Rôles des polymorphismes génétiques dans la détermination de la dose individuelle de la warfarine chez les patients traités avec de l’amiodarone

Introduction : Bien que la pratique de l’usage de la warfarine se soit améliorée au cours de la dernière décennie, aucune recommandation claire basée sur le dosage de l’amiodarone n’a été jusqu’à maintenant standardisée, ce qui représente un grand obstacle pour les cliniciens. La warfarine a un index thérapeutique étroit nécessitant un suivi régulier et un ajustement individuel de la posologie, ceci afin de déterminer la dose thérapeutique, tout en prévenant les effets secondaires qui pourraient être fatals dans certains cas. La variabilité interindividuelle de la réponse à la warfarine dépend de plusieurs facteurs, dont l’âge, le sexe, le poids, l’alimentation et l’interaction médicamenteuse, mais ceux-ci n’expliquent que partiellement les différences de sensibilité à la warfarine. Les polymorphismes des gènes CYP2C9 et VKORC1 jouent un rôle important dans la réponse à la warfarine et expliquent jusqu’à 50% de la variabilité des doses. L’utilisation d’antiarythmiques telle l’amiodarone peut accentuer considérablement l’effet de la warfarine et nécessite généralement une diminution de 30 à 50% de la dose de la warfarine. Aucune étude à ce jour n’a tenté de déterminer l’utilité du génotypage des polymorphismes des gènes CYP2C9 et VKORC1 chez les patients sous traitement combiné de warfarine et amiodarone. Objectif : Notre étude a pour objectif tout d’abord de déterminer si des facteurs génétiques influencent la première dose de stabilisation de la warfarine chez les patients en FA après l’introduction de l’amiodarone. Nous allons également tenter de confirmer l’association préalablement rapportée entre les facteurs génétiques et la première dose de stabilisation de warfarine dans notre population à l’étude. Méthodes : Un devis de cohorte rétrospective de patients qui fréquentaient la clinique d'anticoagulothérapie de l’Institut de cardiologie de Montréal entre le 1er janvier 2007 et le 29 février 2008 pour l’ajustement de leur dose selon les mesures d'INR. Au total, 1615 patients ont été recrutés pour participer à cette étude de recherche. Les critères de sélection des patients étaient les patients avec fibrillation auriculaire ou flutter, ayant un ECG documenté avec l'un de ces deux diagnostics et âgé de moins de 67 ans, en raison d’une moindre comorbidité. Les patients souffrant d’insuffisance hépatique chronique ont été écartés de l’étude. Tous les patients devaient signer un consentement éclairé pour leur participation au projet et échantillon de sang a été pri pour les tests génétiques. La collecte des données a été effectuée à partir du dossier médical du patient de l’Institut de cardiologie de Montréal. Un formulaire de collecte de données a été conçu à cet effet et les données ont ensuite été saisies dans une base de données SQL programmée par un informaticien expert dans ce domaine. La validation des données a été effectuée en plusieurs étapes pour minimiser les erreurs. Les analyses statistiques utilisant des tests de régression ont été effectuées pour déterminer l’association des variants génétiques avec la première dose de warfarine. Résultats : Nous avons identifié une association entre les polymorphismes des gènes CYP2C9 et VKORC1 et la dose de la warfarine. Les polymorphismes génétiques expliquent jusqu’à 42% de la variabilité de dose de la warfarine. Nous avons également démontré que certains polymorphismes génétiques expliquent la réduction de la dose de warfarine lorsque l’amiodarone est ajoutée à la warfarine. Conclusion : Les travaux effectués dans le cadre de ce mémoire ont permis de démontrer l’implication des gènes CYP2C9 et VKORC1 dans la réponse au traitement avec la warfarine et l’amiodarone. Les résultats obtenus permettent d’établir un profil personnalisé pour réduire les risques de toxicité, en permettant un dosage plus précis de la warfarine pour assurer un meilleur suivi des patients. Dans le futur, d’autres polymorphismes génétiques dans ces gènes pourraient être évalués pour optimiser davantage la personnalisation du traitement.

Fatores de risco para óbito precoce em crianças e em adolescentes submetidos a tranplante hepático eletivo no Hospital de Clínicas de Porto Alegre

No próximo ano, completam-se 40 anos desde a primeira tentativa de transplante hepático (TxH) em seres humanos. Há quase 20 anos, o transplante (Tx) tornou-se uma opção terapêutica real para os pacientes portadores de doença hepática terminal. Atualmente, o TxH é o tratamento de escolha para diversas enfermidades hepáticas, agudas ou crônicas. Dos transplantes realizados na Europa ou nos EUA, em torno de 12% dos pacientes são crianças e adolescentes. No Brasil, 20,9% dos pacientes transplantados de fígado em 2001 tinham até 18 anos de idade e, destes, 60,7% tinham 5 anos ou menos. O objetivo do TxH é a manutenção da vida dos pacientes com doença hepática irreversível, e a principal forma de avaliação de sucesso é a sobrevida após o Tx. A primeira semana que se segue ao TxH, apesar dos excelentes progressos dos últimos anos, continua sendo o período mais crítico. A maioria dos óbitos ou das perdas do enxerto ocorrem nas primeiras semanas, em particular, nos primeiros 7 dias de TxH. Diversos fatores de risco para o resultado do TxH podem ser identificados na literatura, porém há poucos estudos específicos do Tx pediátrico. As crianças pequenas apresentam características particulares que os diferenciam do Tx nos adultos e nas crianças maiores. Com o objetivo de identificar fatores de risco para o óbito nos 7 primeiros dias após os transplantes hepáticos eletivos realizados em 45 crianças e adolescentes no Hospital de Clínicas de Porto Alegre entre março de 1995 e agosto de 2001, foi realizado um estudo de caso-controle. Entre os 6 casos (13,3%) e os 39 controles foram comparadas características relacionadas ao receptor, ao doador e ao procedimento cirúrgico e modelos prognósticos. Das variáveis relacionadas ao receptor, o gênero, o escore Z do peso e da estatura para a idade, a atresia de vias biliares, a cirurgia abdominal prévia, a cirurgia de Kasai, a história de ascite, de peritonite bacteriana espontânea, de hemorragia digestiva e de síndrome hepatopulmonar, a albuminemia, o INR, o tempo de tromboplastina parcial ativada e o fator V não foram associados com o óbito na primeira semana. A mortalidade inicial foi maior nas crianças com menor idade (p=0,0035), peso (p=0,0062) e estatura (p<0,0001), bilirrubinemia total (BT) (p=0,0083) e bilirrubinemia não conjugada (BNC) (p=0,0024) elevadas, e colesterolemia reduzida (p=0,0385). Os receptores menores de 3 anos tiveram um risco 25,5 vezes maior de óbito que as crianças maiores (IC 95%: 1,3–487,7). A chance de óbito após o Tx dos pacientes com BT superior a 20 mg/dL e BNC maior que 6 mg/dL foi 7,8 (IC95%: 1,2–50,1) e 12,7 (IC95%: 1,3–121,7) vezes maior que daqueles com níveis inferiores, respectivamente. Das características relacionadas ao doador e ao Tx, as variáveis gênero, doador de gênero e grupo sangüíneo ABO não idênticos ao do receptor, razão peso do doador/receptor, causa do óbito do doador, enxerto reduzido, tempo em lista de espera e experiência do Programa não foram associados com o óbito nos primeiros 7 dias. Transplantes com enxertos de doadores de idade até 3 anos, ou de peso até 12 Kg representaram risco para o óbito dos receptores 6,8 (IC95%: 1,1–43,5) e 19,3 (IC95%: 1,3–281,6) vezes maior, respectivamente. O tempo de isquemia total foi em média de 2 horas maior nos transplantes dos receptores não sobreviventes (p=0,0316). Os modelos prognósticos Child-Pugh, Rodeck e UNOS não foram preditivos do óbito. Os pacientes classificados como alto risco no modelo de Malatack apresentaram razão de chances para o óbito 18,0 (IC95%: 1,2–262,7) vezes maior que aqueles com baixo risco. A mortalidade na primeira semana foi associada a valores elevados do escore PELD. O risco de óbito foi de 11,3 (IC95%: 1,2–107,0) nas crianças com valor do PELD maior que 10. As crianças pequenas e com maior disfunção hepática apresentaram maior risco de óbito precoce. Doador de pequeno porte e prolongamento do tempo de isquemia também foram associados à mortalidade. Somente os modelos de Malatack e PELD foram preditivos da sobrevida.

Investigação das alterações citogenéticas em pacientes pediátricos com leucemia linfóide aguda do Rio Grande do Norte

Leukemia is a heterogeneous group of hematologic malignancies that result from partial or total transformation of the blast cells. The Acute Lymphoblastic Leukemia (ALL) is the most common malignancy in childhood, especially in male, Caucasian children younger than 14 years. Several criteria are adopted to classify ALL, including the cell morphology, cytochemistry, immunophenotyping and cytogenetic analysis. Cytogenetic studies allow a more detailed analysis to detect chromosomal abnormalities of leukemic cells. These modifications will determine the diagnosis, classification, stage characterization, remission assessment and prognosis. In this study were evaluated 30 patients, aged from four months to seventeen years, of both sexes and various ethnicities. The age distribution showed that 67% of patients had between one and ten years (with mean age of XX years old), the most prevalent ethnic was Caucasian (50%) and 57% were males. According to immunophenotype, 93% of patients had B-cells progenitor ALL and 7% early lineage T. Considering the total studied population, the most frequent medical findings were lymphadenopathy (37%), hepatomegaly (77%) and splenomegaly (70%), where one patient could present more than one of these medical findings. Regarding the CBC, the majority of patients had hemoglobin below 10 g / dl (73%), leukocyte count less than 10.000/μL (60%) and platelet count below 150.000/μL (83%). Chromosomal abnormalities were observed in 64% of all patients, where hyperdiploidy was the most common numerical change (67%), followed by hypodiploid (33%). All these data are in agreement with the literature. Moreover, complexes structural and/or number changes not yet described in literature were observed, which indicated poor prognosis. Finally, we concluded that this study demonstrated the importance of cytogenetic study in the diagnosis and identification of prognostic factors in pediatric patients with ALL in Rio Grande do Norte. The results obtained in this study are extremely useful and emphasizes that surveys of this nature must be conducted more frequently in our state

DNA damage in patients infected by Helicobacter pylori

Helicobacter pylori (H. pylori) is considered to predispose carriers to gastric cancer but its role on gastric carcinogenesis is still unknown. The aim of this study was to investigate DNA damage by the comet assay in gastric epithelial cells from antrum and corpus in H. pylori-infected patients with gastritis of different degrees. H. pylori status, gastric histology, and DNA damage were studied in 62 H. pylori-infected and 18 non-infected patients, all of them non-smokers, nonalcoholics, and non-drug users. DNA damage was significantly higher in H. pylori-infected patients presenting gastritis than in non-infected patients with normal mucosa. A direct correlation between the levels of DNA damage and the intensity of gastritis was observed in H. pylori-infected patients. Association between DNA damage and age was also found. The levels of DNA damage were significantly higher in patients older than 50 years than in younger patients with the same degree of gastritis. Our results indicate that H. pylori infection is associated with DNA damage in gastric epithelial cells, which could be a biomarker of risk for gastric cancer in humans.

Salivary cortisol to assess the hypothalamic-pituItary-adrenal axis in healthy children under 3 years old

Objective: To establish reference concentration intervals for salivary cortisol in healthy children, in the morning and in the afternoon, investigating factors that interfere with the concentration measured and the possibility that circadian rhythms are present.Methods: A controlled observational study was carried out with 91 children aged 45 days to 36 months, selected at random and living in Santo Andre, state of São Paulo, Brazil. Inclusion criteria were: healthy, well-nourished, free from fever and corticoid use, subdivided by age group (five subsets) at 6-month intervals. Saliva was collected during home visits in the morning and afternoon. Cortisol was radioimmunoassayed with cortisol 3-oxime-bovine albumin antiserum.Results: the five subsets exhibited higher cortisol concentration during the morning than in the afternoon (p < 0.001), and this difference passed 30% from 1 year of age onwards. Mean concentrations, in nmol/L, were 557.86 (morning) and 346.36 (afternoon). A negative linear correlation was observed between morning concentrations and hours' sleep and frequency of meals (p < 0,05), and in the afternoon with anthropometric measurements (p < 0.05).Conclusions: Reference values for normal salivary cortisol in healthy children were established. At:45 days it was possible to observe circadian rhythms, which reached maturity at 12 months of life. Sleep and food deprivation increased morning cortisol levels.

Immune reconstitution in HIV-1 infected patients treated for two years with highly active antiretroviral therapy

The aim of this paper was to evaluate the immune reconstitution of HIV-1 patients subjected to highly active antiretroviral therapy (HAART) for two years or more according to CD 45RA and CD 45RO cell count; determination of IL-2, IFN-γ, IL-4, IL-10 and TNF-α serum levels; CD 4 + T and CD 8 + T lymphocyte count; and plasma viral load (VL) determination. For this purpose, a cross sectional study was carried out in the Tropical Diseases Area, Botucatu School of Medicine, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil. Between June 2001 and April 2002, 37 HIV-1 infected patients were evaluated, 13 with treatment indication but untreated (G1), 9 subjected to HAART for 5-7 months (G2), and 15 treated for two years or more (G3); both treated groups used medication regularly and without failure. Forty-nine normal individuals were studied as controls (GC-1 and GC-2). There was a tendency (p<0.10) for the predominance of two nucleoside reverse transcriptase inhibitors (NRTI) associated with one non-nucleoside reverse transcriptase inhibitor (NNRTI) regimen in G2; and two NRTI associated with a protease inhibitor (PI) in G3. Statistical differences between groups were seen for CD 45RA (G1<[G3=GC-2]; p<0.05) and CD 45RO (G1[G2=G3]; p<0.001), TNF-α serum determination ([G1>G3; G2=intermediate]>GC-1; p<0.001), IL-2 (G1<[G2=G3=GC-1]; p<0.01), IFN-γ ([G1=GC-1]>[GC-2=G3]; p<0.001), IL-4 and IL-10 ([G1=G2=G3]>GC-1; p<0.001), serum cytokine profiles, with a higher proportion of subtype 2 in G1 and mature subtype 0 in G2 and G3 (p<0.005). There was no statistical difference for CD 8 + T lymphocyte counts (G1=G2=G3; p<0.50). Consistency was seen between positive correlations of profile 1 definer cytokines (IL-2 and IFN-γ), CD 45RA and CD 45RO cells, and CD 4 + T lymphocyte counts and between positive correlations of profile 2 definer cytokines (IL-4 and IL-10) with TNF-α, and VL. The negative correlations were also consistent as they expressed the inverse of the positives. The variables with the highest number of correlations were IL-2, IFN-γ, and VL, followed by CD 45RA and CD 45RO cells, and IL-10. The variables with the lowest number of correlations were CD 4 + T and CD 8 + T lymphocytes. The results express the partial but important immune reconstitution in HIV-1 infected individuals with the interference of HAART and the importance of cytokines especially IL-2 and IFN-γ, and CD 45RA and CD 45RO cells as surrogate markers of this reconstitution.

Fluoride intake from regular and low fluoride dentifrices by 2-3-year-old children: Influence of the dentifrice flavor

This study evaluated the fluoride intake from dentifrices with different fluoride concentrations ([F]) by children aged 24-36 months, as well as the influence of the dentifrice flavor in the amount of fluoride ingested during toothbrushing. Thirty-three children were randomly divided into 3 groups, according to the [F] in the dentifrices: G-A (523 μgF/g), G-B (1,062 μgF/g) and G-C (1,373 μgF/g). Dentifrices A and B are marketed for children, while dentifrice C is a regular product. The amount of F ingested was indirectly obtained, subtracting the amount expelled and the amount left on the toothbrush from the amount initially loaded onto the brush. The results were analyzed by ANOVA, Tukey's test and linear regression analysis (p < 0.05). Children ingested around 60% of the dentifrice loaded onto the brush, but no significant differences were seen among the groups (p > 0.05). Mean daily fluoride intake from dentifrice for G-A, G-B and G-C was 0.022 a, 0.032 a and 0.061 b mg F/kg body weight, respectively (p < 0.01). There was a strong positive correlation (r = 0.86, p < 0.0001) between the amount of dentifrice used and the amount of fluoride ingested during toothbrushing. The results indicate the need for instructing children's parents and care givers to use a small amount of dentifrice (< 0.3 g) to avoid excessive ingestion of fluoride. The use of low-[F] dentifrices by children younger than 6 years also seems to be a good alternative to minimize fluoride intake. Dentifrice flavor did not influence the percentage of fluoride intake.

Componentes da síndrome metabólica preditores da capacidade aeróbica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Quantification of Biological Tissue and Construction of Patient Equivalent Phantom (Skull and Chest) for Infants (1-5 years old)

Our main purpose in this study was to quantify biological tissue in computed tomography (CT) examinations with the aim of developing a skull and a chest patient equivalent phantom (PEP), both specific to infants, aged between 1 and 5 years old. This type of phantom is widely used in the development of optimization procedures for radiographic techniques, especially in computed radiography (CR) systems. In order to classify and quantify the biological tissue, we used a computational algorithm developed in Matlab (R). The algorithm performed a histogram of each CT slice followed by a Gaussian fitting of each tissue type. The algorithm determined the mean thickness for the biological tissues (bone, soft, fat, and lung) and also converted them into the corresponding thicknesses of the simulator material (aluminum, PMMA, and air). We retrospectively analyzed 148 CT examinations of infant patients, 56 for skull exams and 92 were for chest. The results provided sufficient data to construct a phantom to simulate the infant chest and skull in the posterior anterior or anterior posterior (PA/AP) view. Both patient equivalent phantoms developed in this study can be used to assess physical variables such as noise power spectrum (NPS) and signal to noise ratio (SNR) or perform dosimetric control specific to pediatric protocols.

A six-year retrospective study of maxillofacial fractures from radiographic findings in Brazil

Objective: this study aimed to present the causes and prevalence of maxillofacial fractures that occurred in the region of Araraquara-SP – Brazil, during a 6-year period. Methods: information regarding age, gender, etiology, and maxillofacial fracture site, as well as type of radiographic examination were evaluated. Data were gathered from the radiographic examination and radiographic report of the Division of Dentomaxillofacial Radiology, and from charts of the Division of Oral and Maxillofacial Surgery of patients attending for maxillofacial fractures from 2004 to 2009. A descriptive statistical analysis was developed using Epi-Info 3.5.1. Results: from a total of 11,728 patients assisted, 407 patients presented maxillofacial fractures. The prevalent age ranged from 21 to 30 years old. From the total patients, 322 were men and 85 were women (ratio of men to women was 3.8:1). Panoramic radiography (n = 306) was most frequently used to observe maxillofacial fractures. Mandibular fractures were frequent in the body of the mandible (n = 127), followed by symphysis (n = 102), and the prevalent cause was traffic accidents (n = 161). Conclusion: more than half of affected individuals were younger than 40 years of age. The body of the mandible was the most common fracture location visualized by panoramic radiography for traffic accidents and fights.

The effect of chlorhexidine on plaque index and mutans streptococci in orthodontic patients: A pilot study

Aim: The purpose of this study was to assess chlorhexidine effects on plaque index and salivary levels of mutans streptococci (MS) when used as the immersion solution for removable orthodontic appliances and added to their acrylic resin composition. Methods: Forty-five patients (6 to 12 years old) were randomly assigned into three groups with 15 patients each. Group I (control)—without orthodontic appliances disinfection; Group II—removable orthodontic appliances which had been immersed in 0.12% chlorhexidine digluconate overnight (8 hours), and Group III—orthodontic appliances in which 0.12% chlorhexidine digluconate solution had been incorporated into their resin composition. Saliva was collected for quantification of MS and evaluation of plaque index was performed before and after installation of orthodontic appliance at 0, 2, 4, 6, 8, and 10 weeks. Data were analyzed by using analysis of variance. Results: Number of MS colonies in saliva and plaque index showed no statistically differences among groups at the different periods (p > 0.05). Conclusions: It could be concluded that chlorhexidine incorporation into the acrylic resin of removable orthodontic appliances at 0.12% concentration and immersion of the appliance into 0.12% chlorhexidine solution were not effective in reducing plaque index and the number of MS in saliva.

Temporo-spatial gait parameters during street crossing conditions: a comparison between younger and older adults

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Displasia neuronal intestinal: análise de critérios morfológicos e comparação de métodos diagnósticos

Pós-graduação em Patologia - FMB

Association analysis of clinical aspects and vitamin D receptor gene polymorphism with external apical root resorption in orthodontic patients

Introduction: Vitamin D is responsible for the regulation of certain genes at the transcription level, via interaction with the vitamin D receptor, and influences host immune responses and aspects of bone development, growth, and homeostasis. Our aim was to investigate the association of TaqI vitamin D receptor gene polymorphism with external apical root resorption during orthodontic treatment. Methods: Our subjects were 377 patients with Class II Division 1 malocclusion, divided into 3 groups: (1) 160 with external apical root resorption <= 1.43 mm, (2) 179 with external apical root resorption >1.43 mm), and (3) 38 untreated subjects. External apical root resorption of the maxillary incisors was evaluated on periapical radiographs taken before and after 6 months of treatment. After DNA collection and purification, vitamin D receptor TaqI polymorphism analysis was performed by polymerase chain reaction-restriction fragment length polymorphism. Univariate and multivariate analyses were performed to verify the association of clinical and genetic variables with external apical root resorption (P <0.05). Results: There was a higher proportion of external apical root resorption in orthodontically treated patients compared with the untreated subjects. In patients orthodontically treated, age higher than 14 years old, initial size of the maxillary incisor root superior to 30 mm, and premolar extraction were associated with increased external apical root resorption. Genotypes containing the C allele were weakly associated with protection against external apical root resorption (CC + CT x TT [odds ratio, 0.29; 95% confidence interval, 0.07-1.23; P = 0.091]) when treated orthodontic patients were compared to untreated individuals. Conclusions: Clinical factors and vitamin D receptor TaqI polymorphism were associated with external apical root resorption in orthodontic patients. (Am J Orthod Dentofacial Orthop 2012; 142: 339-47)

Prognosis of HIV-1-infected patients up to 5 years after initiation of HAART: collaborative analysis of prospective studies

OBJECTIVE: To estimate the prognosis over 5 years of HIV-1-infected, treatment-naive patients starting HAART, taking into account the immunological and virological response to therapy. DESIGN: A collaborative analysis of data from 12 cohorts in Europe and North America on 20,379 adults who started HAART between 1995 and 2003. METHODS: Parametric survival models were used to predict the cumulative incidence at 5 years of a new AIDS-defining event or death, and death alone, first from the start of HAART and second from 6 months after the start of HAART. Data were analysed by intention-to-continue-treatment, ignoring treatment changes and interruptions. RESULTS: During 61 798 person-years of follow-up, 1005 patients died and an additional 1303 developed AIDS. A total of 10 046 (49%) patients started HAART either with a CD4 cell count of less than 200 cells/microl or with a diagnosis of AIDS. The 5-year risk of AIDS or death (death alone) from the start of HAART ranged from 5.6 to 77% (1.8-65%), depending on age, CD4 cell count, HIV-1-RNA level, clinical stage, and history of injection drug use. From 6 months the corresponding figures were 4.1-99% for AIDS or death and 1.3-96% for death alone. CONCLUSION: On the basis of data collected routinely in HIV care, prognostic models with high discriminatory power over 5 years were developed for patients starting HAART in industrialized countries. A risk calculator that produces estimates for progression rates at years 1 to 5 after starting HAART is available from www.art-cohort-collaboration.org.