Caracterização do eliciador do fungo Mucor ramosissimus e estudo da supressão de sua atividade indutora em soja por fragmentos de polissacarideos pecticos

Orientador : Marcia Regina BragaDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: Mucor ramosissimus Samutsevitsch, um fungo sapróbio isolado do filoplano de rubiáceas nativas tropicais, contém em seus esporos um potente eliciador capaz de induzir a síntese de fitoalexinas em espécies de rubiáceas e em cotilédones de soja. Estudos prévios indicaram que a atividade eliciadora dessa molécula pode ser inibida por ação de fragmentos pécticos (oligogalacturonídeos) derivados de parede celular de plantas. Este trabalho teve por objetivos purificar e caracterizar parcialmente este eliciador, e analisar a possível supressão de sua atividade indutora da síntese de fitoalexinas em cotilédones de soja por ação de oligogalacturonídeos. Parâmetros de crescimento, esporulação e atividade indutora determinadas para o fungo crescido em meio de cultura mínimo demonstraram que o 9° dia de cultivo é o melhor para obtenção de moléculas eliciadoras ativas na indução de gliceolinas. A purificação do eliciador por precipitação etanólica, cromatografias de troca iônica e de afinidade resultou em aumento de sua atividade específica e indicou a presença de ligações do tipo ? em sua estrutura química. O tratamento do eliciador de M. ramosissimus com periodato de sódio aboliu sua capacidade indutora de fitoalexinas em soja, indicando que esta atividade é determinada pela porção glicídica de sua molécula. Análises da composição de açúcares realizadas por HPLC e por GC/MS revelaram a presença de ácido glucurônico e manose em maiores proporções, seguidos de glucose, galactose e traços de fucose. Análises de ligação glicosídica revelaram a ausência de unidades de glucose 3 e 3,6 ligadas, indicando que este eliciador difere estruturalmente do ?-glucano isolado de micélios do fungo Phytophthora sojae. Ensaios de competição, nos quais o eliciador de M. ramosissimus foi co-aplicado com oligogalacturonídeos, revelaram que estes fragmentos parecem inibir diferencialmente as rotas biossintéticas dos metabólitos induzidos em soja pelo eliciador do fungo sapróbio M. ramosissimus quando comparado ao ?-glucano obtido do patógeno P. sojae. Os dados obtidos indicam que as plantas possuem a capacidade de responder a presença de fungos através do reconhecimento de moléculas eliciadoras com estruturas e atividades distintas e que são derivadas tanto de fungos patogênicos como de sapróbios. A atividade dessas moléculas eliciadoras pode ser alterada pela presença de oligogalacturonídeosAbstract: Mucor ramosissimus Samutsevitsch is a saprophytic fungus isolated from the leaf surface of tropical Rubiaceae and that contains in its spores a potent elicitor of phytoalexin production in Rubiaceae species and soybean cotyledons. Previous results indicated that the eliciting activity of this elicitor could be suppressed by pectic fragments from plant cell walls (oligogalacturonides). The aims of this work were to purify and partially characterize the elicitor derived from spores of M. ramosissimus and analyze the suppression of its activity by oligogalacturonides. The 9th day of culture was the best to obtain the elicitor from autoclaved spores, considering growth parameters, sporulation, and inducing activity. The elicitor purification performed by ethanol precipitation and anion exchange and affinity chromatography, resulted in considerable increase in its specific activity, and indicated the presence of ?-linked sugar residues. The eliciting activity was abolished after treatment with sodium periodate, indicating that the sugar moiety is important for its activity. Sugar composition, analyzed by HPLC and GC/MS, revealed the predominance of glucuronic acid and mannose, followed by glucose, galactose and traces of fucose. Glycosyl-linkage analyses of the elicitor revealed the absence of 3 e 3,6-linked glucosyl residues indicating that the M. ramosissimus elicitor presents distinct structure and composition when compared to the ?-heptaglucoside obtained from Phytophthora sojae. The application of M. ramosissimus elicitor together with oligogalacturonides on soybean cotyledons resulted in inhibition of the biosynthetic pathways leading to the synthesis of secondary metabolites, this inhibition being different from that observed when the ?-glucan from P. sojae was used. Our results indicate that plants are able to respond to the presence of fungi by recognizing eliciting molecules with distinct structure and activity derived from pathogenic as well as from saprophytic species. The activity of these eliciting molecules can be altered by the presence of cell wall fragments derived of plant cell wallsMestradoBiologia CelularMestre em Biologia Celular e Estrutura

Mulheres extrativistas e inovação agroecológica: A experiência com o murumuru (Astrocaryum murumuru Mart.)

The work analyzes the development of technical-organizational experiences based on agroecology and its relationship with the sustainable use of natural resources by family farming. The Santo Antônio community was chosen as the research field due to its experience with the collection of oleaginous seeds such as “murumuru” (Astrocarym murumuru Mart.), for the production of biocosmetics. Activity that offers risks, due to the structure and location of the palm tree, is carried out mostly by women; offering economic autonomy in the face of “acai money”, predominantly male. Assuming an important role in the organization of work within the agricultural property and the role of a social agent for the dissemination of new knowledge and agroecological practices, rescuing and preserving a specimen that has already gone to extinction. El trabajo analiza el desarrollo de experiencias técnico-organizativas basadas en la agroecología y su relación con el aprovechamiento sustentable de los recursos naturales por parte de la agricultura familiar. La comunidad de Santo Antônio fue elegida como campo de investigación por su experiencia en la recolección de semillas oleaginosas como el “murumuru” (Astrocarym murumuru Mart.), para la producción de biocosméticos. Actividad que ofrece riesgos, por la estructura y ubicación de la palmera, es realizada en su mayoría por mujeres; ofreciendo autonomía económica frente al “dinero acai”, predominantemente masculino. Asumiendo un rol importante en la organización del trabajo dentro de la propiedad agropecuaria y el rol de agente social para la difusión de nuevos conocimientos y prácticas agroecológicas, rescatando y preservando un espécimen que ya se encuentra en extinción. O trabalho analisa o desenvolvimento de experiências técnico-organizacionais de base agroecológica e sua relação com o uso sustentável dos recursos naturais pela agricultura familiar. A comunidade Santo Antônio foi escolhida como campo de investigação por sua experiência com a coleta de sementes oleaginosas como o “murumuru” (Astrocarym murumuru Mart.), para a produção de biocosméticos. Atividade que oferece riscos, por conta da estrutura e localização da palmeira, é desenvolvida na sua maior parte por mulheres; oferecendo autonomia econômica diante ao “dinheiro do açaí”, preponderantemente masculino. Assumindo importante papel na organização do trabalho dentro da propriedade agrícola e papel de agente social de difusão de novos saberes e práticas agroecológicas, resgatando e preservando um espécime que já caminhou à extinção

The impact of adipose tissue–derived miRNAs in metabolic syndrome, obesity, and cancer

Obesity is a multifactorial and complex condition that is characterized by abnormal and excessive white adipose tissue accumulation, which can lead to the development of metabolic diseases, such as type 2 diabetes mellitus, nonalcoholic fatty liver disease, cardiovascular diseases, and several types of cancer. Obesity is characterized by excessive adipose tissue accumulation and associated with alterations in immunity, displaying a chronic low-grade inflammation profile. Adipose tissue is a dynamic and complex endocrine organ composed not only by adipocytes, but several immunological cells, which can secrete hormones, cytokines and many other factors capable of regulating metabolic homeostasis and several critical biological pathways. Remarkably, adipose tissue is a major source of circulating microRNAs (miRNAs), recently described as a novel form of adipokines. Several adipose tissue–derived miRNAs are deeply associated with adipocytes differentiation and have been identified with an essential role in obesity-associated inflammation, insulin resistance, and tumor microenvironment. During obesity, adipose tissue can completely change the profile of the secreted miRNAs, influencing circulating miRNAs and impacting the development of different pathological conditions, such as obesity, metabolic syndrome, and cancer. In this review, we discuss how miRNAs can act as epigenetic regulators affecting adipogenesis, adipocyte differentiation, lipid metabolism, browning of the white adipose tissue, glucose homeostasis, and insulin resistance, impacting deeply obesity and metabolic diseases. Moreover, we characterize how miRNAs can often act as oncogenic and tumor suppressor molecules, significantly modulating cancer establishment and progression. Furthermore, we highlight in this manuscript how adipose tissue–derived miRNAs can function as important new therapeutic targets

The impact of the adipose organ plasticity on inflammation and cancer progression

Obesity is characterized by chronic and low-grade systemic inflammation, an increase of adipose tissue, hypertrophy, and hyperplasia of adipocytes. Adipose tissues can be classified into white, brown, beige and pink adipose tissues, which display different regulatory, morphological and functional characteristics of their adipocyte and immune cells. Brown and white adipocytes can play a key role not only in the control of energy homeostasis, or through the balance between energy storage and expenditure, but also by the modulation of immune and inflammatory responses. Therefore, brown and white adipocytes can orchestrate important immunological crosstalk that may deeply impact the tumor microenvironment and be crucial for cancer establishment and progression. Recent works have indicated that white adipose tissues can undergo a process called browning, in which an inducible brown adipocyte develops. In this review, we depict the mechanisms involved in the differential role of brown, white and pink adipocytes, highlighting their structural, morphological, regulatory and functional characteristics and correlation with cancer predisposition, establishment, and progression. We also discuss the impact of the increased adiposity in the inflammatory and immunological modulation. Moreover, we focused on the plasticity of adipocytes, describing the molecules produced and secreted by those cells, the modulation of the signaling pathways involved in the browning phenomena of white adipose tissue and its impact on inflammation and cancer

Wound healing: comparative study in hypertensive rats untreated and treated with an angiotensin converting enzime inhibitor

BACKGROUND: We evaluated the influence of captopril on the skin wound healing process of hypertensive rats. METHODS: 111 rats were placed in 4 groups: normotensive control (N=30); hypertensive control (N=30), which received an oral daily dose of saline solution 0,9%; group experiment (N=31) was treated with 7.5mg/kg/day of captopril; and an aferition group (N=20) with 10 hypertensive and 10 normotensive animals in which arterial blood pressure was mesured in the aorta in the last day of the experiment. After 15 days of treatment, an skin incision of 4 cm was made in the animals. Samples of the dorsal wall scar were taken 4, 7 and 14 days after the last procedure. The wounds were excised and divided in 2 pieces. They were sent to tensiontrial and histological analysis. RESULTS: The aferition group showed mean arterial blood pressure of 82.5±7.55 mmHg in the normotensive animals and 150.5± 10.66 mmHg in the hypertensive ones. The resistance analysis showed that the scars of treated and untreated hypertensives were less resistant than those of normotensives rats in the initial days (p<0.05) and that on the 14th day the resistances became similar. There were no diferences among treated and untreated groups. Total collagen had higher density in normotensives rats throughout the study (p<0.05) and there were no diferences among treated and untreated hypertensive rats. Epitelization, inflammatory response and granulation tissue formation were similar in all groups. CONCLUSION: Captopril, doesn't modify the wound healing process in rats, being the differences due to hypertension.OBJETIVO: Reconhecer a interferência do captopril na cicatrização de feridas cutâneas de ratos hipertensos. MÉTODOS: Distribuíram-se 111 ratos em quatro grupos: controle normotenso (N=30); controle hipertenso (N=30), os quais receberam 1 ml/dia de solução de cloreto de sódio a 0.9% por via oral; grupo experimento (N=31), hipertensos que receberam 7,5mg/kg/dia de captopril e um grupo aferição (N=20), 10 hipertensos e 10 normotensos, nos quais aferiu-se a pressão na aorta abdominal, no último dia de experimento. Após 15 dias de medicação, fez-se uma incisão da pele e da tela subcutânea, na região médio-dorsal dos grupos I, II e III, seguida de síntese. Ressecaram-se as cicatrizes de 10 animais de cada grupo, no 4.º, 7.º e 14.º dias após a operação, que divididas em duas partes foram enviadas para a tensiometria e para análise histológica. RESULTADOS: A pressão arterial média de 83,18 ± 7,51 mmHg nos normotensos e 151,36 ± 10,51 mmHg nos hipertensos. As cicatrizes dos hipertensos tratados e não tratados eram menos resistentes que as dos normotensos, nos tempos iniciais (p<0,05) e que ao 14.º dia as resistências se igualaram. Não houve diferença entre o grupo tratado e o não tratado. A densidade de colágeno total foi maior nos normotensos em todos os tempos (p<0,05) e não houve diferença entre hipertensos tratados e não tratados. A epitelização, a reação inflamatória e a formação do tecido de granulação foi semelhante nos três grupos. CONCLUSÕES: O captopril, em ratos, não modifica a cicatrização, ficando as diferenças relacionadas à hipertensão.UNIFESP-EPMPUCPRUniversidade Tuiuti do ParanáThe National Hospital for Nervous Diseases Institute of Neurology Department of NeuropathologyUFSCUNIFESP, EPMSciEL

Eating disorders in the preoperative phase of bariatric surgery

Introdução: A cirurgia bariátrica é considerada o tratamento mais eficaz para a obesidade. Pacientes obesos possuem elevada prevalência de transtornos alimentares. Objetivos:  Avaliar a presença de transtornos alimentares e padrões alimentares em candidatos à cirurgia bariátrica, associando os transtornos alimentares aos dados sociodemográficos e clínicos. Métodos: Estudo retrospectivo de cunho documental, com base em prontuários eletrônicos de pacientes candidatos à cirurgia bariátrica. Foram obtidas variáveis como sexo, idade, estado civil, nível de escolaridade, ocupação, doenças crônicas não transmissíveis, índice de massa corporal (IMC), transtorno e padrão alimentar. Resultados: Dos 281 pacientes avaliados, foi detectado 26,7% de transtornos alimentares, sendo 10,3% transtorno de compulsão alimentar periódica, 6,6% de bulimia nervosa e 5,3% de síndrome do comer noturno. Os padrões alimentares específicos encontrados foram: glutões (46,6%), compulsivos alimentares em períodos de estresse (9,3%), comedores de doces (4%) e beliscadores (1,3%). O IMC variou de 35,38 a 59,03 kg/m² (44,37±5,89), com 100% do grupo apresentando Hipertensão Arterial Sistêmica e 23,3% com Diabetes Mellitus tipo 2 ou dislipidemia. Conclusões: Constatou-se baixa frequência de transtornos alimentares na amostra estudada, sendo o transtorno de compulsão alimentar periódica o mais observado. Doenças crônicas não transmissíveis foram associadas com a presença de transtornos alimentares. Dentre os padrões alimentares observados, os glutões foram os mais frequentes.Introduction: Bariatric surgery is considered the most effective obesity treatment. Obese patients have a high prevalence of eating disorders. Objectives:  Evaluate the occurrence of eating disorders and eating patterns in candidates for bariatric surgery and associate eating disorders with sociodemographic and clinical characteristics. Methods: A retrospective study was conducted using electronic charts of candidates for bariatric surgery. Data were collected on sex, age, marital status, schooling, occupation, non-communicable diseases, body mass index (BMI), eating disorders, and eating patterns. Results: Among the 281 patients evaluated, eating disorders were detected in 26.7%; 10.3% had binge eating disorder, 6.6% had bulimia nervosa, and 5.3% had the night-eating syndrome. The specific eating patterns were overeating (46.6%), binge eating during periods of stress (9.3%), eating sweets (4%), and snaking (1.3%). BMI ranged from 35.38 to 59.03 kg/m² (mean: 44.37 ± 5.89). All individuals (100%) had systemic arterial hypertension, and 23.3% had type 2 diabetes mellitus or dyslipidemia. Conclusions: The frequency of eating disorders was low in the sample studied, the most common of which was binge eating disorder. Non-communicable diseases were associated with eating disorders. Among the eating patterns observed, the most frequent was overeating

Type 1 low z AGN. I. Emission properties

We analyze the emission properties of a new sample of 3,579 type 1 AGN, selected from the SDSS DR7 based on the detection of broad H-alpha emission. The sample extends over a broad H-alpha luminosity L_bHa of 10^40 - 10^44 erg s^-1 and a broad H-alpha FWHM of 1,000 - 25,000 km s^-1, which covers the range of black hole mass 10^6<M_BH/M_Sun<10^9.5 and luminosity in Eddington units 10^-3 < L/L_Edd < 1. We combine ROSAT, GALEX and 2MASS observations to form the SED from 2.2 mic to 2 keV. We find the following: 1. The distribution of the H-alpha FWHM values is independent of luminosity. 2. The observed mean optical-UV SED is well matched by a fixed shape SED of luminous quasars, which scales linearly with L_bHa, and a host galaxy contribution. 3. The host galaxy r-band (fibre) luminosity function follows well the luminosity function of inactive non-emission line galaxies (NEG), consistent with a fixed fraction of ~3% of NEG hosting an AGN, regardless of the host luminosity. 4. The hosts of lower luminosity AGN have a mean z band luminosity and u-z colour which are identical to NEG with the same redshift distribution. With increasing L_bHa the AGN hosts become bluer and less luminous than NEG. The implied increasing star formation rate with L_bHa is consistent with the relation for SDSS type 2 AGN of similar bolometric luminosity. 5. The optical-UV SED of the more luminous AGN shows a small dispersion, consistent with dust reddening of a blue SED, as expected for thermal thin accretion disc emission. 6. There is a rather tight relation of nuL_nu(2 keV) and L_bHa, which provides a useful probe for unobscured (true) type 2 AGN. 7. The primary parameter which drives the X-ray to UV emission ratio is the luminosity, rather than M_BH or L/L_Edd.Comment: 33 pages, 23 figures; accepted for publication in MNRAS; complete versions of tables 1 and B1 can be found at http://physics.technion.ac.il/~stern/PaperData/Type1AGN1