published_or_final_version / Anaesthesiology / Master / Master of Research in Medicine

Thesis (M. Res.(Med.))--University of Hong Kong, 2008. / Includes bibliographical references (p. 46-74)

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

No description available.

The objective of the work presented in this thesis was to explore the role of potential alternatives to platelet transfusions and specifically to investigate whether desmopressin could be used for treatment of thrombocytopenia or platelet dysfunction. Patients with thrombocytopenia or platelet dysfunction are often treated with platelet transfusions to treat or prevent bleeding. However the evidence for the efficacy of platelet transfusion is limited and there is some evidence of harm. I have focused on thrombocytopenic patients with haematological malignancies or critically ill patients, who are amongst the groups most commonly treated with platelet transfusions. The aims of this research were to determine: 1. If levels of Von Willebrand factor (VWF) or other measures of haemostasis are predictive of bleeding in severe thrombocytopenia; 2. Whether VWF compensates for thrombocytopenia in vitro; 3. The evidence for the efficacy of desmopressin in all patients undergoing surgery or invasive procedures; 4. The evidence for desmopressin for platelet dysfunction or thrombocytopenia; 5. If it is feasible to use desmopressin to treat critically ill thrombocytopenic patients in a clinical trial. To identify derangements of haemostasis that may signify candidates for alternatives to platelet transfusions, I analysed blood samples from an observational trial of fifty patients with haematological malignancies and profound thrombocytopenia due to intensive chemotherapy. I used a panel of tests to investigate measures of primary haemostasis, thrombin generation, cross-linked fibrin formation and fibrinolysis. Using multivariable logistic regression, I found no consistent correlation between any measures of haemostasis and the risk of clinically significant bleeding. VWF antigen levels were the best predictor of clinically significant bleeding on the same day (odds ratio 0.31, 95% confidence interval 0.10 to 0.98, p=0.047) but were not predictive of severe bleeding over the 24 hours after the test (odds ratio 0.48, 95% confidence interval 0.10 to 2.34, p=0.36). In a separate set of experiments, I evaluated thrombus formation under flow in thrombocytopenia. This technique was sensitive to the platelet count . Addition of exogenous VWF to thrombocytopenic blood resulted in improvement in thrombus formation, suggesting that agents that affect or influence VWF pathways might have a role. Desmopressin can be used to increase VWF levels, so leading on from my laboratory experiments; I used systematic reviews and meta-analyses to assess whether desmopressin could be used in unselected patients to reduce bleeding peri-operatively. I identified 62 randomised controlled trials. Overall there was no evidence of benefit for administering desmopressin to unselected patients. However further analysis of eleven randomised controlled trials that focused on patients with platelet dysfunction found that desmopressin resulted in transfusion of fewer units of red cells (equivalent to a 25% reduction compared to control), less blood loss (equivalent to a 23% reduction compared to control) and a lower risk of requiring a re-operation due to bleeding (Peto odds ratio 0.39, 95% confidence interval 0.18 to 0.84). There was no evidence for an increase in thrombotic events. There was no randomised controlled trial evidence for perioperative desmopressin for patients with thrombocytopenia. These specific research gaps were addressed by designing new clinical trials. I have commenced a randomised controlled feasibility trial of desmopressin versus placebo for critically ill patients with thrombocytopenia undergoing invasive procedures. This trial is ongoing and is the first randomised trial evaluating peri-procedural desmopressin in thrombocytopenia. The programme of work arising from this research has the potential to benefit a large number of patients by preventing bleeding and reducing exposure to allogeneic blood components such as platelets. The results presented in this thesis are exploratory but are an important step on a path towards larger trials using desmopressin as an alternative, or adjunct to platelet transfusion.

Introduction: This analytic essay is written in completion of a bachelor degree in pharmacy. The focus is in analyzing how different treatments affect the bleeding time. Background: Desmopressin (DDAVP) is a synthetic analogue of the pituitary hormone vasopressin, which controls the reabsorption of water in the kidneys. DDAVP increases the release of von Willebrand's factor and coagulation factor VIII, two major components of the coagulation, which are in imbalance in bleeding disease. Purpose: The purpose of this work is a critical appraisal of clinical trials of DDAVP to evaluate its medical value in bleeding disorders. Method: Since this is a literature study, clinical studies were selected in the PubMed database, which evaluated the effect of DDAVP in bleeding disease and the coagulation of healthy individuals. Result: All nine trials demonstrate an effect of DDAVP on the coagulation system. The hormone has a local effect independent of central influence, the mechanism is not fully clear. DDAVP shortens the bleeding time in bleeding disease, both congenital and acquired, by increasing the release of vWF and FVIII, as well as affecting the adhesiveness of the platelets to strengthen the coagulation. This effect of DDAVP is a blessing for patients suffering from bleeding diseases, but also for patients undergoing major surgery, associated with a risk of major blood loss. Risk of fluid retention, but may be limited if fluid intake is checked throughout the day following the intake of DDAVP.  Conclusion: DDAVP acts as the only therapy form for patients with mild form of haemophilia, and for those with moderate or severe form, DDAVP, in addition to the need for factor concentrate, may also have its place of therapeutic treatment together with tranexamic acid. The conclusion of this survey is that DDAVP has a very high medical value for patients with hemorrhage because it improves coagulation, reduces and stops bleeding without direct side effects, in other words, they raise the quality of life. / Inledning: Som avslutning i utbildningen till receptarie skrivs arbetet utformat som en litteraturstudie. Arbetet innehåller analys av mätning i blödningstid för olika behandlingar. Bakgrund: Desmopressin (DDAVP) är en syntetisk analog till hypofyshormonet vasopressin, vilket styr reabsorberingen av vatten i njurarna. DDAVP ökar frisättning av von Willebrands faktor (vWF) och koagulationsfaktor VIII (FVIII), två viktiga komponenter i koagulationen, vilka är i obalans vid blödarsjuka. Syfte: Detta arbetes syfte är att utvärdera om DDAVP har ett medicinskt värde vid blödarsjuka. Metod: De kliniska studierna valdes ut från databasen PubMed, vilka evaluerade effekten av DDAVP på blödarsjuka samt koagulationen på friska försökspersoner. Resultat: Alla nio artiklarna påvisar effekten av DDAVP hos människa. Hormonet har en lokal verkan oberoende av central påverkan, mekanismen är inte helt klarlagd. DDAVP förkortar blödningstiden hos blödarsjuka, såväl medfödd som förvärvad, genom att öka frisättning av vWF och faktor VIII, samt genom påverkan på trombocyternas adhesivitet, så att koagulationen stärks. Denna effekt av DDAVP är tacksam för blödarsjuka, men även för övriga vårdfall vilka genomgår stora kirurgiska ingrepp med risk för stor blodförlust då effekten kan bli betydande. Det finns en risk för vätskeretention, men den kan begränsas om vätskeintagen kontrolleras dygnet närmast intag av DDAVP. Slutsats: DDAVP agerar som enda terapiform för de med mild form av blödarsjuka, och för de med moderat eller svår form kan förutom behovet av faktorskoncentrat även de DDAVP ha sin givna plats som terapibehandling tillsammans med tranexamsyra. Slutsatsen blir att DDAVP har ett mycket stort medicinskt värde för patienter med blödarobalans eftersom det förbättrar koagulationen, minskar och stoppar blödning utan direkta biverkningar, med andra ord höjer de blödarsjukas livskvalitet.

A doença de von Willebrand (DvW) é uma coagulopatia hereditária, causada por defeitos qualitativos ou quantitativos do fator de von Willebrand. O tratamento e a prevenção das intercorrências da DvW são bastante dispendiosos e, em geral, se baseiam na administração de concentrado de Fator VIII/FvW (CFVIII/FvW) e/ou da Desmopressina (DDAVP). Em muitas situações, o DDAVP é um tratamento eficaz que não expõe os pacientes aos riscos de contaminação viral e apresenta custo inferior quando comparado ao CFVIII/FvW. No entanto, a dificuldade de diagnóstico e classificação da DvW, bem como o baixo número de pacientes que se submetem ao teste para avaliação da resposta ao DDAVP, restringem a indicação do DDAVP como alternativa terapêutica para esses pacientes. O objetivo deste estudo foi avaliar retrospectivamente a indicação, o uso e o custo dos medicamentos no tratamento de pacientes com DvW com DDAVP e CFVIII/FvW no Hemocentro de Belo Horizonte no período entre 2011 a 2013. Este estudo incluiu 124 (24,22%) pacientes com DvW atendidos no hemocentro. Em 18 pacientes (14,52%) o diagnóstico de DvW não pode ser confirmado. Doze pacientes (9,68%) não puderam ser classificados e 73 foram classificados como tipo 1, 19 como tipo 2 e 2 pacientes como tipo 3. Oitenta e um pacientes fizeram o teste de DDAVP, sendo que 87,65% foram considerados responsivos. Nos pacientes tipo 1, a taxa de resposta ao DDAVP foi de 92%. Quase 32% dos pacientes tipo 1 não realizaram o teste. No período avaliado, foram utilizadas 3.794mcg de DDAVP (R$13.165,18) e 1.582.250 UI de CFVIII/FvW (R$1.075.930,00). Vinte por cento dos pacientes responsivos ao DDAVP utilizaram CFVIII/FvW em indicações onde o DDAVP poderia ter sido considerado (69.200UI de CFVIII/FvW versus 131 ampolas de DDAVP). Nos pacientes potencialmente responsivos ao DDAVP 108.700UI de CFVIII/FvW (R$73.916,00) poderiam ter sido substituídas por 247 ampolas de DDAVP (R$3.428,36). A escolha do DDAVP nessas situações poderia representar uma economia de 95,7% do valor gasto no tratamento do grupo de 27 pacientes responsivos e potencialmente responsivos ao DDAVP e 10,6% do valor total gasto para todo o tratamento dos pacientes no período do estudo. Estudos mais complexos de farmacoeconomia serão necessários para avaliar a magnitude da economia gerada com esse uso. O presente estudo mostrou que o DDAVP é uma alternativa terapêutica de menor custo, cuja indicação e utilização podem ser ampliadas no tratamento dos pacientes com DvW. Dessa maneira, a implementação de estratégias visando melhorar o diagnóstico, a classificação da doença, o acesso à testagem quanto à resposta ao DDAVP, bem como a conscientização dos profissionais de saúde e pacientes, quanto ao custo e segurança do DDAVP podem contribuir para o uso racional dos recursos destinados a essa parcela da população. / Von Willebrand disease (VWD) is a hereditary coagulopathy caused by qualitative or quantitative defects on von Willebrand factor. The treatment and the prevention of VWD complications is quite expensive and is generally based on the administration of vWF:FVIII Concentrates and/or Desmopressin (DDAVP). In many situations, DDAVP is an effective treatment that does not expose patients to viral contamination risks and presents a lower cost when compared to vWF:FVIII concentrates. However, the difficulty of diagnosis and classification of VWD, as well as the low number of patients tested to their responsiveness to DDAVP, restrict the use of DDAVP as an alternative treatment for these patients. The aim of this study was to evaluate retrospectively the clinical indications, the use and the cost of treatment of VWD patients with DDAVP and vWF:FVIII concentrates in the Blood Center of Belo Horizonte between 2011 and 2013. This study enrolled 124 (24.22%) VWD patients attended at the Blood Center.For18 (14.52%) patients, the diagnosis of VWD could not be confirmed. Twelve patients (9.68%) could not be classified and 73patients were classified as type 1, 19 as type 2 and 2 as type 3. Eighty-one patients were tested for DDAVP response and 87.65% (n=71) were considered responsive for the treatment. For type 1 VWD patients, the response rate to DDAVP was 92%. Almost 32% of type 1 VWD patients were not tested. In the period evaluated, 3,794mcg of DDAVP (R$ 13,165.18) and 1,582,250 IU of vWF:FVIII concentrates (R$ 1,075,930.00) were used. Between the cases with clinical indication of DDAVP use, 20% patients used vWF:FVIII concentrates (69.200UI of vWF:FVIII versus 131ampoules of DDAVP). In patients with good responsive to DDAVP, 108,700 IU of vWF:FVIII concentrates used (R$ 73,916.00) could be replaced by 247 ampoules of DDAVP (R$ 3,428.36). The choice of DDAVP in these situations could represent an economy of 95.7% of the value spent on the treatment of the 27 responsive and potentially responsive patients to DDAVP and 10.6% of the total value spent for the entire treatment of patients in the study period. More detailed studies of pharmacoeconomics are necessary to assess the magnitude of the economy generated by the use of DDAVP. This study demonstrated that DDAVP is a lower cost therapeutic alternative whose indication and use can be enhanced in the treatment of VWD patients. In this context, adoption of strategies to improve the differential diagnosis, expand the DDAVP responsiveness test, and aware health professionals and patients about the costs and safety use of DDAVP, could contribute to the rational use of resources designated to treatment of VWD.

A doença de von Willebrand (DvW) é uma coagulopatia hereditária, causada por defeitos qualitativos ou quantitativos do fator de von Willebrand. O tratamento e a prevenção das intercorrências da DvW são bastante dispendiosos e, em geral, se baseiam na administração de concentrado de Fator VIII/FvW (CFVIII/FvW) e/ou da Desmopressina (DDAVP). Em muitas situações, o DDAVP é um tratamento eficaz que não expõe os pacientes aos riscos de contaminação viral e apresenta custo inferior quando comparado ao CFVIII/FvW. No entanto, a dificuldade de diagnóstico e classificação da DvW, bem como o baixo número de pacientes que se submetem ao teste para avaliação da resposta ao DDAVP, restringem a indicação do DDAVP como alternativa terapêutica para esses pacientes. O objetivo deste estudo foi avaliar retrospectivamente a indicação, o uso e o custo dos medicamentos no tratamento de pacientes com DvW com DDAVP e CFVIII/FvW no Hemocentro de Belo Horizonte no período entre 2011 a 2013. Este estudo incluiu 124 (24,22%) pacientes com DvW atendidos no hemocentro. Em 18 pacientes (14,52%) o diagnóstico de DvW não pode ser confirmado. Doze pacientes (9,68%) não puderam ser classificados e 73 foram classificados como tipo 1, 19 como tipo 2 e 2 pacientes como tipo 3. Oitenta e um pacientes fizeram o teste de DDAVP, sendo que 87,65% foram considerados responsivos. Nos pacientes tipo 1, a taxa de resposta ao DDAVP foi de 92%. Quase 32% dos pacientes tipo 1 não realizaram o teste. No período avaliado, foram utilizadas 3.794mcg de DDAVP (R$13.165,18) e 1.582.250 UI de CFVIII/FvW (R$1.075.930,00). Vinte por cento dos pacientes responsivos ao DDAVP utilizaram CFVIII/FvW em indicações onde o DDAVP poderia ter sido considerado (69.200UI de CFVIII/FvW versus 131 ampolas de DDAVP). Nos pacientes potencialmente responsivos ao DDAVP 108.700UI de CFVIII/FvW (R$73.916,00) poderiam ter sido substituídas por 247 ampolas de DDAVP (R$3.428,36). A escolha do DDAVP nessas situações poderia representar uma economia de 95,7% do valor gasto no tratamento do grupo de 27 pacientes responsivos e potencialmente responsivos ao DDAVP e 10,6% do valor total gasto para todo o tratamento dos pacientes no período do estudo. Estudos mais complexos de farmacoeconomia serão necessários para avaliar a magnitude da economia gerada com esse uso. O presente estudo mostrou que o DDAVP é uma alternativa terapêutica de menor custo, cuja indicação e utilização podem ser ampliadas no tratamento dos pacientes com DvW. Dessa maneira, a implementação de estratégias visando melhorar o diagnóstico, a classificação da doença, o acesso à testagem quanto à resposta ao DDAVP, bem como a conscientização dos profissionais de saúde e pacientes, quanto ao custo e segurança do DDAVP podem contribuir para o uso racional dos recursos destinados a essa parcela da população. / Von Willebrand disease (VWD) is a hereditary coagulopathy caused by qualitative or quantitative defects on von Willebrand factor. The treatment and the prevention of VWD complications is quite expensive and is generally based on the administration of vWF:FVIII Concentrates and/or Desmopressin (DDAVP). In many situations, DDAVP is an effective treatment that does not expose patients to viral contamination risks and presents a lower cost when compared to vWF:FVIII concentrates. However, the difficulty of diagnosis and classification of VWD, as well as the low number of patients tested to their responsiveness to DDAVP, restrict the use of DDAVP as an alternative treatment for these patients. The aim of this study was to evaluate retrospectively the clinical indications, the use and the cost of treatment of VWD patients with DDAVP and vWF:FVIII concentrates in the Blood Center of Belo Horizonte between 2011 and 2013. This study enrolled 124 (24.22%) VWD patients attended at the Blood Center.For18 (14.52%) patients, the diagnosis of VWD could not be confirmed. Twelve patients (9.68%) could not be classified and 73patients were classified as type 1, 19 as type 2 and 2 as type 3. Eighty-one patients were tested for DDAVP response and 87.65% (n=71) were considered responsive for the treatment. For type 1 VWD patients, the response rate to DDAVP was 92%. Almost 32% of type 1 VWD patients were not tested. In the period evaluated, 3,794mcg of DDAVP (R$ 13,165.18) and 1,582,250 IU of vWF:FVIII concentrates (R$ 1,075,930.00) were used. Between the cases with clinical indication of DDAVP use, 20% patients used vWF:FVIII concentrates (69.200UI of vWF:FVIII versus 131ampoules of DDAVP). In patients with good responsive to DDAVP, 108,700 IU of vWF:FVIII concentrates used (R$ 73,916.00) could be replaced by 247 ampoules of DDAVP (R$ 3,428.36). The choice of DDAVP in these situations could represent an economy of 95.7% of the value spent on the treatment of the 27 responsive and potentially responsive patients to DDAVP and 10.6% of the total value spent for the entire treatment of patients in the study period. More detailed studies of pharmacoeconomics are necessary to assess the magnitude of the economy generated by the use of DDAVP. This study demonstrated that DDAVP is a lower cost therapeutic alternative whose indication and use can be enhanced in the treatment of VWD patients. In this context, adoption of strategies to improve the differential diagnosis, expand the DDAVP responsiveness test, and aware health professionals and patients about the costs and safety use of DDAVP, could contribute to the rational use of resources designated to treatment of VWD.

Introdução: A enurese (EN) é uma condição clínica de etiologia multifatorial com característica de perda de urina intermitente no período noturno que promove uma dificuldade ao convívio social da criança/adolescente e pode ser afetada pela presença de comorbidades e por imaturidade do sistema nervoso na regulação central sobre o funcionamento vesical. O conjunto de eventos promotores do episódio da enurese, desde o enchimento vesical durante o sono até a deflagração do esvaziamento vesical involuntário sem acordar constitui-se no foco principal de abordagem clínico-laboratorial e terapêutica. Objetivo: Estudar o impacto de uma avaliação multidisciplinar do indivíduo com enurese noturna monossintomática (ENM) sobre a eficácia terapêutica dos métodos tradicionalmente utilizados em sua abordagem, assim como comparar, pela polissonografia (PSG), os efeitos da intervenção sobre a estrutura do sono. Método: Estudo prospectivo de crianças de 6 a 17 anos incompletos, com ENM, diagnosticada por avaliação multidisciplinar utilizando anamnese estruturada com enfoque nefropediátrico, exame clínico, diários das eliminações, ultrassonografia de rins e vias urinárias, análises laboratoriais de sangue e urina, exame neurológico, diário do sono, questionários de avaliação do sono e polissonografia, avaliação psicológica de distúrbios do comportamento pelo CBCL e da qualidade de vida pelo PedsQL 4.0 e avaliação fisioterapêutica do equilíbrio. Condições crônicas e genéticas constituíram exclusão do protocolo. O estudo foi aprovado pelo Comitê de Ética em Pesquisa do HCFMUSP de número 0649/10 e apoio da FAPESP (2011/17589-1). Das 140 crianças/adolescentes ingressantes, foram excluídos 58 (41,4%) por comorbidades não tratáveis e/ou não aderência ao protocolo, 82/140 (58,6%) pacientes com ENM foram incluídos para intervenção terapêutica em três grupos de tratamento (alarme, desmopressina e alarme com desmopressina). A resposta à intervenção terapêutica foi avaliada no período pós-intervenção imediato (após seis meses de tratamento) e tardio (após 12 meses da suspensão do tratamento). A PSG foi realizada pré e pós-tratamento para análise comparativa da estrutura do sono. Resultados: Dos 82 pacientes, com idade média de 9,5 anos (? 2,6), 62 eram do sexo masculino (75,6%). Diagnosticou-se antecedente familiar de EN em 91,1% considerando parentes de 1° e 2° graus, constipação em 81,7% e apneia leve/moderada em 40,7%. A avaliação fisioterapêutica realizada na fase pré-intervenção demonstrou presença de alteração no controle do equilíbrio nos pacientes com enurese. A associação entre os fatores reconhecidos na abordagem clínica inicial e a gravidade da enurese (% de episódios/mês) na fase préintervenção, apresentou significância para prematuridade (p=0,03). Previamente à randomização, após abordagem clínica de constipação e/ou terapia comportamental simples, verificou-se cura de 7/82 pacientes. Foram randomizados para tratamento nas três modalidades de intervenção 75 pacientes. Durante a fase de intervenção ocorreram 14/75 (18,7%) desistências, principalmente no grupo alarme (p=0,00). Verificou-se nos 61 pacientes em tratamento, sucesso inicial (resposta completa e parcial) em 56,6% do grupo alarme, 70% do grupo desmopressina e 64% do grupo combinado (p=0,26). Sucesso contínuo (SC) ocorreu em 70% do grupo Alarme, 84,2% do grupo Desmopressina e 100% do grupo combinado (p=0,21). A recidiva ocorreu em 3/20 (15%) pacientes do grupo Alarme e 1/19 (5,2%) do grupo Desmopressina. O sucesso terapêutico se associou a redução nos escores de problemas de comportamento e melhora dos escores de qualidade de vida dos pacientes. A análise comparativa do sono pré e pós-intervenção nos pacientes com e sem alarme demonstrou aumento dos microdespertares (p=0,00), diminuição da eficiência do sono (p=0,02), diminuição de N2 (p=0,00) no grupo alarme. Conclusão: A enurese é um distúrbio multifatorial que exige uma abordagem diagnóstica estruturada. No presente estudo, a abordagem terapêutica utilizando três metodologias de intervenção demonstrou eficácia semelhante. Os benefícios associados ao sucesso terapêutico são amplos, sugere-se, no entanto, que a terapia com alarme possa repercutir negativamente na estrutura do sono / Introduction: Enuresis (NE) is a clinical condition of multifactorial etiology with intermittent nocturnal urine loss characteristic at night that leads to difficulties in child / adolescent social interaction. It can be affected by comorbidities and by immaturity of the central nervous system in the regulation of bladder function. The range of events of the bedwetting episode, from bladder filling until the involuntary triggering of bladder emptying during sleep, constitutes the main focus for clinical-laboratory and therapeutic approach. Objective: To study the impact of a multidisciplinary assessment of the patient with monosymptomatic nocturnal enuresis (MNE) on the therapeutic efficacy of the interventions traditionally used in its approach, and to compare, by polysomnography (PSG), the effects of intervention on sleep structure. Method: Prospective study of children, of 6 to 17 incomplete years, with MNE diagnosed by multidisciplinary assessment, based on: pediatric nephrology - oriented structured history, clinical examination, kidney and urinary tract ultrasound, laboratory tests of blood and urine and daily bladder and intestinal elimination diaries; neurological examination, sleep diary and sleep questionnaires and PSG; psychology evaluation using CBCL and PedsQL 4.0 questionnaires; physiotherapic evaluation of balance. The study was approved by the HCFMUSP Ethics Committee in Research of number 0649/10 and was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Grant # 2011/17589-1. Of the 140 children/adolescents entering the study, 58 (41.4%) were excluded because of non - treatable comorbidities and / or nonadherence to study protocol, 82/140 (58.6%) patients with MNE were included for therapeutic intervention in three treatment groups (alarm, desmopressin and alarm with desmopressin). The response to therapeutic intervention was evaluated in the immediate and late post-intervention period, respectively after six of treatment and after 12 months of posttreatment follow-up. Pre and post intervention PSG were compared to evaluate therapy impact on the structure of sleep. Results: Of 82 patients, 62 were male (75.6%) with mean age of 9.5 years (± 2.6). Family history of NE was diagnosed in 91.1% of first and second degree relatives, constipation in 81.7% and mild/moderate apnea in 40.7%. The physical therapy preintervention evaluation identified an alteration in the balance control of the MNE patients. In the pre-intervention phase, an association of prematurity with severity of enuresis (% of episodes/month) (p = 0.03) was identified. Enuresis cure was achieved, prior to randomization, after clinical approach to constipation and / or simple behavioral therapy, by 7/82 patients, 75 patients were randomized to treatment with three modes of intervention. During the intervention phase, a dropout rate of 14/75 (18.7%) patients was verified, especially in the alarm group (p = 0.00). Initial success (complete and partial response) was observed in 56.6% patients of the alarm group, 70% of the desmopressin group and 64% of the combined group (p = 0.26). Continued success (SC) occurred in 70% patients of the alarm group, 84.2% of desmopressin group and 100% of the combined group (p = 0.21). Recurrence occurred in 3/20 (15%) patients of the alarm group and 1/19 (5.2%) of desmopressin group. Therapeutic success was associated with a reduction in scores for behavioral problems and with improvement of patients\' quality of life scores. The comparative analysis between pre and post-intervention sleep structure parameters, in patients treated with and without alarm, showed increased arousals (p=0.00), decreased sleep efficiency (p = 0.02), decreased N2 (p = 0.00) in the alarm group. Conclusion: Enuresis is a multifactorial disorder that requires a structured diagnostic approach. In the present study, three therapeutic intervention methodologies demonstrated similar efficacy. The benefits associated with treatment success are multiple; the present study data suggest, however, that alarm therapy can have a negative effect on sleep structure

In the last decades NMR spectroscopy has become an invaluable tool both in academic research and in the pharmaceutical industry. This thesis describes applications of NMR spectroscopy in biomedicinal research for structure elucidation of biologically active peptides and peptidomimetics as well as in studies of ligand-protein interactions. The first part of this thesis describes the theory and methodology of structure calculations of peptides using experimental restraints derived from NMR spectroscopy. This methodology has been applied to novel mimetics of the peptide hormones desmopressin and Leu-enkephalin. The results of these studies highlight the complicating issue of conformational exchange often encountered in structural determination of peptides and how careful analysis of experimental data as well as optimization of experimental conditions can enable structure determinations in such instances. Although the mimetics of both desmopressin and Leu-enkephalin were found to adopt the wanted conformations, they exhibited no or very poor biological activity. These results demonstrate the difficulties in designing peptidomimetics without detailed structural information of the receptors. A stereoselective synthetic route towards XxxΨ[CH2O]Ala pseudodipeptides is also presented. Such pseudodipeptides can be used as isosteric amide bond replacements in peptides in order to increase their resistance towards proteolytic degradation. The second part of this thesis describes the study of the interaction between compounds that inhibit pilius assembly, pilicides, and periplasmic chaperones from uropathogenic Escherichia coli. Periplasmic chaperones are key components in assembly of pili, i.e. hair-like protein complexes located on the surface of Escherichia coli that cause urinary tract infections. Detailed knowledge about this interaction is important in understanding how pilicides can inhibit pilus assembly by binding to chaperones. Relaxation-edited NMR experiments were used to confirm the affinity of the pilicides for the chaperones and chemical shift mapping was used to study the pilicide-chaperone interaction surface. These studies show that at least two interaction sites are present on the chaperone surface and consequently that two different mechanisms resulting in inhibition of pilus assembly may exist.