Avaliacoes da infeccao de explantes de feijoeiro por Agrobacterium tumefaciens por meio de microscopia.

Phaseolus vulgaris L. e considerada recalcitrante a transformacao por Agrobacterium tumefaciens. Contudo, alteracoes no meio de co-cultivo, utilizacao de linhagens hipervirulentas de Agrobacterium e de vetores binarios contendo genes vir demostraram que o feijoeiro e susceptivel a essa bacteria. O objetivo do presente trabalho foi estudar o efeito da sonificacao nos tecidos vegetais de feijoeiro, bem como a penetracao da Agrobacterium nas camadas subepidermicas do tecido vegetal, usando a metodologia SAAT ("Sonification-Assisted Agrobacterium-mediated Transformation"). A variedade de feijoeiro utilizada foi a Olathe Pinto, a linhagem de A. tumefaciens foi LBA4404:pTOK.Os embrioes de feijao foram pre-tratados po 14 dias em meio de multibrotacao e, entao submetidos a sonificacao (de 0 ou 60 segundos) na presenca de Agrobacterium. Apos a inoculacao foram co-cultivados por 24 horas em meio liquido seguido de 48 horas em meio solido, ambos, contendo 20 m. L -1 de acetoceringona. Os explantes inoculados foram fixadas em solucao de Karnovsk para avaliacoes em microscopia optica e eletronica de varredura. As analises da microspia demostraram a presenca de rupturas na epiderme, quebras da parede celular e invasao da Agrobacterium nos tecidos subepidermicos. Os reultados demostraram que o metodo SAAT e uma tecnica viavel para a inoculcao de Agrobacterium em explantes de P. vulgaris.bitstream/CPAC-2010/23710/1/bolpd-04.pd

An Overview of the Obese-Asthma Phenotype in Children

Asthma is the most common chronic disease in childhood. Overweight and obesity are included among the comorbidities considered in patients with difficult-to-treat asthma, suggesting a specific phenotype of the disease. Therefore, the constant increase in obesity prevalence in children and adolescents raises concerns about the parallel increase of obesity-associated asthma. The possible correlation between obesity and asthma has been investigated over the last decade by different authors, who suggest a complex multifactorial relationship. Although the particular non-eosinophilic endotype of obesity-related asthma supports the concept that high body weight precedes asthma development, there is ongoing debate about the direct causality of these two entities. A number of mechanisms may be involved in asthma in combination with obesity disease in children, including reduced physical activity, abnormal ventilation, chronic systemic inflammation, hormonal influences, genetics and additional comorbidities, such as gastroesophageal reflux and dysfunctional breathing. The identification of the obesity-related asthma phenotype is crucial to initiate specific therapeutic management. Besides the cornerstones of asthma treatment, lifestyle should be optimized, with interventions aiming to promote physical exercise, healthy diet, and comorbidities. Future studies should clarify the exact association between asthma and obesity and the mechanisms underlying the pathogenesis of these two related conditions with the aim to define personalized therapeutic strategies for asthma management in this population

Carbon partitioning and export in transgenic Arabidopsis thaliana with altered capacity for sucrose synthesis grown at low temperature: a role for metabolite transporters

We investigated the role of metabolite transporters in cold acclimation by comparing the responses of wild-type (WT) Arabidopsis thaliana (Heynh.) with that of transgenic plants over-expressing sucrose-phosphate synthase (SPSox) or with that of antisense repression of cytosolic fructose-1,6-bisphosphatase (FBPas). Plants were grown at 23 degrees C and then shifted to 5 degrees C. We compared the leaves shifted to 5 degrees C for 3 and 10 d with new leaves that developed at 5 degrees C with control leaves on plants at 23 degrees C. At 23 degrees C, ectopic expression of SPS resulted in 30% more carbon being fixed per day and an increase in sucrose export from source leaves. This increase in fixation and export was supported by increased expression of the plastidic triose-phosphate transporter AtTPT and, to a lesser extent, the high-affinity Suc transporter AtSUC1. The improved photosynthetic performance of the SPSox plants was maintained after they were shifted to 5 degrees C and this was associated with further increases in AtSUC1 expression but with a strong repression of AtTPT mRNA abundance. Similar responses were shown by WT plants during acclimation to low temperature and this response was attenuated in the low sucrose producing FBPas plants. These data suggest that a key element in recovering flux through carbohydrate metabolism in the cold is to control the partitioning of metabolites between the chloroplast and the cytosol, and Arabidopsis modulates the expression of AtTPT to maintain balanced carbon flow. Arabidopsis also up-regulates the expression of AtSUC1, and to lesser extent AtSUC2, as down-stream components facilitate sucrose transport in leaves that develop at low temperatures.info:eu-repo/semantics/publishedVersio

Modulating MIOX2 expression in Nicotiana tabacum and impacts on genes involved in cell wall biosynthesis

Cell walls are essential structures for plant development and growth. Apart from its biological functions, the polysaccharides that make cell walls (cellulose, hemicellulose and pectins) are the principal natural fibrous materials, considered the most important renewable resource on earth, used as raw material for many industrial processes among them, for pulp and paper production, charcoal, and biofuels. For all these reasons, the study of molecular composition and biosynthesis of plant cell walls has been a matter of great interest for researchers over the past few years. In this context, a full-length cDNA fragment of Miox2 gene was cloned from Arabidopsis seedlings, using RT-PCR, with an open reading frame of 954 pb and a corresponding protein subunit molecular mass of 37 kDa. The deduced amino acid sequence of the cDNA showed a high degree of homology with myo-Inosytol oxygenases from other organisms. This cDNA was used for genetic transformation of model plants (tobacco), which expressed either antisense or sense RNA. Transgenic homozygous tobacco model plants with either repression or constitutively expressed Miox2 were obtained with the number of copies varying from 1 to 7. Neither, the repression of the endogenous tobacco Miox genes or the constitutive expression of Miox2 gene, caused major impact on plant development, leaf morphology or flowering time. There was however, statistically significant (P<0.05) changes in the arabinan and D-galacturonate contents. These results clearly indicate that the modulation of the myo-Inositol pathway caused no major impact on cell wall polysaccharide biosynthesis.As paredes celulares vegetais são estruturas essenciais para o crescimento e desenvolvimento das plantas. Além de suas diversas funções biológicas, os componentes polissacarídicos constituintes das paredes celulares (celulose, hemiceluloses e pectinas) são de vital importância como fonte natural de fibras, sendo consideradas as fontes principais de recursos renováveis do planeta, utilizados como matéria prima para diversos processos industriais, dentre eles, a produção de papel e celulose, carvão vegetal e biocombustíveis. Todos esses fatores têm despertado grande interesse no estudo da composição e biossíntese das paredes celulares. Neste contexto, um fragmento de cDNA do gene Miox2 foi clonado de plântulas de Arabidopsis, via RT-PCR, com uma região aberta de leitura de 954 pb e sua proteína com massa molecular de 37kDa. A sequência deduzida de aminoácidos do cDNA apresentou alto grau de identidade com mio-Inositol oxigenases de outros organismos. Este cDNA foi usado para transformação genética de plantas modelo (tabaco) que produziram RNA antisense ou sense. Plantas de tabaco homozigotas para o transgene com repressão ou expressão constitutiva do gene Miox2 foram obtidas com um número de cópias do transgene, variando de 1 a 7. A repressão do gene Miox de tabaco endógeno assim como a expressão constitutiva do gene Miox2 de Arabidopsis não causaram alterações no desenvolvimento, morfologia foliar ou tempo de florescimento das plantas. Entretanto, alterações estatisticamente significativas (P<0.05) ocorreram no conteúdo de arabinana e de D-galacturonato. Estes resultados indicam que a modulação do metabolismo do mio-Inositol não causou grandes impactos na biossíntese dos polissacarídeos da parede celular.Fil: Defávari Nascimento, D.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Conti, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Labate, Mônica T. V.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Gutmanis, Gunta. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Bertolo, Ana L. F.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: de Andrade, Alexander. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Bragatto, Juliano. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Pagotto, Luís Otávio. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Damin, Plínio. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Moon, David H.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Labate, Carlos A.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; Brasi

Cardiopulmonary exercise testing reflects similar pathophysiology and disease severity in heart failure patients with reduced and preserved ejection fraction

BACKGROUND: We are unaware of any previous investigation that has compared the relationship of key cardiopulmonary exercise testing (CPX) variables to various measures of pathophysiology between heart failure-reduced ejection fraction (HFrEF) and HF-preserved ejection fraction (HFpEF) cohorts that are well matched with respect to baseline characteristics and their exercise response, which is the purpose of the present study. METHODS: Thirty-four patients with HFpEF were randomly matched to 34 subjects with HFrEF according to age and sex as well as peak oxygen consumption (VO2), ventilatory efficiency (VE/VCO2 slope), and exercise oscillatory ventilation (EOV). In addition to CPX, patients also underwent echocardiography with tissue Doppler imaging (TDI) and assessment of N-terminal pro-B-type natriuretic peptide (NT-proBNP). RESULTS: When matched for age, sex, and CPX variables, the HFrEF and HFpEF cohorts had similar echocardiography with TDI and NT-proBNP values, indicating comparable disease severity. In addition, the correlations between key CPX measures (peak VO2 and VE/VCO2 slope) and echocardiography with TDI and NT-proBNP measures were similar between HFrEF and HFpEF groups. Of note, the correlation between the VE/VCO2 slope and pulmonary artery systolic pressure and NT-proBNP was highly significant in both groups (r\u2009 65\u20090.65, p\u2009<\u20090.01). Moreover, subjects with EOV in both groups had a significantly higher PASP ( 3c47 vs. 3c35\u2009mmHg, p\u2009<\u20090.05). CONCLUSIONS: The results of the current study indicate CPX equally represents disease severity in HFrEF and HFpEF patients. This is a novel finding supporting the key role of CPX in the clinical follow-up of HF patients irrespective of LVEF and cardiac phenotype

Modulating MIOX2 expression in Nicotiana tabacum and impacts on genes involved in cell wall biosynthesis

Cell walls are essential structures for plant development and growth. Apart from its biological functions, the polysaccharides that make cell walls (cellulose, hemicellulose and pectins) are the principal natural fibrous materials, considered the most important renewable resource on earth, used as raw material for many industrial processes among them, for pulp and paper production, charcoal, and biofuels. For all these reasons, the study of molecular composition and biosynthesis of plant cell walls has been a matter of great interest for researchers over the past few years. In this context, a full-length cDNA fragment of Miox2 gene was cloned from Arabidopsis seedlings, using RT-PCR, with an open reading frame of 954 pb and a corresponding protein subunit molecular mass of 37 kDa. The deduced amino acid sequence of the cDNA showed a high degree of homology with myo-Inosytol oxygenases from other organisms. This cDNA was used for genetic transformation of model plants (tobacco), which expressed either antisense or sense RNA. Transgenic homozygous tobacco model plants with either repression or constitutively expressed Miox2 were obtained with the number of copies varying from 1 to 7. Neither, the repression of the endogenous tobacco Miox genes or the constitutive expression of Miox2 gene, caused major impact on plant development, leaf morphology or flowering time. There was however, statistically significant (P<0.05) changes in the arabinan and D-galacturonate contents. These results clearly indicate that the modulation of the myo-Inositol pathway caused no major impact on cell wall polysaccharide biosynthesis.As paredes celulares vegetais são estruturas essenciais para o crescimento e desenvolvimento das plantas. Além de suas diversas funções biológicas, os componentes polissacarídicos constituintes das paredes celulares (celulose, hemiceluloses e pectinas) são de vital importância como fonte natural de fibras, sendo consideradas as fontes principais de recursos renováveis do planeta, utilizados como matéria prima para diversos processos industriais, dentre eles, a produção de papel e celulose, carvão vegetal e biocombustíveis. Todos esses fatores têm despertado grande interesse no estudo da composição e biossíntese das paredes celulares. Neste contexto, um fragmento de cDNA do gene Miox2 foi clonado de plântulas de Arabidopsis, via RT-PCR, com uma região aberta de leitura de 954 pb e sua proteína com massa molecular de 37kDa. A sequência deduzida de aminoácidos do cDNA apresentou alto grau de identidade com mio-Inositol oxigenases de outros organismos. Este cDNA foi usado para transformação genética de plantas modelo (tabaco) que produziram RNA antisense ou sense. Plantas de tabaco homozigotas para o transgene com repressão ou expressão constitutiva do gene Miox2 foram obtidas com um número de cópias do transgene, variando de 1 a 7. A repressão do gene Miox de tabaco endógeno assim como a expressão constitutiva do gene Miox2 de Arabidopsis não causaram alterações no desenvolvimento, morfologia foliar ou tempo de florescimento das plantas. Entretanto, alterações estatisticamente significativas (P<0.05) ocorreram no conteúdo de arabinana e de D-galacturonato. Estes resultados indicam que a modulação do metabolismo do mio-Inositol não causou grandes impactos na biossíntese dos polissacarídeos da parede celular.Fil: Defávari Nascimento, D.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Conti, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Labate, Mônica T. V.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Gutmanis, Gunta. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Bertolo, Ana L. F.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: de Andrade, Alexander. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Bragatto, Juliano. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Pagotto, Luís Otávio. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Damin, Plínio. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Moon, David H.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; BrasilFil: Labate, Carlos A.. Universidade do Sao Paulo. Escola Superior de Agricultura Luiz de Queiroz; Brasi

Self-consistent simulation of plasma scenarios for ITER using a combination of 1.5D transport codes and free-boundary equilibrium codes

Self-consistent transport simulation of ITER scenarios is a very important tool for the exploration of the operational space and for scenario optimisation. It also provides an assessment of the compatibility of developed scenarios (which include fast transient events) with machine constraints, in particular with the poloidal field (PF) coil system, heating and current drive (H&CD), fuelling and particle and energy exhaust systems. This paper discusses results of predictive modelling of all reference ITER scenarios and variants using two suite of linked transport and equilibrium codes. The first suite consisting of the 1.5D core/2D SOL code JINTRAC [1] and the free boundary equilibrium evolution code CREATE-NL [2,3], was mainly used to simulate the inductive D-T reference Scenario-2 with fusion gain Q=10 and its variants in H, D and He (including ITER scenarios with reduced current and toroidal field). The second suite of codes was used mainly for the modelling of hybrid and steady state ITER scenarios. It combines the 1.5D core transport code CRONOS [4] and the free boundary equilibrium evolution code DINA-CH [5].Comment: 23 pages, 18 figure

Vaccines in children with inflammatory bowel disease: Brief review

Incidence of inflammatory bowel diseases (IBDs), including Crohn’s disease (CD) and ulcerative colitis (UC), is increasing worldwide. Children with IBDs have a dysfunctional immune system and they are frequently treated with immunomodulating drugs and biological therapy, which significantly impair immune system functions and lead to an increased risk of infections. Vaccines are essential to prevent at least part of these infections and this explains why strict compliance to the immunization guidelines specifically prepared for IBD patients is strongly recommended. However, several factors might lead to insufficient immunization. In this paper, present knowledge on the use of vaccines in children with IBDs is discussed. Literature review showed that despite a lack of detailed quantification of the risk of infections in children with IBDs, these children might have infections more frequently than age-matched healthy subjects, and at least in some cases, these infections might be even more severe. Fortunately, most of these infections could be prevented when recommended schedules of immunization are carefully followed. Vaccines given to children with IBDs generally have adequate immunogenicity and safety. Attention must be paid to live attenuated vaccines that can be administered only to children without or with mild immune system function impairment. Vaccination of their caregivers is also recommended. Unfortunately, compliance to these recommendations is generally low and multidisciplinary educational programs to improve vaccination coverage must be planned, in order to protect children with IBD from vaccine-preventable diseases

Brivaracetam use in clinical practice: a Delphi consensus on its role as first add-on therapy in focal epilepsy and beyond

Background Antiseizure medications remain the cornerstone of treatment for epilepsy, although a proportion of individuals with the condition will continue to experience seizures despite appropriate therapy. Treatment choices for epilepsy are based on variables related to both the individual patient and the available medications. Brivaracetam is a third-generation agent antiseizure medication.Methods We carried out a Delphi consensus exercise to define the role of brivaracetam in clinical practice and to provide guidance about its use as first add-on ASM and in selected clinical scenarios. A total of 15 consensus statements were drafted by an expert panel following review of the literature and all were approved in the first round of voting by panelists. The consensus indicated different clinical scenarios for which brivaracetam can be a good candidate for treatment, including first add-on use.Results Overall, brivaracetam was considered to have many advantageous characteristics that render it a suitable option for patients with focal epilepsy, including a fast onset of action, favorable pharmacokinetic profile with few drug-drug interactions, broad-spectrum activity, and being well tolerated across a range of doses. Brivaracetam is also associated with sustained clinical response and good tolerability in the long term.Conclusions These characteristics also make it suitable as an early add-on for the elderly and for patients with post-stroke epilepsy or status epilepticus as highlighted by the present Delphi consensus