The percutaneous biopsy is a critical intervention for diagnosis and staging in cancer therapy. Robotic systems can improve the efficiency and outcome of such procedures while alleviating stress for physicians and patients. However, the high complexity of operation and the limited possibilities for robotic integration in the operating room (OR) decrease user acceptance and the number of deployed robots. Collaborative systems and standardized device communication may provide approaches to overcome named problems. Derived from the IEEE 11073 SDC standard terminology of medical device systems, we designed and validated a medical robotic device system (MERODES) to access and control a collaborative setup of two KUKA robots for ultrasound-guided needle insertions. The system is based on a novel standard for service-oriented device connectivity and utilizes collaborative principles to enhance user experience. Implementing separated workflow applications allows for a flexible system setup and configuration. The system was validated in three separate test scenarios to measure accuracies for 1) co-registration, 2) needle target planning in a water bath and 3) in an abdominal phantom. The co-registration accuracy averaged 0.94 ± 0.42 mm. The positioning errors ranged from 0.86 ± 0.42 to 1.19 ± 0.70 mm in the water bath setup and from 1.69 ± 0.92 to 1.96 ± 0.86 mm in the phantom. The presented results serve as a proof-of-concept and add to the current state of the art to alleviate system deployment and fast configuration for percutaneous robotic interventions.

Raman spectroscopy is a powerful tool in the field of biomedicine for disease diagnosis owing to its potential to provide the molecular fingerprint of biological samples. However due to the inherent weak nature of the Raman process, there is a constant quest for enhancing the sensitivity of this technique for enhanced diagnostic efficiency. This thesis focuses on achieving this goal by integrating advanced methods with Raman spectroscopy. Firstly this thesis explores the applicability of a laser based fluorescence suppression technique – Wavelength Modulated Raman Spectroscopy (WMRS) - for suppressing the broad luminescence background which often obscure the Raman peaks. The WMRS technique was optimized for its applications in single cell studies and tissue studies for enhanced sensing without compromising the throughput. It has been demonstrated that the optimized parameter would help to chemically profile single cell within 6 s. A two fold enhancement in SNR of Raman bands was demonstrated when WMRS was implemented in fiber Raman based systems for tissue analysis. The suitability of WMRS on highly sensitive single molecule detection techniques such as Surface Enhanced Raman Spectroscopy (SERS) and Surface Enhanced Resonance Raman Spectroscopy (SERRS) was also explored. Further this optimized technique was successfully used to address an important biological problem in the field of immunology. This involved label-free identification of major immune cell subsets from human blood. Later part of this thesis explores a multimodal approach where Raman spectroscopy was combined with Optical Coherence Tomography (OCT) for enhanced diagnostic sensitivity (>10%). This approach was used to successfully discriminate between ex-vivo adenocarcinoma tissues and normal colon tissues. Finally this thesis explores the design and implementation of a specialized fiber Raman probe that is compatible with surgical environments. This probe was originally developed to be compatible with Magnetic Resonance Imaging (MRI) environment. It has the potential to be used for performing minimally invasive optical biopsy during interventional MRI procedures.

Le muscle strié squelettique est un tissu richement vascularisé. Au delà de l'apport en oxygène et en nutriments, de nouvelles fonctions des vaisseaux ont été récemment identifiées, par le biais des interactions établies entre les cellules du vaisseau (cellules endothéliales) et les cellules du muscle, en particulier les cellules souches musculaires (cellules satellites). Celles-ci interagissent étroitement avec les cellules endothéliales pour leur expansion et leur différenciation, puis avec les cellules péri-endothéliales pour leur auto-renouvellement et leur retour à la quiescence. Les vaisseaux participent ainsi au contrôle de l’homéostasie du muscle squelettique. Grâce à ces interactions, les cellules vasculaires jouent donc un rôle central dans le remodelage tissulaire après un phénomène destructif, survenant par exemple au cours d’un trauma ou d’une myopathie. Pour étudier, les mécanismes de la plasticité vasculaire au cours du remodelage tissulaire, deux situations paradigmatiques de muscle en régénération chez l’enfant : la dermatomyosite juvénile (DMJ) et la dystrophie musculaire de Duchenne (DMD) ont été étudiées. Il existe, dans ces deux pathologies une souffrance musculaire associée à des cycles de nécrose/régénération. Elles se différencient par leur plasticité vasculaire et par leur évolution. En effet, la DMJ, la myopathie inflammatoire la plus fréquente de l’enfant est caractérisée par une vasculopathie avec perte en capillaires. L’évolution peut être favorable avec restitution ad integrum du muscle. La DMD est une myopathie génétique conduisant à une dégradation progressive de la force musculaire associée à une néovascularisation compensatrice. Le volet clinique/histologique incluant une analyse multiparamétrique des critères évolutifs cliniques et de réponse thérapeutique couplée à une réévaluation des données histologiques de la DMJ (analyse morphométrique des muscles DMJ) a permis de montrer qu’il existait des sous groupes phénotypiques homogènes de sévérité différente dans la DMJ. Le degré de sévérité clinique est relié à la gravité de la vasculopathie musculaire Par ailleurs, des marqueurs cliniques et histologiques simples permettant de repérer au diagnostic les patients nécessitant une escalade thérapeutique rapide (CMAS>34, atteinte gastrointestinale, fibrose endomysiale musculaire au diagnostic) ont été identifiés. Le volet cellulaire a permis l’identification in vitro des interactions cellulaires spécifiques et différentielles des myoblastes issues de patients DMD et DMJ sur les cellules endothéliales normales par l’analyse de leur rôle sur la prolifération, migration et différenciation des cellules vasculaires. Dans la DMD, les myoblastes entrainent une réponse angiogénique importante mais non efficace (néovascularisation anarchique). Dans la DMJ, les myoblastes participent efficacement à la reconstruction vasculaire notamment via la sécrétion de facteurs proangiogéniques. Ces résultats ont été renforcés par analyse transcriptomique effectuée à partir de cellules endothéliales et satellites isolées de muscles de patients confirmant le rôle central de la vasculopathie associée à un contexte inflammatoire spécifique lié à l’interféron dans la physiopathologie de la DMJ et montrant dans la DMD une dérégulation de l’homéostasie normale des interactions vaisseau-muscle avec mise en jeu d’un remodelage tissulaire non efficace. Ces données permettent d'identifier de nouvelles fonctions des cellules vasculaires dans le remodelage du muscle strié squelettique au cours des pathologies musculaires de l'enfant, et devraient ouvrir la voie à de nouvelles approches thérapeutiques. / Skeletal muscle is highly vascularised. Beyond oxygen and nutriment supply, new functions for vessels have been recently identified, through the interactions that vessel cells (endothelial cells) establish with muscle cells, particularly with muscle stem cells (satellite cells). These latter closely interact with endothelial cells for their expansion and their differentiation, then with periendothelial cells for their self-renewal and return to quiescence. During skeletal muscle regeneration endothelial cells reciprocally interact with myogenic cells by direct contact or by releasing soluble factors to promote both myogenesis and angiogenesis processes. Skeletal muscle regeneration typically occurs as a result of a trauma or disease, such as congenital or myopathies. To better understand the role of vessel plasticity in tissue remodeling, we took advantage of two muscular disorders that could be considered as paradigmatic situations of regenerating skeletal muscle in the child: Juvenile Dermatomyositis (JDM), the most frequent inflammatory myopathy and Duchenne Muscular Dystrophy (DMD), the most common type of muscular dystrophy. Although these two muscular disorders share, at the tissue level, similar mechanisms of necrosis-inflammation, they differ regarding the vessel domain. In JDM patients, microvascular changes consist in a destruction of endothelial cells assessed by focal capillary loss. This capillary bed destruction is transient. The tissue remodeling is efficient and muscle may progressively recover its function. By contrast, in DMD, despite an increase of vessels density in an attempt to improve the muscle perfusion, the muscle function progressively alters with age. We identified clinical and pathological markers of severity and predictive factors for poor clinical outcome in JDM by computing a comprehensive initial and follow-up clinical data set with deltoid muscle biopsy alterations controlled by age-based analysis of the deltoid muscle capillarization. We demonstrated that JDM can be divided into two distinctive clinical subgroups. The severe clinical presentation and outcome are linked to vasculopathy. Furthermore, a set of simple predictors (CMAS<34, gastrointestinal involvement, muscle endomysial fibrosis at disease onset) allow early recognition of patients needing rapid therapeutic escalation with more potent drugs. We studied in vitro the specific cell interactions between myogenic cells issued from JDM and DMD patients and normal endothelial cells to explore whether myogenic cells participate to the vessel remodeling observed in the two pathologies. We demonstrated that MPCs possessed angiogenic properties depending on the pathological environment. In DMD, MPCs promoted the development of establishment of an anarchic, although strong, EC stimulation, leading to the formation of weakly functional vessels. In JDM, MPCs enhanced the vessel reconstruction via the secretion of proangiogenic factors. This functional analysis was supported by the transcriptomic analysis consistent with a central vasculopathy in JDM including a strong and specific response to an inflammatory environment. On the contrary, DMD cells presented an unbalanced homeostasis with deregulation of several processes including muscle and vessel development with attempts to recover neuromuscular system by MPCs. To summarize, our data should allow the definition of new functions of vessel cells in skeletal muscle remodelling during muscle pathologies of the child that will open the way to explore new therapeutic options and to gain further insights in the pathogenesis of these diseases.

Bildgesteuerte, perkutane Interventionen stellen bei vielen diagnostischen und therapeutischen Fragestellungen eine Alternative zum chirurgischen Vorgehen dar. Hierbei kommen bevorzugt die Sonographie und die Computertomographie (CT) zum Einsatz. Zu den Indikationen für eine gezielte Nutzung der Magnetresonanztomographie (MRT) zählen Befunde, die sich mit anderen Modalitäten nicht ausreichend darstellen lassen, die fehlende Strahlenexposition (CT) sowie Alleinstellungsmerkmale wie der hervorragende native Weichteilkontrast oder die Möglichkeiten zur Darstellung von Temperaturen oder Diffusionsprozessen. Zu den Nachteilen zählen die langen Messzeiten, das starke Magnetfeld sowie die räumliche Enge in den meist röhrenförmigen Geräten, die ein interventionelles Vorgehen oft erschweren. Stereotaktische Führungs- und Navigationshilfen sind kein notwendiger Bestandteil der interventionellen Ausrüstung, ermöglichen jedoch oft eine gezieltere Planung, bessere Visualisierung oder vereinfachte Durchführung, insbesondere gegenüber einer rein kognitiven Einbeziehung der MRT-Informationen. Assistenzsysteme für geschlossene MRT-Geräte sind meist rahmenbasiert und beschränken sich auf bestimmte Regionen, z. B. die Mamma, die Prostata oder das muskuloskelettale System. Diese Arbeit beschreibt hingegen eine leistungsstarke rahmenlose Assistenztechnik (Navigation), die sich praktisch in beliebigen Körperregionen einsetzen lässt. Der Operateur orientiert sich dabei anhand von hochwertigen MRT-Ansichten, die gemäß der in Echtzeit erfassten Nadellage aus einem kurz zuvor erhobenen Referenzdatensatz reformatiert werden. Ausgehend von der Implementierung an einem speziellen offenen MRT-System (0,5 T) werden interventionelle Komponenten und Methoden beschrieben, die erfolgreich auf ein herkömmliches MRT-System (1,5 T) übertragen wurden. Die Einschränkungen des geschlossenen Systems führten dabei zu einer speziellen Registrierungstechnik mit Hilfe einer kompakten, frei positionierbaren Referenzplatte mit resonanten Miniatur-Hochfrequenzspulen (semiaktiv) als MR-Positionsmarker. Im Vordergrund stand die systematische Prüfung der Marker hinsichtlich Signalverhalten und Sicherheit sowie die Zuverlässigkeit und Genauigkeit einer vollautomatischen, bildbasierten 3D-Lokalisation unter experimentellen und klinischen Randbedingungen. Gegenüber herkömmlichen, passiven (Kontrastmittel-) Markern zeichnet sich die semiaktive Technik dadurch aus, dass sie gleichzeitig, auch mehrere, beliebig über das gesamte Messvolumen verteilte Marker, praktisch unabhängig von sämtlichen anatomischen Strukturen lokalisieren kann. Sowohl die Festlegung einer Position (ein Marker) oder einer Ebene (drei Marker) wie auch die navigierte Platzierung einer Nadel zeigten im Experiment ausreichend hohe Genauigkeiten. Auf Basis einer zeitlich optimierten (Subsekunden-) Markerbildgebung konnte experimentell eine robotisch geführte Nadel direkt im MRT bildgebend verfolgt werden, was weitere Anwendungen der Lokalisationstechnik in Aussicht stellt. Navigierte Biopsien an einem Gewebephantom zeigten nach ausschließlich stereotaktischer Positionierung – ohne Kontrollbildgebung – unabhängig vom Erfahrungsgrad der medizinischen Anwender ausreichend hohe Trefferquoten. Gleichzeitig lieferte die Studie wertvolle, auch anwenderspezifische Erkenntnisse über die Bedienbarkeit sowie den Zeitbedarf für einzelne Interventionsschritte. Im Vergleich mit anderen prototypischen oder kommerziellen Systemen zeigte sich die vorgestellte Assistenztechnik – am Beispiel muskuloskelettaler Interventionen – als klinisch flexibel einsetzbar.

Prostate cancer (PCa) is the most common non-skin malignancy of men in developed countries. In spite of treatment with curative intent up to 30-40% of patients have disease recurrence after treatment, resulting from any combination of lymphatic, hematogenous, or contiguous local spread. The concept of early detection of PCa offer benefits in terms of reduced mortality, but at the cost of over-diagnosis and overtreatment of indolent disease. This is largely due to the random nature of conventional biopsies, with a risk of missing significant cancer and randomly hitting indolent disease. In the present thesis, diagnostic performance of MRI DWI and 11C Acetate PET/CT lymph node staging of intermediate and high risk PCa, was investigated, and additionally, predictive factors of regional lymph node metastases were evaluated. Further, additional value of targeted biopsies to conventional biopsies, for detection of clinically significant PCa, was investigated. In paper one and two, 53 and 40 patients with predominantly high risk PCa underwent 11C Acetate PET/CT and 3T MRI DWI, respectively, for lymph node staging, before extended pelvic lymph node dissection (ePLND). The sensitivity and specificity for PET/CT was 38% and 96% respectively. The sensitivity and specificity for MRI DWI was 55% and 90% respectively. In paper three, 53 patients with newly diagnosed PCa were included. All patients underwent multi-parametric MRI, followed by two cognitive targeted biopsies. Five more clinically significant cancers were detected by adding targeted biopsies to conventional biopsies. In paper four the value of quantitative and qualitative MRI DWI and 11C Acetate PET/CT parameters, alone and in combination, in predicting regional lymph node metastases were examined. ADCmean in lymph nodes and T-stage on MRI were independent predictors of lymph node metastases in multiple logistic regression analysis. In conclusion the specificity of diffusion weighted MRI and 11C Acetate PET/CT for lymph node staging was high, although the sensitivity was low. Predictive factors of regional lymph node metastases could be retrieved from diffusion weighted MRI and 11C Acetate PET/CT. By combining targeted biopsies with conventional biopsies the detection rate of clinically significant PCa could be increased.

Le muscle strié squelettique est un tissu richement vascularisé. Au delà de l'apport en oxygène et en nutriments, de nouvelles fonctions des vaisseaux ont été récemment identifiées, par le biais des interactions établies entre les cellules du vaisseau (cellules endothéliales) et les cellules du muscle, en particulier les cellules souches musculaires (cellules satellites). Celles-ci interagissent étroitement avec les cellules endothéliales pour leur expansion et leur différenciation, puis avec les cellules péri-endothéliales pour leur auto-renouvellement et leur retour à la quiescence. Les vaisseaux participent ainsi au contrôle de l’homéostasie du muscle squelettique. Grâce à ces interactions, les cellules vasculaires jouent donc un rôle central dans le remodelage tissulaire après un phénomène destructif, survenant par exemple au cours d’un trauma ou d’une myopathie. Pour étudier, les mécanismes de la plasticité vasculaire au cours du remodelage tissulaire, deux situations paradigmatiques de muscle en régénération chez l’enfant : la dermatomyosite juvénile (DMJ) et la dystrophie musculaire de Duchenne (DMD) ont été étudiées. Il existe, dans ces deux pathologies une souffrance musculaire associée à des cycles de nécrose/régénération. Elles se différencient par leur plasticité vasculaire et par leur évolution. En effet, la DMJ, la myopathie inflammatoire la plus fréquente de l’enfant est caractérisée par une vasculopathie avec perte en capillaires. L’évolution peut être favorable avec restitution ad integrum du muscle. La DMD est une myopathie génétique conduisant à une dégradation progressive de la force musculaire associée à une néovascularisation compensatrice. Le volet clinique/histologique incluant une analyse multiparamétrique des critères évolutifs cliniques et de réponse thérapeutique couplée à une réévaluation des données histologiques de la DMJ (analyse morphométrique des muscles DMJ) a permis de montrer qu’il existait des sous groupes phénotypiques homogènes de sévérité différente dans la DMJ. Le degré de sévérité clinique est relié à la gravité de la vasculopathie musculaire Par ailleurs, des marqueurs cliniques et histologiques simples permettant de repérer au diagnostic les patients nécessitant une escalade thérapeutique rapide (CMAS>34, atteinte gastrointestinale, fibrose endomysiale musculaire au diagnostic) ont été identifiés. Le volet cellulaire a permis l’identification in vitro des interactions cellulaires spécifiques et différentielles des myoblastes issues de patients DMD et DMJ sur les cellules endothéliales normales par l’analyse de leur rôle sur la prolifération, migration et différenciation des cellules vasculaires. Dans la DMD, les myoblastes entrainent une réponse angiogénique importante mais non efficace (néovascularisation anarchique). Dans la DMJ, les myoblastes participent efficacement à la reconstruction vasculaire notamment via la sécrétion de facteurs proangiogéniques. Ces résultats ont été renforcés par analyse transcriptomique effectuée à partir de cellules endothéliales et satellites isolées de muscles de patients confirmant le rôle central de la vasculopathie associée à un contexte inflammatoire spécifique lié à l’interféron dans la physiopathologie de la DMJ et montrant dans la DMD une dérégulation de l’homéostasie normale des interactions vaisseau-muscle avec mise en jeu d’un remodelage tissulaire non efficace. Ces données permettent d'identifier de nouvelles fonctions des cellules vasculaires dans le remodelage du muscle strié squelettique au cours des pathologies musculaires de l'enfant, et devraient ouvrir la voie à de nouvelles approches thérapeutiques. / Skeletal muscle is highly vascularised. Beyond oxygen and nutriment supply, new functions for vessels have been recently identified, through the interactions that vessel cells (endothelial cells) establish with muscle cells, particularly with muscle stem cells (satellite cells). These latter closely interact with endothelial cells for their expansion and their differentiation, then with periendothelial cells for their self-renewal and return to quiescence. During skeletal muscle regeneration endothelial cells reciprocally interact with myogenic cells by direct contact or by releasing soluble factors to promote both myogenesis and angiogenesis processes. Skeletal muscle regeneration typically occurs as a result of a trauma or disease, such as congenital or myopathies. To better understand the role of vessel plasticity in tissue remodeling, we took advantage of two muscular disorders that could be considered as paradigmatic situations of regenerating skeletal muscle in the child: Juvenile Dermatomyositis (JDM), the most frequent inflammatory myopathy and Duchenne Muscular Dystrophy (DMD), the most common type of muscular dystrophy. Although these two muscular disorders share, at the tissue level, similar mechanisms of necrosis-inflammation, they differ regarding the vessel domain. In JDM patients, microvascular changes consist in a destruction of endothelial cells assessed by focal capillary loss. This capillary bed destruction is transient. The tissue remodeling is efficient and muscle may progressively recover its function. By contrast, in DMD, despite an increase of vessels density in an attempt to improve the muscle perfusion, the muscle function progressively alters with age. We identified clinical and pathological markers of severity and predictive factors for poor clinical outcome in JDM by computing a comprehensive initial and follow-up clinical data set with deltoid muscle biopsy alterations controlled by age-based analysis of the deltoid muscle capillarization. We demonstrated that JDM can be divided into two distinctive clinical subgroups. The severe clinical presentation and outcome are linked to vasculopathy. Furthermore, a set of simple predictors (CMAS<34, gastrointestinal involvement, muscle endomysial fibrosis at disease onset) allow early recognition of patients needing rapid therapeutic escalation with more potent drugs. We studied in vitro the specific cell interactions between myogenic cells issued from JDM and DMD patients and normal endothelial cells to explore whether myogenic cells participate to the vessel remodeling observed in the two pathologies. We demonstrated that MPCs possessed angiogenic properties depending on the pathological environment. In DMD, MPCs promoted the development of establishment of an anarchic, although strong, EC stimulation, leading to the formation of weakly functional vessels. In JDM, MPCs enhanced the vessel reconstruction via the secretion of proangiogenic factors. This functional analysis was supported by the transcriptomic analysis consistent with a central vasculopathy in JDM including a strong and specific response to an inflammatory environment. On the contrary, DMD cells presented an unbalanced homeostasis with deregulation of several processes including muscle and vessel development with attempts to recover neuromuscular system by MPCs. To summarize, our data should allow the definition of new functions of vessel cells in skeletal muscle remodelling during muscle pathologies of the child that will open the way to explore new therapeutic options and to gain further insights in the pathogenesis of these diseases.

Distúrbios do metabolismo mineral e ósseo são altamente prevalentes e considerados como causa relevante da morbidade e mortalidade dos pacientes com doença renal crônica. Diversas estratégias diagnósticas e terapêuticas têm sido estudadas nesses doentes; entretanto, pouco valor é dado ao cálcio do dialisato, apesar do impacto que possa exercer sobre o balanço de cálcio durante a hemodiálise. Os fatores determinantes da transferência de cálcio durante o procedimento são ainda controversos. Nesse estudo prospectivo, avaliamos a influência da remodelação óssea sobre o balanço de cálcio em dez pacientes dialíticos em três situações consecutivas: hiperparatireoidismo grave (Pré paratireoidectomia), durante a \"síndrome de fome óssea\" (Fome óssea) imediatamente após a paratireoidectomia e após estabilização clínica (Paratireoidectomia tardia). Durante cada fase os participantes foram submetidos a três sessões randômicas de hemodiálise com diferentes concentrações de cálcio no dialisato: 2,5; 3,0 e 3,5 mEq/L. Todos os pacientes foram submetidos à biópsia óssea para análise histomorfométrica e quantificação de proteínas ósseas no início do estudo. A transferência de cálcio variou grandemente entre os pacientes em cada fase do estudo mesmo usando o mesmo cálcio no dialisato, com valores negativos de medianas no Pré paratireoidectomia e Fome óssea (-161mg e -218mg, respectivamente) e discretamente positivo no Paratireoidectomia tardio (39mg; p < 0,05 versus Pré paratireoidectomia e Fome óssea). Análise de regressão multivariada mostrou que o gradiente de cálcio entre o cálcio iônico sérico inicial e o cálcio do dialisato, a diferença entre o cálcio iônico sérico final e o inicial e a forma não carboxilada da osteocalcina foram preditores independentes da transferencia de cálcio (R2=0.48; p < 0.05). Pelo fato da remodelação óssea também influenciar os níveis séricos de cálcio iônico e suas variações durante a diálise, nesse estudo demonstramos que o esqueleto tem papel fundamental no balanço de cálcio e essas variáveis devem ser consideradas na individualização do cálcio do dialisato de nossos pacientes / Disturbances in mineral and bone metabolism are highly prevalent and are a major cause of morbidity and mortality in chronic kidney disease patients. Different diagnostic and therapeutic strategies have been studied in these patients. However, little attention is paid to the calcium concentration in the dialysate, despite the impact it could exert over calcium balance during dialysis. The variables that determine calcium transfer during hemodialysis are still controversial. In this study, we have prospectively investigated the influence of bone remodeling on calcium balance in ten dialysis patients in three consecutive situations: severe hyperparathyroidism (Pre parathyroidectomy), during \"hungry bone syndrome\" (Hungry bone) right after surgery and after stabilization of clinical status (Late parathyroidectomy). During each phase participants were submitted to 3 random hemodialysis sessions, with different dialysate calcium: 2.5, 3.0 and 3.5 mEq/L. Bone biopsy for hystomorphometric analysis and bone proteins quantification were performed in all patients at baseline. Calcium mass transfer varied widely among patients in each study phase even using the same dialysis calcium with negative median values in Pre parathyroidectomy and Hungry Bone (-161 and -218mg, respectively) and slightly positive in Late parathyroidectomy (39mg; p<0.05 versus Pre parathyroidectomy and Hungry Bone). Multiple regression analysis showed that calcium gradient between initial serum ionic calcium and the dialysate calcium, the difference between final and initial serum ionic calcium and serum undercarboxylated form of osteocalcin were independent predictors of calcium mass transfer (R2=0.48; p<0.05). As bone remodeling also influences the serum levels of ionic calcium and its variance during dialysis, in this study we have added new data by demonstrating that the skeleton plays a key role on calcium balance and these variables must be considered when individualizing calcium dialysate for our patients

INTRODUÇÃO: O refluxo gastroesofágico (RGE) é a passagem involuntária do conteúdo gástrico para a luz do esôfago. É uma condição comum nos lactentes e crianças menores, sendo, na maioria das vezes, considerado fisiológico, no entanto, pode determinar manifestações clínicas e levar à doença do refluxo gastroesofágico (DRGE). O objetivo deste estudo foi avaliar a ultra-sonografia como meio diagnóstico da DRGE, comparando-a com a pHmetria e histopatologia nos pacientes acima de dois anos de idade. MÉTODO: Foram avaliadas 45 crianças, com idade entre dois e 14 anos, com suspeita clínica de RGE por meio da ultra-sonografia, pHmetria e endoscopia digestiva alta com biópsia. As variáveis estudadas foram a presença do refluxo gastroesofágico, a medida do ângulo de His, o comprimento do esôfago intra-abdominal, o número e tempo de duração do RGE, presença de esofagite e hérnia hiatal. Na análise estatística, calculouse a sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo, com p<0,05. Aplicou-se a regressão logística para estimar o desfecho DRGE e esofagite. RESULTADOS: A ultra-sonografia apresentou sensibilidade de 91,7%, especificidade de 61,9%, valor preditivo positivo de 73,3% e valor preditivo negativo de 86,7% para o diagnóstico da DRGE quando comparada à pHmetria. Na análise univariada o ângulo de His mostrou-se o preditor com melhor especificidade para o desfecho DRGE e esofagite, 71,4% e 72,7%, respectivamente. CONCLUSÃO: A ultrasonografia mostrou-se um exame não invasivo, de baixo custo e preciso na avaliação da junção esofagogástrica, a sua alta sensibilidade e boa especificidade quando comparada à pHmetria, permite o seu emprego na avaliação inicial da criança com suspeita de DRGE / INTRODUCTION: Gastroesophageal reflux (GER) is an involuntary passage of the gastric content to the esophagus. Most of the time, it is considered a physiologic condition as it is very common on breast feeding babies and small children, however, it may determine clinical manifestations and lead to gastroesophageal reflux disease (GERD). The aim of this study was to evaluate ultrasonography as a mean of GERD diagnosis, comparing it with pH monitoring and histopathology in patients older than 2 years of age. METHOD: 45 children aged 2 to 14 years old were evaluated, who had been clinically suspected with GER were submitted to ultrasonography, pH monitoring and upper endoscopy with biopsy. Variables for this study were the presence of gastroesophageal reflux, angle of His measurement, length of intra-abdominal esophagus, time duration and frequency of GER, presence of oesophagitis and hiatus hernia. The statistical analysis measured the sensibility, specificity, positive and negative predictive values considering p<0.05. Logistic regression was applied to estimate GERD outcome and oesophagitis. RESULTS: Ultrasonography results showed sensibility of 91.7%, specificity of 61.9%, and positive predictive value of 73.3% and negative predictive value of 86.7% for the diagnosis of GERD when compared to pH monitoring. In the single variable analysis, the angle of His showed to be the predictor with best specificity for GERD and oesophagitis outcome, 71.4% and 72.7%, respectively. CONCLUSION: Ultrasonography showed to be a harmless exam with low costs and precise in the assessment of the esophagi-gastric junction, and its high sensibility and good specificity when compared to pH monitoring allows it to be performed in the early evaluation of children suspected with GERD

Introdução: Apesar de recentes inovações e aprimoramentos no tratamento do Câncer de Bexiga (CaB) não músculo invasivo, o índice de progressão e recorrência continuam altos possivelmente devido a tumores residuais ou não evidenciados na ressecção transuretral de bexiga (RTU), o que profundamente afeta o prognóstico desta doença e evidencia a importância da qualidade desse procedimento padrão não só para o diagnóstico, mas também para o estadiamento e tratamento do tumor de bexiga. A presença de muscular própria no espécime é essencial para conduzir o tratamento, embora esta esteja presente apenas em cerca de 36-51% dos casos. Na sua ausência muitas vezes torna-se necessário um novo procedimento, vindo com isso a morbidade e os gastos de uma nova cirurgia. Dessa forma várias técnicas têm sido propostas com o intuito de melhorar a acurácia da RTUb, o que pode reduzir as ressecções incompletas e o subestadiamento. Objetivo: Avaliar a importância da biópsia de congelação do leito da lesão ressecada no que diz respeito ao estadiamento inicial e controle local da doença. Materiais e Métodos: Estudo prospectivo e randomizado dos pacientes com tumor de bexiga sem tratamento prévio que foram submetidos à RTUb no período de 09/2011 a 08/2013 em uma única instituição (Instituto do Câncer do Estado de São Paulo - ICESP). Esses pacientes foram submetidos à RTUb convencional, conforme o Guideline Europeu (EAU, 2011). No grupo 1, foi realizada biópsia de congelação do leito da lesão após a RTU, onde o cirurgião julgou esta ser possivelmente invasiva, aguardando análise do patologista quanto a presença de muscular própria, se caso esta não estivesse representada, era feita nova biópsia até sua representatividade. O grupo 2 são os controles não sendo submetidos à biópsia de congelação. Foram incluídos apenas os pacientes que tiveram critério e foram submetidos à re-RTUb. Um total de 150 pacientes foram randomizados, tornando-se elegíveis 131, sendo 64 no grupo 1 e 67 no grupo 2. Resultados: Comparando-se os grupos, não houve diferença em relação ao sexo, idade, quantidade e tamanho do tumor. No estudo anatomo-patológico da RTUb houve representatividade muscular em 100% x 58,5%, entre os grupos 1 e 2, respectivamente, com p < 0,001. Na Re-RTUb o índice de tumor residual foi 10,4% x 35,2%, entre os grupos 1 e 2, respectivamente, com p = 0,005. No grupo 1, 15 pacientes foram diagnosticados como pT2 com 100% do diagnóstico na primeira RTUb; no grupo 2, 6 pacientes tiveram diagnóstico de pT2 com apenas 33,3% na primeira RTUb, p=0,003. O tempo cirúrgico médio foi de 50 min no grupo 1 e 42min no grupo 2 (p= 0,037). Não houve diferença em relação à complicações (transfusão e perfuração vesical). Conclusão: A biópsia de congelação melhorou o correto estadiamento e controle local do câncer de bexiga, além de aumentar a acurácia do diagnóstico de doença pT2, podendo permitir o planejamento precoce do tratamento definitivo sem aumentar as complicações / Background and Purpose: Despite recent improvements of bladder cancer treatment, recurrence and progression rates are still high, possibly related to residual or overlooked tumors at the first transurethral resection (TUR), which strongly emphasizes the importance of the quality of this method. In order to improve the effectiveness of the procedure, we sought to evaluate the impact of frozen section during TUR, aiming on increasing muscular layer sample in the specimen, which may minimize incomplete resections and understaging. Patients and Methods: We prospectively included 150 consecutive patients assigned to TUR which were randomized to undergo either frozen section biopsy of the tumor bed during the TUR procedure until muscle was obtained or standard resection procedure (no frozen section). Nineteen patients were excluded after randomization, leaving 131 for analysis. All patients underwent a second TUR performed 4-6 weeks later. Frozen sections and final pathology reports were centralized and all performed by pathologists, the doubtful cases were reviewed by one uropathologist. Exclusion criteria: incomplete resection at first TUR, no criteria for second TUR according to EUA Guideline Update 2011 and previous bladder cancer treatment. (Group w/ biopsy, n = 64; Group control, n=67). Results: Both groups were comparable regarding age, gender, size and number of lesions. The majority of patients had high grade tumor in both groups. In the group where frozen section was obtained, muscle-invasive disease was higher (23% x 3%, p < 0,001). All patients in this group had muscle layer represented in the final pathology at the first TUR, while only 60% of patients in the control group (p < 0.001), including 40,5% of patients with pTa, 81,5% with pT1 e 100% with pT2 and Cis. Ninety percent of patients in the biopsy group had no residual tumor compared to 65% of the control group at second TUR (p=0,002). While all 15 patients in the frozen section group with T2 disease were diagnosed at first TUR, only 2 of 6 patients (33%) in the control group were diagnosed initially. The surgery duration was longer in the study group with mean of 50 min x 42 min (p=0,037) and there were no significant differences regarding complications (perforation and transfusion rates). Conclusion: Our results support the prove of principal that standard TUR with frozen section biopsy of bladder tumor bed increase the disease control and improve the diagnosis of T2 tumors, which may lead to reduced the number of patients in need a second TUR and avoid pT2 disease diagnosis delay, with no more complications

A cromoblastomicose é uma infecção fúngica crônica que acomete pele e tecido subcutâneo. As lesões podem ser classificadas em tumoral, verrucosa, cicatricial e do tipo placa. A resposta imune é principalmente celular e a forma grave da doença correlaciona-se com citocinas de perfil Th2. Nós exploramos populações celulares do tipo T reguladoras e Th17. Foram utilizadas vinte e três biópsias da forma verrucosa obtidas de pacientes com diagnóstico clínico e histopatológico de cromoblastomicose, sem tratamento. Foi realizado o método de imunohistoquímica para detectar Foxp3, CD25, TGF-beta, IL-6, IL-17 e IL-23. A IL-17 predominou sobre os outros marcadores, embora haja número regular de Foxp3. TGF-beta, IL-6 e IL-23 raramente foram visualizados. A constituição de uma resposta imune local com alta expressão de IL-17 e baixa expressão de outras citocinas pode ser, ao menos em parte, uma tentativa de ajudar o sistema imunológico contra infecções fúngicas. Células Foxp3 poderiam ser capazes de interferir na resposta imune eficiente contra fungos, mas também beneficiar o hospedeiro, através da capacidade de reduzir os danos do tecido que seguem uma resposta imune local. Esses elementos celulares podem contribuir para a cronicidade que caracteriza esta doença / Chromoblastomycosis is a chronic fungal infection that affects skin and subcutaneous tissue. Lesions can be classified in tumorous, verrucous, cicatricial and plaque type. The immune response is primarily cellular and the severe form of the disease correlates with a Th2 pattern of cytokines. We intended to explore the populations of regulatory T cells and the Th17 pattern. Twenty-three biopsies of verrucous form were obtained from patients with clinical and histopathological diagnostic of chromoblastomycosis, without treatment. It was performed an immunohistochemistry method to detect Foxp3, CD25, TGF-beta, IL-6, IL-17 and IL-23. IL-17 predominated over the other markers in chromoblastomycosis, although there was a regular number of Foxp3. TGF- beta, IL-6 and IL-23 were rarely visualized. The constitution of a local immune response with high expression of IL-17 and low expression of other cytokines could be at least in part, an attempt to help the immune system against fungal infection. Foxp3 cells could be able to interfere with the efficient immune response against fungi, but also benefit the host, through the ability to reduce the tissue damage that follows a local immune response. They could play a role in chronicity that characterizes this disease