915 resultados para Quantitative real-time PCR
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Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.
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Real-time ultrasonography (RTU) was used to measure the longissimus dorsi muscle (LM) volume in vivo and to predict the carcass composition of rabbits. For this, 63 New Zealand White × Californian rabbits with 2093±63 g live weight were used. Animals were scanned between the 6th and 7th lumbar vertebrae using an RTU equipment with a 7.5 MHz probe. Measurements of LM volume were obtianed both in vivo and on carcass. Regression equations were used for the prediction of carcass composition and LM volume using the LM volume measured obtained with RTU (LMVU) as independent variable. Carcass meat, bone and total dissectible fat weights represented 780, 164 and 56 g/kg of the reference carcass weight, respectively. Regression equations showed a strong relationship between LMVU and the correspondent volume in carcass. Furthermore, LMVU was also useful in predicting the amounts of carcass tissues. It is possible to predict LM volume in the carcass using the LM volume measured in vivo by RTU. The amount of carcass tissues can be predicted by the LM volume measured in vivo by RTU.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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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.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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This paper describes an electronic device conceived to convert common web texts into sequences of corresponding Braille signals, which are immediately reproduced onto an array ( keyboard) of electromechanical actuators. These actuators are reconfigurable in real time, displaying the Braille characters as matrices of points composed by small stems which can be lowered or raised according to the Braille code. The device, together with its conversion software package, can provide direct access to web texts in any personal computer, thus avoiding the use of complicated Braille printers.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Quartz Crystal Microbalance (QCM) was used to monitor the mass changes on a quartz crystal surface containing immobilized lectins that interacted with carbohydrates. The strategy for lectin immobilization was developed on the basis of a multilayer system composed of Au-cystamine-glutaraldehyde-lectin. Each step of the immobilization procedure was confirmed by FTIR analysis. The system was used to study the interactions of Concanavalin A (ConA) with maltose and Jacalin with Fetuin. The real-time binding of different concentrations of carbohydrate to the immobilized lectin was monitored by means of QCM measurements and the data obtained allowed for the construction of Langmuir isotherm curves. The association constants determined for the specific interactions analyzed here were (6.4 +/- 0.2) X 10(4) M-1 for Jacalin-Fetuin and (4.5 +/- 0.1) x 10(2) M-1 for ConA-maltose. These results indicate that the QCM constitutes a suitable method for the analysis of lectin-carbohydrate interactions, even when assaying low molecular mass ligands such as disaccharides. Published by Elsevier B.V.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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A significant part of film production by the coating industry is based on wet bench processes, where better understanding of their temporal dynamics could facilitate control and optimization. In this work, in situ laser interferometry is applied to study properties of flowing liquids and quantitatively monitor the dip coating batch process. Two oil standards Newtonian, non-volatile, with constant refractive indices and distinct flow properties - were measured under several withdrawing speeds. The dynamics of film physical thickness then depends on time as t(-1/2), and flow characterization becomes possible with high precision (linear slope uncertainty of +/-0.04%). Resulting kinematic viscosities for OP60 and OP400 are 1,17 +/- 0,03. St and 9,9 +/- 0,2 St, respectively. These results agree with nominal values, as provided by the manufacturer. For more complex films (a multi-component sol-gel Zirconyl Chloride aqueous solution) with a varying refractive index, through a direct polarimetric measurement, allowing also determination of the temporal evolution of physical thickness (uncertainty of +/- 0,007 microns) is also determined during dip coating.