A palavra DOR é definida como uma percepção consciente do indivíduo de impulsos nociceptivos modulados que originam uma experiência emocional e sensitiva desagradável, associada à lesão tecidual real ou potencial, ou descrita em termos de tal lesão. Considerando-se que a dor é um dos motivos mais comuns que levam um paciente a procurar por atendimento em consultório odontológico, este estudo teve como objetivo quantificar e qualificar a analgesia da musculatura mastigatória e da articulação temporomandibular proporcionada por medicamentos antiinflamatórios não esteroidais, associados ou não a outros agentes terapêuticos. O estudo clínico foi desenvolvido em pacientes que sofriam de algias crônicas na musculatura mastigatória, decorrentes de disfunções temporomandibulares. Foram selecionados, após anamnese e avaliação com a ferramenta RDC/TMD traduzido para a língua Portuguesa (PEREIRA JUNIOR, 2007), 18 voluntários para avaliar o efeito terapêutico (entendendo-se como efeito terapêutico o alívio da sintomatologia dolorosa e do restabelecimento da amplitude dos movimentos bordejantes mandibulares), dos três tratamentos coadjuvantes abaixo-relacionados, sendo dois medicamentos e um placebo para eliminar o efeito psicológico. Os tratamentos avaliados foram: um antiinflamatório não-esteroidal (AINES) Flanaren® (diclofenaco sódico), uma panacéia Sedilax® composta por AINES, miorrelaxante e analgésicos (diclofenaco sódico + carisoprodol + paracetamol + cafeína), ambos produzidos pelo laboratório Teuto® ; e um placebo, que consistia de pílulas preenchidas com 110 g de amido de milho, produzidas pela Faculdade de Ciências Farmacêuticas de Ribeirão Preto - USP. A administração de cada medicamento consistia de 1 unidade a cada 12 horas, durante um período de 10 dias, precedido e sucedido por avaliações de dor dos pacientes. Foi estabelecido um período de washout de 11 dias entre cada troca de tratamento. A avaliação dos tratamentos medicamentosos foi desenvolvida com diferentes ferramentas, como o McGill Pain Questionnaire (VAROLI; PEDRAZZI, 2006), para qualificar e quantificar dor não provocada, a escala visual analógica para dor à palpação, escala numérica para a quantificação da dor durante o tratamento, além de mensurações de amplitude de movimentos excursivos mandibulares. Foram colhidas também informações sobre possíveis efeitos colaterais indesejáveis relacionados aos tratamentos. O projeto foi submetido e aprovado pelo Comitê de Ética em Pesquisa envolvendo Seres Humanos da Faculdade de Odontologia de Ribeirão Preto da USP, Processo n.2006.1.558.58.0, Caae n.0022.0.138.000-06. Os resultados mostraram que a analgesia para a dor em repouso foi melhor com a utilização do Flanaren® e para a dor à palpação, igual para ambos os tratamentos. Os medicamentos Sedilax® e Flanaren® reduziram significantemente a dor após três dias de tratamento, enquanto o placebo, após oito dias. Não foram observadas melhoras na amplitude dos movimentos limítrofes da mandíbula. Também não foram observados efeitos colaterais significantes estatisticamente. Concluiu-se que o tratamento utilizando o diclofenaco sódico como adjuvante reduziu a dor em repouso; todos os tratamentos promoveram analgesia à dor à palpação, mas tanto o diclofenaco isolado como associado agiram no terceiro dia e o placebo, apenas no oitavo. Nenhum efeito colateral observado foi estatisticamente significante. / The word PAIN is defined as a conscious perception of modulated nociceptive input from an unpleasant emotional and sensitive experience, associated to a real or potential, or described in terms of such lesion. Considering that pain is one of main reasons which motivate patients to search for dental treatment, the aim of this study was quantify and qualify analgesia in masticatory muscles and temporomandibular joints by administration of non steroidal anti-inflammatory drugs, isolated or associated to other therapeutic agents. This clinical trial has been developed treating patients who had been suffering with chronic pain in masticatory muscles due to temporomandibular disorders. Eighteen volunteers were selected after anamnesis exam and assessment using RDC/TMD translated to Portuguese (PEREIRA JUNIOR, 2007), to evaluate the therapeutics effect (pain relief and maximum eccentric jaw movement recovery) of three adjuvant treatment: two medicines and one placebo, to eliminate psychological effects. Assessed treatments were: a non steroidal anti-inflammatory Flanaren® (sodium diclofenac), a panacea composed by an anti-inflammatory, muscle relaxant and analgesics (sodium diclofenac + carisoprodol + acetaminophen + caffeine), both produced by pharmaceutical laboratory Teuto® ; and a placebo, that were pills filled by 110 g of corn starch, produced by Faculty of Pharmaceutical Sciences of Ribeirão Preto USP. The dosage of all medicines was one pill every 12 hours, during 10 days, preceded and succeeded by patients` pain assessment. An 11 days washout period among each therapy has been established. The assessment of drug therapies were done using distinct instruments, as McGill Pain Questionnaire (VAROLI; PEDRAZZI, 2006), to qualify and quantify unprovoked pain; visual analogue scale for pain on palpation, numerical scale to quantify pain during treatment, and measurement of range of motion during maximum eccentric jaw movements. It has been obtained information about side effects related to treatments. The research project was submitted and approved by Ethics in Research Committee of Faculty of Dentistry of Ribeirão Preto USP, Lawsuit n.2006.1.558.0, CAAE n. 0022.0.138.000-06. Data analysis has shown that relief of unprovoked pain was better using Flanaren® , and reduction of pain on palpation was equal in all treatments. Both, diclofenac alone, also diclofenac associate to other drugs, reduced significantly pain after three days of treatment, while placebo, after eight days. It has not been observed increase of range of motion during maximum jaw excursive movements, neither statistically significant side-effect. It has been concluded that treatment using diclofenac as an adjuvant reduced unprovoked pain; all therapies relief pain on palpation, but it was observed on third day for diclofenac and diclofenac associated and on eighth day for placebo. There was not any statistically significant side effect.

A palavra DOR é definida como uma percepção consciente do indivíduo de impulsos nociceptivos modulados que originam uma experiência emocional e sensitiva desagradável, associada à lesão tecidual real ou potencial, ou descrita em termos de tal lesão. Considerando-se que a dor é um dos motivos mais comuns que levam um paciente a procurar por atendimento em consultório odontológico, este estudo teve como objetivo quantificar e qualificar a analgesia da musculatura mastigatória e da articulação temporomandibular proporcionada por medicamentos antiinflamatórios não esteroidais, associados ou não a outros agentes terapêuticos. O estudo clínico foi desenvolvido em pacientes que sofriam de algias crônicas na musculatura mastigatória, decorrentes de disfunções temporomandibulares. Foram selecionados, após anamnese e avaliação com a ferramenta RDC/TMD traduzido para a língua Portuguesa (PEREIRA JUNIOR, 2007), 18 voluntários para avaliar o efeito terapêutico (entendendo-se como efeito terapêutico o alívio da sintomatologia dolorosa e do restabelecimento da amplitude dos movimentos bordejantes mandibulares), dos três tratamentos coadjuvantes abaixo-relacionados, sendo dois medicamentos e um placebo para eliminar o efeito psicológico. Os tratamentos avaliados foram: um antiinflamatório não-esteroidal (AINES) Flanaren® (diclofenaco sódico), uma panacéia Sedilax® composta por AINES, miorrelaxante e analgésicos (diclofenaco sódico + carisoprodol + paracetamol + cafeína), ambos produzidos pelo laboratório Teuto® ; e um placebo, que consistia de pílulas preenchidas com 110 g de amido de milho, produzidas pela Faculdade de Ciências Farmacêuticas de Ribeirão Preto - USP. A administração de cada medicamento consistia de 1 unidade a cada 12 horas, durante um período de 10 dias, precedido e sucedido por avaliações de dor dos pacientes. Foi estabelecido um período de washout de 11 dias entre cada troca de tratamento. A avaliação dos tratamentos medicamentosos foi desenvolvida com diferentes ferramentas, como o McGill Pain Questionnaire (VAROLI; PEDRAZZI, 2006), para qualificar e quantificar dor não provocada, a escala visual analógica para dor à palpação, escala numérica para a quantificação da dor durante o tratamento, além de mensurações de amplitude de movimentos excursivos mandibulares. Foram colhidas também informações sobre possíveis efeitos colaterais indesejáveis relacionados aos tratamentos. O projeto foi submetido e aprovado pelo Comitê de Ética em Pesquisa envolvendo Seres Humanos da Faculdade de Odontologia de Ribeirão Preto da USP, Processo n.2006.1.558.58.0, Caae n.0022.0.138.000-06. Os resultados mostraram que a analgesia para a dor em repouso foi melhor com a utilização do Flanaren® e para a dor à palpação, igual para ambos os tratamentos. Os medicamentos Sedilax® e Flanaren® reduziram significantemente a dor após três dias de tratamento, enquanto o placebo, após oito dias. Não foram observadas melhoras na amplitude dos movimentos limítrofes da mandíbula. Também não foram observados efeitos colaterais significantes estatisticamente. Concluiu-se que o tratamento utilizando o diclofenaco sódico como adjuvante reduziu a dor em repouso; todos os tratamentos promoveram analgesia à dor à palpação, mas tanto o diclofenaco isolado como associado agiram no terceiro dia e o placebo, apenas no oitavo. Nenhum efeito colateral observado foi estatisticamente significante. / The word PAIN is defined as a conscious perception of modulated nociceptive input from an unpleasant emotional and sensitive experience, associated to a real or potential, or described in terms of such lesion. Considering that pain is one of main reasons which motivate patients to search for dental treatment, the aim of this study was quantify and qualify analgesia in masticatory muscles and temporomandibular joints by administration of non steroidal anti-inflammatory drugs, isolated or associated to other therapeutic agents. This clinical trial has been developed treating patients who had been suffering with chronic pain in masticatory muscles due to temporomandibular disorders. Eighteen volunteers were selected after anamnesis exam and assessment using RDC/TMD translated to Portuguese (PEREIRA JUNIOR, 2007), to evaluate the therapeutics effect (pain relief and maximum eccentric jaw movement recovery) of three adjuvant treatment: two medicines and one placebo, to eliminate psychological effects. Assessed treatments were: a non steroidal anti-inflammatory Flanaren® (sodium diclofenac), a panacea composed by an anti-inflammatory, muscle relaxant and analgesics (sodium diclofenac + carisoprodol + acetaminophen + caffeine), both produced by pharmaceutical laboratory Teuto® ; and a placebo, that were pills filled by 110 g of corn starch, produced by Faculty of Pharmaceutical Sciences of Ribeirão Preto USP. The dosage of all medicines was one pill every 12 hours, during 10 days, preceded and succeeded by patients` pain assessment. An 11 days washout period among each therapy has been established. The assessment of drug therapies were done using distinct instruments, as McGill Pain Questionnaire (VAROLI; PEDRAZZI, 2006), to qualify and quantify unprovoked pain; visual analogue scale for pain on palpation, numerical scale to quantify pain during treatment, and measurement of range of motion during maximum eccentric jaw movements. It has been obtained information about side effects related to treatments. The research project was submitted and approved by Ethics in Research Committee of Faculty of Dentistry of Ribeirão Preto USP, Lawsuit n.2006.1.558.0, CAAE n. 0022.0.138.000-06. Data analysis has shown that relief of unprovoked pain was better using Flanaren® , and reduction of pain on palpation was equal in all treatments. Both, diclofenac alone, also diclofenac associate to other drugs, reduced significantly pain after three days of treatment, while placebo, after eight days. It has not been observed increase of range of motion during maximum jaw excursive movements, neither statistically significant side-effect. It has been concluded that treatment using diclofenac as an adjuvant reduced unprovoked pain; all therapies relief pain on palpation, but it was observed on third day for diclofenac and diclofenac associated and on eighth day for placebo. There was not any statistically significant side effect.

Cellular senescence a specialized form of growth arrest that contributes to the pathogenesis of several aging-related disorders including cancer. While by definition tumor cells are considered immortalized, they can undergo senescence when exposed to conventional and targeted cancer therapy. Therapy-Induced Senescence (TIS) represents a fundamental response to therapy and impacts its outcomes. However, TIS has been considered a positive therapeutic goal since senescent tumor cells are expected to enter a state of permanent growth abrogation. In this work we examined the hypothesis that a subpopulation of senescent cells can re-acquire proliferative potential after a state of senescent dormancy, indicating that senescence is not obligatorily an irreversible process. Our observations indicate that H460 non-small cell lung cancer cells induced into senescence by exposure to etoposide, and enriched based on β-galactosidase staining and size, were shown to recover reproductive capacity, which was accompanied by resolution of the DNA-damage-response (downregulation of p53 and p21Cip1 induction), attenuation of the Senescence-associated Secretory Phenotype (SASP). To overcome the reservation that the newly dividing cells may not have been derived from the senescent population and in an effort to establish that escape from TIS is feasible, tumor cells induced into senescence by chemotherapy were enriched for senescence by flow cytometry; the subsequent division of senescent cells was demonstrable utilizing both real-time, live microscopy and High Speed Live Cell Interferometry (HSLCI). Furthermore, sorted senescent cells were observed to form tumors when implanted in immune deficient mice and with a significant delay in immunecompetent mice. As chemotherapy induced senescent cells have been identified in patient tumors, it is reasonable to propose that tumor cells that escape from senescence could contribute to disease recurrence. In addition, therapy-induced senescence could prove to reflect one form of tumor dormancy. Recently, ABT263 has been used as a senolytic drug, effectively eliminating senescent cells from aging-related animal models. Here, we utilize ABT263 in a two-hit approach to eliminate senescent tumor cells that persistent after exposure to chemotherapy. ABT263 results in the killing of senescent tumor cells in a concentration-dependent manner and shifts the response towards apoptotic cell death. Furthermore, sequential administration of ABT263 interferes with the ability of senescent tumor cells to recover growth potential. These results indicate that senescent tumor cells can contribute to cancer relapse by acquiring proliferative properties and that senolytic therapy allows for the clearance of dormant senescent tumor cells and will potentially decrease cancer recurrence rates.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The study concerning the lifelong dynamic of undifferentiated cell populations present in the organism expanded the therapeutics frontiers aiming to healing several kinds of wounds and diseases. Different cell populations have been used for this purpose, as those derived from pre-implantation embryos, called embryonic stem cells (ESC); those derived from pluripotent somatic cells niches, responsible for organic regeneration throughout life, named adult stem cells (ASC); and cells derived from the bone marrow mononuclear fraction (BMNC), which has ASC and hematopoietic lineage cells at different stages of differentiation. Important barriers have been overcome aiming to elucidate benefits and risks inherent to cell therapy. However, the knowledge derives from an evolution process fraught with doubts, and there are many things to be unveiled for safe exploration of different kinds of cell therapy, mainly when allogeneic cells are used. Toxoplasmosis is an opportunistic zoonosis caused by the protozoan Toxoplasma gondii, which has a worldwide distribution and affects about a third of the human population. In the acute infection, T. gondii tachyzoites spread up and multiply themselves into host cells. Immune response leads to evasion of the protozoan in tissue cysts of (semi-latent) bradyzoites, mantaining the chronic infection of the host. Immunosuppression, as used in conditioning for allogeneic cells transplantation, is a risk factor for toxoplasmosis reactivation in chronically infected hosts. The studies presented in this thesis were performed aiming to contribute with: a. methods used for molecular detection of allogeneic cells after systemic transplantation by endovenous injection, allowing to mapping the organic distribution and the fate of these cells in recipient body; and b. understanding about interrelationships among conditioning (pharmacological immunosuppression) for transplantation, allogeneic cell therapy with BMNC and toxoplasmosis reactivation in chronically infected hosts. The study described in the Chapter 1 shows a nested-PCR test with increased sensitivity and assurance for molecular diagnosis of presence or absence of male mice cells in body fluids and tissues of female recipients after intravenous transplantation. This method, based on detection of specific DNA fragment from the Y chromosome, was effective for cells from two distinct populations: BMNC and in vitro cultured cells derived from adipose tissue. Therefore, this method can be suitable for other cell types. The study presented in the Chapter 2 shows the effects caused by an immunosuppressive pharmacological regimen with high doses of busulfan and cyclophosphamide (PCT), followed by allogeneic BMNC transplantation in a model of chronic toxoplasmosis in mice. This study was divided in three subsequent steps: I. experimental infection with type II cystogenic strain of T. gondii (ME-49) and characterization of acute and chronic toxoplasmosis; II. evaluation of the pharmacological immunosuppression effects in female Balb/c; and III. evaluation of the effects resulting from transplantation of male mice BMNC to recipient females. The pharmacological immunosuppression caused worsening of the general clinical condition of the animals and high mortality rates. This effect occurred remarkably earlier in animals chronically infected with T. gondii, showing the high risk of the combination between persistent toxoplasmosis and pharmacological immunosuppression in conditioning for allogeneic transplantation. This study showed the safety of the allogeneic BMNC transplantation in female Balb/c independently of the infection by T. gondii, provided that they have not been subjected to conditioning regimen before transplantation. In the Chapter 3, a pharmacological immunosuppressive regimen, with reduced doses of busulfan and cyclophosphamide (PCTrd), was tested as an alternative to minimize conditioning regimen inherent risks in mice chronically infected with T. gondii and submitted to allogeneic transplantation of BMNC. This study was also divided in three sequential steps: I. oral infection of female Balb/c with tissue cysts of T. gondii (cystogenic VEG strain) and monitoring over a 120 days period (chronic toxoplasmosis); II. evaluation of conditioning regimen effects using reduced doses of busulfan and cyclophosphamide (PCTrd); III. endovenous transplantation of allogeneic BMNC from male donors to the female Balb/c. The results showed the safety of the PCTrd for use in female Balb/c chronically infected with T. gondii. Reduced doses of busulfan and cyclophosphamide (PCTrd) resulted in lower incidence and intensity of clinical signs and absence of mortality, in contrast to results that were observed with high doses (PCT) recommended for myeloablation. Transplantation of allogeneic bone marrow-derived mononuclear cells from male donors was safe in the short term for female Balb/c mice submitted to PCTrd regardless of persistent infection by T. gondii. / O estudo da dinâmica das populações celulares indiferenciadas, presentes no organismo ao longo da vida, ampliou a fronteira de alternativas para o tratamento de vários tipos de lesões e de doenças. Diversas populações celulares têm sido usadas para isso, como as derivadas de embriões em fase de pré-implantação, denominadas células-tronco embrionárias (CTE); as derivadas de nichos de células somáticas pluripotentes, responsáveis pela regeneração dos tecidos orgânicos ao longo da vida, chamadas de células-tronco adultas (CTA); e as células da fração mononuclear da medula óssea (CFMO), que conta com CTA e células da linhagem hematopoiética em variados estágios de diferenciação. Importantes obstáculos têm sido superados no que se refere ao conhecimento de benefícios e de riscos inerentes à terapia celular. Contudo, a construção do conhecimento é permeada de dúvidas e há muito a ser desvendado para que se possam explorar com segurança as potencialidades das terapias celulares, especialmente usando células alogênicas. A toxoplasmose é uma zoonose oportunista causada pelo protozoário Toxoplasma gondii, que apresenta distribuição mundial e acomete cerca de um terço da população humana. Na fase aguda da infecção, taquizoítos de T. gondii disseminam-se e multiplicam-se nas células hospedeiras. A resposta imune induz a evasão do protozoário em cistos teciduais de bradizoítos (forma de semilatência), mantendo o hospedeiro cronicamente infectado. A imunossupressão, como a utilizada no pré-condicionamento para transplante de células alogênicas, é um fator de risco de reativação da toxoplasmose em portadores crônicos. Na presente tese, são apresentados estudos realizados com objetivo de contribuir com: a. métodos de detecção molecular de células alogênicas após o transplante sistêmico pela via endovenosa, permitindo o mapeamento da distribuição e do destino destas células no organismo receptor; e b. compreensão das inter-relações envolvendo pré-condicionamento (imunossupressão farmacológica) para o transplante, terapia celular alogênica com CFMO e reativação da toxoplasmose em portadores crônicos. No Capítulo 1 demonstra-se uma nested-PCR que aumenta a sensibilidade e a segurança do diagnóstico molecular de presença ou ausência de células de camundongos machos em fluidos e tecidos de receptoras fêmeas após transplante endovenoso. O método, que envolve a detecção de fragmento de DNA específico do cromossomo Y, foi eficaz para células de duas populações distintas: CFMO e células cultivadas in vitro derivadas do tecido adiposo. Portanto, o método pode ser útil para outros tipos celulares. O estudo apresentado no Capítulo 2 demonstra os efeitos de um protocolo de imunossupressão farmacológica com altas doses de busulfan e ciclofosfamida (PCT), seguido do transplante alogênico de CFMO em um modelo de toxoplasmose crônica em camundongos. Esse estudo foi dividido em três etapas subsequentes: I. infecção experimental com cepa cistogênica do tipo II de T. gondii (ME-49) e caracterização da toxoplasmose aguda e crônica; II. avaliação dos efeitos da imunossupressão farmacológica nas fêmeas Balb/c; e III. avaliação dos efeitos do transplante de CFMO de camundongos machos para as fêmeas receptoras. A imunossupressão farmacológica provocou agravamento do quadro clínico geral dos animais e altas taxas de mortalidade. Esse efeito foi notavelmente precoce nos animais cronicamente infectados por T. gondii, demonstrando o alto risco da combinação entre toxoplasmose persistente e imunossupressão farmacológica na preparação para o transplante alogênico. Demonstrou-se a segurança do transplante alogênico de CFMO em fêmeas Balb/c infectadas ou livres de infecção por T. gondii, desde que não tenham sido submetidas ao protocolo de pré-condicionamento para o transplante. No Capítulo 3, um regime de imunossupressão farmacológica com doses reduzidas de busulfan e ciclofosfamida (PCTdr) foi testado como alternativa para minimizar os riscos inerentes ao pré-condicionamento em camundongos cronicamente infectados com T. gondii e submetidos ao transplante alogênico de CFMO. Esse estudo também foi dividido em três etapas sequenciais: I. infecção oral de fêmeas Balb/c com cistos teciduais de T. gondii (cepa VEG, cistogênica) e monitoramento durante um período de 120 dias (toxoplasmose crônica); II. avaliação dos efeitos do regime de pré-condicionamento, com doses reduzidas de busulfan e ciclofosfamida (PCTdr); III. transplante alogênico endovenoso de CFMO de doadores machos para as fêmeas Balb/c. Os resultados demonstraram a segurança do PCTdr para uso em Balb/c fêmeas cronicamente infectadas com T. gondii. Doses reduzidas de busulfan e ciclofosfamida (PCTdr) resultaram em menor incidência e intensidade de sinais clínicos e ausência de mortalidade, em oposição ao ocorrido com as altas doses (PCT) recomendadas para mieloablação. O transplante alogênico de células mononucleares da medula óssea de doadores machos foi seguro no curto prazo em fêmeas Balb/c submetidas ao PCTdr, independentemente de infecção persistente pelo T. gondii.

Trauma-induced coagulopathy (TIC) is a recently described condition which traditionally has been diagnosed by the common coagulation tests (CCTs) such as prothrombin time/international normalized ratio (PT/INR), activated partial thromboplastin time (aPTT), platelet count, and fibrinogen levels. The varying sensitivity and specificity of these CCTs have led trauma coagulation researchers and clinicians to use Viscoelastic Tests (VET) such as Thromboelastography (TEG) to provide Targeted Thromboelastographic Hemostatic and Adjunctive Therapy (TTHAT) in a goal directed fashion to those trauma patients in need of hemostatic resuscitation. This review describes the utility of VETs, in particular, TEG, to provide TTHAT in trauma and acquired non-trauma-induced coagulopathy.

Diese Schrift faßt in zwei Abschnitten eigene Beiträge zur Einführung der lokalisierten Magnetresonanzspektroskopie (MRS) und der funktionellen Magnetresonanztomografie (fMRT) in die diagnostische Medizin zusammen. Im ersten Teil wird beschrieben, wie die biochemischen Metabolite N-Azetylaspartat, Kreatin und Phosphokreatin, cholin-enthaltende Verbindungen und Laktat durch die Einführung der stimulierten Echo-Akquisitionsmethode (STEAM) als Lokalisationstechnik in die diagnostische Magnetresonanzspektroskopie in definierten Hirnregionen gesunder Versuchspersonen nichtinvasiv zugänglich gemacht und erstmals in Form von In-vivo-Konzentrationen quantifiziert werden konnten. Daraufhin wird gezeigt, wie die Weiterentwicklung der robusten STEAM-Technik zu kurzen Echozeiten das Signal-zu-Rauschverhältnis und damit die Messung kleinerer Untersuchungsvolumina erheblich verbesserte. Zudem wurde dadurch die Erkennung und Quantifizierung weiterer Metabolite wie z. B. des myo- und scyllo-Inosits, des Glutamats und Glutamins, des N-Azetylaspartylglutamats und der Glukose ermöglicht. Diese Methode setzte damit zusammen mit der verwendeten linearen Kombinationsmethode (LCModel) zur Konzentrationsbestimmung den spektralen Qualitätsstandard des gesamten letzten Jahrzehnts. Ferner werden die parallelen Pionierarbeiten zu Hirnerkrankungen fokaler und generalisierter Art beleuchtet. Diese Anwendungen der lokalisierten STEAM-Protonenspektroskopie in Einzelvolumentechnik zur Messung umschriebener Prozesse umfassen zerebrale Tumore und Infarkte, Plaques der multiplen Sklerose sowie andere entzündliche und degenerative Läsionen. Auch die lokalisierte STEAM-Phosphorspektroskopie und nichtzerebrale Anwendungen wie die lokalisierte Protonenspektroskopie von Faserbündeln des Skelettmuskels und der Niere bauen weitgehend auf diesem Fortschritt in der Methode auf. Zusätzlich werden Anwendungen bei generalisierten Erkrankungen gestreift, speziell angeborenen Stoffwechselerkrankungen des Kindesalters wie mitochondrialen und lysosomalen Defekten, Stoffwechselentgleisungen bei Diabetes mellitus und Leberzirrhose, psychiatrischen Erkrankungen wie der Alzheimer-Demenz. Die weitere Verbreitung dieser Erkenntnisse in die klinische Diagnostik wird entscheidend von der Beachtung des hier eingeführten Qualitätsmaßstabs und der darauf aufbauenden absoluten Metabolitquantifizierung abhängen. Der zweite Teil dieser Arbeit faßt ausgehend von funktionellen protonenspektroskopischen Untersuchungen des visuellen Kortex bei photischer Aktivierung Fortschritte zusammen, die bei der Entwicklung und Anwendung der suszeptibilitätsempfindlichen MR-Tomografie zur Messung physiologischer Hirnaktivierung mit dem Modell der visuellen Stimulation erzielt wurden. Während die Belastung des Energiestoffwechsels im angeregten striatären Kortex anhand abgesunkener Gewebespiegel von Glukose und angestiegener Laktatkonzentrationen mithilfe der zeitaufgelösten Spektroskopie beobachtet werden konnte, gelang die Demarkierung der Ausdehnung der Hirngewebeaktivierung mithilfe der T2*-gewichteten FLASH-MRT, die begleitende Verminderungen des paramagnetischen Desoxyhämoglobins im funktionell aktiven Gewebe mit Anstiegen der Bildsignalintensität wiedergibt. Schließlich werden Untersuchungen beschrieben, die die Empfindlichkeit dieses endogenen, sauerstoffspiegelabhängigen Suszeptibilitätskontrastes für die Wirkung verschiedener Medikamente bzw. pharmakologischer Stimulantien zeigen, die direkt oder indirekt über bestimmte vaskuläre Rezeptoren wirken. Diese Untersuchungen befördern wiederum ein neues Gebiet der Bildgebung, die pharmakologische MRT. / This work has two main parts that summarize pioneering contributions to localized magnetic resonance spectroscopy (MRS), functional magnetic resonance tomography (fMRI), and the introduction of these modalities into diagnostic medicine. First, it is described how biochemical metabolites such as the intracellular pools of N-acetylaspartate, creatine and phosphocreatine, choline-containing compounds, and lactate have been made accessible to noninvasive detection and to the quantification of their respective concentrations in vivo in defined cerebral regions of healthy subjects by utilizing the stimulated echo-acquisition mode (STEAM) localization technique. Then it is shown that further development of the robust STEAM technique to short echo times not only increased the signal-to-noise of the measurement, thereby providing access to smaller volumes-of-interest, but also allowed for the detection and quantification of additional metabolites such as myo- and scyllo-inositol, glutamate, glutamine, N-acetylaspartylglutamate, and glucose. Thus, together with the adoption of the linear combination method (LCModel) for concentration calculation, this method has set the standard for spectroscopic state-of-the-art in the field well over the last decade. Moreover, pioneering achievements have been highlighted with regard to applications in brain diseases of focal and generalized nature. Pertinent applications of localized single-volume STEAM proton spectroscopy to circumscribed processes include cerebral tumors, cerebral infarction, multiple sclerosis plaques, and other inflammatory and degenerative lesions. Also localized STEAM phosphorus spectroscopy and non-cerebral applications including localized proton spectroscopy of skeletal muscle and kidney largely depend on the short-echo time STEAM technique. In addition, applications in generalized disorders have been explored, which include inborn errors of metabolism in childhood, such as mitochondrial and lysosomal defects, metabolic disturbances in diabetes mellitus and liver cirrhosis, and psychiatric diseases such as Alzheimer's dementia. The further utilization of these novel methods in clinical diagnostics will heavily depend on quality measures and the mastering of a true quantification procedure as demonstrated. Second, this work summarizes achievements made in developing and applying both proton MR spectroscopy and susceptibility-sensitized MR imaging to measure physiologic brain activation using visual stimulation as a model. Whereas metabolic stress, brought upon the bioenergetic steady state in the responding striate cortex, was detected by decreased parenchymal glucose and increased lactate using time-resolved spectroscopy, mapping the extent of parenchymal activation was found to be possible by increases of image intensity in T2*-weighted FLASH MRI made sensitive to concomitant decreases of paramagnetic deoxyhemoglobin in the functionally active tissue. Finally, studies are described, which show the sensitivity of this endogenous, susceptibility-sensitive contrast, now generally known as BOLD effect, to various drugs or pharmacologic stimuli acting either directly or indirectly on vascular receptors. These latter studies open up again a new field of imaging, dubbed pharmacologic MRI.

This research used a focused ion beam in order to fabricate record small nano-magnetic structures, investigate the properties of magnetic materials in the rarely studied range of nanometer size, and exploit their extraordinary characteristics in medicine and nano-electronics. This study consists of two parts: (i) Fabrication and study of record small magnetic tunnel junctions (ii) Introduction of a novel method for detection of magnetoelectric nanoparticles (MENs) in the tissue. A key challenge in further scaling of CMOS devices is being able to perform non-volatile logic with near zero power consumption. Sub-10-nm nanomagnetic spin transfer torque (STT) magnetic tunneling junctions (MTJs) have the potential for a universal memory that can address this key challenge. The main problem is to decrease the switching current density. This research studied these structures in sub-10-nm size range. In this range, spin related excitations consume considerably smaller amounts of energy as compared to the larger scale. This research concluded that as predicted a decrease in switching current superior to that of the linear scaling will happen in this size range. Magneto-electric nanoparticles (MENs) can be used to directly couple intrinsic electric-field-driven processes with external magnetic fields for controlling neural activity deep in the brain. These particles have been proven to be capable of inducing deep brain stimulation non-invasively. Furthermore, these magneto-electric nano-particles can be used for targeted drug delivery and are contenders to replace conventional chemotherapy. The circulatory system can deliver a drug to almost every cell in the body; however, delivering the drug specifically into the tumor cell and then releasing it on demand remains a formidable task. Nanomedicine can accomplish this, but ensuring that the drug is released at an appropriate rate once at the target site is an important task. In order to have a complete understanding of the behavior of these MENs when injected into the body, a comprehensive bio-distribution study was performed. This study introduced a novel spectroscopy method for tracing the nanoparticles in the bloodstream. This study investigated the post injection distribution of the MENs in vital organs throughout a period of two months.

Les procédures impliquant des aiguilles sont la plus importante source de douleur chez les enfants. Considérant qu’il est impossible d’éliminer complètement la douleur ressentie lors de procédures douloureuses, il est de la responsabilité professionnelle de l’infirmière d’assurer sa gestion à l’aide d’interventions non-pharmacologiques et/ou pharmacologiques. Les interventions actuellement disponibles pour le soulagement de la douleur procédurale des enfants nécessitent beaucoup de temps ou de personnel, ce qui représente des barrières à leur implantation dans les services d’urgence. Par conséquent, l’utilisation d’une intervention non-pharmacologique rapide et facile d’utilisation pourrait permettre de pallier ces contraintes. À cet effet, un dispositif combinant le froid et la vibration (Buzzy) a été développé dans l’objectif d’optimiser le soulagement de la douleur procédurale pédiatrique. Ce dernier repose sur deux mécanismes permettant la modulation du message douloureux, soit la théorie du portillon et les contrôles inhibiteurs nociceptif diffus. Le dispositif Buzzy semble donc être une avenue prometteuse pour surmonter les obstacles reliés à la gestion de la douleur lors de procédures impliquant des aiguilles dans les services d’urgence. Le but principal de cette étude était de déterminer si un dispositif combinant le froid et la vibration (dispositif Buzzy) était considéré comme étant non-inférieur à un anesthésique topique (lidocaïne liposomale 4%) pour la gestion de la douleur d’enfants lors de procédures impliquant des aiguilles dans les services d’urgence. Cet essai clinique randomisé de non-infériorité parallèle à deux groupes a été mené dans le service d’urgence d’un centre hospitalier universitaire pédiatrique (CHU Sainte-Justine, Montréal, Canada). Un total de 352 enfants âgés entre 4 et 17 ans a été recruté et 346 ont été randomisés, soit au groupe expérimental (dispositif Buzzy=172) ou au groupe contrôle (anesthésique topique=174). Pour les participants ayant été assignés au groupe expérimental, le dispositif Buzzy a été appliqué à environ 5 cm au-dessus du site d’insertion et a été maintenu en place tout au long de la procédure. Pour les participants ayant été assignés au groupe contrôle, un anesthésique topique (lidocaïne liposomale 4%) a été appliqué au niveau du site d’insertion 30 minutes avant la procédure. Selon l’analyse en intention de traiter, la différence moyenne de douleur procédurale entre les deux groupes à l’étude (dispositif Buzzy : 3,92  3,13; anesthésique topique : 3,27  3,02) était de 0,64 (95%IC -0,10 à 1,26) sur l’échelle de mesure Color Analogue Scale (0-10). Ceci démontre donc que le dispositif Buzzy n’est pas considéré comme étant non-inférieur à l’anesthésique topique. En ce qui concerne la détresse procédurale, aucune différence statistiquement significative n’a été décelée avec les deux échelles de mesures utilisées (Procedure Behavior Check List : p=0,104 ; Children’s Fear Scale : p=0,421). Il n’y avait également aucune différence statistiquement significative en ce qui concerne le succès de la procédure au premier essai (p=0,489) ainsi que la mémoire de la douleur 24 heures après la procédure (p=0,346). De plus, aucun effet secondaire n’a été enregistré pour les participants ayant utilisé le dispositif Buzzy et seulement un évènement est survenu chez un participant ayant reçu l’anesthésique topique. Enfin, les parents des deux groupes ont rapporté être satisfaits avec l’intervention reçu par leur enfant (p=0,257) et la majorité des infirmières (65,0%) ont manifesté une préférence pour le dispositif Buzzy. Le dispositif Buzzy n’est pas considéré comme étant non-inférieur à un anesthésique topique pour la gestion de la douleur d’enfants lors de procédures impliquant des aiguilles. Toutefois, considérant que les anesthésiques topiques sont sous-utilisés dans les services d’urgence, le dispositif Buzzy représente une alternative intéressante à cette méthode pharmacologique nécessitant un temps d’application. Par conséquent, le dispositif Buzzy pourrait être davantage transférable à la pratique et les cliniciens pourraient possiblement l’intégrer plus facilement dans leur routine quotidienne. / Needle-related procedures are considered as the most important source of pain in children in hospital settings. It is now recognized that even minor procedures can result in physiological, psychological and emotional consequences. For these reasons, management of children’s pain and distress through pharmacological and/or non-pharmacological intervention is essential. Healthcare professionals working in the Emergency Department (ED) setting face particular challenges regarding procedural pain management related to their work environment. Time constraints, heavy workload, and busy environment represent barriers to the use of available interventions for pain management during needle-related procedures. Therefore, the use of a rapid, easy-to-use intervention could improve procedural pain management practices in the context of the ED. As such, the Buzzy device, which is a bee-shaped device combining vibration and cold, seems to be a promising alternative. The theoretical bases of this device are the Gate Control Theory and the diffuse noxious inhibitory controls, both generating the modulation of pain. The primary objective of this study was to determine if a device combining cold and vibration (Buzzy device) was considered as non-inferior (no worse) to a topical anesthetic (liposomal lidocaine 4% cream) for pain management in children undergoing needle-related procedures in the ED. This randomized, controlled, non-inferiority trial with two parallel groups was conducted in the ED of a university pediatric tertiary hospital center (CHU Sainte-Justine, Montreal, Canada). A total of 352 participants were enrolled and 346 were randomized to either experimental (Buzzy device=172) or control (topical anesthetic=174) groups. For the participants allocated to the experimental, the Buzzy device was applied 5 cm above the insertion site group just before the needle-related procedure and was maintained in place throughout the procedure. For the participants allocated to the control group, a topical anesthetic cream (liposomal lidocaine 4%) was applied at the insertion site 30 minute before the needle-related procedure. Using an intention-to-treat analysis, the mean difference in procedural pain scores between the experimental group (3,92  3,13) and the control group (3,27  3,02) was 0.64 (95%CI -0,10 to 1,26) using the Color Analogue Scale, showing that the Buzzy device was not non-inferior to the topical anesthetic. Regarding the procedural distress, no significant difference was found between groups using the both measuring scales (Procedure Behavior Check List: p=0,104; Children’s Fear Scale: p=0,421). In addition, no significant difference was also found between groups regarding the success of the procedure at first attempt (p=0,489) and the memory for pain 24 hours after the procedure (p=0,346). Parents of both groups were satisfied with the received interventions (p=0,257) and the majority of nurses (65,0%) preferred the Buzzy device over the topical anesthetic. No adverse events occurred in the cold and vibrating group and one adverse event was reported in the topical anesthetic cream group. The non-inferiority of the cold and vibrating device over a topical anesthetic was not demonstrated for pain management of children during needle-related procedure in the ED. However, considering that topical anesthetics are underused in the ED setting, the Buzzy device seems to be a promising alternative as it is a rapid, low-cost, easy-to-use and reusable intervention. Consequently, it could be more likely to be translated into clinical practice and adopted by clinicians for routine use.

Excessive alcohol consumption is one of the most important lifestyle factors affecting the disease burden in the Western world. The results of treatment in daily practice are modest at best. The aim of the RESCueH programme is to develop and evaluate methods, which are as practice-near as possible, and therefore can be implemented quickly and easily in everyday clinical practice. It is the first clinical alcohol programme to be transatlantic in scope, with implementation in treatment centers located in Denmark, Germany and the US. The RESCueH programme comprises 5 randomized controlled trials, and the studies can be expected to result in (1) more patients starting treatment in specialized outpatient clinics, (2) a greater number of elderly patients being treated, (3) increased patient motivation for treatment and thus improved adherence, (4) more patients with stable positive outcomes after treatment and (5) fewer patients relapsing into harmful drinking. The aim of this paper is to discuss the rationale for the RESCueH programme, to present the studies and expected results.

Background: Up to 55% of patients administered ketamine, experience an emergence phenomena (EP) that closely mimics schizophrenia and increases their risk of injury. While genetics accounts for about 50% of severe adverse drug reactions, no studies have investigated genetic association of ketamine-induced EP in healthy patients. Ketamine is metabolized by CYP 2B6 enzymes and CYP 2B^8^ allele significantly alter ketamine metabolism. In addition, ketamine exerts most of its effects by inhibiting the N-methyl-D-aspartate receptor (NMADR), and NMDAR genes (GRIN2B) are associated with learning and memory impairment and schizophrenia. Purpose: To investigate the relationship between CYP2B6*6 and GRIN2B single nucleotide polymorphisms (SNPs) and ketamine-induced emergence phenomena (EP). Methods: This cross-sectional pharmacogenetic study recruited 75 patients having minor orthopedic, hand, foot, anorectal surgeries from two outpatient surgical centers. EP was measured with the Clinician Administered Dissociative State Scale (CADSS). DNA was genotyped using standard Taqman assays and protocols. Genetic association of CYP2B6*6 and GRIN2B (rs1019385 & rs1806191) SNPs and ketamine induced EP occurrence and severity were tested using multivariate logistic and linear regression, adjusting for age, ketamine dose, duration of anesthesia, and time since ketamine administration. Results: Forty-seven patients (63%) received ketamine and were genotyped. Nineteen EP cases were identified (CADSS > 4), leaving 28 non-EP controls. For our population, CADSS has an internal consistency reliability Cronbach’s alpha of 0.82, and could reliably distinguish ketamine from non-ketamine cases. Occurrence and severity of EP were not associated with CYP2B6*6 or GRIN2B (p > 0.1). Models removing genotype and containing age, ketamine dose, duration of v anesthesia, and time since ketamine administration significantly predicted EP occurrence (p = 0.001) and severity (p = 0.007). Presence and severity of EP did not affect patient satisfaction with care. Discussion: Younger age, higher dose and longer duration of anesthesia significantly predicted EP occurrence and severity among our sample. This study provides effect size estimates useful for the design of adequately powered future genetic association studies. The feasibility of recruitment from patients undergoing elective, outpatient surgeries and ease of post-operative EP assessment with CADSS supports our approach. However, the small sample size may have limited about ability to determine significant differences. Conclusion: Fully powered studies are needed to investigate this important phenomena. Determining factors for anesthesia-related EP symptoms may reduce risks and costs associated with this adverse medication effect.