Water-insoluble exopolysaccharides (I-EPS) are a virulence factor for dental biofilms. It has already been demonstrated that mango pulp induces the secretion of glucan-hydrolytic enzymes in the fungus Trichoderma harzianum, and that they have an effect on I-EPS from young biofilms. Aim Evaluate the effect of mango peel as an enzyme inducer in T. harzianum, and the effect of enzymes secreted on mature biofilms. Methods Fractions of the peel (PL) and ethanol-precipitated pulp (PP) of Tommy Atkins mangoes were sterilized and added to a culture medium containing T. harzianum for induction of hydrolytic enzymes. After 192 h, the culture medium was centrifuged and the supernatant (enzyme extract) was used as treatment on S. mutans biofilms (n=9): a) NaCl 0.9 %; b) 0.12 % chlorhexidine digluconate; and c) extract of enzymes induced by PL or PP. Acidogenicity, bacterial viability, quantification of insoluble polysaccharides, and three-dimensional analysis of the biofilm by scanning electron microscopy (SEM) was performed. Data were analyzed by ANOVA followed by the Tukey test (α=5 %). Results The hydrolytic enzymes did not alter the metabolism or bacterial viability of the biofilm (p<0.05). Although the images obtained by SEM suggest some degree of matrix degradation, the quantification of I-EPS for the PL and PP groups did not differ from the control group (p>0.05), suggesting a slight effect on the disorganization of the mature S. mutans biofilm. Conclusion The results suggest that mango peel fraction can induce secretion of mutanase by T. harzianum, however in an insufficient amount to generate significant degradation on cariogenic biofilm. Waterinsoluble Water IEPS I EPS (I-EPS glucanhydrolytic glucan T (PL ethanolprecipitated ethanol precipitated (PP 19 h S n=9 n9 n 9 (n=9) 09 0 0. b 012 12 0.1 digluconate c Acidogenicity polysaccharides threedimensional three dimensional (SEM performed α=5 α5 α 5 (α= %. . %) p<0.05. p005 p p<0.05 05 (p<0.05) p>0.05, p>0.05 , (p>0.05) 1 n= (n=9 01 α= (α p00 p<0.0 (p<0.05 p>0.0 (p>0.05 (n= p0 p<0. (p<0.0 p>0. (p>0.0 (n p<0 (p<0. p>0 (p>0. p< (p<0 p> (p>0 (p< (p> (p

Abstract Proteases are enzymes that act in the hydrolysis of proteins and have several industrial applications. Moreover, proteases have gained prominence as enzymes for the generation of bioactive peptides from the hydrolysis of different protein sources. Milk is the most studied protein source to obtain peptides due to its nutritional and physiological effects and has been studied as complementary therapeutic approaches for the cancer treatment, interacting specifically with cancer cells, consequently fewer side effects. The ability of Eupenicillium javanicum metalloprotease to generate whey-derived peptides with antioxidant activity has already been demonstrated. For this reason, we thus hypothesized that whey-derived peptides from Eupenicillium javanicum metalloprotease hydrolysis could also have a potential against melanoma cell lines. In this study, B16F10 melanoma cells were treated for 72 h with whey-derived peptides and the effects on cell viability were determined. Moreover, the protein profiles of the treated and nontreated cells were compared in proteomic assay and mass spectrometry analyzes. Whey-derived peptides impaired about 62% cell viability, and proteomic approach associated this behavior to modulate proteins involved in proliferation, energy, apoptosis, metastatic and malignancy rates. This study describes the relevance of microbial enzymes in generation of whey-derived peptides with biological activity against melanoma cells.

Abstract Bioprocess studies have been highlighted due to the importance of physiological processes and industrial applications of enzymes. The potential of peptidase production from Aspergillus section Flavi using different amino acids as a supplemental nitrogen source was investigated. A production profile revealed that amino acids had positive effects on peptidase production when compared to the control without amino acids. Optimal production (100 U/mL) was obtained with Arginine amino acid in 96 h of fermentation. Extracellular peptidase from Aspergillus section Flavi was identified in submerged bioprocesses by in situ activity. Biochemical studies revealed that the maximum activities of the enzyme extract were obtained at pH 6.5 and a temperature of 55°C. The inhibition by EDTA and PMSF suggests the presence of more than one peptidase while the Ni2+ and Cu2+ had a negative influence on the enzyme activity. When the crude extract was reversibly immobilized on ionic supports, DEAE-Agarose and MANAE-Agarose the derivative showed different profiles of thermal and pH stabilities. Hence, this study revealed the basic properties and biochemical characteristics that allowed the production improvement of this class of enzyme. Moreover, with known properties stabilization and immobilization process is required to further explore its biotechnological capacities.

Abstract Background: Lachesis muta rhombeata is one of the venomous snakes of medical importance in Brazil whose envenoming is characterized by local and systemic effects which may produce even shock and death. Its venom is mainly comprised of serine and metalloproteinases, phospholipases A2 and bradykinin-potentiating peptides. Based on a previously reported fractionation of L. m. rhombeata venom (LmrV), we decided to perform a subproteome analysis of its major fraction and investigated a novel component present in this venom. Methods: LmrV was fractionated through molecular exclusion chromatography and the main fraction (S5) was submitted to fibrinogenolytic activity assay and fractionated by reversed-phase chromatography. The N-terminal sequences of the subfractions eluted from reversed-phase chromatography were determined by automated Edman degradation. Enzyme activity of LmrSP-4 was evaluated upon chromogenic substrates for thrombin (S-2238), plasma kallikrein (S-2302), plasmin and streptokinase-activated plasminogen (S-2251) and Factor Xa (S-2222) and upon fibrinogen. All assays were carried out in the presence or absence of possible inhibitors. The fluorescence resonance energy transfer substrate Abz-KLRSSKQ-EDDnp was used to determine the optimal conditions for LmrSP-4 activity. Molecular mass of LmrSP-4 was determined by MALDI-TOF and digested peptides after trypsin and Glu-C treatments were analyzed by high resolution MS/MS using different fragmentation modes. Results: Fraction S5 showed strong proteolytic activity upon fibrinogen. Its fractionation by reversed-phase chromatography gave rise to 6 main fractions (S5C1-S5C6). S5C1-S5C5 fractions correspond to serine proteinases whereas S5C6 represents a C-type lectin. S5C4 (named LmrSP-4) had its N-terminal determined by Edman degradation up to the 53rd amino acid residue and was chosen for characterization studies. LmrSP-4 is a fibrinogenolytic serine proteinase with high activity against S-2302, being inhibited by PMSF and benzamidine, but not by 1,10-phenantroline. In addition, this enzyme exhibited maximum activity within the pH range from neutral to basic and between 40 and 50 °C. About 68% of the LmrSP-4 primary structure was covered, and its molecular mass is 28,190 Da. Conclusions: Novel serine proteinase isoforms and a lectin were identified in LmrV. Additionally, a kallikrein-like serine proteinase that might be useful as molecular tool for investigating bradykinin-involving process was isolated and partially characterized.

Resumen: El texto propone una reflexión sobre el cine de animación de los estudios Disney a partir del análisis del largometraje Pinocho (Pinocchio), lanzado originalmente en 1940 y dirigido por Ben Sharpsteen y Hamilton Luske. El artículo se orienta principalmente por una lectura de las imágenes de la película, con el objetivo de interpretar el estilo gráfico de los dibujos Disney y los repertorios iconográficos y de composición imagética referenciados por las animaciones. El texto se cierra con una reflexión sobre la relación de los dibujos Disney con la tradición narrativa hollywoodiana y su efectivo valor como producción de caricaturas y escenarios.

Abstract: The text proposes a reflection on the animated films of Disney studios based on the analysis of the feature film Pinocchio, originally released in 1940 and directed by Ben Sharpsteen and Hamilton Luske. The article is mainly oriented by a reading of the images of the film, with the aim of interpreting the graphic style of the Disney drawings and the iconographic and imaginative repertoires referenced by the animations. The text closes with a reflection on the relationship of the Disney drawings with the Hollywood narrative tradition and its effective value as a production of cartoons and staging.

Resumo: O texto propõe uma reflexão sobre os filmes de animação dos estúdios Disney, a partir da análise do longa-metragem Pinóquio (Pinocchio), originalmente lançado em 1940 e dirigido por Ben Sharpsteen e Hamilton Luske. O artigo orienta-se principalmente pela leitura das imagens do filme, com o objetivo de interpretar o estilo gráfico dos desenhos da Disney e os repertórios iconográficos e imagéticos referenciados pelas animações. O texto termina com uma reflexão sobre a relação dos desenhos da Disney com a tradição narrativa hollywoodiana e seu efetivo valor como produção de caricaturas e encenações.

RESUMO Objetivo: Realizar uma revisão sobre a Síndrome de Prader-Willi (SPW) com base nas publicações mais recentes e fornecer recomendações ao pediatra geral para diagnóstico precoce e seguimento. Fonte de dados: Artigos publicados nas bases Pubmed e SciELO. A pesquisa não foi limitada a um período e incluiu todos os artigos das bases de dados. Síntese dos dados: A SPW é uma síndrome genética rara, resultante da perda do imprinting gênico expresso no cromossomo paterno 15q11-q13, sendo caracterizada por alterações endocrinológicas, como deficiência de hormônio de crescimento, obesidade, insuficiência adrenal central, hipotireoidismo, hipogonadismo, além de alterações comportamentais e déficit intelectual. Há outras comorbidades associadas, como distúrbios de sono, escoliose, constipação, problemas dentários e alterações de coagulação. O protocolo de seguimento da SPW do Instituto da Criança da Universidade de São Paulo se baseia em quarto pilares principais: dieta, exercício físico, terapia com hormônio de crescimento humano recombinante (rhGH) e manejo comportamental e cognitivo. A dieta deve ser restrita a 900 kcal/dia, de acordo com a Pirâmide Alimentar do Prader-Willi, e o exercício físico deve ser diário, aeróbico e postural. A terapia com rhGH é fortemente recomendada pela literatura científica internacional e deve ser iniciada assim que for realizado o diagnóstico da síndrome. O manejo do comportamento é realizado com estratégias para estabelecer rotina e regras. Conclusões: Se a SPW se tornar mais familiar ao pediatra geral, o diagnóstico e o tratamento começarão mais precocemente, o que irá melhorar a qualidade de vida e os cuidados desses pacientes.

ABSTRACT Objective: To carry out a review about Prader-Willi Syndrome based on the most recent data about the subject and to give recommendation for the general pediatricians for early diagnoses and follow-up. Data sources: Scientific articles in the PubMed and SciELO databases. The research was not limited to a specific time period and included all articles in such databases. Data synthesis: The Prader-Willi Syndrome (PWS) is a rare genetic disorder resulting from the loss of imprinted gene expression within the paternal chromosome 15q11-q13. PWS is characterized by endocrine abnormalities, such as growth hormone (GH) deficiency, obesity, central adrenal insufficiency, hypothyroidism, hypogonadism and complex behavioral and intellectual difficulties. PWS individuals also may present other comorbidities, such as sleep disorders, scoliosis, constipation, dental issues and coagulation disorders. The follow-up protocol of the Children’s Institute at Universidade de São Paulo is based on four main pillars: diet, exercise, recombinant human growth hormone (rhGH) therapy and behavioral and cognitive issues. The diet must include a caloric restriction of 900 kcal/day, according to the Prader-Willi Eating Pyramid and exercise plan is focused on daily aerobic exercises and postural therapy. The rhGH therapy is highly recommended by the international scientific literature and must be started as soon as the diagnostic is made. The management of behavioral issues is based on strategies to establish routine and rules. Conclusions: If the general pediatrician becomes more familiar with PWS, the diagnosis and treatment will start earlier, which is essential to improve the quality of life and care for these individuals.

Abstract Background: L-amino acid oxidases isolated from snake venoms (SV-LAAOs) are enzymes that have great therapeutic potential and are currently being investigated as tools for developing new strategies to treat various diseases, including cancer and bacterial infections. The main objective of this study was to make a brief evaluation of the enzymatic stability of two Bothrops LAAOs, one isolated from Bothrops jararacussu (BjussuLAAO-II) and the other from Bothrops moojeni (BmooLAAO-I) venoms. Methods and results: The enzymatic activity and stability of both LAAOs were evaluated by microplate colorimetric assays, for which BjussuLAAO-II and BmooLAAO-I were incubated with different L-amino acid substrates, in the presence of different ions, and at different pH ranges and temperatures. BjussuLAAO-II and BmooLAAO-I demonstrated higher affinity for hydrophobic amino acids, such as Phe and Leu. The two enzymes showed high enzymatic activity in a wide temperature range, from 25 to 75 °C, and presented optimum pH around 7.0. Additionally, Zn2+, Al3+, Cu2+ and Ni2+ ions negatively modulated the enzymatic activity of both LAAOs. As to stability, BjussuLAAO-II and BmooLAAO-I showed high enzymatic activity for 42 days stored at 4°C in neutral pH solution. Moreover, the glycan portions of both LAAOs were analyzed by capillary electrophoresis, which revealed that BjussuLAAO-II presented two main glycan portions with relative masses of 7.78 and 8.13 CGU, while BmooLAAO-I showed three portions of 7.58, 7.94 and 8.37 CGU. Conclusions: Our results showed that, when stored properly, BjussuLAAO-II and BmooLAAO-I present enzymatic stability over a long time period, which is very important to allow the use of these enzymes in pharmacological studies of great impact in the medical field.

Abstract Background: Lachesis muta rhombeata (Lmr) is the largest venomous snake in Latin America and its venom contains mainly enzymatic components, such as serine and metalloproteases, L-amino acid oxidase and phospholipases A2. Metalloproteases comprise a large group of zinc-dependent proteases that cleave basement membrane components such as fibronectin, laminin and collagen type IV. These enzymes are responsible for local and systemic changes, including haemorrhage, myonecrosis and inflammation. This study aimed the isolation and enzymatic characterization of the first metalloprotease (Lmr-MP) from Lmr venom (LmrV). Methods and results: Lmr-MP was purified through two chromatographic steps and submitted to enzymatic characterization. It showed proteolytic activity on azocasein with maximum activity at pH 7.0-9.0. It was inhibited by EDTA (a metal chelator that removes zinc, which is essential for enzymatic activity) and no effect was observed with PMSF, iodoacetic acid or pepstatin (inhibitors of serine, cysteine and aspartyl proteases, respectively). Ca2+, Mg2+ and Ba2+ ions increased its activity, while Al3+, Cu2+, Ni2+ and Zn2+ inhibited it. Additionally, ZnCl2 showed a dose dependent inhibition of the enzyme. Lmr-MP activity was also evaluated upon chromogenic substrates for plasma kallikrein (S-2302), plasmin and streptokinase-activated plasminogen (S-2251) and Factor Xa (S-2222) showing the highest activity on S-2302. The activity in different solutions (5 mM or 50 mM ammonium bicarbonate, pH 7.8; 0.1% trifluoroacetic acid + 50% acetonitrile; phosphate buffer saline, pH 7.4; 50 mM sodium acetate, pH 4.0 or ammonium acetate pH 4.5) was also evaluated and the results showed that its activity was abolished at acidic pHs. Its molecular mass (22,858 Da) was determined by MALDI-TOF and about 90% of its primary structure was verified by high-resolution mass spectrometry using HCD and ETD fragmentations and database search against the sequence of closely related species. It is a novel enzyme which shared high identity with other snake venom metalloproteases (svMPs) belonging to the P-I group. Conclusion: The purification procedure achieved a novel pure highly active metalloprotease from LmrV. This new molecule can help to understand the metalloproteases mechanisms of action, the Lachesis envenoming, as well as to open new perspectives for its use as therapeutic tools.

RESUMO Objetivou-se avaliar o impacto do apoio social sobre os sintomas de mulheres brasileiras com fibromialgia (FM). Trata-se de um estudo observacional descritivo que selecionou 66 mulheres que atendiam aos critérios do Colégio Americano de Reumatologia (ACR) de 1990. O apoio social foi medido com o Social Support Survey (MOS-SSS), a funcionalidade com o Questionário do Impacto da Fibromialgia (FIQ), a depressão com o Inventário de Depressão de Beck (BDI), a ansiedade com a Escala de Ansiedade de Hamilton (HAS), a afetividade com o Positive and Negative Affect Schedule (Panas) e foi feita algometria para registrar o limiar da dor à pressão (LDP) e a tolerância álgica à pressão (TAP) nos 18 pontos recomendados pelo ACR. Os pacientes foram divididos nos grupos apoio social normal (ASN) ou ruim (ASR); o ASR foi definido como uma pontuação nos MOS-SSS abaixo do percentil 25 da amostra total. Usou-se o teste de Mann-Whitney ou o teste t não pareado para comparar variáveis intergrupos e o de Fisher para as variáveis categóricas. Usaram-se a análise de covariância e o teste de correlação de Pearson. Não houve diferença nas variáveis sociodemográficas entre os grupos ASN e ASR. Observaram-se diferenças entre os grupos ASN e ASR para todas as quatro subcategorias de apoio social e pontuação total do MOS-SSS. Encontraram-se diferenças significativas entre o ASN e o ASR na depressão (p = 0,007), afeto negativo (p = 0,025) e LDP (p = 0,016). A subcategoria apoio afetivo mostrou correlação positiva entre a dor e o afeto positivo no grupo ASR. A subcategoria interação social positiva mostrou uma correlação negativa entre o FIQ e o estado de depressão. Portanto, o apoio social parece contribuir para a melhoria na saúde mental e física na FM.

ABSTRACT We aimed to assess the impact of social support on symptoms in Brazilian women with FM. An observational, descriptive study enrolling 66 women who met the 1990 American College of Rheumatology (ACR) criteria. Social support was measured by the Social Support Survey (MOS-SSS), functionality was evaluated using the Fibromyalgia Impact Questionnaire (FIQ), depression was assessed using the Beck Depression Inventory (BDI), anxiety was measured using the Hamilton Anxiety Scale (HAS), affectivity was measured by Positive and Negative Affect Schedule (PANAS), and algometry was carried out to record pressure pain threshold (PPth) and tolerance (PPTo) at 18 points recommended by the ACR. Patients were divided into normal (NSS) or poor social support (PSS) groups with PSS defined as having a MOS-SSS score below the 25th percentile of the entire sample. Mann-Whitney or Unpaired t-test were used to compare intergroup variables and Fisher's for categorical variables. Analysis of covariance and Pearson correlation test were used. No differences in sociodemographic variables between PSS and NSS were found. Differences between NSS and PSS groups were observed for all four subcategories of social support and MOS-SSS total score. Significant differences between NSS and PSS on depression (p = 0.007), negative affect (p = 0.025) and PPTh (p = 0.016) were found. Affectionate subcategory showed positive correlation between pain and positive affect in PSS. Positive social interaction subcategory showed a negative correlation between FIQ and depression state. Therefore social support appears to contribute to ameliorate mental and physical health in FM.

Resumo A Federação Internacional de Diabetes (IDF-2015) estima a existência no Brasil de 30.900 menores de 15 anos portadores de diabetes mellitus tipo 1 (DM1), com previsão de aumento de 3,0% ao ano. Esta revisão buscou artigos de metanálise e atualização em diabetes infantil com o objetivo de alertar para a necessidade do planejamento de estratégias de enfrentamento deste que tende a ser um sério problema de saúde pública para os próximos anos. O DM1 é considerado uma doença imunomediada de transmissão complexa, influenciada por fatores genéticos e ambientais determinantes da destruição gradual das células beta pancreáticas produtoras de insulina. A positividade sorológica dos autoanticorpos associados ao DM1 e a alteração de testes metabólicos que avaliam a secreção de insulina e o estado glicêmico podem ser utilizados como critérios de previsão da reserva funcional de células beta e do início clínico da doença. O tratamento do DM1 sintomático é complexo, e a manutenção do bom controle metabólico é ainda a única estratégia efetiva de preservação das células beta ainda funcionantes. Tempo de duração da doença e hiperglicemia são fatores de risco para a instalação das complicações vasculares crônicas, que afetam negativamente a qualidade de vida e a sobrevida desses indivíduos. Torna-se necessária a formação de equipes de saúde preparadas para fornecer a melhor informação possível em diabetes infantil, através de programas de educação continuada, com potencial de capacitar esses jovens e suas famílias para o autocuidado.

Summary The International Diabetes Federation (IDF-2015) estimates the existence of 30,900 children under 15 years old with type 1 diabetes mellitus (DM1) in Brazil, and an increase of 3.0% per year is expected. This review focused on meta-analysis and pediatric diabetes update articles in order to draw attention to the need of planning coping strategies to support this serious public health problem in coming years. DM1 is considered an immuno-mediated disease with a complex transmission influenced by genetic and environmental factors responsible for a gradual destruction of the insulin producing pancreatic beta cells. Seroconversion to DM1-associated autoantibodies and abnormalities in metabolic tests that assess insulin secretion and glucose tolerance can be used as predictive criteria of beta cells functional reserve and the onset of the clinical disease. Symptomatic DM1 treatment is complex and the maintenance of good metabolic control is still the only effective strategy for preserving beta cell function. Disease duration and hyperglycemia are both risk factors for the onset of chronic vascular complications that negatively affect the quality of life and survival of these patients. In this regard, health teams must be trained to provide the best possible information on pediatric diabetes, through continuing education programs focused on enabling these young people and their families to diabetes self-management.

BackgroundCrotalus durissus terrificus venom (CdtV) is one of the most studied snake venoms in Brazil. Despite presenting several well known proteins, its L-amino acid oxidase (LAAO) has not been studied previously. This study aimed to isolate, characterize and evaluate the enzyme stability of bordonein-L, an LAAO from CdtV.Methods The enzyme was isolated through cation exchange, gel filtration and affinity chromatography, followed by a reversed-phase fast protein liquid chromatography to confirm its purity. Subsequently, its N-terminal amino acid sequence was determined by Edman degradation. The enzyme activity and stability were evaluated by a microplate colorimetric assay and the molecular mass was estimated by SDS-PAGE using periodic acid-Schiff staining and determined by mass spectrometry.Results The first 39 N-terminal amino acid residues exhibited high identity with other snake venom L-amino acid oxidases. Bordonein-L is a homodimer glycoprotein of approximately 101 kDa evaluated by gel filtration. Its monomer presents around 53 kDa estimated by SDS-PAGE and 58,702 Da determined by MALDI-TOF mass spectrometry. The enzyme exhibited maximum activity at pH 7.0 and lost about 50 % of its activity after five days of storage at 4 °C. Bordonein-L’s activity was higher than the control when stored in 2.8 % mannitol or 8.5 % sucrose.Conclusions This research is pioneering in its isolation, characterization and enzyme stability evaluation of an LAAO from CdtV, denominated bordonein-L. These results are important because they increase the knowledge about stabilization of LAAOs, aiming to increase their shelf life. Since the maintenance of enzymatic activity after long periods of storage is essential to enable their biotechnological use as well as their functional studies.

The aim of this study was to evaluate peptidase production by Aspergillus terreus in solid-state bioprocess and evaluate its parameters. The best conditions were 5.0 g of wheat bran as substrate, incubation temperature 30°C, inoculum 2.0x105spores/g and 75% saline volume, with production reaching 677 U/mL (5400 U/g culture medium) after 72 h of fermentation. Biochemical characterization of the crude enzymatic extract showed the optimum pH and temperature of 6.5 and 55°C, respectively. The stability at different temperatures and pH values showed that the extract could endure different pH. The evaluation of the ions influence and inhibitors proved that the enzyme required an ion for better activity, which was corroborated with the inhibition of EDTA and PMSF, characterizing serine and/or metallo peptidase. The extract was also tested for specific activities and showed promising results for keratinolytic and collagenolytic activities (0.252 and 0.165 OD/mL, respectively).

Relatamos um caso de hipoplasia adrenal congênita (HAC) e hipogonadismo hipogonadotrófico (HH) causado por uma nova mutação do gene DAX1. Paciente do sexo masculino com 19 meses de idade, hiperpigmentação e desenvolvimento inadequado foi encaminhado ao nosso serviço. Antecedente familiar de três irmãos falecidos por falência da adrenal, e um primo materno portador de insuficiência adrenal. Excluída a hipótese de adrenoleucodistrofia. A RM demonstrou hipófise e hipotálamo normais. Os níveis de hormônios plasmáticos mostraram alta concentração de ACTH (até 2.790 pg/mL) e baixos níveis de androstenediona, DHEA-S, 11-deoxicortisol e cortisol. Aos 14 anos de idade, o paciente ainda era pré-púbere, com peso de 43,6 kg (SDS: -0,87) e altura de 161 cm (SDS: -0,36), proporcionado. O teste do GnRH mostrou níveis basais e máximos de LH e FSH, respectivamente, iguais a 0,6/2,1 e < 1,0/< 1,0 U/L. A análise molecular identificou uma nova mutação que consiste da deleção do códon 372 (AAC; asparagina) no éxon 1 do gene DAX1. Essa mutação não foi encontrada em 200 alelos de indivíduos normais. A análise no site PredictProtein indicou que essa alteração, localizada no domínio de ligação do DAX1, pode danificar a proteína. Nossa hipótese é que essa nova mutação (p.Asp372del) do gene DAX1 pode levar a uma alteração na função da proteína DAX1 e está provavelmente envolvida no desenvolvimento da HAC e HH nesse paciente. Arq Bras Endocrinol Metab. 2012;56(8):496-500

We report a case of adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism (HH) due to a novel DAX1 mutation. A 19-month-old boy with hyperpigmentation and failure to thrive came to our service for investigation. Three brothers of the patient had died due to adrenal failure, and a maternal cousin had adrenal insufficiency. Adrenoleukodystrophy was excluded. MRI showed normal pituitary and hypothalamus. Plasma hormone evaluation revealed high ACTH (up to 2,790 pg/mL), and low levels of androstenedione, DHEA-S, 11-deoxycortisol, and cortisol. At 14 years of age the patient was still prepubescent, his weight was 43.6 kg (SDS: -0.87) and his height was 161 cm (SDS: -0.36), with normal body proportions. In the GnRH test, basal and maximum values of LH and FSH were respectively 0.6/2.1 and < 1.0/< 1.0 U/L. Molecular investigation identified a novel mutation that consists of a deletion of codon 372 (AAC; asparagine) in exon 1 of DAX1. This mutation was not found in a study of 200 alleles from normal individuals. Prediction site analysis indicated that this alteration, located in the DAX1 ligand-binding domain, may damage DAX1 protein. We hypothesize that the novel (p.Asp372del) DAX1 mutation might be able to cause a disruption of DAX1 function, and is probably involved in the development of AHC and HH in this patient. Arq Bras Endocrinol Metab. 2012;56(8):496-500

INTRODUÇÃO: O Acidente Vascular Encefálico (AVE) constitui uma das principais causas de internações e mortalidade, causando, em cerca de 90% dos sobreviventes, algum tipo de deficiência, seja ela parcial ou total. Os comprometimentos funcionais variam de um indivíduo para o outro e o desempenho das habilidades de atividades de vida diária (AVD) são fortemente prejudicados. OBJETIVOS: Avaliar a independência funcional de indivíduos na fase crônica após AVE e verificar a sua relação com a realização de tratamento fisioterapêutico. MATERIAIS E MÉTODOS: A amostra constou de 69 hemiparéticos crônicos com média de idade de 64-65 anos. Para avaliar a funcionalidade, utilizou-se a Medida de Independência Funcional (MIF) e foram determinados os efeitos "chão" e "teto". A análise estatística incluiu o teste de normalidade de Kolmogorov-Smirnov, média e desvio-padrão, e o teste de Mann-Whitney. RESULTADOS: Segundo o domínio motor da MIF, o item "controle de esfíncteres (fezes)" apresentou o maior número de indivíduos realizando de forma totalmente independente (88,4%) e o item "subir e descer escadas" foi o que obteve menor escore. Todos os itens do domínio cognitivo obtiveram médias superiores a 6,4 pontos. Foi encontrado elevado efeito teto para ambos os domínios, motor e cognitivo. Apenas a dimensão transferências diferiu significativamente entre os que faziam e os que não faziam fisioterapia (p = 0,01). CONCLUSÃO De modo geral, não houve relação entre a independência funcional e a realização da fisioterapia. Entretanto, o elevado efeito teto pode ter interferido nos resultados, sugerindo limitação da MIF em discriminar os indivíduos avaliados.

INTRODUCTION: Stroke is a leading cause of hospitalization and mortality, causing some type of disability in approximately 90% of survivors, whether partial or total. The functional impairment varies from one individual to the other and the performance skills of activities of daily living (ADLs) are strongly affected. OBJECTIVES: To assess the functional independence of individuals in the chronic phase after stroke and to determine its relation with the treatment by physiotherapy. MATERIALS AND METHODS: The sample consisted of 69 chronic hemiparetic individuals with a mean age of 64-65 years old. In order to evaluate the functionality, the Functional Independence Measure (FIM) was used and the floor and ceiling effects were determined. Statistical analysis included the normality test of Kolmogorov-Smirnov, mean and standard deviation and the Mann-Whitney test. RESULTS: According to the FIM motor domain, the item "sphincter control (feces)" had the highest number of individuals performing totally independently (88.4%) and the item "going up and down the stairs" showed the lowest scores. All items in the cognitive domain had means higher than 6.4 points. Ceiling effect for both motor and cognitive areas was found. Only the transfer dimension differed significantly between those who did and those who did not do physical therapy (p = 0.01). CONCLUSION: Overall, there was no relation between functional independence and physical therapy. However, the ceiling effect may have influenced the results, suggesting MIF restriction in discriminating the evaluated individuals.

OBJETIVO: Estabelecer um padrão de crescimento tumoral (volume) em ratos Wistar submetidos ao modelo C6 de glioblastoma multiforme por meio de imagens de ressonância magnética para posterior verificação de redução de volume tumoral com a terapia de magnetohipertermia. MÉTODOS: Para o modelo C6, utilizamos ratos Wistar, machos, jovens, pesando entre 250 e 300 g. Após anestesiados (cetamina 55 mg/kg e xilazina 11 mg/kg) foram injetadas estereotaxicamente células tumorigênicas linhagem C6 suspensas em meio de cultura (10(5) células em 10 µL) no córtex frontal direito (coordenadas a partir do bregma: anteroposterior = 2,0 mm; látero-lateral = 3,0 mm; profundidade = 2,5 mm) com uma seringa Hamilton. No Grupo Controle, houve a injeção do meio de cultura sem as células. Posteriormente, foram feitas imagens mediante a técnica de imagem por ressonância magnética em 14, 21 e 28 dias após a injeção em um escâner de imagem por ressonância magnética 2.0 T (Bruker BioSpec, Germany). Para o exame, os animais foram anestesiados com cetamina 55 mg/kg e xilazina 11 mg/kg. Multifatias coronais foram adquiridas utilizando uma sequência spin-echo padrão com os seguintes parâmetros: TR/TE = 4,000 ms/67,1 ms, FOV = 3,50, Matrix 192, slice thickness = 0,4 mm e slice separation = 0 mm. RESULTADOS: A análise das imagens de ressonância magnética do tumor possibilitou a clara visualização da massa tumoral, sendo possível ainda estabelecer parâmetros de volume tumoral nos diferentes dias analisados. O volume de 14 dias após a indução do foi de 13,7 ± 2,5 mm³. Aos 21 dias, o volume alcançado foi de 31,7 ± 6,5 mm³ e, aos 28 dias, a massa tumoral atingiu 122,1 ± 11,8 mm³. CONCLUSÃO: Estes resultados mostraram a possiblidade de avaliação do volume tumoral no modelo C6 em ratos, o que possibilitará, no futuro, a aplicação da terapia de magnetohipertermia bem como verificação de seus resultados.

OBJECTIVE: The objective was to establish a pattern of tumor growth of the C6 model of glioblastoma multiform in Wistar rats via magnetic resonance imaging (MRI) for the subsequent verification of tumor volume reduction due to magnetic hyperthermia therapy. METHODS: Young male Wistar rats weighing between 250 and 300 g were used for the C6 model. After the rates were anesthetized (55 mg/kg ketamine and 11 mg/kg xylazine), C6 lineage tumorigenic cells suspended in culture medium (10(5) cells in 10 µl) were stereotaxically injected into the right frontal cortex (bregma coordinates: 2.0 mm anteroposterior, 3.0 mm laterolateral, and 2.5 mm depth) of the rats using a Hamilton syringe. For the control group, the rats were injected with culture medium without cells. MRI scans were performed at 14, 21, and 28 d after the injection using a 2.0 T MRI scanner (Bruker BioSpec, Germany). The animals were anesthetized with 55 mg/kg ketamine and 11 mg/kg xylazine before being examined. Coronal multilayers were acquired using a standard spin echo sequence with the following parameters: repetition/echo time = 4.000 ms/67.1 ms, field of view = 3.50, matrix = 192, slice thickness = 0.4 mm, and slice separation = 0 mm. RESULTS: The MRI analysis enabled a clear visualization of the tumor mass, and it was possible to establish the tumor volume parameters on the various days that were examined. The volume at 14 d after induction was 13.7 ± 2.5 mm³. On days 21 and 28, the tumor volumes were 31.7 ± 6.5 mm³ and 122.1 ± 11.8 mm³, respectively. CONCLUSION: These results demonstrated that it is possible to evaluate the C6 model tumor volume in rats, which will allow for the future implementation and verification of magnetic hyperthermia therapy.