Global methylation in 'Valencia' orange seedlings associated with rootstocks and Huanglongbing.

Citrus farming is one of the main activities that contributed to the Brazilian trade balance, with citrus seedling being the most important input in the formation of orchards to guarantee high productivity and fruit quality, which fundamentally depends on the chosen genetics. The present study aimed to analyze the existence of epigenetic variability in 'Valencia' orange plants on rootstocks, associated or not with HLB, through the quantification of the global methylation of its genome, in order to support works on genetic improvement and crop production. For this purpose, this work was carried out in greenhouse in a completely randomized experimental design, with 5 treatments and 6 replicates per treatment, each seedling being considered a replicate, namely: T1 = "Valencia" orange grafted onto "Rangpur" lemon, inoculated with HLB; T2 = "Valencia" orange grafted onto "Swingle" citrumelo, inoculated with HLB; T3 = "Valencia" orange grafted onto "Rangpur" lemon, without HLB inoculation ; T4 = "Valencia" orange grafted onto "Swingle" citrumelo, without HLB inoculation ; T5 = "Valencia" orange in free standing. The DNA was extracted from leaves and the ELISA test (Enzyme-Linked Immunosorbent Assay) was carried out, based on the use of receptors sensitive to 5-mC., to measure the relative quantification of global methylation between genomic orange DNAs . Since the control treatment (T5) consists of "Valencia" orange in free standing, it could be inferred that both the normal grafting technique in the seedling formation process and the inoculation of buds infected with HLB are external factors capable of changing the methylation pattern in the evaluated plants, including the DNA demethylation process, causing an adaptive response in association with the expression of genes previously silenced by genome methylation.

Correlation of genome methylation of fig tree accessions with natural nematode and rust incidence.

Commercial fig tree cultivation in Brazil involves a single cultivar, 'Roxo-de-Valinhos'. The use of a single cultivar results in serious diseases and related problems. The aim of this study was to characterize fig accessions by analyzing the natural root-knot nematode and leaf rust incidence in relation to the epigenomic profile of the plant, since epigenetic variations affect plant-pathogen interactions. All plants were attacked by nematodes, indicating susceptibility; Meloidogyne incognita was the root-knot nematode species involved. Joint analysis of data showed that methylation and leaf rust incidence were correlated when observed in the same phenological phase, presenting initial evidence of the same factorial pressure loads in genotypes, suggesting similar behavior within these genotypes.

Effects of gibberellic acid concentration and fruit maturation stage on seed germination and vigor of pitahaya seedlings.

With increased interest in cultivation, the study of white-fleshed pitahaya (Selenicereus undatus (Haw.) D.R. Hunt, Cactaceae family) seedling production is of fundamental importance in the search for novel techniques to increase cultivation and guarantee homogeneous and productive orchards. The present study investigated the influence of various gibberellic acid (GA3) concentrations and fruit maturation stages on seed germination and vigor of white-fleshed pitahaya seedlings, considering the physiological quality of seedlings produced to support genetic breeding and conservation programs of the species. White-fleshed pitahaya seeds at two maturation stages (physiologically ripe and maintained at 10 °C in Biochemical Oxygen Demand incubators for three months) were treated with varying GA3 concentrations of 0, 50, 100, and 500 mg/L. We observed the influence of fruit storage on seedling germination, emergence, and growth as a function of GA3 concentration. According to the results, seeds extracted from ripe white-fleshed pitahaya fruits grown under the conditions tested here required GA3 application to increase seedling emergence and vigor, with optimal doses in the 150-300-mg/L range. In the case of pitahaya fruits intended for storage for future seed removal and maintained under the same sowing conditions, the application of higher doses of GA3 was necessary when compared to the previous condition, with a minimum dose of 500 mg/L GA3. The present study shows that the maturation stage of white-fleshed pitahaya fruits intended for seed removal influences the quality of seedlings; therefore, the use of seeds extracted from ripe pitahaya fruits without fermentation is more appropriate for the purpose.

Pulmonar ARFI elastography and ultrasonography of canine fetal hydrops: case report / Elastografia ARFI e ultrassonografia pulmonar de fetos com hidropsia fetal canina: relato de caso

This case report describes diagnostic and prognostic applicability of pulmonary acoustic radiation force impulse elastography and ultrasonography in canine hydrops fetalis. We also explore these methods' potential in prediction of postnatal respiratory dysfunction. Two pregnant bitches (English bulldog [case 1] and French bulldog [case 2]) were referred for sonographic evaluation in their last week of pregnancy. Ultrasound showed that in each bitch, one fetus presented with lung alterations (hyperechogenicity, irregular surface, and pleural effusion) and anasarca. The other fetuses of the litter were normal, and they were observed as light gray and dark blue on pulmonary elastography. Their shear-wave velocity was 0.75m/s. Fetuses with hydrops were observed as medium gray and dark blue, and the average shear-wave velocities were 1.05m/s (case 1) and 1.12m/s (case 2). Findings were compatible with increased lung rigidity. Six neonates of English bulldog and two of French bulldog showed no signs of clinical abnormalities during neonatal assessment. One puppy in each gestation presented with anasarca and respiratory distress, and died approximately 24 hours after birth. Novel ultrasound techniques (elastography) for assessing pulmonary tissues in abnormal fetuses in veterinary obstetrics can promote early, safe, and non-invasive diagnosis of canine prenatal and neonatal alterations.(AU)

Este relato de caso descreve a aplicabilidade diagnóstica e prognóstica da elastografia ARFI e ultrassonografia pulmonar em fetos caninos com hidropisia, como um método potencial para predizer a disfunção respiratória pós-natal nesses conceptos. Duas cadelas gestantes (Buldogue Inglês - caso 1 e Buldogue Francês - caso 2) foram encaminhadas para avaliação ultrassonográfica na última semana de gestação. Pela ultrassonografia foram observadas, em cada cadela, um feto apresentando alterações pulmonares (hiperecogenicidade, superfície irregular e derrame pleural) e anasarca. Outros fetos não apresentaram anormalidades. Os fetos normais apresentaram elastograma pulmonar cinza-claro e azul-escuro e velocidade de cisalhamento de 0,75m/s. Os fetos com hidropisia apresentaram tonalidades cinza-média e azul-escura e velocidade de cisalhamento de 1,05m/s (caso 1) e 1,12m/s (caso 2). Esses achados são compatíveis com o aumento da rigidez pulmonar. Seis neonatos Buldogue Inglês e dois Buldogues Franceses não mostraram sinais de anormalidades clínicas na avaliação neonatal. Um filhote de cada gestação apresentou anasarca e dificuldade respiratória, vindo a óbito cerca de 24 horas após o nascimento. O uso das novas técnicas de ultrassonografia (elastografia) para avaliação de tecidos pulmonares em feto anormal, em obstetrícia veterinária, pode promover o diagnóstico precoce, seguro e não invasivo de alterações pré-natais e neonatais em conceptos caninos.(AU)

Search for methylation-sensitive amplification polymorphisms in mutant figs.

Fig (Ficus carica) breeding programs that use conventional approaches to develop new cultivars are rare, owing to limited genetic variability and the difficulty in obtaining plants via gamete fusion. Cytosine methylation in plants leads to gene repression, thereby affecting transcription without changing the DNA sequence. Previous studies using random amplification of polymorphic DNA and amplified fragment length polymorphism markers revealed no polymorphisms among select fig mutants that originated from gamma-irradiated buds. Therefore, we conducted methylation-sensitive amplified polymorphism analysis to verify the existence of variability due to epigenetic DNA methylation among these mutant selections compared to the main cultivar 'Roxo-de-Valinhos'. Samples of genomic DNA were double-digested with either HpaII (methylation sensitive) or MspI (methylation insensitive) and with EcoRI. Fourteen primer combinations were tested, and on an average, non-methylated CCGG, symmetrically methylated CmCGG, and hemimethylated hmCCGG sites accounted for 87.9, 10.1, and 2.0%, respectively. MSAP analysis was effective in detecting differentially methylated sites in the genomic DNA of fig mutants, and methylation may be responsible for the phenotypic variation between treatments. Further analyses such as polymorphic DNA sequencing are necessary to validate these differences, standardize the regions of methylation, and analyze reads using bioinformatic tools.

Genetic characterization of fig tree mutants with molecular markers.

The fig (Ficus carica L.) is a fruit tree of great world importance and, therefore, the genetic improvement becomes an important field of research for better crops, being necessary to gather information on this species, mainly regarding its genetic variability so that appropriate propagation projects and management are made. The improvement programs of fig trees using conventional procedures in order to obtain new cultivars are rare in many countries, such as Brazil, especially due to the little genetic variability and to the difficulties in obtaining plants from gamete fusion once the wasp Blastophaga psenes, responsible for the natural pollinating, is not found in Brazil. In this way, the mutagenic genetic improvement becomes a solution of it. For this reason, in an experiment conducted earlier, fig plants formed by cuttings treated with gamma ray were selected based on their agronomic characteristics of interest. We determined the genetic variability in these fig tree selections, using RAPD and AFLP molecular markers, comparing them to each other and to the Roxo-de-Valinhos, used as the standard. For the reactions of DNA amplification, 140 RAPD primers and 12 primer combinations for AFLP analysis were used. The selections did not differ genetically between themselves and between them and the Roxo-de-Valinhos cultivar. Techniques that can detect polymorphism between treatments, such as DNA sequencing, must be tested. The phenotypic variation of plants may be due to epigenetic variation, necessitating the use of techniques with methylation-sensitive restriction enzymes.

[Oral health: a challenge for the health care team]. / Saúde oral: um desafio para a equipe de saúde.

The objective of this study is to reveal the knowledge of nursing and medicine students from a university in Rio Grande do Sul regarding the importance of promoting dental health. A questionnaire with three semi-structured questions was answered by 26 students from the nursing school, and by 32 students from the medical school. Through the method of triangulation it was concluded that these students had deficient knowledge regarding dental health. Since a small number of the population in Brazil has access to dental clinics, we consider important that nursing and medicine professionals know how to promote dental care among children.

Staphylococcus aureus meningitis in children: a review of 30 community-acquired cases.

In spite of the steady increase in the incidence of Staphylococcus aureus infections, it remains a relatively uncommon cause of meningitis. To our knowledge, no series of community-acquired S. aureus meningitis (CASAM) restricted to children has been published. So far in this retrospective study we report our experience with CASAM in children, hospitalized from 1983 to 1998 at Nossa Senhora da Glória Children's Hospital (HINSG). During the sixteen-year study period, 2,319 new cases of acute pyogenic meningitis were diagnosed at HINSG. Community-acquired S. aureus was identified as the causative agent in 30 patients (1.3 percent). The predominantly spinal localization of the agent is stressed. In contrast with publications which analyze adults, it has a better prognosis.

Poliomielite no Espirito Santo,Brasil epidemia de 1979. / Poliomyelitis in Espirito Santo, Brazil; epidemic in 1979

A evolucao da poliomielite foi revista desde 1965, com uma analise de 107 pacientes, originarios da epidemia de 1979. O surto teve inicio em janeiro, atingiu o maximo em abril, e o controle efetivo so foi obtido em dezembro. Laboratorialmente foi confirmado o poliovirus tipo 1, na maioria dos casos. Como medida de controle realizaram-se quatro campanhas em massa, vacinando as criancas entre dois meses e cinco anos, com a vacina oral viva (Sabin trivalente)