874 resultados para Forms of expression
Resumo:
Edgardo Antonio Vigo fue un artista plástico que utilizó distintas técnicas y formas de expresión, como la poesía visual, las performances urbanas, el arte correo, la escritura de manifiestos y otros textos, así como la edición de revistas, entre otras actividades. Vigo produjo su poética en clave de rebeldía de su tiempo, especialmente entre los '60 y '70, pero su rebelión transcurrió también por fuera del estereotipo más consolidado para la época. La obra de Vigo retoma algunos procedimientos de las vanguardias, como la utilización del objeto ya hecho, el uso de espacios alternativos de producción y difusión de sus obras, el abandono casi completo del formato de cuadro, entre otros. Interesa pensar en este trabajo de qué modo esta apropiación de técnicas y procedimientos previos, se combina con otra ruptura dirigida no sólo al sistema artístico, sino también al orden social. En este sentido, la obra de Vigo piensa su tiempo. ;Así como tematizó ciertos acontecimientos de relevancia política de los ámbitos nacional e internacional, también realizó una operación novedosa en su obra: la utilización del discurso judicial-administrativo. Se analizan en este trabajo algunos de sus usos en acciones artísticas. Concluimos en que se trató de una materia que incorporaría en su poética para desnaturalizarla de su lugar original, aristocrático y privatista. En este sentido, toma un aspecto no menor del funcionamiento del orden social disociándolo de su lugar normal y, al ofrecerlo a todos a través de acciones artísticas, permite una apropiación descentrada, fuera de los límites impuestos por su naturaleza
Resumo:
A presente tese investiga as manifestações de misticismo no Protestantismo brasileiro, especialmente em seus primórdios, tendo como campo de observação a atividade missionária. Este trabalho registra-se em meio aos estudos que abordam as relações entre cultura, religião e modernidade, principalmente quanto ao campo simbólico. Trata-se de pesquisa exploratória fundamentada no método fenomenológico. Para tanto, utiliza-se a redução fenomenológica e a redução eidética conforme as orientações de Edmund Husserl. Esta tese procura trazer contribuições para esclarecer elementos presentes na construção, transformação e difusão do Protestantismo, considerando a mística protestante como fenômeno religioso. A experiência mística é analisada do ponto de vista individual e coletivo, uma vez que este trabalho privilegia uma perspectiva sociológica em interface com a Psicologia, a Antropologia e, menos centralmente, a Filosofia e a História. Como pesquisa qualitativa e delimitada por seu objeto de estudo, o misticismo pode ser inicialmente apontado como relação direta entre o sagrado e o fiel, reconhecida desta forma individualmente e pelo grupo a que pertence.Tomou-se como referência teórica, entre outros autores, as idéias do sociólogo Roger Bastide, procurando confirmar ou refutar sua hipótese que um dos significados do misticismo pudesse representar a emergência do sagrado selvagem. Esta expressão designa a presença de elementos simbólicos que se manifestam de modo espontâneo ou explosivo nos rituais religiosos. Processos como a secularização e a laicização, decorrentes da institucionalização e modernidade da religião, fazem manifestar o sagrado selvagem, revelando os resíduos das produções sócio -culturais aparentemente ocultas ou mesmo latentes.Os instrumentos utilizados para a realização da pesquisa são trechos de obras literárias características do Protestantismo como A Peregrina e O Peregrino de autoria de John Bunyan, bem como jornais e periódicos evangélicos brasileiros referentes à segunda metade do século XIX: O Estandarte, Puritano e Expositor Cristão, que transmitiam concepções do Puritanismo e do Pietismo. Trechos de diários dos missionários e textos autobiográficos de pastores complementam a análise. O trabalho situa-se historicamente no período antecedente ao da Reforma, seu desenvolvimento e desdobramentos. Priorizaram-se especialmente os movimentos de reavivamento inglês e americano. em suas respectivas influências no imaginário e nas práticas religiosas brasileiras. Após cuidadosa leitura, seleção e organização por unidade de sentido dos dados analisados, firmou-se a hipótese de que a mística protestante é um elemento constitutivo do Protestantismo, representando modalidades de significação social, entre as quais destacam-se as mediações culturais entre: razão/emoção; ingularização/institucionalização; fragmentação/unidade; tradição/inovação; e padronização/automia, expressando relações com as formas de poder, mudança social e divergência de interesses sócio-econômicos.Verificou-se a existência, principalmente nas camadas socialmente menos privilegiadas da população, de indícios relevantes de aproximação cultural entre alguns elementos de práticas religiosas do Pentecostalismo vinculadas ao campo simbólico africano e indígena. Sugere-se que novas pesquisas sejam realizadas, revelando os significados das formas de expressão místicas no Protestantismo brasileiro e suas relações com o Pentecostalismo. Essas investigações podem clarificar os processos de justaposição de elementos culturais distintos nas práticas religiosas brasileiras, como afirmados por Roger Bastide, e corroborados por esta pesquisa.
Resumo:
O estudo realizado nesta pesquisa tem como foco aprofundar as discussões e reflexões sobre o processo criativo e sua importância na relação pedagógica a partir das práticas de professores de Artes. Quando abordamos o processo de criar do aluno e a experimentação no contexto escolar, que possibilidades, limites e obstáculos se manifestam? Trata-se de uma investigação de cunho qualitativo, realizada com base teórica em pesquisa bibliográfica, tendo como referenciais: Paulo Freire (2007) e sua discussão sobre a importância da relação-pedagógica para a construção da autonomia do aluno; Ana Mae Barbosa (2008) e seus estudos sobre a Arte-educação no Brasil e a formação do professor de Arte; os Parâmetros Curriculares Nacionais de Arte (1998) com ênfase na recomendação sobre a articulação dos três eixos metodológicos: criação/produção em Arte, fruição estética, e reflexão; Porcher (1982) e sua discussão sobre a importância da Educação Artística e Albano (1998) com o seu relato sobre a experiência vivida entre a artista Tarsila do Amaral e o seu aprendiz Tuneu. A pesquisa de campo foi desenvolvida com a realização de entrevistas com três professoras de Arte e três alunos (um aluno de cada professora), de duas escolas estaduais da Rede Pública de Ensino de São Bernardo do Campo e uma escola estadual da Rede Pública de Ensino de Diadema. Para o tratamento dos dados coletados nas entrevistas foi utilizada a Metodologia de Análise de Conteúdo proposta por Franco (2008). A análise dos dados demonstrou que o processar de ideias, a experimentação e todo o processo criativo do aluno passa quase sempre desapercebido na sua importância enquanto prática pedagógica desenvolvida no contexto escolar nas aulas de Arte. Os dados apresentados na pesquisa, quanto à formação do professor, indicam um grau elevado formativo, mas que, quando comparado à prática pedagógica, percebemos lacunas sobre a importância do papel da relação pedagógica no processo criativo do aluno. Apesar disto se apresentar como uma realidade na pesquisa, não poderíamos deixar de mencionar que, mesmo tendo uma pequena representação, observamos uma prática metodológica que cristaliza positivamente o processo criativo do aluno como forma de construção da autonomia, onde o aluno é presenteado com uma relação pedagógica que promove o experimentar, o criar e todas as formas de se expressar, de se fazer atuante, de expressar sua visão de mundo.
Resumo:
Since the emergence of the first demands for actions that were intended to give greater attention to culture in Brazil, came the first discussions which concerned the way the Brazilian government could have a positive influence in encouraging the culture, as is its interaction with the actors interested and involved with the cause. During the military dictatorship, there were programs which relied on the direct participation of the State to ensure that right, from the viewpoint of its support and implementation of public resources in developing the "cultural product" to be brought to society in its various forms of expression - all this, funded by the government. It is an example of "EMBRAFILMES" and "Projeto Seis e Meia", continued until the present day in some regions of the country, though maintained by entities not directly connected with the administration or the government. However, it was from the period of democratization and the end of the dictatorship that the Brazilian government began to look at the different culture, under its guarantee to the society. Came the first incentive laws, led by "Lei Sarney" Nº 7.505/86, which was culture as a segment which could receive foreign assistance in order to assist the government in fulfilling its public duty. After Collor era and the end of the embargo through the encouragement of culture incentive laws, consolidated the incentive model proposed in advance of Culture "Lei Sarney" and the federal laws, state and local regimentares as close to this action. This applies to the Rouanet Law (Lei Rouanet), Câmara Cascudo Law (Lei Câmara Cascudo) and Djalma Maranhão Cultural Incentive Law (Lei de Incentivo à Cultura Djalma Maranhão), existing in Natal and Rio Grande do Norte. Since then, business entities could help groups and cultural organizations to keep their work from the political sponsorship under control and regiment through the Brazilian state in the form of their Cultural Incentive Law. This framework has contributed to the strengthening of NGOs and with the consolidation of these institutions as the linchpin of Republican guaranteeing the right to access to culture, but corporate social responsibility was the one who took off in the segment treated here, through the actions of Responsibility Cultural enterprises arising from the Cultural Organizations. Therefore, in the face of this discourse, this study ascertains the process of encouraging the Culture in Rio Grande do Norte from the Deviant Case Analysis at the Casa da Ribeira, the main Cultural Organization that operates, focused action in Natal in order to assess the relationships established between the same entity and the institutions which are entitled to maintain the process of encouraging treated in this study - Enterprise, from the viewpoint of corporate sponsorship and Cultural Responsibility and State in the form of the Laws Incentive Funds and Public Culture Incentive
Resumo:
This work intends to describe and analyze the parties of forró that happen in Serra da Gameleira, in São Tomé/RN. Serra is a divided social space: groups of different ethnic origins live together in Gameleira de Baixo, Salgadinho (or Gameleira de Cima) and Chaves Belas. They are approximately two hundred families that live exclusively from agriculture. We try to understand how the parties inform about the social organization, the ethnic composition of the families that live there and the past of Serra, through the genealogy offorró players. In the discussion, we identify the festive places: in the total, we have Five houses of forró that function regularly one of them has been described. The private and public spaces inside them are intimate related, with no clear limits between the house of forró and the residence. Each house of forró has an owner, that regularly makes the parties, mobilizing a big part of the inhabitants, and provoking the straitening of the social relations. Observing the festive sociability between different social segments, the forró appears like na element that minimizes social conflicts, providing news ways of association and cooperation in the space of Serra da Gameleira. For the collection of facts, we used the ethnographic method, through the direct observation, interviewing and documentary research. The local history is recounted following the routes of oral memory and historical documents analysis. In the end of the analysis, we concluded that music and party are elements that aggregate the different groups that live in the location and determine forms of expression of what is seen as a traditional culture
Resumo:
Since the emergence of the first demands for actions that were intended to give greater attention to culture in Brazil, came the first discussions which concerned the way the Brazilian government could have a positive influence in encouraging the culture, as is its interaction with the actors interested and involved with the cause. During the military dictatorship, there were programs which relied on the direct participation of the State to ensure that right, from the viewpoint of its support and implementation of public resources in developing the "cultural product" to be brought to society in its various forms of expression - all this, funded by the government. It is an example of "EMBRAFILMES" and "Projeto Seis e Meia", continued until the present day in some regions of the country, though maintained by entities not directly connected with the administration or the government. However, it was from the period of democratization and the end of the dictatorship that the Brazilian government began to look at the different culture, under its guarantee to the society. Came the first incentive laws, led by "Lei Sarney" Nº 7.505/86, which was culture as a segment which could receive foreign assistance in order to assist the government in fulfilling its public duty. After Collor era and the end of the embargo through the encouragement of culture incentive laws, consolidated the incentive model proposed in advance of Culture "Lei Sarney" and the federal laws, state and local regimentares as close to this action. This applies to the Rouanet Law (Lei Rouanet), Câmara Cascudo Law (Lei Câmara Cascudo) and Djalma Maranhão Cultural Incentive Law (Lei de Incentivo à Cultura Djalma Maranhão), existing in Natal and Rio Grande do Norte. Since then, business entities could help groups and cultural organizations to keep their work from the political sponsorship under control and regiment through the Brazilian state in the form of their Cultural Incentive Law. This framework has contributed to the strengthening of NGOs and with the consolidation of these institutions as the linchpin of Republican guaranteeing the right to access to culture, but corporate social responsibility was the one who took off in the segment treated here, through the actions of Responsibility Cultural enterprises arising from the Cultural Organizations. Therefore, in the face of this discourse, this study ascertains the process of encouraging the Culture in Rio Grande do Norte from the Deviant Case Analysis at the Casa da Ribeira, the main Cultural Organization that operates, focused action in Natal in order to assess the relationships established between the same entity and the institutions which are entitled to maintain the process of encouraging treated in this study - Enterprise, from the viewpoint of corporate sponsorship and Cultural Responsibility and State in the form of the Laws Incentive Funds and Public Culture Incentive
Resumo:
This work intends to describe and analyze the parties of forró that happen in Serra da Gameleira, in São Tomé/RN. Serra is a divided social space: groups of different ethnic origins live together in Gameleira de Baixo, Salgadinho (or Gameleira de Cima) and Chaves Belas. They are approximately two hundred families that live exclusively from agriculture. We try to understand how the parties inform about the social organization, the ethnic composition of the families that live there and the past of Serra, through the genealogy offorró players. In the discussion, we identify the festive places: in the total, we have Five houses of forró that function regularly one of them has been described. The private and public spaces inside them are intimate related, with no clear limits between the house of forró and the residence. Each house of forró has an owner, that regularly makes the parties, mobilizing a big part of the inhabitants, and provoking the straitening of the social relations. Observing the festive sociability between different social segments, the forró appears like na element that minimizes social conflicts, providing news ways of association and cooperation in the space of Serra da Gameleira. For the collection of facts, we used the ethnographic method, through the direct observation, interviewing and documentary research. The local history is recounted following the routes of oral memory and historical documents analysis. In the end of the analysis, we concluded that music and party are elements that aggregate the different groups that live in the location and determine forms of expression of what is seen as a traditional culture
Resumo:
LaAdministración Pública manifiesta su voluntad de diferentes maneras conocidas como Acto Administrativo, Actos de Simple Administración, Hecho Administrativo, Contrato Administrativo, Acto Normativo; todas estas formas de manifestación de voluntad son parte de la rama del Derecho Administrativo, las cuales han sido estudiadas por diversos doctrinarios; en este caso en concreto el estudio de esta monografía se centra en el Hecho Administrativoque no es más que toda actividad material traducida en actuaciones técnicas o físicas que son ejecutadas por medio de quien representa a la Administración Pública. En la primera parte de este proyecto monográfico el objetivo es recoger las diversas formas de manifestación de voluntad de la Administración Pública, para poder estudiarlas y analizarlas según la doctrina, y dentro de nuestro ordenamiento jurídico. Para así, una vez identificadas cada una de estas figuras, proceder al estudio del Hecho Administrativo. Una vez estudiado e identificado al Hecho Administrativo en nuestro ordenamiento jurídico; analizaremos su importancia dentro del campo práctico del Derecho Administrativo, para lo que nos remitiremos a un caso en la vía administrativa, como lo es la muerte de la hija de la ex ministra Guadalupe Larriva; y a un caso en la vía jurisdiccional, en que el señor Iván Vásquez demanda a la Federación Deportiva del Azuay, la impugnación de un Hecho Administrativo.
Resumo:
ResumenLa historia de los derechos humanos en Costa Rica muestra una característica muy particular,por cuanto desde mucho antes de haberse promulgado la Declaración Universal de los Derechos Humanos en 1948 ya se daban pasos en su búsqueda. Desde el siglo XlX, aunque fuera de forma clandestina o artesanal, se empezaron a hacer sentir organizaciones y distintas formas de expresión a través de las cuales denunciar las malas condiciones de vida y alcanzar algunos beneficios. En medio de circunstancias adversas y poco apropiadas los movimientos fueron aumentando de intensidad al punto de que ya para 1930 cobran más vigor, lo que se evidencia con un movimiento obrero más organizado y la conformación del Partido Comunista. La lucha por lo que hoy conocemos como derechos humanos logra alcanzar su mayor esplendor en los años de 1940 gracias a las circunstancias coyunturales que favorecieron la presencia de un gobierno reformista como el del presidente Rafael Ángel Calderón Guardia y una activa participación de la Iglesia católica a través de Mons. Víctor Sanabria. De esta forma en Costa Rica se obtuvo el paquete de garantías sociales: un código de trabajo que regulara todo lo pertinenteal trabajo y la seguridad social con la creación de la CCSS. En un tiempo prudencial y con una amplia participación social los y las costarricenses alcanzaron derechos económicos, sociales, políticos y culturales.Palabras clave: derechos humanos, legislación social, código de trabajo, movimiento obrero, Partido Comunista, Iglesia católica.AbstractThe history of Human rights in Costa Rica shows a particular characteristic because long before the promulgation of the Universal Declaration of Human Rights in 1948, steps towards its search were taken in the country. Since the nineteenth century, although in a clandestine or rustic manner, different organizations and forms of expression through which expose the poor conditions of life and reach some benefits began to be felt. In the midst of adverse and inappropriate circumstances, the movements were increasing in intensity to the point that they got stronger by 1930, as evidenced with a more organized labor movement and the formation f the Communist Party. The fight we know today as human rights reaches its greatest prominence in the 1940’s thanks to temporary circumstances favoring the presence of a reformist government as that of President Rafael Angel Calderon Guardia’s, and an active participation of the Catholic Church by Bishop Víctor Sanabria. In this way Costa Rica obtained the social security package: a working code to regulate all matters related to labor and social security with the creation of the CCSS. In an appropriate manner and with broad social participation, costa ricans reached economic, social, political an cultural rights.Keywords: human rights, social legislation, Labor Code, labor movement, the Communist Party, Catholic Church.
Resumo:
Although human toxocariasis ranks among the most common zoonotic infections worldwide, it remains relatively unknown to the public. The causal agents are the nematode parasites Toxocara canis and T. cati, whose definitive hosts are dogs and cats, respectively. When embryonated eggs are accidentally ingested by humans, larvae hatch in the small intestine, penetrate the intestinal wall and migrate, via the bloodstream, to the liver, lungs, muscles, eye and central nervous system. Although most human infections are asymptomatic, two well-defined clinical syndromes are classically recognised: visceral larva migrans (a systemic disease caused by larval migration through major organs) and ocular larva migrans (a disease limited to the eyes and optic nerves). Two less-severe syndromes have recently been described, one mainly in children (covert toxocariasis) and the other mainly in adults (common toxocariasis). Here, the current laboratory diagnosis, epidemiology and main clinical features of both the systemic and ocular forms of human toxocariasis are reviewed. New developments in serological diagnosis are described, the available seroprevalence data are analysed, and the results of relevant clinical studies that have been published over the last decade are explored, to provide an updated overview of this neglected but highly prevalent human infection.
Resumo:
Maize streak disease is a severe agricultural problem in Africa and the development of maize genotypes resistant to the causal agent, Maize streak virus (MSV), is a priority. A transgenic approach to engineering MSV-resistant maize was developed and tested in this study. A pathogen-derived resistance strategy was adopted by using targeted deletions and nucleotide-substitution mutants of the multifunctional MSV replication-associated protein gene (rep). Various rep gene constructs were tested for their efficacy in limiting replication of wild-type MSV by co-bombardment of maize suspension cells together with an infectious genomic clone of MSV and assaying replicative forms of DNA by quantitative PCR. Digitaria sanguinalis, an MSV-sensitive grass species used as a model monocot, was then transformed with constructs that had inhibited virus replication in the transient-expression system. Challenge experiments using leafhopper-transmitted MSV indicated significant MSV resistance - from highly resistant to immune - in regenerated transgenic D. sanguinalis lines. Whereas regenerated lines containing a mutated full-length rep gene displayed developmental and growth defects, those containing a truncated rep gene both were fertile and displayed no growth defects, making the truncated gene a suitable candidate for the development of transgenic MSV-resistant maize. © 2007 SGM.
Resumo:
Expression vectors were constructed for Trichoderma reesei using the promoters, secretion signals and the modular structure of the efficiently expressed and secreted cellulase enzymes EGL2 (Cel5A) and CBH2 (Cel6A) as a prelude to establishing a platform where a gene of interest can be expressed under several promoters simultaneously. The designs featured (i) EGL2sigpro (egl2 promoter and secretion signal), (ii) EGL2cbmlin (egl2 promoter, secretion signal, EGL2 cellulose binding module and linker), (iii) CBH2sigpro (cbh2 promoter and secretion signal) and (iv) CBH2cbmlin (cbh2 promoter, secretion signal, CBH2 cellulose binding module and linker). Recombinant vectors were introduced individually into the high protein-secreting T. reesei RUT-C30 strain to generate single-promoter transformants expressing the Dictyoglomus thermophilum xynB gene that encodes a thermophilic xylanase enzyme (XynB). Ten transformants producing XynB representing each of the four different types of vectors were selected for further testing and the highest XynB production was achieved from a transformant containing 1–2 copies of the EGL2cbmlin vector. Best xylanase producers did not show any particular pattern in terms of the number of gene copies and their mode of integration into the chromosomal DNA. Transformants generated with the cbmlin-type vectors produced multiple forms of XynB which were decorated with various N- and O-glycans. One of the O-glycans was identified as hexuronic acid, whose presence had not been observed previously in the glycosylation patterns of T. reesei.
Resumo:
Thraustochytrids have become of considerable industrial and scientific interest in the past decade due to their health benefits. They have been proven to be the principle source in marine and estuarine fish diets with high percentage of long chain (LC) or polyunsaturated fatty acids (PUFA). Therefore, the oil extracted from fish for human document.forms[0].elements[13].select();consumption is rich in PUFA with high omega-3 fatty acid content. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) of all of the omega-3 fatty acids, are considered beneficial essential oils for humans with a wide range of health benefits. These include brain and neural development in infants, general wellbeing of adults and drug delivery through precursor molecules. They have become one of the most extensively studied organisms for industrial oil preparations as PUFA extraction from fish becomes less profitable. Many forms of these Thraustochytrid oils are being trialled for human consumption all over the world. In Australia, there has been little research performed on these organisms in the past ten years. A few Australian studies have been conducted in the form of comparative studies related to PUFA production within the related genera, but not focussed on their identification or cellular and genomic characterisation. Therefore, the main aim of this study was to investigate the morphological and genetic characteristics of Australian Thraustochytrids in order to aid in their identification and characterisation, as well as to better understand the effect of environmental conditions in the regulation of PUFA production. It was also noted that there was a knowledge gap in the preservation and total genomic DNA extraction of these organisms for the purposes of scientific research. The cryopreservation of these organisms for studies around the world follows existing generic methods. However, it is well understood that many of these generic methods attract not only high costs for chemicals, but also uses considerable storage space and other resources, all of which can be improved with new or modified approaches. In this context, a simple and inexpensive bead preservation method is described, without compromising the storage shelf life. We also describe, for the first time, the effects of culture age on the successful cryopreservation of Thraustochytrids. It was evident in the literature that DNA and RNA extractions for molecular and genetic studies of Thraustochytrids follow the classical phenol-chloroform extraction methods. It was also observed that modern protocols failed to avoid the use of phenol-chloroform rather than improving preparation and cell disruption. In order to provide a high quantity and quality DNA extraction, a modified protocol has been introduced that employs the use of modern commercial extraction kits and standard laboratory equipment. Thraustochytrids have been shown to be highly conserved in their 18S rDNA gene sequences, which is used as the current standard for identification. It was demonstrated that the 18S rDNA gene sequence limits the recognition of closely related genera or within the genera from each member. Therefore, it was proposed that another profile, such as a randomly amplified polymorphic DNA (RAPD) based profiling system, be tested for use in the characterisation of Thraustochytrids. The RAPD profiles were shown to provide a unique DNA fingerprint for each isolate and small variations in their genome were able to be detected. This method involved the use of a minimum number of standard arbitrary primers and with an increase in the number of different primers used, a very high discrimination between organisms could be achieved. However, the method was not suitable for taxonomic purposes because the results did not correlate with other taxonomic features such as morphology. Another knowledge gap was found with respect to Australian Thraustochytrid growth characteristics, in that these had not been recorded and published. In order to rectify this, a record of colony and microscopic features of 12 selected isolates was performed. The results of preliminary studies indicated that further microbiological and biochemical studies are needed for full characterisation of these organisms. This information is of great importance to bio-prospecting of new Thraustochytrids from Australian ecosystems and would allow for their accurate identification, and so permit the prediction of their PUFA capability by comparison with related genera/species. It was well recognized that environmental stress plays a role in the PUFA production and is mainly due to the reactive oxygen species as abiotic stress (Chiou et al., 2001; Okuyama et al., 2008; Shabala et al., 2009; Shabala et al., 2001). In this aspect, this study makes the first attempt towards better understanding of this phenomenon by way of the use of real-time PCR for the detection of environmental effects on the regulation of PUFA production. Three main environmental conditions including temperature, pH and oxygen availability were monitored as stress inducers. In summary, this study provides novel approaches for the preservation and handling of Thraustochytrids, their molecular biological features, taxonomy, characterisation and responses to environmental factors with respect to their oil production enzymes. The information produced from this study will prove to be vital for both industrial and scientific investigations in the future.
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Guanylyl cyclase C (GC-C) is a membrane-associated form of guanylyl cyclase and serves as the receptor for the heat-stable enterotoxin (ST) peptide and endogenous ligands guanylin, uroguanylin, and lymphoguanylin. The major site of expression of GC-C is the intestinal epithelial cell, although GC-C is also expressed in extraintestinal tissue such as the kidney, airway epithelium, perinatal liver, stomach, brain, and adrenal glands. Binding of ligands to GC-C leads to accumulation of intracellular cGMP, the activation of protein kinases G and A, and phosphorylation of the cystic fibrosis transmembrane conductance regulator (CFTR), a chloride channel that regulates salt and water secretion. We examined the expression of GC-C and its ligands in various tissues of the reproductive tract of the rat. Using reverse transcriptase and the polymerase chain reaction, we demonstrated the presence of GC-C, uroguanylin, and guanylin mRNA in both male and female reproductive organs. Western blot analysis using a monoclonal antibody to GC-C revealed the presence of differentially glycosylated forms of GC-C in the caput and cauda epididymis. Exogenous addition of uroguanylin to minced epididymal tissue resulted in cGMP accumulation, suggesting an autocrine or endocrine activation of GC-C in this tissue. Immunohistochemical analyses demonstrated expression of GC-C in the tubular epithelial cells of both the caput epididymis and cauda epididymis. Our results suggest that the GC-C signaling pathway could converge on CFTR in the epididymis and perhaps control fluid and ion balance for optimal sperm maturation and storage in this tissue.
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Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .