Identification of suitable internal control genes for expression studies in Coffea arabica under different experimental conditions

Background: Quantitative data from gene expression experiments are often normalized by transcription levels of reference or housekeeping genes. An inherent assumption for their use is that the expression of these genes is highly uniform in living organisms during various phases of development, in different cell types and under diverse environmental conditions. To date, the validation of reference genes in plants has received very little attention and suitable reference genes have not been defined for a great number of crop species including Coffea arabica. The aim of the research reported herein was to compare the relative expression of a set of potential reference genes across different types of tissue/organ samples of coffee. We also validated the expression profiles of the selected reference genes at various stages of development and under a specific biotic stress.Results: The expression levels of five frequently used housekeeping genes (reference genes), namely alcohol dehydrogenase (adh), 14-3-3, polyubiquitin (poly), beta-actin (actin) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) was assessed by quantitative real-time RT-PCR over a set of five tissue/organ samples (root, stem, leaf, flower, and fruits) of Coffea arabica plants. In addition to these commonly used internal controls, three other genes encoding a cysteine proteinase (cys), a caffeine synthase (ccs) and the 60S ribosomal protein L7 (rpl7) were also tested. Their stability and suitability as reference genes were validated by geNorm, NormFinder and BestKeeper programs. The obtained results revealed significantly variable expression levels of all reference genes analyzed, with the exception of gapdh, which showed no significant changes in expression among the investigated experimental conditions.Conclusion: Our data suggests that the expression of housekeeping genes is not completely stable in coffee. Based on our results, gapdh, followed by 14-3-3 and rpl7 were found to be homogeneously expressed and are therefore adequate for normalization purposes, showing equivalent transcript levels in different tissue/ organ samples. Gapdh is therefore the recommended reference gene for measuring gene expression in Coffea arabica. Its use will enable more accurate and reliable normalization of tissue/organ-specific gene expression studies in this important cherry crop plant.

Th1/Th2 cytokines' expression and production by propolis-treated mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chronic caffeine intake increases androgenic stimuli, epithelial cell proliferation and hyperplasia in rat ventral prostate

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Priority areas for the conservation of Atlantic forest large mammals

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Friction losses in valves and fittings for liquid food products

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito do teor de soro, açúcar e de frutooligossacarídeos sobre a população de bactérias lácticas probióticas em bebidas fermentadas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversidade de ácaros (Acari, Arachnida) em seringueiras (Hevea brasiliensis Muell. Arg., Euphorbiaceae) na região noroeste do Estado de São Paulo, Brasil

The mites of three rubber tree cultures (Cedral, Pindorama and Taquaritinga) in order to determine the abundance of populations, the richness, the diversity and the degree of similarity among the communities was studied. Twenty one species were found, five of which were common to the three cultures. The richness and the abundance were greatest at the beginning of the dry season. The composition of communities differed probably as consequence of the kind of neighboring vegetation to each area, and because of the acaricid pulverization on the culture of Taquaritinga, reductng the richness of mite species in that area. The influence of neighboring vegetation can be shown by the occurrence of Iphiseiodes zuluagui Denmark & Muma, 1972, a common species to citrus trees, on neighboring rubber trees in Taquaritinga, and Tenuipalpus heveae Baker, 1945, a common species on rubber trees, on a coffee culture neighbor to the rubber trees of Pindorama. This data suggests that mites move among neighbor cultures, and can be an importam factor towards pest management and control. The diversity was small on the three cultures, as a result of the occurrence of one dominam species on each area, Calacarus heveae Feres, 1992 or T. heveae, which are considered pests of the rubber tree. The small diversity and the occurrence of dominant species are patterns expected in monocultures, systems with small environmental heterogeneity.

In vitro evaluation of dental bleaching effectiveness using hybrid lights activation

OBJETIVO: Avaliar se fontes de luz aumentam a eficácia do peróxido de hidrogênio na técnica de clareamento profissional. METODOLOGIA: Foram empregados 60 dentes incisivos bovinos, com dimensões coronárias e radiculares padronizadas a partir do limite amelo-cementário, sendo descartada a porção lingual. Os corpos-de-prova (cp) foram limpos em ultra-som por 20 min e a dentina condicionada com H3PO4 a 38% por 15 s, sendo os (cp) imersos em solução de café solúvel a 25% por duas semanas. A dentina foi impermeabilizada com esmalte e os (cp) divididos em 5 grupos, sendo a cor inicial mensurada através do espectofotômetro-EasyShade (VITA). Todos os (cp) receberam três aplicações por 10 min do gel clareador Opalescence Xtra-Boost (Ultradent) conforme segue: Grupo 1 - controle, não recebeu fotoativação, Grupo 2 - ativado com luz halôgena, Grupo 3 - ativado com LED azul/LASER, Grupo 4 - ativado com LED verde/LASER e Grupo 5 - ativado com LED vermelho. Após o clareamento foi mensurada a variação de cor E, a*, b*e L* e as referentes à escala de cor Vita Clássico. Os dados foram submetidos à análise de variância, teste de Tukey e de Dunn (α=5%). RESULTADOS: A diferença geral da cor foi reduzida quando se empregou LED Azul e Luz Halógena, sendo que o desempenho do peróxido de hidrogênio a 38% foi intensificado dependendo da fonte de luz utilizada. A avaliação quantitativa de cor, obtida por espectrofotômetro e pela escala de cor Vita Clássico, foram coincidentes. CONCLUSÃO: O tipo de fonte de luz empregada interfere na eficácia do agente clareador.

Assessment of the effectiveness of light-emitting diode and diode laser hybrid light sources to intensify dental bleaching treatment

Objective. To evaluate the effectiveness of the color change of hybrid light-emitting diode (LED) and low-intensity infrared diode laser devices for activating dental bleaching and to verify the occurrence of a color regression with time. Material and methods. A total of 180 specimens obtained from human premolars were immersed in a coffee solution for 15 days for darkening and then divided into eight experimental groups (n = 20 in each) as follows: G1, bleaching without light; G2, bleaching with halogen light; G3, bleaching with a blue LED (1000 mW/470 nm) and a laser device (120 mW/795 nm) simultaneously; G4, bleaching with an LED emitting blue light (1000 mW/470 nm); G5, bleaching with a blue LED (800 mW/470 nm) and a laser device (500 mW/830 nm) simultaneously; G6, bleaching with a blue LED device (800 mW); G7, bleaching with a green LED (600 mW/530 nm) and a laser device (120 mW/795 nm) simultaneously; and G8, bleaching with a green LED (600 mW). Three measurements were performed (at baseline and 14 days and 12 months after bleaching) using a Vita Easyshade spectrophotometer. The data were submitted to two-way ANOVA and a Tukey test. Results. All groups showed significantly higher Delta E values than Group G1, with the exception of Group G8. Variations in the Delta E values at 14 days were significant when compared with those obtained at baseline and after 12 months. Conclusions. Light activation of the bleaching gel provided faster and more intense bleaching than use of the bleaching gel without light activation. Combinations of low-intensity diode lasers are ineffective as a bleaching gel activator. Color regression was observed after 12 months of storage.

Influence of surfactants on the effectiveness of bleaching gels

This study evaluated the influence of surfactants on the effectiveness of 35% hydrogen peroxide (HP) and 10% carbamide peroxide (CP) bleaching gels. One hundred and forty bovine teeth were used, which were stained by immersion in a coffee, red wine, and tobacco mixture for 7 days. At the end of this process, the color measurement at baseline was taken with the Vita Easyshade spectrophotometer. The teeth were divided into seven groups: (a) negative control (NC), (b) positive control for HP (PC-35), (c) HP + Tween 20 (T20-35), (d) HP + laurel sodium sulfate (LSS-35), (e) positive control for CP (PC-10), (f) CP + Tween 20 (T20-10), and (g) CP + laurel sodium sulfate (LSS-10). Group NC was kept in artificial saliva for 21 days. Groups PC-35, T20-35, and LSS 35 received three applications of bleaching gel for 10 min; the process was repeated after 7 days. Groups PC-10, T20-10, and LSS-10 received the gel for 8 h per day for 14 days. After the bleaching process, the final color was measured. The analysis of variance and Tukey tests showed statistically significant differences for the parameters of a dagger L, a dagger b, and a dagger E of the HP gels with surfactant and positive control group (PC-35). Within the limits of this in vitro study, the addition of surfactants to HP bleaching gel increased the bleaching effectiveness.

The Effectiveness of Low-Intensity Red Laser for Activating a Bleaching Gel and Its Effect in Temperature of the Bleaching Gel and the Dental Pulp

Statement of the Problem: The effectiveness of low-intensity red laser for activating a bleaching gel and its effect in pulp temperature was not investigated in dental literature. Purpose: The objective of this study was to assess the effectiveness of low-intensity red laser for activating a bleaching gel, as well as its effect in temperature of the bleaching gel and the dental pulp. Materials and Methods: Forty extracted bovine teeth were immersed in a solution of coffee 14 days for darkening. The initial colors were recorded by spectrophotometric analysis. The specimens were randomly distributed into two groups (N = 20): the control, which did not receive light and the experimental group that received light from an appliance fitted with three red light-emitting laser diodes (? = 660 nm). A green-colored, 35% H2O2based bleaching gel was applied for 30 minutes, and changed three times. After bleaching, the colors were again measured to obtain the L*a*b* values. Color variation was calculated (?E) and the data submitted to the non-paired t-test (5%). To assess temperature, 10 human incisors were prepared, in which one thermocouple was placed on the bleaching gel applied on the surface of the teeth and another inside the pulp chamber. Results: There was a significant difference between the groups (p = 0.016), and the experimental group presented a significantly higher mean variation (7.21 +/- 2.76) in comparison with the control group (5.37 +/- 1.76). There was an increase in pulp temperature, but it was not sufficient to cause damage to the pulp. Conclusion: Bleaching gel activation with low-intensity red laser was capable of increasing the effectiveness of bleaching treatment and did not increase pulp temperature to levels deleterious to the pulp. CLINICAL SIGNIFICANCE The application of a low-intensity red laser was effective for activating a bleaching gel with green dye, without any deleterious increases in pulpal temperature. (J Esthet Restor Dent 24:126134, 2012)

The Influence of Chemical Activation on Tooth Bleaching Using 10% Carbamide Peroxide

The aim of this study was to assess the influence of manganese gluconate, a chemical activator of bleaching agents, at a concentration of 0.01% on the efficiency of a 10% carbamide peroxide-based bleaching agent. Forty bovine incisors were immersed in a 25% instant coffee solution for seven days and randomly divided into two groups. Group 1 was the control group and consisted of 10% carbamide peroxide-based bleaching gel only. Group 2 consisted of 10% carbamide peroxide-based bleaching gel and 0.01% manganese gluconate. Three readings of color were taken using the Vita Easy-shade spectrophotometer: the initial reading, a reading at seven days, and a reading at 14 days. Total color variation was calculated by Delta E*Lab. Data were submitted to the statistical t-test (5%), which showed that after seven days group 2 had a significant increase in the degree of tooth bleaching compared with group 1. The mean values (+/-SD) were 16.33 (+/-3.95) for group 1 and 19.29 (+/-4.97) for group 2. However, the results for group 1 and group 2 were similar after 14 days. Adding 0.01% manganese gluconate to 10% carbamide peroxide bleaching gel increased the degree of tooth bleaching after a seven-day treatment and did not influence the resulting shade after 14 days.

In Vitro Assessment of Chemical Activation Efficiency During In-office Dental Bleaching

Purpose: This study compared five types of chemical catalyzing agents added to 35% hydrogen peroxide gel, with regard to their capacity of intensifying in-office dental bleaching results.Methods: One-hundred and twenty bovine incisors were used, of which the crowns and roots were cut in the incisor-apical direction, to acquire the dimensions of a human central incisor. The specimens were sectioned in the mesiodistal direction by means of two longitudinal cuts, the lingual halves being discarded. The vestibular halves received prophylaxis with a bicarbonate jet, ultrasound cleaning and acid etching on the dentinal portion. Next, the specimens were stored in receptacles containing a 25% instant coffee solution for two weeks. After the darkening period, initial measurement of the shade obtained was taken with the Easy Shade appliance, which allowed it to be quantified by the CIELab* method. The samples were divided into six groups, corresponding to the chemical activator used: a) none (CON); b) ferric chloride (CF); c) ferrous sulphate (SF); d) manganese gluconate (GM); e) manganese chloride (CM); f) mulberry root extract (RA). Each group received three 10-minute applications of the gels containing the respective activating agents. Next, a new shade measurement was made.Results: The Analysis of Variance and Tukey tests (alpha=5%) showed statistically significant differences for the shade perception values (p=0.002). Groups GM, CM and RA showed significantly higher means than the control group.Conclusion: The presence of some chemical activators is capable of resulting in a significant increase in tooth shade variation.

Chemical and antifungal investigations of six Lippia species (Verbenaceae) from Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The activity of amino acids produced by Paenibacillus macerans and from commercial sources against the root-knot nematode Meloidogyne exigua

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)