Optimization of the Sybr Green real time PCR for the detection of Human Herpes Virus type 6 (HHV-6)

HHV-6 is the etiological agent of Exanthem subitum which is considered the sixth most frequent disease in infancy. In immuno-compromised hosts, reactivation of latent HHV-6 infection may cause severe acute disease. We developed a Sybr Green Real Time PCR for HHV-6 and compared the results with nested conventional PCR. A 214 pb PCR derived fragment was cloned using pGEM-T easy from Promega system. Subsequently, serial dilutions were made in a pool of negative leucocytes from 10-6 ng/µL (equivalent to 2465.8 molecules/µL) to 10-9 (equivalent to 2.46 molecules/µL). Dilutions of the plasmid were amplified by Sybr Green Real Time PCR, using primers HHV3 (5' TTG TGC GGG TCC GTT CCC ATC ATA 3)'and HHV4 (5' TCG GGA TAG AAA AAC CTA ATC CCT 3') and by conventional nested PCR using primers HHV1 (outer): 5'CAA TGC TTT TCT AGC CGC CTC TTC 3'; HHV2 (outer): 5' ACA TCT ATA ATT TTA GAC GAT CCC 3'; HHV3 (inner) and HHV4 (inner) 3'. The detection threshold was determined by plasmid serial dilutions. Threshold for Sybr Green real time PCR was 24.6 molecules/µL and for the nested PCR was 2.46 molecules/µL. We chose the Real Time PCR for diagnosing and quantifying HHV-6 DNA from samples using the new Sybr Green chemistry due to its sensitivity and lower risk of contamination.

Contribuição ao estudo do problema alimentar da Amazônia

Salientando os diversos aspectos a atender, para a solução racional do problema da alimentação na Amazônia, aliás já fixados em 1941 pela Comissão que traçou as linhas gerais de um plano de saneamento dessa vasta região, aludem os A.A. às realizações já empreendidas dentro do programa traçado, e que versaram apenas sôbre os hábitos alimentares de um grande núcleo de população e sôbre o valor nutritivo de alguns elementos pouco conhecidos da fauna e da flora locais. Abordam, à guisa de ensaio, neste trabalho, o ponto concernente ao planejamento de regimes adequados, que se adaptem tanto às exigências, como as possibilidades regionais. Frisam, então, de início, as bases racionais a que devem eles obedecer respeito não só à redução do total de calorias, fornecidas, nos seus 2/3, por hidratos de carbono e ao qual se subordinam as cotas das três principais vitaminas do complexo B,* como também a restrição, igualmente indicada, da taxa de proteínas; respeito, ainda, as cotas recomendáveis das vitaminas A e C e de cálcio, dando aí especial atenção ao detalhe da sua aproveitabilidade. Referem, de passagem, à conveniência de não se descurar do problema do ferro alimentar, em face das endemias reinantes na região e das dificuldades para fazer, artificialmente, o enriquecimento marcial dos regimes, já que, para instituí-los, partem do principio de ser vantajoso lançar mão de recursos de produção local, sem ficar em marcada dependência de grandes centres distribuidores regionais. Mostrando as dificuldades para a utilização, na escala desejada, da carne e leite de vaca, como artigos básicos de regime — e apontam, entre os percalços, os inerentes ao transporte e conservação desses alimentos — apresentam uma tabela básica, para o adulto em trabalho moderado, a qual lhe fornece 2.600 calorias diárias e obedece aos pontos fundamentais já aludidos. Nela figuram: os peixes, cujas variedades de pequeno porte poderão, com vantagem, ser consumidas fritas ou torradas, com espinhas; o amendoim; as verduras de produção econômica na Amazônia, incluídas na lista as ramas de batata doce, da mandioca e do inhame; essas raízes e tubérculos feculentos, de parceria com o cara; a farinha de mandioca, de grande uso na região; frutas, em que e, alias, rica a flora local; melado ou rapadura, como boa fonte de açucarados, cálcio e ferro; gorduras de origem animal e vegetal. Detêm-se, a propósito de cada um desses alimentos, sôbre o seu valor nutritivo e as possibilidades reais de produção local ou regional. Enumeram, por fim, vários outros, a que, similarmente, será possível recorrer, em maior ou menor escala — criação de animais domésticos, caças, carne e ovos de tartaruga, arroz, raízes, brotos de palmeiras, feijão de vara, castanhas de sapucaia, do caju e do Para — numa demonstração de ser possível a Amazônia valer-se, de muito, a si própria, no tocante à alimentação das suas populações.

Genetic diversity of a brazilian wine grape germplasm collection based on morphoagronomic traits

The objectives of this study were to evaluate the performance of cultivars, to quantify the variability and to estimate the genetic distances of 66 wine grape accessions in the Grape Germplasm Bank of the EMBRAPA Semi-Arid, in Juazeiro, BA, Brazil, through the characterization of discrete and continuous phenotypic variables. Multivariate statistics, such as, principal components, Tocher's optimization procedure, and the graphic of the distance, were efficient in grouping more similar genotypes, according to their phenotypic characteristics. There was no agreement in the formation of groups between continuous and discrete morpho-agronomic traits, when Tocher's optimization procedure was used. Discrete variables allowed the separation of Vitis vinifera and hybrids in different groups. Significant positive correlations were observed between weight, length and width of bunches, and a negative correlation between titratable acidity and TSS/TTA. The major part (84.12%) of the total variation present in the original data was explained by the four principal components. The results revealed little variability between wine grape accessions in the Grape Germplasm Bank of Embrapa Semi-Arid.

Agronomic and molecular traits of the 'Italia Muscat' clone in Sao Francisco River Valley

The objective of this study was to evaluate agronomic and molecular traits of the 'Italia Muscat' clone and compare it with the cv. 'Italia', providing information to support the cultivation of 'Italia Muscat' this cultivar in the São Francisco River Valley. Agronomic characteristics of both clones were evaluated for two seasons in 2004. The characteristics were phenology, bud break (%), bud fertility (%), yield (kg) mass of bunches (g), length and width of bunches (cm), mass of berries (g), length and diameter of berries (mm), TSS (ºBrix), ATT (% titratable acidity) and TSS/TTA. Molecular analysis of seven SSR markers were carried out. The clone 'Italia Muscat' showed larger berries, mass of bunches and better TSS/TA ratio than 'Italia'. The molecular analysis resulted in the same allelic profile in both clones, highlighting the need to use a larger number of microsatellite markers or other molecular technique to allow their discrimination. Based on their morpho-agronomic characteristics, 'Italia Muscat' seems to be a good table grape cultivar alternative for grape growers of São Francisco River Valley.

Ciclização do lapachol induzida por sais de tálio III

This work describes the cyclization of lapachol (1) induced by thallium triacetate (TTA) and thallium trinitrate (TTN) in several solvents using magnetic stirring and under microwave irradiation. alpha-Xyloidone (2) - dehydro-alpha-lapachone - was obtained as the main product in these reactions in 20 75% yield. However, rhinacanthin-A (4) was isolated as main product in a 40% yield, using TTA and acetic anhydride:water (1:1) as solvent, and dehydro-iso-alpha-lapachone (3) in 21% yield, using TTA and dichloromethane as solvent. The reaction time decreased drastically under microwave conditions, but the yields of these reactions were not the expected.

Mapas conceptuales como herramienta pedagógica en la enseñanza de la química orgánica

Organic Chemistry is a branch of Chemistry involving the study of the carbon atom, its compounds and reactions. Numerous investigations carried out in the field of teaching and learning processes indicate that knowledge of a conceptual and explanatory type is not conveyed in a conventional manner, but rather each individual is constructed. Therefore, the proposal for this study was devised to help students achieve significant sustainable learning in the area of reactivity in organic chemistry, using the Concept Maps described by Novak (1998) as a teaching tool.

A NOVEL METHOD FOR THE DETERMINATION OF TRACE THORIUM BY DISPERSIVE LIQUID-LIQUID MICROEXTRACTION BASED ON SOLIDIFICATION OF FLOATING ORGANIC DROP

In this study, dispersive liquid-liquid microextraction based on the solidification of floating organic droplets was used for the preconcentration and determination of thorium in the water samples. In this method, acetone and 1-undecanol were used as disperser and extraction solvents, respectively, and the ligand 1-(2-thenoyl)-3,3,3-trifluoracetone reagent (TTA) and Aliquat 336 was used as a chelating agent and an ion-paring reagent, for the extraction of thorium, respectively. Inductively coupled plasma-optical emission spectrometry was applied for the quantitation of the analyte after preconcentration. The effect of various factors, such as the extraction and disperser solvent, sample pH, concentration of TTA and concentration of aliquat336 were investigated. Under the optimum conditions, the calibration graph was linear within the thorium content range of 1.0-250 µg L-1 with a detection limit of 0.2 µg L-1. The method was also successfully applied for the determination of thorium in the different water samples.

Adaptation of "Niagara Rosada" grape must to winemaking by partial cluster dehydration

This study aimed to verify the influence of partial dehydration of "Niagara Rosada" grape clusters in physicochemical quality of the pre- fermentation must. In Brazil, during the winemaking process it is common to need to adjust the grape must when the physicochemical characteristics of the raw material are insufficient to produce wines in accordance with the Brazilian legislation for classification of beverages, which establishes the minimum alcohol content of 8.6 % for the beverage to be considered wine. Therefore, given that the reduction in the water content of grape berries allows the concentration of chemical compounds present in its composition, especially the concentration of total soluble solids, we proceeded with the treatments that were formed by the combination of two temperatures (T1-37.1ºC and T2-22.9 ºC) two air speeds (S1: 1.79 m s-1 and S2: 3.21 m s-1) and a control (T0) that has not gone through the dehydration treatment. Analysis of pH, Total Titratable Acidity (TTA) were performed in mEq L-1, Total Soluble Solids (TSS) in ºBrix, water content on a dry basis and Concentration of Phenolic Compounds (CPC) in mg of gallic acid per 100g of must. The average comparison test identified statistically significant modifications for the adaptation of must for winemaking purposes, having the treatment with 22.9 ºC and air speed of 1.79 m s-1 shown the largest increase in the concentration of total soluble solids, followed by the second best result for concentration of phenolic compounds.

Diversidade e estrutura comunitária de cerrado sensu stricto em afloramentos rochosos no parque estadual dos Pireneus, Goiás

(Diversidade e estrutura comunitária de cerrado sensu stricto em afloramentos rochosos no Parque Estadual dos Pireneus, Goiás). Este estudo foi realizado no Parque Estadual dos Pireneus e tem como objetivo analisar a composição florística e a estrutura comunitária do componente lenhoso do cerrado sensu stricto sobre afloramentos de rochas e comparar os resultados com outros locais. A área localiza-se entre as coordenadas 15°48'42"-47" S e 48°52'40"-63" W, a 1.310 m de altitude média. Dez parcelas de 20 m × 50 m (1.000 m²) foram alocadas aleatoriamente onde mediu-se todos os indivíduos lenhosos com diâmetro do tronco igual ou superior a 5 cm, tomado a 30 cm do solo. O estudo registrou 65 espécies, pertencentes a 51 gêneros e 35 famílias. As famílias mais representativas quanto ao número de espécies foram: Myrtaceae (oito spp.), Fabaceae (sete spp.) e Melastomataceae (seis spp.). As doze primeiras espécies em VI foram responsáveis por mais de 50% da dominância e densidade totais. A densidade foi de 1.105 indivíduos ha-1 e a área basal 11,03 m² ha-1. O índice de diversidade de Shannon (H') foi de 3,65 nats ind-1e encontra-se dentro da faixa de variação de estudos em cerrado sensu stricto. Os agrupamentos das parcelas do parque gerados pela análise de classificação mostraram que este cerrado é composto por espécies características do local como Schwartzia adamantium (Cambess.) Bedell ex Giraldo-Cañas, Alchornea triplinervia (Spreng.) Müll. Arg. e Clusia burchellii Engl., espécies generalistas das fisionomias de cerrado e também espécies mais comuns em mata de galeria, e apresenta estrutura similar a esta vegetação em outras condições edáficas.

Comparação da vegetação arbustivo-arbórea de uma área de cerrado rupestre na Chapada dos Veadeiros, Goiás, e áreas de cerrado sentido restrito do Bioma Cerrado

Foram determinadas a composição florística e a estrutura da vegetação arbustivo-arbórea, incluindo monocotiledôneas, em uma comunidade de cerrado rupestre, no município Alto Paraíso de Goiás, Goiás. Os objetivos desse estudo foram: 1. Comparar os aspectos florísticos e estruturais da área estudada com aqueles de outros 14 estudos com comunidades arbustivo-arbóreas, com e sem a presença de monocotiledôneas, 2. Avaliar os padrões fitogeográficos das 15 áreas comparadas. Foram amostradas 71 espécies, sendo cinco espécies de monocotiledôneas (três Velloziaceae e duas Arecaceae). Considerando as monocotiledôneas, a comunidade inventariada foi a mais densa (1.977 indivíduos ha-1) e apresentou a segunda maior área basal (11,25/ m²/ ha-1), entre as áreas comparadas. No entanto, sem as monocotiledôneas a densidade e área basal reduziram para 892 indivíduos ha-1 e 7,55/ m²/ ha-1, respectivamente. Os índices de diversidade (H'/†=/ 2,81) e equabilidade (J'/†=/ 0,66) foram baixos com a presença das monocotiledôneas, mas se elevaram, com a exclusão das espécies desse grupo (H/ =/ 3,63, J'/†=/ 0,86). Os aumentos nos valores de riqueza, densidade e área basal, com a inclusão das monocotiledôneas indicaram elevada importância das famílias Velloziaceae e Arecaceae na comunidade de cerrado rupestre estudada. A flora foi representada predominantemente por espécies de cerrado sentido restrito sobre solos profundos. No entanto, cinco das dez espécies com maiores VI são consideradas habitat especialistas de cerrado rupestre (Vellozia variabilis Mart. ex Schult. f., Wunderlichia cruelsiana Taub., Schwartzia adamantium (Cambess.) Bedell ex Giraldo-Cañas, Hyptis pachyphylla Epling e Vellozia tubiflora (A. Rich.) Kunth). Além do mais, foi observada baixa similaridade florística entre a área de estudo e as áreas comparadas, sugerindo particularidade florística do cerrado rupestre de Alto Paraíso de Goiás. Os padrões fitogeográficos, revelados por análises de agrupamento (UPGMA) e ordenação (DCA), indicaram influência da altitude, da proximidade geográfica entre áreas e de suas localizações em relação aos biomas adjacentes ao Cerrado (Floresta Amazônica, Mata Atlântica e Caatinga).

Ripening pattern of guava cv. Pedro Sato

Guava is a fruit with high respiration rates and a very short shelf life. Since information on its respiration pattern is contradictory, the objective was to study the changes occurring in the fruit during ripening and to relate them to the respiration behavior of this fruit. Guavas were picked at the half-ripe stage and stored for 8 days at 22 ± 1 ºC and 78 ± 1% relative humidity. The analyses conducted were: peel and pulp coloration, firmness, total soluble solids (TSS), total titratable acidity (TTA), and ethylene production. According to the results, it was verified that the parameters analyzed apparently do not coincide and are ethylene-independent. There was an accentuated ethylene production during ripening, starting from the 4th day. The ethylene synthesis continued increasing up to the 8th day, when the fruits were already decomposing. It was observed that the firmness decreased sharply in the first three days of ripening, and the skin and pulp color changed during ripening. The TSS, total soluble solids, and the TTA, total titratable acidity, practically did not change during the ripening, even with the increased ethylene production. It can be concluded that guava is a fruit that presents characteristics of climacteric and non-climacteric fruits.

Chemical and physical characterization of mume fruit collected from different locations and at different maturity stages in São Paulo State

Prunus mume is widely studied due to its health benefits regarding increase of blood fluidity and consequent improvement of the cardiovascular system and the prevention or even the fight against different types of cancer. However, in Brazil this culture is found only among oriental descendants. The present study aimed to characterize mume fruit collected from three different locations in the State of São Paulo regarding general aspects such as pH, total titratable acidity (TTA), total soluble solids (TTS), pectin content and yield of pulp and chemical characteristics: total phenolic compounds (TPC) and antioxidant capacity. Mume fruit were collected unripe and analyzed until maturation about 88 days after flowering. Fruit collected in Botucatu came from a commercial mume fruit producer and had average weight of 16.9 g, while in fruit from other locations weight varied from 5.7-6.9 g. TSS ranged from 9.5 to 10.0 Brix, total solids was 10.2-12.2% and pH showed values between 2.5 and 2.7 for all locations. TTA expressed in citric acid decreased from 4.0-5.7 g (100g- 1) at unripe stage to 2.0-3.8 g (100g- 1) in mature-stage fruit. Pectin content decreased from 11.2 to 10.8% during fruit maturation, TPC content was 147-226 mg catechin (g- 1) on a dry matter basis and the antioxidant capacity was 96-169 µMol Trolox (g- 1) on a dry matter basis or 21-34 µMol Trolox (g- 1) on a wet matter basis.