This study estimates the association between dietary factors and spontaneous pancreatitis in dogs. A case-control study was conducted using 198 dogs with a clinical diagnosis of pancreatitis and 187 control dogs with a diagnosis of renal failure without clinical evidence of pancreatitis. Information on signalment, weight, body condition, dietary intake, medical history, diagnostic tests performed, concurrent diseases, treatment, length of hospital stay, and discharge status was extracted from medical records for dogs admitted to the Texas A&M University Small Animal Clinic (TAMU SAC) during January 2000 to December 2005. Information on dietary intake, signalment, weight, medical, surgical and environmental history was collected for the same dogs through a telephone questionnaire conducted from November 2006 through January 2007. Descriptive statistics were calculated, tabular analyses performed, and logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI). Based on information extracted from the medical records, ingesting unusual food (OR=4.3; CI=1.7 to 10.7), ingesting table food (OR=1.5; CI=1.0 to 2.2), or exposure to both of these dietary factors (OR=2.1; CI=1.3 to 3.2) increased the odds of pancreatitis. Collected through the telephone questionnaire, ingesting unusual food (OR=6.1; CI=2.2 to 16.5), ingesting table scraps the week before diagnosis (OR=2.2; CI=1.2 to 3.8) or regularly throughout life (OR=2.2; CI=1.2 to 4.0), and getting into the trash (OR=13.2; CI=2.1 to undefined) increased the odds of pancreatitis. Multivariable modeling estimated the associations of exposure to one or more dietary factors reported through the telephone questionnaire (OR=2.6; CI=1.4 to 5.0), overweight (OR=1.3; CI=0.7 to 2.5), year of diagnosis (OR=3.5; CI=1.9 to 6.5), neuter status (OR=3.6; CI=1.4 to 9.5), non-neuter surgery (OR=21.1; CI=3.3 to 133.9) and an interaction term between neuter status and non-neuter surgery (OR=0.1; CI=0.01 to 0.4). Dietary factors increase the odds of spontaneous pancreatitis in dogs.

Na realização da colangiopancreatografia endoscópica retrógrada a cateterização da papila duodenal maior é passo fundamental na obtenção do acesso biliar profundo e correlaciona -se com complicações biliopancreáticas das quais a pancreatite aguda pós-CPER é a mais comum. Os objetivos deste trabalho foram: a) comparar o índice de sucesso na canulação seletiva da via biliar com uso do canulótomo e canulótomo com fio guia; b) comparar, entre ambos os grupos, as dosagens séricas de amilase, lipase e proteína C reativa; c) avaliar a incidência de pancreatite nos grupos em estudo. No período de julho de 2002 a outubro de 2003 foram realizadas 341 CPER em três Instituições de nível terciário, destas foram randomizados prospectivamente e de maneira consecutiva 300 pacientes para cateterização papilar com canulótomo (Grupo I) e canulótomo com fio guia (Grupo II). Os procedimentos endoscópicos foram realizados pelo autor nas três Instituições. Procedeu-se a caracterização do perfil técnico-laboratorial e avaliação da incidência de pancreatite através de métodos clínicolaboratoriais e imagenológicos, para ambos os grupos. Todos os pacientes do estudo foram mantidos internados por 24 horas após a CPRE. A cateterização inadvertida do ducto pancreático foi semelhante para os dois grupos (p= 0,161). A fistulopapilotomia foi mais freqüente no grupo I (p= 0,011), porém apresentou significativamente menor incidência de pancreatite aguda no grupo II (p= 0,041). As dosagens séricas de amilase coletadas quatro, 12 e 24 horas após CPER foram significativamente maior no grupo I (p= 0,0087; p= 0,045; p= 0,0474; respectivamente). As dosagens séricas de lipase e proteína C-reativa após a CPER foram similares para ambos os grupos. O tempo de manipulação pancreática apresentou elevação similar nas dosagens séricas de amilase após a CPRE, porém todas as dosagens de lipase coletadas após a CPER foram significativamente maior no grupo I para a categorização de um a cinco minutos (p= 0,025; p= 0,032; p= 0,049). O número de cateterizações pancreáticas categorizadas em uma a cinco vezes apresentou elevação significativamente maior no grupo I, para as amostras de amilase, lipase e proteína C-reativa coletadas quatro, 12 e 24 horas após a CPER (amilase: p=0,006; p= 0,0023; p= 0,0095/lipase: p= 0,13; p= 0,018; p= 0,028 / PC-R: p= 0,005; p= 0,01; p= 0,01). As papilotomias realizadas no grupo II apresentaram significativamente maior elevação das dosagens séricas de amilase coletadas 12 e 24 horas após a CPER (p= 0,033; p= 0,049). As dosagens séricas de lipase e proteína C-reativa apresentaram elevações similares tanto na papilotomia como na fistulopapilotomia. A pancreatite aguda pós-CPER foi significativamente maior no grupo I (p= 0,037). Conclusões: a) O acesso biliar através do cateter com fio guia proporcionou maior índice de sucesso na canulação biliar seletiva; b) No perfil laboratorial estudado a dosagem de amilase se mostrou com diferença significante na comparação entre os grupos estudados. O mesmo não ocorreu nas dosagens de lipase e PC-R; c) O uso do fio guia foi um fator de prevenção na ocorrência da pancreatite aguda pós-CPRE / During the endoscopic retrograde cholangiopancreatography (ERCP) the main step is the cannulation of major duodenal papilla to obtain deep bile duct access, and it is correlated to pancreaticobiliary complications being acute pancreatitis the most frequent. The aims were: a) compare the rate of success to achieve selective cannulation of common bile duct using a single cannula and cannula with guide-wire; b) compare the amylase, lypase and Creactive protein serum level between the groups; c) evaluate the incidence of pancreatitis in the groups. From July 2002 to October 2003 there were performed 341 ERCP on three institutions of tertiary level. From them, 300 patients were randomized, on a prospective and consecutive fashion to major duodenal papilla cannulation using single cannula (Group I) and cannula with guide wire (Group II). The author himself performed all the endoscopic procedures on the three institutions. The characterization of technicallaboratory profile and evaluation of the incidence of pancreatitis were proceeded by clinical-laboratory and image methods to both groups. All patients were hospitalized by 24 hours after ERCP. The cannulations of pancreatic duct were similar to both groups (p=0,161). The fistulosphincterotomy was more frequent in group I (p=0,011), but group II presented significant lower incidence of acute pancreatitis (p=0,041). The amylase serum were collected 4, 12 and 24 hours after ERCP and were significantly higher in group I (p=0,0087; p=0,045; p=0.0474, respectively). The lypase and C-reactive protein after ERCP were similar to both groups. The time of pancreatic manipulation presented similar elevation of amylase serum after ERCP, therefore all lypase serum after ERCP were significantly higher in group I for the categorization of 1 to 5 minutes (p=0,025; p=0,032;p=0,049). The number of pancreatic cannulations categorized in 1 to 5 times presented significant higher elevation in group I, to the samples of amylase, lypase and C-reactive protein serum collected 4, 12 and 24 hours after ERCP (amylase: p=0,006; p=0,0023; p=0,0095/ lypase: p=0,13; p=0,018;p=0,028/ C-RP: p=0,005; p=0,01; p=0,01). The endoscopic papillotomy performed in group II presented significant higher elevation of amylase serum collected at 12 and 24 hour post ERCP (p=0,033;p=0,049). The lypase and C-reactive protein serum presented similar elevation such as in papillotomy as in fistulosphincterotomy. The acute pancreatitis post ERCP were significantly higher in group I (p=0,037). Conclusion: a) The biliar access by cannula with guide wire offered a higher success to selective biliar cannulation; b) the laboratory profile of amylase serum showed a significant difference between the groups. It did not occur with lypase and C-reactive protein serum levels; c) the use of guide wire was a preventing factor of acute pancreatitis post ERCP

Le traitement de la leucémie lymphoblastique aiguë (LLA) de l’enfant, une affection d'origine maligne des cellules progénitrices lymphoïdes, s’est considérablement amélioré au cours des dernières décennies. En effet, le taux de succès du traitement a dépassé 90% dans des conditions favorables. Cependant, des toxicités liées au traitement peuvent être fatales et entrainer l’interruption ou la cessation du traitement. L'allergie, la pancréatite et la thrombose sont des complications fréquentes du traitement de la LLA et sont associées à l'utilisation de l'asparaginase (ASNase), tandis qu’une toxicité fréquente due à la vincristine (VCR) induit la neuropathie périphérique (VIPN). Étant donné que l’ajustement du schéma posologique afin d’augmenter l'efficacité et diminuer la toxicité est un processus sensible, ceci demeure un défi majeur dans plusieurs protocoles de traitement. La pharmacogénétique étudie comment des altérations de la composante génétique peuvent influer sur la variabilité interindividuelle observée dans la réponse au traitement. Une meilleure compréhension de la base moléculaire de cette variabilité pourrait améliorer considérablement les résultats du traitement, en permettant la personnalisation de ce dernier en fonction du profil génétique du patient. Des études récentes suggèrent l’avantage d’appliquer l’analyse de l’exome à la découverte de variants associés à des traits humains complexes ainsi qu’à des phénotypes de réactions médicamenteuses. L'objectif de notre travail était d'utiliser les données de séquençage pour réaliser des études d'association à l'échelle de l'exome, y compris des étapes de filtrage et de validation, afin d'identifier de nouveaux variants génétiques susceptibles de moduler le risque de développer des complications associées à ASNase et à VIPN. Douze SNP étaient associés à des complications due à l’ASNase dans la cohorte initiale, dont 3 étaient associés à une allergie, 3 à une pancréatite et 6 à une thrombose. Parmi ceux-ci, les variants rs3809849, rs11556218 et rs34708521 des gènes MYBBP1A, IL16 et SPEF2 respectivement ont été associés à des complications multiples et leur association à une pancréatite a été répliquée dans une cohorte de validation indépendante. En ce qui concerne la VCR, trois variantes ont été associées à la modulation du risque de VIPN: rs2781377 dans SYNE2, rs10513762 dans MRPL47 et rs3803357 dans BAHD1. Nous démontrons également le puissant effet combiné de la présence de plusieurs variants de risque pour chacune des toxicités étudiées et fournissons des modèles de prédiction du risque pour la pancréatite et le VIPN basés sur la méthode d’évaluation du risque génétique pondérée et qui ont été validés à l’interne. De plus, étant donné une association du polymorphisme du gène MYBBP1A avec de multiples issus de traitement, nous avons cherché à comprendre comment cette altération génétique se traduit par des variabilités de réponse aux traitements à l’ASNase. En utilisant la technique CRISPR-CAS9 pour induire l'inactivation de gènes dans des lignées cellulaires cancéreuses PANC1 (pancréatiques) nous avons testé la différence de viabilité entre les cellules inactivées et les cellules du type sauvage à la suite de la suppression du gène et du traitement par ASNase. Nos résultats suggèrent un rôle fonctionnel de ce gène dans la modulation de la viabilité, de la capacité de prolifération et de la morphologie des cellules knock-out, ainsi que dans leur sensibilité à l'ASNase, et plaident en outre pour que le gène influence l’issus du traitement de la LLA par ASNase. Le présent travail démontre que l’utilisation de l’approche de séquençage de l’exome entier dans le contexte d’une étude d’association à l’échelle de l’exome est une stratégie valide « sans hypothèse » pour identifier de nouveaux marqueurs génétiques modulant l’effet du traitement de la LLA de l’enfant, et souligne l’importance de l'effet synergique de la combinaison des locus à risque. / Treatment of childhood acute lymphoblastic leukemia (ALL), a malignant disorder of lymphoid progenitor cells has improved significantly over the past decades and treatment success rates have surpassed 90% in favorable settings. However, treatment-related toxicities can be life-threatening and cause treatment interruption or cessation. Allergy, pancreatitis and thrombosis are common complications of ALL treatment associated with the use of asparaginase (ASNase), while vincristine-induced peripheral neuropathy (VIPN) is a frequent toxicity of vincristine (VCR). It is a sensitive process and a constant struggle to adjust the dosing regimen to ensure maximum efficacy and minimum toxicity. Pharmacogenetics studies show alterations in the genetic component between individuals can influence the observed variability in treatment response. A better understanding of the molecular basis of this variability in drug effect could significantly improve treatment outcome by allowing the personalization of ALL treatment based on the genetic profile of the patient. Emerging reports suggest the benefit of applying exome analysis to uncover variants associated with complex human traits as well as drug response phenotypes. Our objective in this work was to use available whole-exome sequencing data to perform exome-wide association studies followed by stepwise filtering and validation processes to identify novel variants with a potential to modulate the risk of developing ASNase complications and VIPN. Twelve SNPs were associated with ASNase complications in the discovery cohort including 3 associated with allergy, 3 with pancreatitis and 6 with thrombosis. Of those, rs3809849 in MYBBP1A, rs11556218 in IL16 and rs34708521 in SPEF2 genes were associated with multiple complications and their association with pancreatitis was replicated in an independent validation cohort. As for VCR, three variants were associated with modulating the risk of VIPN: rs2781377 in SYNE2, rs10513762 in MRPL47 and rs3803357 in BAHD1. We also demonstrate a strong combined effect of harbouring multiple risk variants for each of the studied toxicities, and provide internally-validated risk-prediction models based on the weighted genetic risk score method for pancreatitis and VIPN. Furthermore, given the association of the polymorphism in MYBBP1A gene with multiple treatment outcomes, we aimed at understanding how this genetic alteration translates into differences in ASNase treatment response through cell-based functional analysis. Using CRISPR-CAS9 technology we produced gene knockout of PANC1 (pancreatic) cancer cell-lines and tested the difference in viability between the knockouts and wild-type cells following gene deletion and ASNase treatment. Our results suggest a functional role of this gene in modulating the viability, proliferation capacity and the morphology of the knockout cells as well as their sensitivity to ASNase and further advocates the implication of the gene in influencing the outcome of ALL treatment with ASNase. The present work demonstrates that using whole-exome sequencing data in the context of exome-wide association study is a successful “hypothesis-free” strategy for identifying novel genetic markers modulating the effect of childhood ALL treatment and highlights the importance of the synergistic effect of combining risk loci.