Maternal obesity in mice exacerbates the allergic inflammatory response in the airways of male offspring

It was previously demonstrated that non-allergen-sensitized rodents born to mothers exposed to a high-fat diet (HFD) spontaneously develop lower respiratory compliance and higher respiratory resistance. In the present study, we sought to determine if mice born to mothers consuming HFD would exhibit changes in inflammatory response and lung remodeling when subjected to ovalbumin (OVA) sensitization/challenge in adult life. Mice born to dams consuming either HFD or standard chow had increased bronchoalveolar lavage (BAL) levels of IL-1 beta, IL-4, IL-5, IL-10, IL-13, TNF-alpha and TGF-beta 1 after challenge with OVA. IL-4, IL-13, TNF-alpha and TGF-beta 1 levels were further increased in the offspring of HFD-fed mothers. Mice born to obese dams also had exacerbated values of leukocyte infiltration in lung parenchyma, eosinophil and neutrophil counts in BAL, mucus overproduction and collagen deposition. The programming induced by maternal obesity was accompanied by increased expression of miR-155 in peripheral-blood mononuclear cells and reduced miR-133b in trachea and lung tissue in adult life. Altogether, the present data support the unprecedented notion that the progeny of obese mice display exacerbated responses to sensitization/challenge with OVA, leading to the intensification of the morphological changes of lung remodeling. Such changes are likely to result from long-lasting changes in miR-155 and miR-133b expression1112CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçãosem informação2017/20742-2; 2016/22722-6; 2015/18997-7; 2013/07607-8; 2017/15175-

Energy Metabolism in H460 Lung Cancer Cells: Effects of Histone Deacetylase Inhibitors

BACKGROUND: Tumor cells are characterized by accelerated growth usually accompanied by up-regulated pathways that ultimately increase the rate of ATP production. These cells can suffer metabolic reprogramming, resulting in distinct bioenergetic phenotypes, generally enhancing glycolysis channeled to lactate production. In the present work we showed metabolic reprogramming by means of inhibitors of histone deacetylase (HDACis), sodium butyrate and trichostatin. This treatment was able to shift energy metabolism by activating mitochondrial systems such as the respiratory chain and oxidative phosphorylation that were largely repressed in the untreated controls. METHODOLOGY/PRINCIPAL FINDINGS: Various cellular and biochemical parameters were evaluated in lung cancer H460 cells treated with the histone deacetylase inhibitors (HDACis), sodium butyrate (NaB) and trichostatin A (TSA). NaB and TSA reduced glycolytic flux, assayed by lactate release by H460 cells in a concentration dependent manner. NaB inhibited the expression of glucose transporter type 1 (GLUT 1), but substantially increased mitochondria bound hexokinase (HK) activity. NaB induced increase in HK activity was associated to isoform HK I and was accompanied by 1.5 fold increase in HK I mRNA expression and cognate protein biosynthesis. Lactate dehydrogenase (LDH) and pyruvate kinase (PYK) activities were unchanged by HDACis suggesting that the increase in the HK activity was not coupled to glycolytic flux. High resolution respirometry of H460 cells revealed NaB-dependent increased rates of oxygen consumption coupled to ATP synthesis. Metabolomic analysis showed that NaB altered the glycolytic metabolite profile of intact H460 cells. Concomitantly we detected an activation of the pentose phosphate pathway (PPP). The high O(2) consumption in NaB-treated cells was shown to be unrelated to mitochondrial biogenesis since citrate synthase (CS) activity and the amount of mitochondrial DNA remained unchanged. CONCLUSION: NaB and TSA induced an increase in mitochondrial function and oxidative metabolism in H460 lung tumor cells concomitant with a less proliferative cellular phenotype

Mielomeningocele e anomalias associadas: uma série de casos e revisão sistemática

A mielomeningocele é uma malformação congênita grave do sistema nervoso central, representando um dos tipos mais complexos de defeitos do tubo neural. A condição é marcada pela exposição das meninges e, em alguns casos, da medula espinhal, através de uma abertura na coluna vertebral, desafiando tanto o prognóstico do paciente quanto as estratégias de tratamento. O presente estudo visa explorar os avanços recentes no diagnóstico, intervenções cirúrgicas e desfechos neurológicos associados à mielomeningocele, com foco particular na eficácia e segurança das abordagens atuais. Para isso, foi realizada uma revisão sistemática da literatura de 2016 a 2024 nas bases de dados PubMed (Medline), Cochrane Library e SciELO, aplicando critérios de inclusão e exclusão rigorosos para selecionar estudos que abordassem esses aspectos. Três estudos chave foram analisados, destacando-se pelas técnicas de diagnóstico antenatal, pela utilização de intervenções cirúrgicas inovadoras, como o fechamento fetal, e pelos cuidados pós-operatórios visando melhorias nos desfechos neurológicos. Os resultados indicam uma tendência positiva na melhoria da qualidade de vida dos pacientes, com avanços significativos no controle e na prevenção de complicações a longo prazo. No entanto, foi observada a necessidade de uma abordagem multidisciplinar no tratamento, integrando cuidados neurocirúrgicos, ortopédicos e reabilitativos. A mielomeningocele, apesar dos desafios, tem testemunhado progressos notáveis nas últimas décadas, sugerindo um futuro mais promissor para os pacientes afetados. Ainda assim, são necessários mais estudos para consolidar essas abordagens e otimizar as estratégias de tratamento na prática clínica

ATLANTIC-PRIMATES: a dataset of communities and occurrences of primates in the Atlantic Forests of South America

Primates play an important role in ecosystem functioning and offer critical insights into human evolution, biology, behavior, and emerging infectious diseases. There are 26 primate species in the Atlantic Forests of South America, 19 of them endemic. We compiled a dataset of 5,472 georeferenced locations of 26 native and 1 introduced primate species, as hybrids in the genera Callithrix and Alouatta. The dataset includes 700 primate communities, 8,121 single species occurrences and 714 estimates of primate population sizes, covering most natural forest types of the tropical and subtropical Atlantic Forest of Brazil, Paraguay and Argentina and some other biomes. On average, primate communities of the Atlantic Forest harbor 2 ± 1 species (range = 1–6). However, about 40% of primate communities contain only one species. Alouatta guariba (N = 2,188 records) and Sapajus nigritus (N = 1,127) were the species with the most records. Callicebus barbarabrownae (N = 35), Leontopithecus caissara (N = 38), and Sapajus libidinosus (N = 41) were the species with the least records. Recorded primate densities varied from 0.004 individuals/km 2 (Alouatta guariba at Fragmento do Bugre, Paraná, Brazil) to 400 individuals/km 2 (Alouatta caraya in Santiago, Rio Grande do Sul, Brazil). Our dataset reflects disparity between the numerous primate census conducted in the Atlantic Forest, in contrast to the scarcity of estimates of population sizes and densities. With these data, researchers can develop different macroecological and regional level studies, focusing on communities, populations, species co-occurrence and distribution patterns. Moreover, the data can also be used to assess the consequences of fragmentation, defaunation, and disease outbreaks on different ecological processes, such as trophic cascades, species invasion or extinction, and community dynamics. There are no copyright restrictions. Please cite this Data Paper when the data are used in publications. We also request that researchers and teachers inform us of how they are using the data. © 2018 by the The Authors. Ecology © 2018 The Ecological Society of Americ

Isolation, characterization and biological activity of a lectina from brazilian variety of lima beans seed (Phaseolus lunatus var. cascavel)

Lectins are proteins with the ability to interact specifically and reversibly with carbohydrates. They are present in all forms of life, performing vital functions for the maintenance of these beings. The legume lectins have numerous biological activities that can be used for therapy. The aim of this study was to isolate a lectin of Phaseolus lunatus seeds var. cascavel (PLUN), as well as to characterize it and check anti-hemolytic, antioxidant, antitumor and gastroprotective activities. The presence of lectins and an indication of their carbohydrate inhibitor was determined using the hemagglutination activity against human type ABO red blood cells. Protein concentration was determined by Bradford method using bovine serum albumin as standard. The isolation of PLUN was performed by size exclusion chromatography on Sephadex G-100. The weight and the purity of lectin was estimated by EGPA (polyacrylamide gel electrophoresis) under reducing and non-reducing conditions. The characterization was performed PLUN checking their resistance to pH and temperature variations, the presence front integrity of urea, DTT (dithiothreitol), beta-mercaptoethanol, EDTA (ethylenediaminetetra acetic acid), as well as the action of the trypsin and chymotrypsin enzymes . The characterization of glycoproteins was carried out by electrophoretic and staining for determination of total carbohydrates. The anti-hemolytic activity was performed using human ABO red blood cells, the antioxidant activity was determined by the molybdenum reduction methods (total antioxidant) and capture of the DPPH (2,2-diphenyl-1-picryl-hidrazila), and ABTS (2,2' sulfonic -azinobis-3-ethylbenzthiazoline-6 acid), antitumor activity was determined by the MTT (3- [4,5-dimethyl-thiazol-2-yl] -2,5-diphenyltetrazolium bromide) against cells melanoma A375 and the gastroprotective the protection induced gastropathy ethanol. The hemagglutinating activity indicated the presence of lectins only able to agglutinate erythrocytes of type A. This was inhibited only by lectin-carbohydrate N-acetyl-D-galactosamine at a concentration of 25 mM. The molecular exclusion was enough to isolate the lectin, where the peak I obtained from chromatography was active against type A red blood cells (PLUN). The estimated weight by EGPA indicated that it has about 128 kDa. The final purification yield was 80%, and this purified protein about 28 times. Thermostability and pH indicated variations in resistance to 80°C and pH between 2 and 11. Only the 8 M urea was able to inhibit the lectin. The 250 mM EDTA was inhibitory effect on hemagglutination activity was recovered in the presence of manganese ions. PLUN is a glycoprotein, with approximately 2% of carbohydrates in its composition. The lectin has no anti-hemolytic effect for no human blood type. PLUN has antioxidant action with AAEAA (μMAA/g) of 418.20, 326 and 82.9 for total antioxidant activity, ABTS radical capture and capture of DPPH radical, respectively. The IC50 (mg/ml) stood at 1.22, 0.21 and 4.39 for the tests cited above, respectively. The lectin has antitumor activity against melanoma derived cells at doses of 100 and 50 mg/ml, reducing up to 83% tumor cells, and gastroprotective action in a concentration of 1000 mg / kg, reducing up to 63% of the injured area stomach.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqLectinas são proteínas com a capacidade de interagir de forma específica e reversível com carboidratos. Estão presentes em todas as formas de vida desempenhando funções vitais. As lectinas de leguminosas apresentam inúmeras atividades biológicas que podem ser utilizadas pela terapêutica. O objetivo desse estudo foi isolar uma lectina de sementes de Phaseolus lunatus var. cascavel (PLUN), bem como caracterizá-la e verificar as atividades antihemolítica, antioxidante, antitumoral e gastroprotetora. A presença de lectinas e a indicação do seu carboidrato inibidor foi determinada utilizando a atividade hemaglutinante. A concentração proteica foi determinada pelo método de Bradford utilizando a albumina sérica bovina como padrão. O isolamento de PLUN foi realizado por cromatografia de exclusão em Sephadex G-100. O peso e a pureza da lectina foram estimados por EGPA (eletroforese em gel de poliacrilamida). A caracterização de PLUN foi realizada verificando sua resistência às variações de temperatura e pH, integridade frente à presença de ureia, ditiotreitol, beta-mercaptoetanol, EDTA, tripsina e quimiotripsina. A caracterização para glicoproteínas foi realizada pela coloração eletroforética e pela dosagem de carboidratos totais. A atividade antihemolítica foi realizada utilizando hemácias humanas ABO, a atividade antioxidante foi determinada pelos métodos de redução do molibdênio (antioxidante total) e captura dos radicais DPPH e ABTS, a atividade antitumoral foi determinada pelo ensaio do MTT contra células de melanoma A375 e a gastroprotetora pela proteção a gastropatia induzida por etanol. A atividade hemaglutinante indicou a presença de lectinas capazes de aglutinar apenas hemácias do tipo A. Essa lectina foi inibida apenas pelo carboidrato n-acetil-D-galactosamina em uma concentração de 25 mM. A exclusão molecular foi suficiente para isolar a lectina, onde o pico I obtido da cromatografia foi ativo contra hemácias tipo A (PLUN). A estimativa do peso por EGPA indicou que esta apresenta aproximadamente 128 kDa. O rendimento final da purificação ficou em 80%, sendo essa proteína purificada cerca de 28 vezes. A termoestabilidade e as variações de pH indicaram resistência em até 80ºC e em pH entre 2 e 11. Apenas a ureia 8 M foi capaz de inibir a lectina. O EDTA 250 mM teve efeito inibidor sobre a atividade hemaglutinante que foi recuperada em presença de íons manganês. PLUN é um glicoproteína, apresentando aproximadamente 2% de carboidratos em sua composição. A lectina não apresenta efeito antihemolítico para nenhum tipo sanguíneo humano. PLUN apresenta ação antioxidante com AAEAA (μMAA/g) de 418,20, 326 e 82,9 para a ação antioxidante total, captura do radical ABTS e captura do radical DPPH, respectivamente. As IC50 (mg/mL) ficaram em 1,22, 0,21 e 4,39 para os ensaios acima citados, respectivamente. A lectina apresenta atividade antitumoral contra células oriundas de melanoma em doses de 100 e 50 μg/mL, reduzindo em até 83% as células tumorais, e ação gastroprotetora em uma concentração de 1000 μg/kg, reduzindo em até 63% a área lesada do estômago. Os resultados indicam uma lectina com considerada atividades antioxidante, antitumoral e gastroprotetora. Mais estudos devem ser realizados para verificar possíveis mecanismos moleculares envolvidos nessas observações, bem como a determinação da estrutura tridimensional da proteína

Maternal obesity in mice programs the intensification of allergic response inflammation in the offspring

Orientador: Gabriel Forato AnhêTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: A obesidade materna durante a gestação e a lactação representa uma condição que pode acarretar inúmeras doenças na vida pós-natal da descendência à curto e longo prazo. Vias metabólicas, imunológicas e estruturais são modificadas negativamente na nova vida em formação sob a ação do excesso de gordura por parte da progenitora, incluindo as doenças de cunho respiratório, como a asma, que podem ser precipitadas na infância e na vida adulta. Esse trabalho se propôs a avaliar, utilizando modelos murinos de obesidade e asma experimental, se a presença de obesidade por parte da progenitora durante a gestação/lactação pode resultar no agravamento da resposta inflamatória alérgica da descendência masculina na vida adulta. Para isso foram utilizados camundongos C57Bl/6 que foram submetidos à engorda por dieta hiperlipídica intermitente por 6 semanas e durante o cruzamento, gestação e lactação. As proles de animais machos foram avaliadas para critérios inflamatórios na vida adulta comparando-se com proles de mães que ingeriram dieta normolipídica pelo mesmo período. A obesidade materna induziu na sua prole o aumento da migração leucocitária para o lavado broncoalveolar, estimulando o aumento do número de eosinófilos e neutrófilos. O mesmo resultado foi observado no tecido pulmonar, que além da leucocitose tecidual, demonstrou sofrer uma maior destruição da camada epitelial, hipertrofia e hiperplasia do músculo liso e espessamento da camada subepitelial. Os níveis de IgE, IL-4 e IL-13 também permaneceram elevados nesse grupo. Não foram observadas diferenças nos níveis de IL-5 e IL-10. Os filhos de obesas apresentaram um maior remodelamento pulmonar com aumento dos níveis de colágeno e TGB-?1, bem como, exacerbação da produção de muco e do número de células caliciformes. A elevação dos níveis de TNF-? e NF-?B fosforilado e a redução da expressão da proteína A20 podem indicar uma possível ativação do complexo inflamassoma. Não foram notadas diferenças significativas nos níveis de IL-1?. A expressão do miRNA-133b demonstrou estar diminuída na traquéia e no tecido pulmonar das proles de obesas, diferentemente do miRNA-155 que demonstrou estar elevado nos linfócitos periféricos do mesmo grupo. O conjunto de dados nos revelam que a obesidade materna pode comprometer severamente sua descendência, atuando na migração e ativação celular, na expressão de proteínas inflamatórias e regulando negativamente na expressão gênicaAbstract: Maternal obesity during gestation and lactation is a condition that can lead to numerous diseases in the postnatal life of the offspring in the short and long term. Metabolic, immunological and structural pathways are negatively modified in the new life in formation under the action of excess fat on the parent, including respiratory diseases, such as asthma, which may be precipitated in childhood and adulthood. The aim of this study was to evaluate, using murine models of obesity and experimental asthma, whether the presence of obesity by the mother during gestation/lactation may result in an aggravation of the allergic inflammatory response of the male offspring in adult life. C57B1/6 mice were submitted to fattening by an intermittent hyperlipidic diet for 6 weeks and during crossing, gestation and lactation. The offspring of male animals were evaluated for inflammatory criteria in adult life comparing with offspring of mothers who ate normolipidic diet for the same period. Maternal obesity induced in the offspring the increase of the leukocyte migration to the bronchoalveolar lavage, stimulating the increase in the number of eosinophils and neutrophils. The same result was observed in the lung tissue, which in addition to tissue leucocytosis, showed a high destruction of the epithelial layer, hypertrophy and smooth muscle hyperplasia and thickening of the subepithelial layer. Levels of IgE, IL-4 and IL-13 also remained elevated in this group. No differences were observed in IL-5 and IL-10 levels. The obese children presented a high pulmonary remodelling with increased levels of collagen and TGB-?1, as well as exacerbation of mucus production and number of goblet cells. Elevation of phosphorylated TNF-? and NF-?B levels and reduction of A20 protein expression may indicate a possible activation of the inflammasome pathway. No significant differences in IL-1? levels were noted. The expression of miRNA-133b has been shown to be decreased in the trachea and lung tissue of obese offspring, and the miRNA-155 which has been shown to be elevated in peripheral lymphocytes of the same group. The data set reveals that maternal obesity can severely compromise their offspring, acting on migration and cellular activation, on the expression of inflammatory proteins, and negatively regulating gene expressionDoutoradoFarmacologiaDoutor em Farmacologia2015/18997-7CAPESFAPES

Sex differences in the influence of obesity on a murine model of allergic lung inflammation

Despite the overwhelming evidence showing the influence of sex or obesity in the development of respiratory diseases in humans and animals, the mechanisms by which these combined two factors influence allergic asthma are not well understood. We have investigated the interaction between sex and weight gain in an experimental model of lung allergic inflammation induced by chicken egg ovalbumin (OVA) in mice. Animals were fed a high-fat diet for 8 weeks and then sensitized and challenged with OVA. Our results demonstrate that in comparison with males, high-fat diet (HFD) allergic female mice exhibit a reduction in the number of leucocytes in the lung lumen when challenged with OVA and, in contrast, an accumulation of these cells in the lung tissue. In addition, we also observed that allergic HFD female mice presented a robust lung remodelling in comparison with HFD males, evidenced by higher deposition of collagen in the airways and TGF-beta in lung fluid. Measuring epithelial adhesion molecule expression, we observed that female mice presented a significantly lower expression of CD103 than males in BAL cells, regardless of the diet. Similarly, HFD female mice express lower levels of EpCAM in lung tissue in comparison with males and lean females. Levels of A20/TNFAIP3 expression in lung tissue demonstrated that HFD female mice express lower levels of these regulatory factors than all the other groups. However, this reduction was not accompanied by an increase in activated NF-kappa B. Our results present evidence that the interaction between sex and weight gain alters the progression of allergic asthma in mice with females developing airway remodelling at a much earlier stage than males. These data may contribute to a better understanding of the clinical differences in the development and severity of allergic asthma observed between men and women of reproductive age502256266FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2015/18997-7; 2016/22722-

Acute appendicitis mimicking acute scrotum: a rare complication of a common abdominal inflammatory disease

Introduction: Acute appendicitis is the most common surgical disease of the abdomen in clinical practice, affecting mainly young adults. It has a wide variety of clinical presentations, due to the anatomical variation of the cecal appendix. Its presentation as acute scrotum and scrotal abscess is quite rare and atypical, occurring mainly in young male patients with patent processus vaginalis. Case presentation: An 18-years-old male patient attended the emergency unit complaining of diffuse abdominal pain, fever and hyporexia for four days followed by inflammatory signs in the scrotum. He was taken to the operation room after diagnosis of scrotal and abdominal sepsis. During scrotum exploration, pus was found inside the right hemiscrotum coming down from the groin and communicating with the abdominal cavity. The laparotomy found perforated appendicitis and peritonitis leading to the scrotal abscess. The abscess was drained, appendectomy was performed and the scrotal and abdominal cavity were washed with saline solution. Despite postoperative complications such as pneumonia and intra-abdominal abscess, the reported patient recovered and was discharged in the 44th postoperative day. Conclusion: Acute appendicitis can mimic acute scrotum and surgeons must have a high index of suspicion of this complication for diagnosing. This unusual clinical presentation may be challenging and can delay the diagnosis leading to perforated peritonitis. Resumo: Introdução: A apendicite aguda é a doença cirúrgica mais comum do abdome na prática clínica, afetando principalmente adultos jovens. Tem uma grande variedade de apresentações clínicas, devido à variação anatômica do apêndice cecal. Sua apresentação como escroto agudo e abscesso escrotal é bastante rara e atípica, ocorrendo principalmente em pacientes jovens do sexo masculino com túnica vaginalis patente. Apresentação do caso: um paciente do sexo masculino de 18 anos de idade compareceu à unidade de emergência queixando dor abdominal difusa, febre e hiporexia por quatro dias que se seguiram de sinais inflamatórios no escroto. Ele foi levado para centro cirúrgico após o diagnóstico de sepse de origem escrotal e abdominal. Durante a exploração da bolsa escrotal, secreção purulenta foi encontrada do lado direito oriunda do canal inguinal e comunicando-se com a cavidade abdominal. Durante a laparotomia observou-se sinais de apendicite aguda perfurada e peritonite levando ao abscesso escrotal. O abscesso foi drenado, a apendicectomia foi realizada e as cavidades escrotal e abdominal foram lavadas com solução salina. Apesar de complicações pós-operatórias como pneumonia e abscesso intra-abdominal, o paciente recuperou-se bem, recebendo alta hospitalar no 44° dia pós-operatório. Conclusão: A apendicite aguda pode simular o escroto agudo e os cirurgiões devem ter um alto grau de suspeição dessa complicação para o diagnóstico. Esta apresentação clínica incomum pode ser desafiadora e retardar o diagnóstico levando à peritonite por perfuração. Keywords: Acute appendicitis, Appendectomy, Acute abdomen, Spermatic cord torsion, Acute scrotum, Palavras-chave: Apendicite, Apendicectomia, Abdomen agudo, Torção do cordão espermático, Escroto agud