Resistência extrema a duas estirpes do Potato virus Y (PVY) de batata transgênica, cv. Achat, expressando o gene da capa protéica do PVYO

The coat protein (CP) gene of the potato virus Y strain “o” (PVYO) was introduced into potato, cultivar Achat, via Agrobacterium tumefaciens-mediated transformation. Sixty three putative transgenic lines were challenged against the Brazilian strains PVY-OBR and PVY-NBR. An extremely resistant phenotype, against the two strains, was observed in one line, denominated 1P. No symptoms or positive ELISA results were observed in 16 challenged plants from this line. Another clone, named as 63P, showed a lower level of resistance. Southern blot analysis showed five copies of the CP gene in the extremely resistant line and at least three copies in the other resistant line. The stability of the integrated transgenes in the extreme resistant line was examined during several in vitro multiplications over a period of three years, with no modification in the Southern pattern was observed. The stability of the transgenes, the absence of primary infections and the relatively broad spectrum of resistance suggest that the extremely resistant line obtained in this work can be useful for agricultural purposes.O gene da capa protéica (CP) do Potato virus Y estirpe “o”, foi introduzido em batata cultivar Achat, via Agrobacterium tumefaciens. Sessenta e três linhas possivelmente transgênicas foram desafiadas com as estirpes brasileiras PVY-OBR e PVY-NBR. Uma linha apresentou extrema resistência às duas estirpes inoculadas, e foi denominado clone 1P. Não foram observados sintomas sistêmicos de infecção e as plantas foram negativas em Elisa. Outra linha, denominada clone 63P, mostrou algum nível de resistência. Análises por Southern blot indicaram a presença de pelo menos cinco cópias do gen CP no clone 1P e pelo menos três cópias no clone 63P. A estabilidade do gene introduzido no clone 1P foi avaliada durante três anos, após várias multiplicações in vitro. Não foram observadas mudanças no padrão do Southern blot. A estabilidade do transgene, na ausência de infecções primárias e relativo largo espectro de resistência sugerem que o clone 1P pode ser utilizado para fins comerciais.Fil: Romano, Eduardo. Embrapa Recursos Genéticos; BrasilFil: Ferreira, Adriana T.. Embrapa Hortaliças; BrasilFil: Dusi, André N.. Embrapa Hortaliças; BrasilFil: Proite, Karina. Embrapa Recursos Genéticos; BrasilFil: Buso, Jose A.. Embrapa Hortaliças; BrasilFil: Avila, Antonio C.. Embrapa Hortaliças; BrasilFil: Nishijima, Marta L.. Embrapa Hortaliças; BrasilFil: Nascimento, Adriana S.. Embrapa Hortaliças; BrasilFil: Bravo Almonacid, Fernando Felix. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Mentaberry, Alejandro Nestor. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Monte, Damares. Embrapa Recursos Genéticos; BrasilFil: Campos, Magnolia A.. Embrapa Recursos Genéticos; BrasilFil: Melo, Paulo Eduardo. Embrapa Hortaliças; BrasilFil: Cattony, Monica K.. No especifica;Fil: Torres, Antonio C.. Embrapa Hortaliças; Brasi

Efeito do alumínio e de Glomus leptotichum Shenck e Smith no desenvolvimento de cultivares de Leucaena leucocephala (Lam.) de Wit tolerante e intolerante ao alumínio

A greenhouse experiment was carryed out to evaluate the mycorrhiza x toxic Al interaction in the substrate using a factorial design 2x2x2. Two cultivars of Leucaena leucocephala; a Al-tolerant (plant 49 - Estrada do Bongue) and another Al-intolerant (NO 749), were grown in washed sand and sterilized with addition of nutrient solution. The mycorrhiza was established with the inoculation of Glomus leptotichum Shenck e Smith and a control plot without VA fungi. It were evaluated two Al levels, 0 and 9 ppm, added to nutrient solution. The plants were harvested at 65 days after seedlings transplantation and shoot height, shoot dry matter weight, root colonization and shoot concentration of N, P, K,Ca, Mg and Al were determined. The following results can be formulated from the experiment herein analyzed: a), there was a plant growth increased and uptake of nutrient promoted by Glomus leptotichum, b) the Al-intolerant cultivar showed greater mycorrhizal dependence and were more benefit, presenting greater shoot dry matter weight and shoot nutrients acumulation. In these experimental conditions there was not significant difference between the Al levels used and also there was not interference in the cultivars growth.Com o objetivo de avaliar a interação micorrízica X Al tóxico no substrato efetuou-se um experimento fatorial 2x2x2, em casa de vegetação, no Departamento de Genética da ESALQ/USP, em Piracicaba. Foram empregadas duas cultivares de Leucaena leucocephala, uma Al - tolerante (Planta 49 - Estrada do Bongue) e outra Al-intolerante (NO749), cultivadas em areia lavada e esterilizada, com adição de solução nutritiva. A micorriza foi estabelecida com a inoculação de Glomus teptotichum Shenck e Smith no substrato, mantendo-se como testemunha a ausência do fungo. Dois níveis de Al foram avaliados, 0 e 9 ppm, adicionados à solução nutritiva. Aos 65 dias após o transplante das plântulas procedeu-se à colheita e foram determinados o peso da metária seca da parte aérea, altura de planta, porcentagem de colonização radicular, teor e quantidade acumulada de N, P, K, Ca, Mg e Al na parte aérea. Observou-se um acentuado efeito promotor da micorriza em Leucaena, o que geroulhe incrementos tanto no desenvolvimento da planta como na absorção de nutrientes. A cultivar intolerante ao Al tóxico mostrou-se mais dependente da micorriza e inclusive mais beneficiada,apresentando os maiores acréscimos na produção de matéria seca e acúmulo de nutrientes na parte aérea. Nas condições do experimento não houve acentuada diferença entre as doses de Al empregadas e, apesar do maior acúmulo do elemento em plantas micorrizadas, não foi observado interferência no desenvolvimento das cultivares

Pluronic® F127 Thermoresponsive Viscum album Hydrogel: Physicochemical Features and Cellular In Vitro Evaluation

Viscum album L., popularly known as mistletoe, is well known for its anti-cancer properties, and the pharmaceutical application of hydroalcoholic dry extracts is still limited due to its low solubility in aqueous media, and physicochemical instability. The Pluronic® F127 is an amphiphilic polymer, which permits the solubilization of lipophilic and hydrophilic compounds. In this investigation, physicochemical features of hydrogel containing V. album dry extract (VADE-loaded-hydrogel) were performed by: dynamic light scattering (DLS), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and transmission electron microscopy (TEM). VADE-loaded-hydrogel presented nanometer-size micelles with volume distribution ranging from 10.58 nm to 246.7 nm, and a polydispersity index of 0.441. The sample thermal analyses (TG and DSC) showed similar decomposition curves; however, the thermal events indicated an increase in thermal stability in relation to the presence of the extract. In addition to these interesting pharmaceutical features, IC50 values of 333.40 µg/mL and >1000 µg/mL were obtained when tumor (SCC-25) and non-tumor (L929) cells were incubated with VADE-loaded-hydrogel, respectively. The optical and ultrastructural cellular analysis confirmed the tumor selectivity since the following alterations were detected only in SCC-25 cells: disorganization of plasmatic membrane; an increase of cytoplasmatic vacuole size; alteration in the cristae mitochondrial shape; and generation of amorphous cellular material. These results emphasize the promising antitumoral potential of VADE-loaded-hydrogel as an herbal drug delivery system via in vitro assays

Metabolic syndrome, dyslipidemia, hypertension and type 2 diabetes in youth: from diagnosis to treatment

Overweight and obesity in youth is a worldwide public health problem. Overweight and obesity in childhood and adolescents have a substantial effect upon many systems, resulting in clinical conditions such as metabolic syndrome, early atherosclerosis, dyslipidemia, hypertension and type 2 diabetes (T2D). Obesity and the type of body fat distribution are still the core aspects of insulin resistance and seem to be the physiopathologic links common to metabolic syndrome, cardiovascular disease and T2D. The earlier the appearance of the clustering of risk factors and the higher the time of exposure, the greater will be the chance of developing coronary disease with a more severe endpoint. The age when the event may occur seems to be related to the presence and aggregation of risk factors throughout life

Confiabilidade da informação sobre antecedente de varicela em crianças pré-escolares

OBJECTIVE: To verify how reliable is the information provided by parents about the history of varicella in their children. METHODS: 204 parents of previously healthy children attending two municipal day-care centers of São Paulo city were interviewed between August 2003 and September 2005. A standardized form was filled out with information regarding age, sex, history of varicella and other diseases, drug use and antecedent of immunization, After medical history, physical examination and checking of immunization records, 5 ml of blood were collected for ELISA (in house) varicella test. Exclusion criteria were: age less than 1 year or more than 60 months, previous immunization against chickenpox, presence of co-morbidities or recent use of immunosuppressive drugs. Data were filed in a data bank using the Excel 2003 Microsoft Office Program and stored in a PC computer. The exact Fisher test was employed to calculate sensibility, specificity, positive and negative predictive values of history of varicella informed by children's parents. RESULTS: The age of the children varied from 12 to 54 months (median, 26 months; 49 (24%) children had positive history of varicella, 155 (76%) a negative or doubtful history. The predictive positive and negative values of the information were 90% and 93%, respectively (p = 0.0001). CONCLUSIONS: The degree of reliability of information about history of varicella informed by parents of children attending day care centers was high and useful to establish recommendations on varicella blocking immunization in day-care centers.OBJETIVOS: Verificar o grau de confiabilidade da informação fornecida pelos pais de crianças atendidas em creches sobre o antecedente de varicela. MÉTODOS: Os pais de 204 crianças previamente saudáveis matriculadas em duas creches da cidade de São Paulo foram entrevistados entre Agosto de 2003 e Setembro de 2005 para preenchimento de um questionário padronizado com informações sobre idade, sexo, antecedente de varicela e outras doenças, uso de medicamentos e história vacina. Após anamnese, exame físico e verificação da carteira de vacinação, foram colhidos 5 ml de sangue para realização do teste ELISA (in house) para varicela. Os critérios de exclusão foram: idade < 1 ano ou acima de 60 meses, vacinação prévia contra varicela, presença de comorbidades e uso recente de imunossupressores. Os dados foram registrados num banco de dados do programa Excel 2003 Microsoft Office e armazenados em microcomputador. Utilizou-se o teste exato de Fisher para calcular a sensibilidade, especificidade e valores preditivo positivo e negativo das informações fornecidas pelos pais sobre o antecedente de varicela das crianças. RESULTADOS: A idade das crianças variou entre 12 e 54 meses (mediana, 26 meses); 49 crianças (24%) tinham antecedente positivo para varicela e 155 (76%) negativo ou duvidoso. Os valores preditivo positivo e negativo da informação foram de 90% e 93%, respectivamente (p = 0.0001). CONCLUSÕES: O grau de confiabilidade das informações sobre varicela informadas pelos pais de crianças atendidas em creches foi elevado e útil para estabelecer recomendações para vacinação de bloqueio de surtos de varicela em creches

Polar lipids of commercial Ulva spp. of different origins: profiling and relevance for seaweed valorization

Macroalgae of the genus Ulva have long been used as human food. Local environmental conditions, among other factors, can have an impact on their nutrient and phytochemical composition, as well as on the value of the seaweed for food and non-food applications. This study is the first to initiate a comparison between commercial Ulva spp. from different European origins, France (FR, wild-harvested Ulva spp.), and Portugal (PT, farm-raised Ulva rigida), in terms of proximate composition, esterified fatty acids (FA), and polar lipids. The ash content was higher in PT samples, while FR samples had higher levels of proteins, lipids, and carbohydrates and other compounds. The profile of esterified FA, as well as FA-containing polar lipids at the class and species levels were also significantly different. The FR samples showed about three-fold higher amount of n-3 polyunsaturated FA, while PT samples showed two-fold higher content of monounsaturated FA. Quantification of glycolipids and phospholipids revealed, respectively, two-fold and three-fold higher levels in PT samples. Despite the differences found, the polar lipids identified in both batches included some lipid species with recognized bioactivity, valuing Ulva biomass with functional properties, increasing their added value, and promoting new applications, namely in nutraceutical and food markets.UIDB/50011/2020+UIDP/50011/2020, UID/QUI/00062/2019, UIDB/50006/2020, UIDB/50017/2020+UIDP/50017/2020, LISBOA-01-0145-FEDER-402-022125, POCI-01-0145-FEDER-030962, BPD/UI51/5041/2017, BPD/UI51/5042/2018; EC/H2020/727892/EUinfo:eu-repo/semantics/publishedVersio

The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody

Hemolytic Uremic Syndrome (HUS) associated with Shiga-toxigenic Escherichia coli (STEC) infections is the principal cause of acute renal injury in pediatric age groups. Shiga toxin type 2 (Stx2) has in vitro cytotoxic effects on kidney cells, including human glomerular endothelial (HGEC) and Vero cells. Neither a licensed vaccine nor effective therapy for HUS is available for humans. Recombinant antibodies against Stx2, produced in bacteria, appeared as the utmost tool to prevent HUS. Therefore, in this work, a recombinant FabF8:Stx2 was selected from a human Fab antibody library by phage display, characterized, and analyzed for its ability to neutralize the Stx activity from different STEC-Stx2 and Stx1/Stx2 producing strains in a gold standard Vero cell assay, and the Stx2 cytotoxic effects on primary cultures of HGEC. This recombinant Fab showed a dissociation constant of 13.8 nM and a half maximum effective concentration (EC50 ) of 160 ng/mL to Stx2. Additionally, FabF8:Stx2 neutralized, in different percentages, the cytotoxic effects of Stx2 and Stx1/2 from different STEC strains on Vero cells. Moreover, it significantly prevented the deleterious effects of Stx2 in a dose-dependent manner (up to 83%) in HGEC and protected this cell up to 90% from apoptosis and necrosis. Therefore, this novel and simple anti-Stx2 biomolecule will allow further investigation as a new therapeutic option that could improve STEC and HUS patient outcomes.Fil: Luz, Daniela. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Gomez, Fernando Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Ferreira, Raissa L.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Melo, Bruna S.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Guth, Beatriz E. C.. Universidade de Sao Paulo; BrasilFil: Quintilio, Wagner. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Moro, Ana Maria. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Presta, Agostina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Sacerdoti, Flavia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Ibarra, Cristina Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Chen, Gang. University of Toronto; CanadáFil: Sidhu, Sachdev S.. University of Toronto; CanadáFil: Amaral, María Marta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Fontes Piazza, Roxane María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentin

Soil biochemistry and microbial activity in vineyards under conventional and organic management at Northeast Brazil.

The São Francisco Submedium Valley is located at the Brazilian semiarid region and is an important center for irrigated fruit growing. This region is responsible for 97% of the national exportation of table grapes, including seedless grapes. Based on the fact that orgThe São Francisco Submedium Valley is located at the Brazilian semiarid region and is an important center for irrigated fruit growing. This region is responsible for 97% of the national exportation of table grapes, including seedless grapes. Based on the fact that organic fertilization can improve soil quality, we compared the effects of conventional and organic soil management on microbial activity and mycorrhization of seedless grape crops. We measured glomerospores number, most probable number (MPN) of propagules, richness of arbuscular mycorrhizal fungi (AMF) species, AMF root colonization, EE-BRSP production, carbon microbial biomass (C-MB), microbial respiration, fluorescein diacetate hydrolytic activity (FDA) and metabolic coefficient (qCO2). The organic management led to an increase in all variables with the exception of EE-BRSP and qCO2. Mycorrhizal colonization increased from 4.7% in conventional crops to 15.9% in organic crops. Spore number ranged from 4.1 to 12.4 per 50 g-1 soil in both management systems. The most probable number of AMF propagules increased from 79 cm-3 soil in the conventional system to 110 cm-3 soil in the organic system. Microbial carbon, CO2 emission, and FDA activity were increased by 100 to 200% in the organic crop. Thirteen species of AMF were identified, the majority in the organic cultivation system. Acaulospora excavata, Entrophospora infrequens, Glomus sp.3 and Scutellospora sp. were found only in the organically managed crop. S. gregaria was found only in the conventional crop. Organically managed vineyards increased mycorrhization and general soil microbial activity