Estudo mutacional em tumor de Wilms por sequenciação de nova geração

O Tumor de Wilms (TW) é o tumor renal mais comum da infância, com uma incidência de 1 em ~10000 crianças. Esta patologia é de etiologia genética complexa e diversificada. No entanto, cerca de um terço dos doentes apresenta mutações somáticas associadas aos genes WT1, CTNNB1, TP53 e/ou AMER1. Assim, foi desenvolvido um painel de amplicões destes 4 genes para a identificação de mutações num grupo de doentes portugueses com TW, através de uma metodologia baseada na sequenciação de nova geração. As bibliotecas de DNA foram preparadas a partir de amostras de sangue periférico e tumor de 36 doentes com TW e sequenciadas no MiSeq. Foram identificadas alterações somáticas em 7 dos 36 (19,4%) doentes estudados. Conclui-se que a sequenciação de um painel de genes é um método rápido para a deteção de mutações somáticas quando desenhado com cuidado de forma a serem evitados problemas de perda de cobertura.

Avaliação das alterações volumétricas da cavidade nasal decorrentes da expansão rápida de maxila assistida cirurgicamente

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

Influência do volume vesical na diminuição da dose recebida na bexiga e no PTV em doentes com tumor de reto tratados com belly-board

Introdução: O cancro retal continua a ser um dos principais problemas de saúde a nível mundial, sendo a toxicidade gastro-intestinal e génito-urinária os efeitos tardios da radioterapia mais reportados. A utilização da Belly-Board para minimizar essa toxicidade, reduzindo o volume de bexiga e intestino delgado irradiados é recomendada. No entanto, o protocolo mais adequado para o volume vesical nestes doentes é ainda tema de controvérsia. Objetivo: Avaliar a influência do volume vesical na dose recebida na bexiga e no PTV, em doentes com tumor de reto, posicionados em decúbito ventral, com belly-board. Materiais e Métodos: 38 doentes com tumor de reto tratados no CHBM, agrupados em dois grupos: o 1º grupo, com 19 doentes que realizaram tratamento com bexiga cheia e o 2º grupo, com 19 doentes que realizaram tratamento com bexiga vazia. Os dados foram obtidos através dos HDV’s e foram comparadas as doses máximas no PTV e a percentagem de volume de bexiga que recebe 50Gy. Foi utilizado o teste estatístico U-Mann Whitney com um nível de significância de 0,05. A hipótese de pesquisa deste estudo propõe que os dois grupos diferem significativamente entre si e a hipótese nula propõe que os dois grupos não diferem significativamente entre si, para ambas as variáveis. Resultados: Não se observaram diferenças estatisticamente significativas entre os grupos no que diz respeito à dose máxima no PTV. No que se refere à percentagem de volume de bexiga que recebe 50Gy verificaram-se diferenças estatisticamente significativas, tendo o grupo de doentes que realizaram tratamento com bexiga cheia apresentado valores mais baixos. Conclusões: Este estudo demonstrou o benefício da utilização do protocolo de bexiga cheia em doentes com tumor de reto tratados com belly-board, na diminuição da percentagem de volume de bexiga que recebe 50Gy.

Prognóstico de pacientes com tumor de Wilms unilateral no Rio de Janeiro, 1990-2000

OBJETIVO: Analisar a sobrevida e os principais fatores prognósticos entre os pacientes com tumor de Wilms unilateral. MÉTODOS: A coorte de estudo incluiu 132 casos de tumor de Wilms unilateral em menores de 15 anos de idade matriculados em serviço de oncologia pediátrica, de janeiro de 1990 a dezembro de 2000. Curvas de sobrevida foram confeccionadas utilizando-se o método de Kaplan-Meier e fatores prognósticos foram analisados pelo modelo de riscos proporcionais de Cox. RESULTADOS: A estimativa de sobrevida global em cinco anos foi 84,6%. As probabilidades de sobrevida para os estádios I, II, III e IV foram de 100%; 94,2%; 83,2% e 31,3%, respectivamente. A taxa de sobrevida para os pacientes com: histologia favorável foi de 89,4%, para aqueles com anaplasia focal 66,7 % e com anaplasia difusa 40%. Todos os pacientes com doença em estádio IV e anaplasia difusa foram a óbito (n=4). Todos os pacientes com doença em estádio I, independente da histologia, permaneceram vivos até o final do período de seguimento. CONCLUSÕES: Entre as variáveis escolhidas para o modelo final apenas o estadiamento e a histologia permaneceram associados ao elevado risco de óbito enquanto que os casos na faixa etária entre 24 e 47 meses apresentaram melhor prognóstico que os demais. Esses resultados mostram a importância do diagnóstico em fases iniciais da doença e que a histologia é fundamental para orientar a terapia adequada.

Topological Complexity and Predictability in the Dynamics of a Tumor Growth Model with Shilnikov's Chaos

Dynamical systems modeling tumor growth have been investigated to determine the dynamics between tumor and healthy cells. Recent theoretical investigations indicate that these interactions may lead to different dynamical outcomes, in particular to homoclinic chaos. In the present study, we analyze both topological and dynamical properties of a recently characterized chaotic attractor governing the dynamics of tumor cells interacting with healthy tissue cells and effector cells of the immune system. By using the theory of symbolic dynamics, we first characterize the topological entropy and the parameter space ordering of kneading sequences from one-dimensional iterated maps identified in the dynamics, focusing on the effects of inactivation interactions between both effector and tumor cells. The previous analyses are complemented with the computation of the spectrum of Lyapunov exponents, the fractal dimension and the predictability of the chaotic attractors. Our results show that the inactivation rate of effector cells by the tumor cells has an important effect on the dynamics of the system. The increase of effector cells inactivation involves an inverse Feigenbaum (i.e. period-halving bifurcation) scenario, which results in the stabilization of the dynamics and in an increase of dynamics predictability. Our analyses also reveal that, at low inactivation rates of effector cells, tumor cells undergo strong, chaotic fluctuations, with the dynamics being highly unpredictable. Our findings are discussed in the context of tumor cells potential viability.

Reliability of nasal auscultation and their relation to middle ear condition – a clinical interest to pediatrics respiratory physiotherapy

Background: Acute respiratory infections are usual in children under three years old occurring in upper respiratory tract, having an impact on child and caregiver’s quality of life predisposing to otitis media or bronchiolitis. There are few valid and reliable measures to determine the child’s respiratory condition and to guide the physiotherapy intervention. Aim: To assess the intra and inter rater reliability of nasal auscultation, to analyze the relation between sounds’ classification and middle ear’s pressure and compliance as well as with the Clinical Severity Score. Methods: A cross-sectional observational study was composed by 125 nursery children aged up to three years old. Tympanometry, pulmonary and nasal auscultation and application of Clinical Severity Score were performed to each child. Nasal auscultation sounds’ were recorded and sent to 3 blinded experts, that classified, as “obstructed” and “unobstructed”, with a 48 hours interval, in order to analyze inter and intra rater reliability. Results: Nasal auscultation revealed a substantial inter and intra rater reliability (=0,749 and evaluator A - K= 0,691; evaluator B - K= 0,605 and evaluator C - K= 0,724, respectively). Both ears’ pressure was significantly lower in children with an "unobstructed" nasal sound when compared with an “obstructed” nasal sound (t=-3,599, p<0,001 in left ear; t=-2,258, p=0,026 in right ear). Compliance in both ears was significantly lower in children with an "obstructed" nasal sound when compared with “unobstructed” nasal sound (t=-2,728, p=0,007 in left ear; t=-3,830, p<0,001 in right ear). There was a statistically significant association between sounds’ classification and tympanograms types in both ear’s (=11,437, p=0,003 in left ear; =13,535, p=0,001 in right ear). There was a trend to children with an "unobstructed" nasal sound that had a lower clinical severity score when compared with “obstructed” children. Conclusion: It was observed a good intra and substantial inter reliability for nasal auscultation. Nasal auscultation sounds’ classification was related to middle ears’ pressure and compliance.

Optimização das fugas pela boca durante o CPAP nasal: banda submentoniana versus máscara facial

Introdução: O CPAP nasal é o tratamento de eleição para os pacientes com Síndrome da Apneia Obstrutiva do Sono (SAOS). Com a máscara nasal podem ocorrer fugas de ar pela boca, que podem por em causa a aderência do paciente ao tratamento devido muitas vezes ao desconforto que provocam, ao aumento do trabalho respiratório e por afectarem a qualidade do sono. Objectivos: Este estudo tem como principal objectivo verificar a eficácia da banda submentoniana e da máscara facial na correcção das fugas pela boca em pacientes com SAOS. Métodos e Participantes: Uma amostra de conveniência de 15 pacientes (8 homens) com SAOS e a fazerem CPAP com máscara nasal, foi recrutada. Foram divididos em dois grupos A e B, onde no grupo A se colocou a banda submentoniana e no grupo B se alterou a interface para máscara facial. Medidas e Resultados: As variáveis avaliadas neste estudo foram as fugas, o IAH, o percentil 95 da pressão de tratamento, a Sa,O2 e os efeitos adversos das duas intervenções. O nível de fugas reduziu no grupo A de 38±11,27 para 24,55±14,30L/min (p=0,002) e no grupo B de 34,34±16,50 para 18,51±16,22L/min (p=0,008). O IAH aumentou no grupo B de 2,60±2,13 para 4,41±3,88 (p=0,016). Relativamente ao percentil 95 da pressão de tratamento aumentou nos dois grupos (Grupo A de 10,15±2,63 para 12,08±2,45cmH2O (p=0,008) e no Grupo B 10,51±1,88 para 12,64±1,29cmH2O (p=0,008)). A Sa,O2 mínima aumentou e o tempo<90% diminui no grupo A com p=0,008, p=0,031, respectivamente. Quanto ao uso auto-reportado verificaram-se poucos efeitos adversos, salientando-se apenas a facilidade de colocação da banda submentoniana, a secura da boca nos dois grupos, a pressão no queixo provocada pela banda e a dor no dorso do nariz provocada pela máscara facial. Conclusão: Ambas as estratégias provaram ser mais eficazes a reduzir a fuga que a máscara nasal. Foram bem toleradas e com poucos efeitos adversos.

Characterization of the glycosylation of human tumor cells

Ovarian cancer is within the most lethal gynecological malignancies in woman. Therefore, many investigators study its biological aspects with the purpose of discovering more rapid diagnostic methods and efficient treatment. Resembling many other tumors, in ovarian cancer, aberrant glycosylation occurs with the appearance of novel or altered carbohydrate structures. These can be terminal motifs, such as the Lewis determinants, or entire carbohydrate sequences, which have been related to tumorigenesis and its outcome.(...)

Activation of effector immune cells promotes tumor stochastic extinction: A homotopy analysis approach

In this article we provide homotopy solutions of a cancer nonlinear model describing the dynamics of tumor cells in interaction with healthy and effector immune cells. We apply a semi-analytic technique for solving strongly nonlinear systems – the Step Homotopy Analysis Method (SHAM). This algorithm, based on a modification of the standard homotopy analysis method (HAM), allows to obtain a one-parameter family of explicit series solutions. By using the homotopy solutions, we first investigate the dynamical effect of the activation of the effector immune cells in the deterministic dynamics, showing that an increased activation makes the system to enter into chaotic dynamics via a period-doubling bifurcation scenario. Then, by adding demographic stochasticity into the homotopy solutions, we show, as a difference from the deterministic dynamics, that an increased activation of the immune cells facilitates cancer clearance involving tumor cells extinction and healthy cells persistence. Our results highlight the importance of therapies activating the effector immune cells at early stages of cancer progression.

Metal ions modulate the folding and stability of the tumor suppressor protein S100A2

Febs Journal (2009)276:1776-1786

Plasma levels of tumor necrosis factor-alpha in patients with visceral leishmaniasis (Kala-Azar). Association with activity of the disease and clinical remisson following antimonial therapy

Evaluation of TNF-alpha in patients with Kala-azar has drawn increasing interest due to its regulatory role on the immune system, in addition to its cachetizing activity. The objective of this study was to examine the association between plasma levels of TNF-alpha, measured by immunore-activity (ELISA) and bioactivity (cytotoxicity assay with L-929 cells), and clinical manifestations of visceral leishmaniasis. Plasma samples from 19 patients with Kala-azar were obtained before, during and at the end of antimonial therapy. TNF-alpha determinations was done by using the cytotoxicity assay (all patients) and the enzyme-linked immunoassay (ELISA - 14 patients). A discrepancy between results obtained by ELISA and cytotoxicity assay was observed. Levels of circulating TNF-alpha, assessed by ELISA, were higher in patients than in healthy controls, and declined significantly with improvement in clinical and laboratory parameters. Plasma levels before treatment were 124.7 ± 93.3 pg/ml (mean ± SD) and were higher than at the end of therapy 13.9 ± 25.1 pg/ml (mean ± SD) (p = 0.001). In contrast, plasma levels of TNF-alpha evaluated by cytotoxicity assay did not follow a predicted course during follow-up. Lysis, in this case, might be not totally attributed to TNF-alpha. The discrepancy might be attributed to the presence of factor(s) known to influence the release and activity of TNF-alpha.

Tumor-like lesion due to Chagas' disease in a patient with lymphocytic leukemia

A 73 year-old white male, living in the interior of the state of Mato Grosso do Sul, in central Bra­zil, after an initial diagnosis of sinusitis was transferred to the neurology service with a 3-day evolution of intracranial hypertension. Exams showed lymphocytic leukemia and a tumor-like lesion, either an expanding inflammatory process such as an abscess or a neoplasm. Treatment with Ceftriaxone and Decadron was started and intracranial hypertension was controlled. Methotrexate was injected on the occasion of the next puncture considering a possible leukemia infiltration. Flagellate forms of T. cruzi were observed in the CSF and treatment with Benznidazole was started. After 4 days the CSF presented fractionated forms of trypomastigotes. The protein level was 27%. Signs of intracranial hypertension ceased. Tomography and magnetic resonance images showed an important reduction of the tumor-like lesion. The clinical condition of the patient improved.

Presence of Circulating Levels of Interferon-g, Interleukin-10 and Tumor Necrosis Factor-a in Patients with Visceral Leishmaniasis

Experimental murine L. major infection is characterized by the expansion of distinct CD4+ T cell subsets. The Th1 response is related to production of IFN-g and resolution of infection, whereas Th-2 response with production of IL-4 and IL-10 and dissemination of infection. The objective of this study was to measure the circulating levels of IFN-g, IL-10 and TNF-a in patients with visceral leishmaniasis (VL) before, during and at the end of therapy and to examine the association between cytokine levels and activity of VL. Fifteen patients with VL were evaluated. The cytokine determinations were done by using the enzyme-linked immunoassay (ELISA) before, during and at the end of therapy. At baseline, we detected circulating levels of IFN-g in 13 of 15 patients (median = 60 pg/ml); IL-10 in 14 of 15 patients (median = 141.4 pg/ml); and TNF-a in 13 of 14 patients (median = 38.9 pg/ml). As patients improved, following antimonial therapy, circulating levels of IL-10 showed an exponential decay (y = 82.34 e–0,10367x, r = –0.659; p < 0.001). IFN-g was no longer detected after 7/14 days of therapy. On the other hand, circulating levels of TNF-a had a less pronounced decay with time on therapy, remaining detectable in most patients during the first seven days of therapy (y = 36.99-0.933x, r = –0.31; p = 0.05). Part of the expression of a successful response to therapy may, therefore, include reduction in secretion of inflammatory as well as suppressive cytokines. Since IL-10 and IFN-g are both detected prior to therapy, the recognized cellular immune depression seen in these patients may be due to biological predominance of IL-10 (type 2 cytokine), rather than lack of IFN-g (type 1 cytokine) production.

Mycotic bovine nasal granuloma

A case of mycotic bovine nasal granuloma in a 10 year-old Jersey cow, produced by Drechslera halodes is presented. Histopathological sections showed abundant hyaline and pigmented extra and intracellular fungal structures together with a polymorphic cellular granuloma formed by neutrophils, lymphocytes, plasmocytes, histiocytes and giant cells of the Langhans type. It is the first case of mycotic bovine nasal granuloma recognized in Uruguay although this disease seems to be frequent according to the opinion of veterinarian specialists. Another similar clinical case also in a Jersey cow from the same dairy house with an intense cellular infiltrate rich in eosinophils without granulomatous image, together with extracellular hyaline and fuliginous fungal forms, is also referred for comparative purposes. Geotrichum sp. was isolated. The need of an early diagnosis and treatment of the disease is stressed.