Estudo do genoma do vírus causador da mionecrose infecciosa em camarões e desenvolvimento de métodos para detecção de diferentes variantes

Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites

Análise do efeito de polimorfismos não-sinônimos em genes de reparo de DNA da via BER na resposta inflamatória da meningite

In vitro and in animal models, APE1, OGG1, and PARP-1 have been proposed as being involved with inflammatory response. In this work, we have investigated if the SNPs APE1 Asn148Glu, OGG1 Ser326Cys, and PARP-1 Val762Ala are associated to meningitis and also developed a system to enable the functional analysis of polymorphic proteins. Patients with bacterial meningitis (BM), aseptic meningitis (AM) and controls (non-infected) genotypes were investigated by PIRA-PCR or PCR-RFLP. DNA damages were detected in genomic DNA by Fpg treatment. IgG and IgA were measured from plasma and the cytokines and chemokines were measured from cerebrospinal fluid samples using Bio-Plex assays. The levels of NF-κB and c-Jun were measured in CSF by dot blot assays. A significant (P<0.05) increase in the frequency of APE1 148Glu allele in BM and AM patients was observed. A significant increase in the genotypes Asn/Asn in control group and Asn/Glu in BM group was also found. For the SNP OGG1 Ser326Cys, the genotype Cys/Cys was more frequent (P<0.05) in BM group. The frequency of PARP-1 Val/Val genotype was higher in control group (P<0.05). The occurrence of combined SNPs increased significantly in BM patients, indicating that these SNPs may be associated to the disease. Increasing in sensitive sites to Fpg was observed in carriers of APE1 148Glu allele or OGG1 326Cys allele, suggesting that SNPs affect DNA repair activity. Alterations in IgG production were observed in the presence of SNPs APE1Asn148Glu, OGG1Ser326Cys or PARP-1Val762Ala. Reductions in the levels ofIL-6, IL-1Ra, MCP-1/CCL2and IL-8/CXCL8 were observed in the presence of APE1148Glu allele in BM patients, however no differences were observed in the levels of NF-κB and c-Jun considering genotypes and analyzed groups. Using APE1 as model, a system to enable the analysis of cellular effects and functional characterization of polymorphic proteins was developed using strategies of cloning APE1 cDNA in pIRES2-EGFP vector, cellular transfection of the construction obtained, siRNA for endogenous APE1 and cellular cultures genotyping. In conclusion, we obtained evidences of an effect of SNPs in DNA repair genes on the regulation of immune response. This is a pioneering work in the field that shows association of BER variant enzymes with an infectious disease in human patients, suggesting that the SNPs analyzed may affect immune response and damage by oxidative stress level during brain infection. Considering these data, new approaches of functional characterization must be developed to better analysis and interactions of polymorphic proteins in response to this context

Marcadores genéticos associados com a resposta inflamatória na meningite bacteriana

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Tornar-se, ser e viver do professorado: entre regularidades e variações identitárias

The main aim of this research has been to analyze the identity patterns of the teacher s staff of fundamental education public schools in the Metropolitan Area of Natal-RN. It sets out from the hypothesis that being a teacher within this context grows out of the regularities of a specific habitus, which, according to Bourdieu, develops into mental schemes of thought and action within a specific social group. This habitus forms the basis on which is built the social representation of being a teacher prevailing in the group, as well as the symbolic differences that typify its identity variations. Three data sources have been fundamental in building up this thesis: (a) formative essays of students graduating from a Higher Teacher s Formation Course, as well as observing some of the public defense of these essays during field work; (b) a questionnaire aimed at classifying economically, socially, and culturally a sample of public teachers of the Natal-RN county; and (c) submitting a sub-sample of this group to the process of Multiple Classification Procedures (MCP). The analysis of data was done according to the multidimensional, non-parametric statistical procedures of both the Category Content Analysis and Enunciation Analysis methods. The results of the analysis took into account an ample set of variables, its associations and implications, the cultural and social profile of the population under scrutiny, their life styles, as well as the strategies they developed in the process of becoming a teacher, and the social representation of being a teacher . We came to the conclusion that the social identity of the teachers corps, or as we prefer to say it being a teacher , is a result of a set of regularities produced by the habitus that gives social shape and meaning to the existence of the group proper. We note the existence of identity variations caused by the variables (a) educational level; and (b) mode of action in fundamental education (if these are the first or last grades where the subjects operate). However, these variations will not break the power of the regularities that give shape, meaning, and social visibility to the group. The social representation of being a teacher points to the tensions, ambiguities, and trends inherent to common sense, as well as to a strong tendency to reassign a new meaning to being a teacher. Our thesis, therefore, is that the identity configuration of the teachers corps under scrutiny is characterized by an integrative synthesis, by-product of a habitus that is superimposed, and at the same time co-exists with different identity variations

Representação social sobre educação inclusiva por professores de Cruzeiro do Sul - Acre

Nowadays the discussion about providing a quality education for all students is more and more recurrent, including for those who have special educational necessities. The discussion increases regarding that it is not enough to include these students in the regular school, but also to provide conditions of learning and development. This requirement implies changes in the educational system, as well as in the teachers‟ everyday; and these changes manifest themselves through the complexity of the functions which are assigned to the teachers and the school. The news that are generated from the perspective of the inclusive education asks for new formative models for teacher‟s performance, once he tries to (re)build knowledge, knowings and doings among troubles and conflicts as a way to decrease the impact caused by the necessary transformations. This research approached this context in order to understand the social representation about the inclusive education from teachers who work in the regular public school system in the Municipality of Cruzeiro do Sul, State of Acre. To approach the symbolic content, we elected the Multiple Classification Procedure (MCP) as the methodological approach. For that, it was necessary to apply the Technique of Free-Association of Words (TFAW) to 60 participants what provided data to the first step of the chosen methodological procedure. The criterion of choice of the participants took into account if they dealt with student who has special educational necessities and with public schools. Later, we applied MCP to 50 teachers from specialized course in inclusive education subgroup 01 and 30 teachers who has no specialized formation subgroup 02. The collected data from this step was examined through multidimensional and content analysis for a better understanding of their symbolic dimensions. The results from the multidimensional analysis showed that the subject inclusive education‟ for subgroup 01 involved the following facets: circulating discourses that meant the naming of the characteristics that teachers think indispensable to inclusive education; teachers in relation to the inclusive practice that was focused on the relation between teaching and included student, and repercussion to the student that showed the advantages that the inclusive education provided to the student who has special educational necessities. Subgroup 02 dealt with the following facets: characteristics of the included student that approached teachers‟ view on this student; negative aspects that regarded the naming of the obstacles in the achievement of inclusive education; and teacher‟s relations to the inclusive education that approach professional, affective, and formative elements. The content analysis revealed four categories: disagreeing concepts; conception of inclusive education; dimension of the teacher‟s inclusive doing, and difficulties and resistance to carry out the inclusion of a student who has special educational necessities in the regularschool. Both analyses multidimensional and content one showed that the constitution of elements in a social representation of inclusive education was a mixture formed by the characteristics of this sort of education and the school integration, materialized on the figure of the different‟ student. The representational field of the subject inclusive education‟ was associated to the social representation of the student who has special educational necessities, making clear the deficiency/ difference, and causing the difficulty of the teachers in achieving what they say about the phenomenon

Como vão se formando os professores em física e química: embates entre o ser, o ter e o fazer na formação de licenciandos da UFRN sob a perspectiva da teoria das representações sociais

The purpose of this work is to approach and understand the Social Representations (SR) (MOSCOVICI, 2003) about Physics and Chemistry from people who are major in these courses, as well as their Social Representations about teaching . We took as principle that approaching these representations it would be possible to relate their symbolic contents, in order to show how people who are following the first segments of bachelor degree courses in Physics and Chemistry become teachers, taking into account a psychosocial view. Two source of data was used during this research: Free-association Technique FA (ABRIC, 1994); and Multiple Classification Procedure (MCP) (ROAZZI, 1995). The analytical treatment of the collected data from FA was done according to the proposition of Grize, Vergés and Silem (1987 apud ABRIC, 1994, p. 66). MCP data were analyzed through MSA (Multidimensional Scalogram Analysis) and SSA (Singular Spectrum Analysis) methods associated with the Facet Theory (BILSKY, 2003). The discourses of MCP discussing groups at the moment of explanations were studied by Content Analysis as it was proposed by Bardin (1977) and Franco (2005). Indicative of an approach to the relations with knowledge (CHARLOT, 2000), the connections which aroused from the analyses showed that the group of future majors in Physics thought that this scientific field was based on a rationalist conception, influencing the idealization sense of the phenomena to be explained by Physics. Thus, Physics as school content started to require the student of the fundamental and high school to think abstractly as a cognitive skill of learning. The identifying elements observed in the relations between SR about Physics and Teaching aroused from the antagonism between future majors and their teacher, as well as from the speculation between their fundamental and high school students and themselves, mainly when they had to face the act of teaching due to the obstacles imposed by the own educational system, and by the weakness of the initial preparation. The group of future majors in Chemistry, through its discourses, showed these relations when they conceived empiricist Chemistry and said that teaching was the way of transmission of this knowledge, and didactics of Chemistry teaching was the direction to learning through pedagogic methods in order to lead the students to discoveries. The psychosocial contents which were built and showed from the symbolic relations in the studied SR achieved the relation of identity. This relation revealed identifying elements for these people, resulting from the traffic between their condition as students of Chemistry, and as teachers regarding their work, what placed the current relational contents in the teaching space, named as Knowledge changing and Adaptability . In order to study emerging questions in the discussing environment about formation and teaching professionalization, we focused the psychosocial view on this traffic and managed to observe epistemological practical and pedagogic obstacles that limited a configuration of the teaching work as a professional activity, especially from the particular conditions which led the relations of senses to Physics , Chemistry and Teaching ; and Chemistry and Physics as it was seen in this research. Generally speaking, we noted that these obstacles can denounce such obstacles concerning to the pedagogic doings which mainly impair the learning process of fundamental and high school students

Dimensões didática, afetiva e formativa de docência que tecem as representações sociais entre licenciandos da UFPI

This study intends to analyze the social representation of teaching for students of first years of undergraduate courses in Education, Letters and Biology. The field of this research was the Federal University of Piaui - Campus of Picos in 2009. To reach the objective proposed above, was used the theory of social representations to the seizure of the elements which constitute such representations according to Moscovici and colleagues, considering the contribution of Abric with his Theory of Central Nucleus and Wagner with the Theory of Sociogenesis. Data were collected in two phases: first, through the Technique of Free Association of Words (TFAW) of which 100 subjects evoked representations of their teaching through the inductor terms 'to teach', 'student' and 'teacher'. For those data we used the EVOC software that promoted to detect the elements of the core and to conclude that the social representation of teaching is one of the work performed by a master / teacher who transmits, directed to an apprentice who learns, confined to the school involving the student with all his virtues to be smart, interested and dedicated, and teacher to be friend, wise bearer of knowledge and also intelligent. Then, using the Multiple Classification Procedure (MCP) only 10 subjects made ratings of 25 more evoked words in the first phase, for the analysis of data from the MPC with the use of the SPSS software we used Multidimensional Statistical Analysis (MSA) for the Free Classification we found three dimensions of Social Representation of Teaching: The Didactic, that focuses on teacher and student being superimposed, meaning the inseparability of these elements, the Affective, which presents the elements inherent in teachers with love as the strong point of this dimension, and the Formative, that is as ambiguous as ambivalent because it sees the teacher as a professional directed to help students get an education; for Directed Classification with Similarity Structure Analysis (SSA), we learn that teaching is a profession that materializes in the classroom, which is extremely true because the action happens in this teaching space, supposing a caring, loving, cheerful, capable, apt, patient, partner, responsible, dedicated, committed educator, who has wisdom and knows how to teach and help students through dialogue to there study and learning. All of it s necessary to occur the result of teaching, which is education in a disciplinary manner. The results point to a very traditional social representation of what is the role of teachers, the role of students and the role of the relationship itself between teaching and learning through the act of teaching. With this statement, we can confirm that the structure of Social Representation of Teaching for the investigated subjects reflects the sociogenetic conditions that engendered them, and that these conditions permeate their structural organization, in particular time-context in which social representation was captured

Ativação local da rota da quinurenina por moduladores inflamatórios durante a meningite bacteriana.

Activation of the kynurenine (KYN) pathway (KP) by modulators of immune system has been observed during several neurological diseases. Here we assessed the association of chemo-/cytokine levels with the concentration of KP metabolites in cerebrospinal fluid (CSF) and plasma samples from patients with bacterial meningitis (BM). All samples were collected from 42 patients diagnosed with acute bacterial meningitis (ABM), aseptic meningitis, tuberculous meningitis and patients without infection neurological disorders. CSF and plasma concentration of metabolites from the KP was assessed by high pressure liquid chromatography (HPLC) and cytokines and chemokines by Bio-plex 200 suspension array system. Concentrations of the KP metabolites KYN and kynurenic acid (KYNA) were significantly higher in CSF of patients with ABM compared to other groups. Tryptophan (TRP), anthranilic acid (AA), 3-hydroxykynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) did not show statistical significance, although some of them presented a good accumulation during ABM. The expression of TNF-alpha, IL-6, IL-1beta, IFN-gamma, IL-10, IL-1 receptor antagonist (IL-1Ra), MIP-1alpha, MIP-1beta, MCP-1 and G-CSF was about 100-fold higher in CSF from ABM patients than other infected groups. In all CSF and plasma samples, the concentration of IL-2, IL-12(p70), IL-4, IL-8 and GM-CSF was not significant. ABM still showed significant concentrations of IL-6, IL-10, IL-1Ra and MCP-1 in plasma samples. Based on the comparison of KP metabolites concentrations between plasma and CSF samples we conclude that the activation of the tryptophan pathway upon BM occurs within the brain. This increase in KP metabolites is most due to activation of the KP by molecules as IFN-gamma and TNF-alpha in response to infection.

A memória de curto prazo e a síndrome de down: a relação entre contextos de desenvolvimento

Down syndrome (DS) is one of the most frequent causes of intellectual disability, affecting one in every 600 to 1000 live births. Studies have demonstrated that people with DS have a lower capacity for short-term memory (STM) and working memory (WM), which affects their capability to learn new words and to follow spoken instructions, specially when they involve multiple information or consecutive orders/orientations. It seems that the basis of the learning process, as it happens with language and mathematics comprehension and reasoning, relies in the STM and WM systems. Individuals with DS are increasingly included in mainstream education, and yet, very few researches have been conducted to investigate the influence of memory development and the type of enrollment (regular school and special school). This study investigated the relationship between the type of school enrollment with the performance on STM tests and also, the relationship of this performance with early stimulation (ES). The tests used in the first research were the digit span, free recall, word recognition and subtests of the Wechsler Intelligence Scale for Children Third Edition (WISC-III). Individuals enrolled in the regular schools group had higher scores on the digit span test and the subtests of the WISC-III. In the free recall and recognition tests, no differences were found. This study indicates that the type of enrollment might influence the memory development of individuals with DS and clearly points the need for future investigations. In the second research, the tests used were the digit span, free word recall and subtests of the WISC-III. The test results showed better performance by adults that received ES before six months of age. The studies showed improvement in STM both in people who attended or were attending regular school, as well as those who benefited from ES before six months of age. However, some issues still need to be better understood. What is the relation between this stimulation with the individual s education? Since ES may reflect a greater family involvement with the individual, what is the role of emotional components derived from this involvement in the cognitive improvement? These and other questions are part of the continuity of this study

Estampa temporal na memória de reconhecimento em estudantes universitários

Several studies have shown that there is a circadian modulation of explicit memory. This modulation can occur independently in each one of the mnemonic processes. The aim of this study was to evaluate the influence of time of training on short-term memory (STM) and long-term memory (LTM), using a recognition task. Moreover, a possible circadian modulation in retrieving was investigated when this process matched the acquisition hour (time stamp). The chronotype variable was also considered. Fifty-seven undergraduate students aging between 18 and 25 years (21,72 ± 2,14; 28 ♂) participated in this study. In the training phase (acquisition) the subjects heard a ten word list. Following this, they answered a recognition test to evaluate STM and one week later they answered a recognition test to evaluate LTM. In each chronotype, the subjects were divided in groups according to the training hour, part of them in the morning and the other in the afternoon. One week later some of the subjects in each group underwent LTM testing in the morning and others in the afternoon. When the subjects performances were analyzed together, independently of the chronotypes, a training hour effect was found in the LTM. The subjects trained in the afternoon had better performance. No time of day effect was found in the STM and in retrieving from the LTM. However, the morning types who were trained and tested in the same hour had a better performance in the LTM when compared to morning types trained and tested in different hours. This effect did not occur when the other chronotypes were analyzed. The circadian modulation seems to occur at least in two different ways. First, there is a circadian modulation in the acquisition/consolidation processes, with a better performance occuring in the afternoon. Secondly, there is a modulation in the retrieval mnemonic process, called time stamp phenomenon. This phenomenon, that occurred in the morning types, is showed for the first time in humans

Estudo do genoma do vírus causador da mionecrose infecciosa em camarões e desenvolvimento de métodos para detecção de polimorfismos

Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites

Análise do efeito de polimorfismos não-sinônimos em genes de reparo de DNA da via BER na resposta inflamatória da meningite

In vitro and in animal models, APE1, OGG1, and PARP-1 have been proposed as being involved with inflammatory response. In this work, we have investigated if the SNPs APE1 Asn148Glu, OGG1 Ser326Cys, and PARP-1 Val762Ala are associated to meningitis and also developed a system to enable the functional analysis of polymorphic proteins. Patients with bacterial meningitis (BM), aseptic meningitis (AM) and controls (non-infected) genotypes were investigated by PIRA-PCR or PCR-RFLP. DNA damages were detected in genomic DNA by Fpg treatment. IgG and IgA were measured from plasma and the cytokines and chemokines were measured from cerebrospinal fluid samples using Bio-Plex assays. The levels of NF-κB and c-Jun were measured in CSF by dot blot assays. A significant (P<0.05) increase in the frequency of APE1 148Glu allele in BM and AM patients was observed. A significant increase in the genotypes Asn/Asn in control group and Asn/Glu in BM group was also found. For the SNP OGG1 Ser326Cys, the genotype Cys/Cys was more frequent (P<0.05) in BM group. The frequency of PARP-1 Val/Val genotype was higher in control group (P<0.05). The occurrence of combined SNPs increased significantly in BM patients, indicating that these SNPs may be associated to the disease. Increasing in sensitive sites to Fpg was observed in carriers of APE1 148Glu allele or OGG1 326Cys allele, suggesting that SNPs affect DNA repair activity. Alterations in IgG production were observed in the presence of SNPs APE1Asn148Glu, OGG1Ser326Cys or PARP-1Val762Ala. Reductions in the levels ofIL-6, IL-1Ra, MCP-1/CCL2and IL-8/CXCL8 were observed in the presence of APE1148Glu allele in BM patients, however no differences were observed in the levels of NF-κB and c-Jun considering genotypes and analyzed groups. Using APE1 as model, a system to enable the analysis of cellular effects and functional characterization of polymorphic proteins was developed using strategies of cloning APE1 cDNA in pIRES2-EGFP vector, cellular transfection of the construction obtained, siRNA for endogenous APE1 and cellular cultures genotyping. In conclusion, we obtained evidences of an effect of SNPs in DNA repair genes on the regulation of immune response. This is a pioneering work in the field that shows association of BER variant enzymes with an infectious disease in human patients, suggesting that the SNPs analyzed may affect immune response and damage by oxidative stress level during brain infection. Considering these data, new approaches of functional characterization must be developed to better analysis and interactions of polymorphic proteins in response to this context