em Hospital Universitário de referência terciária. As pacientes foram submetidas a CAF e reavaliadas com coleta de colpocitologia e colposcopia com três, seis, nove e 12 meses após o procedimento. As taxas de efetividade da CAF consideraram a evolução da doença cervical ao longo do seguimento desses dois grupos de pacientes por um ano. Resultados:Após o seguimento de 12 meses foram observadas situações de cura (citologia e colposcopia normais), melhora (citologia e/ou comcolposcopia com sinais de infecção HPV ou LIEBG) ou de piora da LIEAG inicial (lesão microinvasora/invasora). Sendo assim, foram obtidos para as pacientes portadoras do HIV: 56,7% de cura; 32,4% de melhora e nenhum caso de piora após o tratamento inicial. Para as pacientes não portadoras do HIV os resultados foram: 75,8% de cura; 13,8% demelhora e 1,7% de piora, indicando desfecho clínico mais favorável entre as pacientes não portadoras do HIV (X 2 , p= 0,02). As taxas de estenose de canal cervical entre as pacientes portadoras do HIV (13,5%) não foram estatisticamente diferentes daquelas observadas entre as pacientes não portadoras do HIV (10,5%). A excisão completa da LIE que motivou a CAF ocorreu em 69 (71,2%) pacientes. Entretanto, 21 (21,7%) mulheres tiveram excisão incompleta com margens da peça cirúrgica comprometida pela LIE. Dessas 21 pacientes que apresentaram margens da peça cirúrgica comprometidas apenas cinco tiveram necessidade de novo tratamento. Também foram submetidas a novo tratamento três das mulheres que tiveram margens cirúrgicas livres da peça excisada e uma paciente com a margemcirúrgica carbonizada. Conclusões: Não foi encontrada diferença estatisticamente significativa entre os dois grupos estudados em relação ao número de complicações após a realização da CAF e também na necessidade de novo tratamento no caso de mulheres portadoras do HIV. Entretanto, o desfecho clínico favorável (cura e melhora) para essa população estudada foi significativamente melhor para as pacientes não portadoras do HIV que para as infectadas por esse vírus após um ano de seguimento pós-CAF. A recidiva da LIEAG é mais freqüente em pacientes com margens cirúrgicas comprometidas, independente da presença da infecção pelo HIV. / Introduction: It is known that the high prevalence of HPV infection in the sexually active population is associated with the development of high and low-grade squamous intraepithelial lesions (HSIL and LSIL). Loop electrosurgical excision procedure (LEEP) has contributed to the treatment of SIL, replacing invasive and expensive procedures. The population contaminated with HPV consists of immunocompetent and immunodepressed patients, especially HIV-infected patients, who may respond differently to this surgery Objectives:To assess the effectiveness of LEEP in the treatment of HSIL in the uterine cervix and the rate of late complications of this therapeutic modality in patients infected or not with HIV and to determine whether HIV infection predisposes to the persistence of HSIL after LEEP. Patients and Methods: This was a prospective longitudinal observational study conducted on 97 patients with HSIL and treated with LEEP who were divided into two groups: 38 HIV-infected and 59 non-HIV-infected patients attended at a tertiary reference University Hospital. The patients were submitted to LEEP and re-evaluated bycolpocytology and colposcopy at three, six, nine and 12 months after the procedure.The rates of LEEP effectiveness were determined on the basis of the evolution of cervical disease along a one year follow-up of the two groups of patients Results:After a follow-up of 12 months, situations of cure (normal cytology and colposcopy), improvement (cytology and/or colposcopy with signs of HPV infection or HSIL) or of worsening of the original HSIL (microinvasive/invasive lesion) were observed. On this basis, the followingresults were obtained for HIV-infected patients: 55.2% of cure, 34.3% of improvement and no case of worsening after the initial treatment. For non-HIV-infected patients, the results were: 75.4% of cure, 14.1% of improvement and 1.7% of worsening, indicating a more favorable clinical outcome among non-HIV-infected patients (X 2 , p= 0.02). The rates of cervical canal stenosis among HIV-infected patients (13.5%) did not differ significantly from those among non-HIV-infected patients (10.5%). Complete excision of the SIL that motivated LEEP occurred in 69 (71.2%) patients. However, in 21 (21.7%) women, excision was incomplete, with SIL involvement of the margins of the surgical piece. Of these 21 patients with compromised surgical piece margins, only five required new treatment. Three of the women whose surgical margins were freeof disease and one patient with a carbonized surgical margin were also submitted to new treatment. Conclusions:No statistically significant difference was detected between the two groups regarding the number of complications after LEEP or also regarding the need for new treatment among HIV-infected women. However, a favorable clinical outcome (cure and improvement) for this population was significantly better for non-HIV-infected patients than for HIV-infected patients after one year of post-LEEP follow-up. HSIL recurrence was more frequent among patients with involved surgical margins regardless of the presence of HIV infection.

Introdução: Avaliar a relação do custo e da efetividade da radiofrequência (RF) em relação ao bloqueio seqüencial na Síndrome Dolorosa Complexa Regional-tipo I (SDCR-I). Métodos: 15 pacientes com dor somática e autonômica cervical foram submetidos a bloqueio torácico simpático, combinado com bloqueio facetário cervical bilateral por 4 semanas. Esta seqüência foi repetida quando a dor atingiu VAS 4 cm, e este período foi definido como tempo de analgesia. Posteriormente, os mesmos pacientes foram submetidos a um bloqueio teste seguido de modulação de RF do gânglio simpático torácico e ablação de ramos facetários medianos cervicais. Os pacientes atuaram como seu próprio controle relacionado à analgesia, atividades rotineiras e padrão de sono. Resultados: 13 pacientes completaram o estudo. O tempo de analgesia após a sequência de 4 bloqueios foi de 4 ± 1 mês e o custo anual R$ 15.000,00. O tempo de analgesia após RF foi de 13 ± 2 meses (p <0,001) e os custos foram reduzidos em 26% no primeiro ano e 34% -38% nos anos seguintes, com extrapolação. A qualidade de vida melhorou para ambos os tratamentos (p> 0,05). Não houve efeitos adversos. Discussão: A RF resultou em analgesia de 13 meses comparada a 4 meses após os bloqueios clássicos e melhora na capacidade física e no padrão de sono. Além disso, a RF foi rentável e reduziu as taxas em 23% durante a avaliação do primeiro ano, seguida de uma redução de custos de 32% a 36% nos anos seguintes, com extrapolação. / Objectives: To evaluate cost-effectiveness of radiofrequency (RF) compared to sequential block in Complex Regional Pain Syndrome-I (CRPS). Methods: 15 patients with cervical somathic and autonomic pain were submitted to a 4- weekly sympathetic thoracic block, combined to bilateral cervical facetary block. This sequence was repeated when pain reached VAS 4-cm, and this period was defined as time of analgesia. Thereafter, same patients were submitted to a test block followed by RF modulation of thoracic sympathetic ganglion and ablation of facetary cervical median branches. Patients acted as their own control related to analgesia, routine activities, sleep pattern and costs. Results: 13 patients completed the study. The analgesia time after the 4-block sequence was 4±1 months and the annual costs USA$5000. Analgesia time after RF was 13±2 months (p<0.001) and costs were reduced by 26% in the first year and 34%-38% in the following years extrapolation. Quality of life improved for both treatments (p> 0.05). There were no adverse effects. Discussion: RF resulted in 13-month compared to 4-month analgesia after the classical 4- weekly blocks, and improved physical capacity and sleep pattern. Besides that, RF was costeffective, and reduced rates by 23% during the first-year evaluation, followed by 32%-36% cost reduction in following years, by extrapolation.

A ifosfamida é um pró-fármaco que apresenta um átomo de fósforo quiral, disponível na clínica como mistura racêmica dos enantiômeros(+)-(R)-ifosfamida e (-)-(S)-ifosfamida para a utilização na quimioterapia. O objetivo do presente estudo foi o de avaliar a disposição cinética dos enantiômeros da ifosfamida em plasma de pacientes portadoras de câncer de colo do útero. As pacientes investigadas (n=6) receberam 2,5 g/m2 de ifosfamida racêmica administrada como infusão de 12 horas, sendo coletadas amostras de sangue imediatamente antes da administração e em 6, 10, 11, 12, 13, 14, 16, 18, 20 e 22 horas após a administração do fármaco. Os enantiômeros da ifosfamida foram quantificados por LC-MS/MS, sendo separados na coluna OD-R em aproximadamente 14 min empregando como fase móvel mistura de acetonitrila e água (20:80) adicionada de 0,2% de ácido fórmico. O método é linear no intervalo de 1-100 ?g de cada enantiômero/mL de plasma a partir de extrações de alíquotas de 25 ?L de plasma, compatíveis com a aplicação em farmacocinética de infusão de curta duração da ifosfamida em pacientes com câncer de colo do útero.A disposição cinética da ifosfamidaéenantiosseletiva, com observação de maiores valores de AUC (437,31 vs349,18 h.?g/mL) e menores valores de clearance(4,17 vs5,22 L/h) para o enantiômero(+)-(R)-ifosfamida. / The prodrugifosfamide has a chiral phosphorus atom, and is available clinically as a racemic mixture of the enantiomers (+)-(R)-ifosfamide and (-)-(S)-ifosfamide for use in chemotherapy. The aim of this study was to evaluate the kinetic disposition of the enantiomers of ifosfamide in plasma of patients with cancer of the cervix. The investigated patients (n = 6) received 2.5 g/m2 of racemic ifosfamide administered as infusion of 12 hours and blood samples were collected immediately before administration and at 6, 10, 11, 12, 13, 14, 16, 18, 20 and 22 hours after drug administration. The enantiomers of ifosfamide were quantified by LC-MS/MS and were separated in an OD-R column in about 14 min using as mobile phase a mixture of acetonitrile and water (20:80) plus 0.2% of formic acid. The method is linear within the range of 1-100 mg of each enantiomer/mL of plasma from extractions of 25 mL aliquots of plasma, suitable for the application in pharmacokinetics of short duration infusion of ifosfamide in patients with cervical cancer. The kineticdisposition of ifosfamide is enantioselective, with observation of higher values of AUC (437.31 vs 349.18 h.?g/mL) and lower values of clearance (4.17 vs 5.22 L/h) for the enantiomer (+)-(R)-ifosfamide.

O desenvolvimento do câncer cervical depende da exposição ao HPV, fator necessário, mas não suficiente. Outras bactérias, tais como ureaplasmas, têm sido associadas como cofatores. O objetivo deste estudo foi avaliar a presença de ureaplasmas em mulheres com lesão cervical, e observar alterações em PHK causadas pela infecção por ureaplasmas. 140 swabs vaginais foram coletados. O material foi submetido a PCR para a detecção de HPV, Mollicutes, U. urealyticum, U. parvum e seus sorotipos, e outras bactérias de importância ginecológica; e qPCR para U. urealyticum e U. parvum. Também foi realizada a infecção de ureaplasmas em PHK transformados com HPV. As células foram contadas e realizou-se a dosagem das citocinas IL1-&beta;, IL-6 e TNF-&alpha;. HPV, Mollicutes, U. parvum, sorotipos 1 e 6 de U. parvum, T. vaginalis e G. vaginalis, além de alguns fatores socioeconômicos, foram associados com lesão cervical. Verificou-se maior carga de U. parvum entre mulheres com lesão. Houve diminuição do número de células e maior liberação de IL-6 e TNF-&alpha; nos grupos infectados. Com os resultados obtidos neste estudo, foi possível verificar uma associação entre os ureaplasmas e HPV no início das lesões cervicais, contudo mais estudos precisam ser realizados para aprimorar essa hipótese. / The development of cervical cancer depends on the exposure to HPV, necessary factor, but not enough. Other bacteria, such as ureaplasmas, have been associated as cofactors. The aim of this study was to evaluate the presence of ureaplasmas in women with cervical injury, and observe changes in PHK infected by ureaplasmas. 140 vaginal swabs were collected. The material was subjected to PCR for detection of HPV, Mollicutes, Ureaplasma urealyticum, U. parvum (and serotypes) and other bacteria gynecological importance; qPCR for U. urealyticum and U. parvum was made. PHK transformed by HPV was infected by ureaplasma. Cells were counted and it was done titration of IL1-&beta;, IL-6 and TNF-&alpha;. HPV, Mollicutes, U. parvum, serotypes 1 and 6 U. parvum, T. vaginalis and G. vaginalis, and some socioeconomic factors were associated with cervical injury. Besides this, it was detected higher load U. parvum among women with injury. There was decrease in cell number and increased release of IL-6 and TNF-&alpha; in infected groups. With the results of this study, we found an association among HPV and ureaplasmas at the beginning of cervical lesions, but more studies are needed to enhance this hypothesis.

Thesis advisor: Junona Moroianu / During the papillomavirus replication cycle, the L2 minor capsid protein enters the nucleus in the initial phase after uncoating of the incoming virions and in the productive phase when L2 together with L1 major capsid protein mediate the encapsidation of the newly replicated viral genome. L2 proteins of both high risk HPV16 L2 and low risk HPV11 L2 have two nuclear localization signals (NLSs): one at the N-terminus (nNLS) and one at the C terminus (cNLS). The purpose of these experiments is to determine the minimal mutations necessary to inhibit the function of the NLSs. In this study, subcellular localization of enhanced green fluorescent protein (EGFP) fusions with full length L2 and L2 mutants lacking either the cNLS (EGFP-L2ΔC), nNLS (EGFP-L2ΔN), or both NLSs (EGFP-L2ΔNΔC) was analyzed in HeLa cell transfection assays. Full length HPV16 L2 and HPV11 L2 proteins localize to the nucleus. For both HPV16 and 11 L2, each NLS could independently mediate nuclear import in vivo. EGFP fusions were also made with mutated nNLS (EGFP-L2ΔCSbN) or mutated cNLS (EGFP-L2ΔNSbC). Transfected HeLa cells were examined by fluorescence microscopy and quantitative studies were done. In both HPV16 and 11 L2 proteins, mutation of basic residues in either NLS inhibited its nuclear import ability. / Thesis (BS) — Boston College, 2008. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: Biology. / Discipline: College Honors Program.