Functional XPB/RAD25 redundancy in Arabidopsis genome: characterization of AtXPB2 and expression analysis

The xeroderma pigmentosum complementation group B (XPB) protein is involved in both DNA repair and transcription in human cells. It is a component of the transcription factor IIH (TFIIH) and is responsible for DNA helicase activity during nucleotide (nt) excision repair (NER). Its high evolutionary conservation has allowed identification of homologous proteins in different organisms, including plants. In contrast to other organisms, Arabidopsis thaliana harbors a duplication of the XPB orthologue (AtXPB1 and AtXPB2), and the proteins encoded by the duplicated genes are very similar (95% amino acid identity). Complementation assays in yeast rad25 mutant strains suggest the involvement of AtXPB2 in DNA repair, as already shown for AtXPB1, indicating that these proteins may be functionally redundant in the removal of DNA lesions in A. thaliana. Although both genes are expressed in a constitutive manner during the plant life cycle, Northern blot analyses suggest that light modulates the expression level of both XPB copies, and transcript levels increase during early stages of development. Considering the high similarity between AtXPB1 and AtXPB2 and that both of predicted proteins may act in DNA repair, it is possible that this duplication may confer more flexibility and resistance to DNA damaging agents in thale cress. (C) 2004 Elsevier B.V. All rights reserved.

Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer

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

Effect of heat exposure on the thermoregulatory responses of selected naked neck chickens

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

Compreensão do típico ideal de técnicos de enfermagem acerca da sistematização da assistência de enfermagem

This study aimed to understand the typical ideal of the nursing technician about the systematization of nursing care in the light of the theoretical framework of Alfred Schutz. It is a comprehensive phenomenological research, using the theoretical framework of Alfred Schutz. For the unveiling of the phenomenon (the typical ideal of the nursing technician about the systematization of nursing care), the search process was configured from the proposed guiding principles for a research methodology based on the work itself of Schutz held by Zeferino (2010) in his PhD. For data collection, we used the focus group technique, counting on the collaboration of thirteen practical nurses working in a university hospital in Rio Grande do Norte, who responded positively to the inclusion criteria: working in the study hospital, performing care direct to patients. Forty-four subjects showed interest in participating, being held a draw for selection of the research sample, consisting of 14 professionals, one of whom did not attend the gathering of data collection. The focus group, entitled "What I think about the systematization of nursing care", took place on February 15th, 2013, totaling 101 minutes. It was performed according to the Experiential Education Humanescent using building posters as projective technique, from the key question: "What is the systematization of nursing care for you?". In order to understand some of the biographical situation of the participants, a questionnaire was administered to study participants. From the agreement of the subjects, the focus group was recorded and photographed with the cooperation of one reporter and two other employees. We used Microsoft Word 2010 to perform the transcript of the meeting and Microsoft Excel 2010 for synthesizing the results via a spreadsheet. The study followed the ethical and legal principles that govern scientific research on humans, recommended in Resolution nº 196/96, it was approved by Opinion Embodied Ethics Committee in Research of UFRN (Federal University of Rio Grande do Norte), nº 98 424, of August 31th, 2012, CAAE No. 05906912.0.0000.5537. The analysis of the nursing staff speeches, along with the contemplation of their posters and their written descriptions, allowed from the guiding principles of Zeferino (2010), in light of the reference of Alfred Schutz, unveiling the typical ideal of nursing technicians about the systematization of nursing care, passing four themes: typing of the concept of systematization of nursing care; benefits, which resulted in the reasons to believe in the positivity of this working tool; experienced problems, revealing the world of everyday life of nursing professionals, and possibilities for improvement. It was concluded that the nursing technicians are unaware of the systematization of nursing care. However, they typify a very positive perception about the same, especially with regard to improvements that may foster care

Transcription profiling of signal transduction-related genes in sugarcane tissues

A collection of 237,954 sugarcane ESTs was examined in search of signal transduction genes. Over 3,500 components involved in several aspects of signal transduction, transcription, development, cell cycle, stress responses and pathogen interaction were compiled into the Sugarcane Signal Transduction (SUCAST) Catalogue. Sequence comparisons and protein domain analysis revealed 477 receptors, 510 protein kinases, 107 protein phosphatases, 75 small GTPases, 17 G-proteins, 114 calcium and inositol metabolism proteins, and over 600 transcription factors. The elements were distributed into 29 main categories subdivided into 409 sub-categories. Genes with no matches in the public databases and of unknown function were also catalogued. A cDNA microarray was constructed to profile individual variation of plants cultivated in the field and transcript abundance in six plant organs (flowers, roots, leaves, lateral buds, and 1(st) and 4(th) internodes). From 1280 distinct elements analyzed, 217 (17%) presented differential expression in two biological samples of at least one of the tissues tested. A total of 153 genes (12%) presented highly similar expression levels in all tissues. A virtual profile matrix was constructed and the expression profiles were validated by real-time PCR. The expression data presented can aid in assigning function for the sugarcane genes and be useful for promoter characterization of this and other economically important grasses.

Respostas Fisiológicas Associadas à Termotolerância em Pintos de Corte de Duas Linhagens por Exposição a Altas Temperaturas

Com o objetivo de se avaliar o comportamento fisiológico das aves, bem como, se ocorre a expressão da proteína do choque térmico (Hsp 70) no fígado e cérebro, como conseqüência da exposição ao estresse por calor, em quatro dias sucessivos (2 a 5 dias de vida), 20 pintos de corte de duas linhagens Pescoço pelado (Na/na) e Hubbard-Petersen, foram ou não estressados à temperatura de 36-37°C, durante cinco horas. Foram colhidas amostras de fígado e cérebro, ao final do período de estresse, e analisadas por Western Blotting, para determinação dos níveis de Hsp 70. Os resultados não mostraram associação entre perda de peso corporal e as demais características. Aumentos da variação de temperatura cloacal apresentaram aumentos nos níveis de Hsp70 no tecido hepático e cerebral. Concluiu-se que a concentração de Hsp70 no cérebro, apresentou-se correlacionada com a variação de temperatura cloacal, quando se usou o condicionamento ao calor em pintos de corte, embora não se possa afirmar que seria indicativo de termotolerância de aves.

Segurança da informação no correio eletrônico com base na ISO/IEC 17799 :um estudo de caso em uma instituição superior com foco no treinamento

The electronic mail service is one of the most Internet services that grow in the corporate environment. This evolution is bringing several problems for the organizations, especially to information that circulates inside of the corporate net. The lack of correct orientation to the people, about the usage and the security importance of these resources, is leaving breaches and causing misusage and overuse of service, for example. In recent literature, it starts to coming out several ideas, which has helped to rganizations how to plain and how to implement the information security system to the electronic mail in computer environment. However, these ideas are still not placed in practice in many companies, public or private. This dissertation tries to demonstrate the results of a research that has like goal, identify the importance that user training has over the information security policy, through a case study inside of private superior education institute in this state. Besides, this work had by basic orientation the ISO/IEC 17799, which talk about People Security. This study was developed over a proposed model to this research, which looked for offer conditions to guide the institution studied, how to plan better a information security policy to the electronic mail. Also, this research has an exploratory and descreptive nature and your type, qualitative. Firstly, it was applied na questionary to the information technology manager, as better way to get some general data and to deepen the contact which still then, it was being kept through e-mail. Thereupon this first contact, eleven interviews were done with the same manager, beside one interview with twenty-four users, among employees e students. After that to collect and transcript the interviews, were review with the manager all informations given, to correct any mistakes and to update that informations, to then, start the data analyze. The research suggests that the institution has a pro attitude about the information security policy and the electronic mail usage. However, it was clear that answers have their perception about information security under a very inexperient way, derived of a planning lack in relation to training program capable to solve the problem

A linguagem da reflexão de uma professora de Língua Inglesa: um estudo de caso sob a perspectiva do sistema de avaliatividade

Research about teacher education, carried out in the area of Applied Linguistics (AL), reveals the importance of reflective practices in the professional development of teachers. With the aim of contributing to this area, we present this case study conducted at a technical school in Natal, RN. The corpus of the study is formed mainly from the teacher‟s discourse, generated during a stimulated recall session and the instruments used to collect the data: an initial questionnaire, a video recording of a class and the text transcript of the stimulated recall session. The central objective is to understand the way in which the reflection-on-action (SCHÖN, 1983, 1987) can contribute to raise the awareness of an English as a Foreign Language teacher (EFL) about her actions in the classroom. With this proposal, we begin our discussion presenting the origins, the presuppositions and characteristics of the concept of reflection according to Schön (1983, 1987), and supported by other authors (PERRENOUD, 2002; GÓMEZ, 1995; IMBERNÓN, 2009, among others); of critical reflection (LISTON e ZEICHNER, 1993; PIMENTA, 2002; DUTRA e MELLO, 2004, among others); and of the process of critical reflection (SMYTH, 1992). To evidence the reflections that emerge in the teacher‟s discourse, we found support in the theories and methods of Systemic Functional Grammar (SFG), which was initially proposed by Halliday (1985, 1994), Halliday and Hasan (1989), Halliday and Matthiessen (2004) and followers, such as, Eggins (1994), Thompson (1996), among others. We focus mainly on the subsystem of Attitude, an integral component of the system of discourse resources, Appraisal, presented by Martin (2000), Martin and Rose (2003, 2007), Martin and White (2005). The results reveal that the actions of the teacher in the classroom reflect not only her professional experiences, but also her values and concepts about teaching/learning languages. The results also show the teacher‟s awareness of the need for changes in her practices. Faced with these findings, we believe that this study reveals important concepts that can direct teacher educators to rethink new ways of approaching teacher training courses. In addition, it also reveals the importance of discourse analysis based on a systemic functional approach.

Splice variants of the human ZC3H14 gene generate multiple isoforms of a zinc finger polyadenosine RNA binding protein

The human ZC3H14 gene encodes an evolutionarily conserved Cys(3)His zinc finger protein that binds specifically to polyadenosine RNA and is thus postulated to modulate post-transcriptional gene expression. Expressed sequence tag (EST) data predicts multiple splice variants of both human and mouse ZC3H14. Analysis of ZC3H14 expression in both human cell lines and mouse tissues confirms the presence of multiple alternatively spliced transcripts. Although all of these transcripts encode protein isoforms that contain the conserved C-terminal zinc finger domain, suggesting that they could all bind to polyadenosine RNA, they differ in other functionally important domains. Most of the alternative transcripts encode closely related proteins (termed isoforms 1, 2. 3, and 3short) that differ primarily in the inclusion of three small exons, 9, 10, and 11, resulting in predicted protein isoforms ranging from 82 to 64 kDa. Each of these closely related isoforms contains predicted classical nuclear localization signals (cNLS) within exons 7 and 11. Consistent with the presence of these putative nuclear targeting signals, these ZC3H14 isoforms are all localized to the nucleus. In contrast, an additional transcript encodes a smaller protein (34 kDa) with an alternative first exon (isoform, 4). Consistent with the absence of the predicted cNLS motifs located in exons 7 and 11, ZC3H14 isoform 4 is localized to the cytoplasm. Both EST data and experimental data suggest that this variant is enriched in testes and brain. Using an antibody that detects endogenous ZC3H14 isoforms 1-3 reveals localization of these isoforms to nuclear speckles. These speckles co-localize with the splicing factor, SC35, suggesting a role for nuclear ZC3H14 in mRNA processing. Taken together, these results demonstrate that multiple transcripts encoding several ZC3H14 isoforms exist in vivo. Both nuclear and cytoplasmic ZC3H14 isoforms could have distinct effects on gene expression mediated by the common Cys(3)His zinc finger polyadenosine RNA binding domain. (C) 2009 Elsevier B.V. All rights reserved.

An interaction between two RNA binding proteins, Nab2 and Pub1, links mRNA Processing/Export and mRNA stability

mRNA stability is modulated by elements in the mRNA transcript and their cognate RNA binding proteins. Poly(U) binding protein 1 (Pub1) is a cytoplasmic Saccharomyces cerevisiae mRNA binding protein that stabilizes transcripts containing AU-rich elements (AREs) or stabilizer elements (STEs). In a yeast two-hybrid screen, we identified nuclear poly(A) binding protein 2 (Nab2) as being a Pub1-interacting protein. Nab2 is an essential nucleocytoplasmic shuttling mRNA binding protein that regulates poly(A) tail length and mRNA export. The interaction between Pub1 and Nab2 was confirmed by copurification and in vitro binding assays. The interaction is mediated by the Nab2 zinc finger domain. Analysis of the functional link between these proteins reveals that Nab2, like Pub1, can modulate the stability of specific mRNA transcripts. The half-life of the RPS16B transcript, an ARE-like sequence-containing Pub1 target, is decreased in both nab2-1 and nab2-67 mutants. In contrast, GCN4, an STE-containing Pub1 target, is not affected. Similar results were obtained for other ARE- and STE-containing Pub1 target transcripts. Further analysis reveals that the ARE-like sequence is necessary for Nab2-mediated transcript stabilization. These results suggest that Nab2 functions together with Pub1 to modulate mRNA stability and strengthen a model where nuclear events are coupled to the control of mRNA turnover in the cytoplasm.

The contribution of 700,000 ORF sequence tags to the definition of the human transcriptome

open reading frame expressed sequences tags (ORESTES) differ from conventional ESTs by providing sequence data from the central protein coding portion of transcripts. We generated a total of 696,745 ORESTES sequences from 24 human tissues and used a subset of the data that correspond to a set of 15,095 full-length mRNAs as a means of assessing the efficiency of the strategy and its potential contribution to the definition of the human transcriptome. We estimate that ORESTES sampled over 80% of all highly and moderately expressed, and between 40% and 50% of rarely expressed, human genes. In our most thoroughly sequenced tissue, the breast, the 130,000 ORESTES generated are derived from transcripts from an estimated 70% of all genes expressed in that tissue, with an equally efficient representation of both highly and poorly expressed genes. In this respect, we find that the capacity of the ORESTES strategy both for gene discovery and shotgun transcript sequence generation significantly exceeds that of conventional ESTs. The distribution of ORESTES is such that many human transcripts are now represented by a scaffold of partial sequences distributed along the length of each gene product. The experimental joining of the scaffold components, by reverse transcription-PCR, represents a direct route to transcript finishing that may represent a useful alternative to full-length cDNA cloning.

Rapid test for the evaluation of the activity of the prodrug hydroxymethylnitrofurazone in the processing of Trypanosoma cruzi messenger RNAs

No fully effective treatment has been developed since the discovery of Chagas' disease by Carlos Chagas in 1909. Since drug-resistant Trypanosoma cruzi strains are occurring and the current therapy is effectiveness in the acute phase but with various adverse side effects, more studies are needed to characterize the susceptibility of T. cruzi to new drugs. Many natural and/or synthetic substances showing trypanocidal activity have been used, even though they are not likely to be turned into clinically approved drugs. Originally, drug screening was performed using natural products, with only limited knowledge of the molecular mechanism involved in the development of diseases. Trans-splicing, which is unusual RNA processing reaction and occurs in nematodes and trypanosomes, implies the processing of polycistronic transcription units into individual mRNAs; a short transcript spliced leader (SL RNA) is trans-spliced to the acceptor pre-mRNA, giving origin to the mature mRNA. In the present study, permeable cells of T. cruzi epimastigote forms (Y, BOL and NCS strains) were treated to evaluate the interference of two drugs (hydroxymethylnitrofurazone - NFOH-121 and nitrofurazone) in the trans-splicing reaction using silver-stained PAGE analysis. Both drugs induced a significant reduction in RNA processing at concentrations from 5 to 12.5 µM. These data agreed with the biological findings, since the number of parasites decreased, especially with NFOH-121. This proposed methodology allows a rapid and cost-effective screening strategy for detecting drug interference in the trans-splicing mechanism of T. cruzi.

Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of estrogen receptor alpha and beta and progesterone receptor expression and correlation with clinicopathologic factors and proliferative marker Ki-67 in breast cancers

To elucidate the molecular profile of hormonal steroid receptor status, we analyzed ER-alpha, ER-beta, and PGR mRNA and protein expression in 80 breast carcinomas using reverse transcriptase polymerase chain reaction (RT-PCR), quantitative RT-PCR, and immunohistochemical analysis. Qualitative analysis revealed positive expression of ER-alpha, ER-beta, and PGR mRNA in 48%, 59%, and 48% of the breast carcinomas, respectively. ER-alpha, ER-beta, and PGR transcript overexpression was observed in 51%, 0%, and 12% of the cases, respectively, whereas moderate or strong protein expression was detected in 68%, 78%, and 49% of the cases, respectively. Tumor grade was negatively correlated with transcript and protein levels of ER-alpha (P = .0169 and P = .0006, respectively) and PGR (P = .0034 and P = .0005, respectively). Similarly, proliferative index Ki-67 was negatively associated with transcript and protein levels of ER-alpha (P = .0006 and P < .0001, respectively) and PGR (P = .0258 and P =. 0005, respectively). These findings suggest that ER-alpha and PGR expression are associated with well-differentiated breast tumors and less directly related to cell proliferation. A significant statistical difference was observed between lymph node status and ER-beta protein expression (P = .0208). In ER-alpha-negative tumors, we detected a correlation between ER-beta protein expression and high levels of Ki-67. These data suggest that ER-beta could be a prognostic marker in human breast cancer. (C) 2008 Elsevier B.V. All rights reserved.

Tumor microenvironmental genomic alterations in juvenile nasopharyngeal angiofibroma

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