Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient

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

Influence of early quantitative food restriction on long bone growth at different environmental temperatures in broiler chickens

1. This investigation was carried out to study the influence of early quantitative food restriction (40% of ad libitum intake from 7 to 14 d of age) and environmental rearing temperature (thermoneutral, cold and hot) on long bone (tibia, femur and humerus) development in broiler chickens.2. Tibia, femur and humerus were obtained at the ages of 14, 21, 28, 35 and 42 d; and the variables studied were: length and width (mm, expressed as absolute values), bone breaking strength (kgf) and bone weight/bone length index (BW/BL, mg/mm).3. All bone variables increased with bird age. Food restriction reduced bone length and width but did not affect bone weight. High environmental rearing temperature reduced bone length and width at 42 d of age. Bone strength was not affected by environmental temperature but the calculated BW/BL index was reduced by heat exposure.4. The findings suggest that food restriction and high environmental rearing temperature reduce long bone growth; bone breaking strength was affected by bird age but not by food restriction nor rearing ambient temperature.

Comparison of the quantification of caffeine in human plasma by gas chromatography and ELISA

In the present study we evaluated the precision of the ELISA method to quantify caffeine in human plasma and compared the results with those obtained by gas chromatography. A total of 58 samples were analyzed by gas chromatography using a nitrogen-phosphorus detector and routine techniques. For the ELISA test, the samples were diluted to obtain a concentration corresponding to 50% of the absorbance of the standard curve. To determine whether the proximity between the I50 of the standard curve and that of the sample would bring about a more precise result, the samples were divided into three blocks according to the criterion of difference, in modulus, of the I50 of the standard curve and of the I50 of the sample. The samples were classified into three groups. The first was composed of 20 samples with I50 up to 1.5 ng/ml, the second consisted of 21 samples with I50 ranging from 1.51 to 3 ng/ml, and the third of 17 samples with I50 ranging from 3.01 to 13 ng/ml. The determination coefficient (R² = 0.999) showed that the data obtained by gas chromatography represented a reliable basis. The results obtained by ELISA were also reliable, with an estimated Pearson correlation coefficient of 0.82 between the two methods. This coefficient for the different groups (0.88, 0.79 and 0.49 for groups 1, 2 and 3, respectively) showed greater reliability for the test with dilutions closer to I50.

Chicken serologic response to Salmonella enterica serotype Typhimurium assessed by Elisa

This study evaluated two enzyme-linked immunosorbent assays (ELISA) in the detection of chicken serologic response against Salmonella enterica sorotype Typhimurium. The assays have used as detecting antigen the soluble bacterial proteins of a non-flagellated strain of Salmonella Typhimurium (AgTM), and antibody conjugated to peroxidase or alkaline phosphatase. According to the results, optimal dilutions of antigen (concentration 5.49 mg/mL) and serum samples in both assays were 1:20,000 and 1:1,000, respectively. In such conditions, the ELISA/AgTM was able to detect serological response to Salmonella Typhimurium. Cross-reactions to Salmonella serotypes Gallinarum and Pullorum were seen, but not with other serotypes such as Enteritidis.

Detection of anti-Toxoplasma gondii antibodies in experimentally and naturally infected non-human primates by Indirect Fluorescence Assay (IFA) and indirect ELISA

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

An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against Leishmania chagasi in dogs

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

A quantitative nature of somatic embryogenesis in soybean

The objectives of this research were to investigate the genetic parameters associated with the in vitro formation of somatic embryos in soybean and to determine the effect of light intensity on the embryogenic capability of F-1, F-2, and backcross (RC1P1 and RC1P2) progenies derived from crosses between embryogenic (IAS-5 and Embrapa-1) and nonembryogenic (Parana) cultivars. Immature cotyledons (4-6 mm in length) derived from the parental lines, F-1, F-2, RC1P1, and RC1P2 were grown for 90 d on the inductive N10 medium, after which the number of somatic embryos was recorded. Chi-square tests for goodness of fit showed that the genetic component of the somatic embryogenesis trait is controlled in a quantitative manner by approximately 10 genes. A normal distribution for somatic embryo formation in the F-2 generations was observed reinforcing the quantitative nature of the trait. Variation in light intensity (8-12 and 27-33 mu mol m(-2) s(-1)) had no effect on somatic embryo formation in the parental material tested.

Selection and validation of reference genes for gene expression studies by reverse transcription quantitative PCR in Xanthomonas citri subsp citri during infection of Citrus sinensis

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

Quantitative and qualitative characteristics of the non-carcass components and the meat of lambs fed sunflower seeds and vitamin E

The objective of this investigation was to evaluate the weight and percentage of the non-carcass components and the mineral content (macro minerals and trace minerals), crude protein, ether extract, moisture and vitamin E of the heart, liver, tongue, lungs, reticulum, kidneys and meat from the longissimus dorsi of lambs in feedlot finishing. Thirty-two non-castrated Ile de France male lambs, fed diets containing sunflower seeds and vitamin E from 15 to 32 kg of body weight were allotted in a completely randomized design in a 2 × 2 factorial arrangement. The weight of the gastrointestinal tract was higher in the lambs fed diets containing vitamin E (10%). No difference was observed in the liver as to the mineral matter, crude protein, ether extract, moisture (2.01; 20.03; 2.39 and 74.78 g/100 g, respectively), the macro minerals and trace minerals, except iron. In the tongue, lungs, reticulum, kidneys and meat there was no in fluence of diets in the studied variables. The liver and the meat presented different values of crude protein (20.01 and 18.34 g/100 g, respectively), and the heart (1.03 mg/100 g) showed a higher content of vitamin E. High contents of manganese, zinc and copper were observed in the liver. The evaluated non-carcass components were nutritionally equal to the sheep meat, once, in addition to their high yield in relation to the body weight at slaughter, the non-carcass components are sources of nutrients.

Quantitative control of Lettuce mosaic virus fitness and host defence inhibition by P1-HCPro

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

Quantitative study of the myenteric plexus of the stomach of rats with streptozotocin-induced diabetes

A proposta deste trabalho foi estudar as alterações morfológicas e quantitativas dos neurônios do plexo mientérico do estômago de ratos com diabetes induzido por estreptozootocina e estabelecer uma comparação com animais não diabéticos. Amostras do corpo do estômago foram submetidas a preparados de membrana corados pelo método da NADH-diaforase e a cortes histológicos corados por hematoxilina-eosina. Observou-se que o diabetes provoca significante redução no número de neurônios.

Mapping of quantitative trait loci controlling tick [Riphicephalus (Boophilus) microplus] resistance on bovine chromosomes 5, 7 and 14

Differences in domestication and selection processes have contributed to considerable phenotypic and genotypic differences between Bos taurus and Bos indicus cattle breeds. of particular interest in tropical and subtropical production environments are those genetic differences between subspecies that underlie the phenotypic extremes in tolerance and susceptibility to parasite infection. In general, B. taurus cattle are more susceptible to ectoparasites than B. indicus cattle in tropical environments, and much of this difference is under genetic control. To identify genomic regions involved in tick resistance, we developed a B. taurus x B. indicus F-2 experimental population to map quantitative trait loci (QTL) for resistance to the Riphicephalus (Boophilus) microplus tick. About 300 individuals were measured for parasite load in two seasons (rainy and dry) and genotyped for 23 microsatellite markers covering chromosomes 5, 7 and 14. We mapped a suggestive chromosome-wide QTL for tick load in the rainy season (P < 0.05) on chromosome 5. For the dry season, suggestive (P < 0.10) chromosome-wide QTL were mapped on chromosomes 7 and 14. The additive effect of the QTL on chromosome 14 corresponds to 3.18% of the total observed phenotypic variance. Our QTL-mapping study has identified different genomic regions controlling tick resistance; these QTL were dependent upon the season in which the ticks were counted, suggesting that the QTL in question may depend on environmental factors.

O Uso de operações linguístico-discursivas da crítica genética na reescritura de textos

As a professor in Curso de Licenciatura em Letras, from Campus Avançado Profa Maria Eliza Albuquerque Maia (CAMEAM),do Estado do Estado do Rio Grande do Norte (UERN), in the town of Pau do Ferros, in the state of Rio Grande do Norte, we had the chance to carry out several writing activities , as well as guiding re-writing activities for the texts produced. From this experience, we started looking at the need of reflecting upon the writing process in higher education. Thus, we aim at analyzing, in this research, the methodology used in the moment of carrying out the writing practices activities in higher education, investigating, in particular, the rewriting practices, concerning the operations used for carrying out such activities, as well as the sense effects produced from the alterations which were made in the texts. Our theoretical foundation is grounded on a conception of text as a verbal action , what reveals a socio-interactional view of the language (MARCUSCHI, 2008; SAUTCHUK, 2003). As the production of written texts, our research focus, we assume that, for this activity, we deal with distinct figures (active writer and internal reader), so that we can, apart from writing, reflecting upon our writing and, this way, deciding about operations which are carried out to make the alterations which are necessary to the rewriting of our texts (SAUTCHUK, 2003). Still about the theoretical foundations used in this research, we made use of the theories from the Textual Analysis of Discourse (TAD) which discusses the belief on the evidence on the existence of the texts, which is opposite to the fixist view of textuality which believes that the texts exist by themselves. (ADAM, 2008; [2005]2010). Under this perspective, we have also adopted, the concepts which come from genetics criticism which is concerned about the relation between text and genesis, using as objects documents which bring traits of the text in progress, on the ground that the text is the result of work in progress, and the writing practice, on the other hand, as an activity in a continuous movement (HAY, [1975]2002; DE BIASI, [2000]2010; GRÉSILLON, 1989; [1990]2008; [1992]2002; SALLES, 2008a). The methodology in this research is an ethnography-based one, an approach which focuses on the process, as well as is meaning-based. To understand the objectives proposed in our research, we made use of different procedures of collecting data which include an ethnographic study, such as: observation, note-taking, document analysis. The data which were analyzed were collected during the semester of 2008.2, in a first term classroom of Curso de Letras from CAMEAM, when we were able to collect twenty-one written texts and all of them were rewritten based on rewriting activities, what provides a corpus of forty-two texts which will be analyzed based on the linguistics operations identified by Generative Grammar and adopted by Lebrave and Grésillon (2009). From these analyses, we were able to confirm that writing is a process, and rewriting has become an extremely important activity for this process. Still due to these data, we observed that substitution was the most used operation by text authors. We believe that this result is justified by the fact that the substitution, according to what proposes the Genetic Criticism, constitutes the source of all erasure, from which one can easily make a change in writing. Regarding the operations of addition and deletion, we found that they were used in quantitative terms, almost equivalently, which can be explained when we see that the two operations require, by the author of the text, different strategies from those used for the replacement, what includes , respectively, adding or removing a segment. Finally, we found out that the shift operation was the least used, since it works with a segment that will not be replaced, added or deleted, but transferred to another place of text, which requires a greater ability of the author to perform this operation and not compromising the meaning of his/her writing. As a result, we hope to contribute to the reflection on the teaching of writing, considering, in a particular way, those with a Bachelor in Arts. Our analysis will contribute to the teaching of Portuguese language, specifically for activities that guide the production of texts in order to explore with students the ability to rewrite their own text

Analytical Validation of Quantitative High-Performance Liquid Chromatographic Methods in Pharmaceutical Analysis: A Practical Approach

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

Plasmid-based controls to detect rpoB mutations in Mycobacterium tuberculosis by quantitative polymerase chain reaction-high-resolution melting

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