Resposta celular Th1 /Th2 na doença periodontal experimental, em ratos: um estudo imuno-histoquímico

Host response plays a major role in the pathogenesis of periodontal disease. Mediators such as inflammatory cytokines which are secreted during the immune response to bacterial challenges have ambiguous functions that may or may not lead to protection of the attacked tissue. In this context, experimental evidence suggests that T-helper 1 (Th-1) and T-helper 2 (Th-2) mediated responses are potentially important during the disease process. The aims of this study therefore were to further clarify the role played by Th2 cells during different time points of the active phase of periodontal disease, as well as, to investigate whether there was any evidence of a Th1 response in the periodontal disease microenvironment. Experimental periodontitis was induced in 30 Wistar male rats by placing cotton ligatures around the mandibular first molars. The rats were then randomly divided into two groups. Group1 (G1=15) and Group 2 (G2=15). In G1 the ligatures were maintained for 2 days, whereas in G2 the ligatures were left for 15 days, a time point that corresponds to the advanced stage of periodontal disease The contra-lateral teeth served as controls (no ligatures). Immunohistochemical investigation for the presence in gingival tissue of Th2 specific transcription factor (GATA3) and the subunit of the IFN-γ receptor was carried out after the disease induction period. Light microscopy analysis revealed a decrease in the expression of GATA-3 as bone loss progressed. On the other hand, although IFN-γ R1 was detected at an early stage of the active phase of disease its expression remained unaltered during the remaining period of the study. These results indicate that the Th2 response have a protective role during the pathogenesis of periodontal disease and that the progression of the periodontal disease is related with the unbalance of the responses Th1/Th2

Estudo da detecção do DNA do papiloma vírus humano (HPV) e da expressão imuno-histoquímica de proteína do ciclo celular no carcinoma epidermóide oral

Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral cavity and human papillomavirus (HPV) may have an important role in its development. The aim of this experiment was to investigate the HPV DNA and viral types in 90 cases of OSCC. Moreover, a comparative analysis between the cases of OSSC with and without HPV DNA was performed by using cell cycle markers p21 and pRb in order to detect a possible correlation of these proteins and HPV infection. DNA was extracted from paraffin embedded tissue and amplified by PCR (polymerase chain reaction) with primers PCO3+ e PCO4+ for a fragment of human β-globin gene. After this procedure, PCR for HPV DNA detection was realized using a pair of generic primers GP5+ e GP6+. Immunohistochemical study was performed by streptoavidin-biotin technique and antibodies against p21 and pRb proteins were employed. Eighty-eight cases were positive for human β-globin gene and HPV DNA was found in 26 (29.5%) of then. It could not be detected significant correlation between HPV and age, sex and anatomical sites of the lesion. The most prevalent viral type was HPV 18 (80.8%). Regarding the immunohistochemical analysis, it was detected significant association between HPV presence and pRb immunoexpression (p=0,044), nevertheless, the same was not observed in relation to p21 protein (p =0,416). It can be concluded that the low detection of HPV DNA in OSCC by the present experiment suggests a possible role of the virus in the development and progression in just a subset of this disease

Plasticidade de células tronco mesenquimais de medula óssea de ratos na presença de meio condicionado do nervo facial e do fator de crescimento fibroblástico 2

A number of evidences show the influence of the growth of injured nerve fibers in Peripheral Nervous System (PNS) as well as potential implant stem cells (SCs) to make it more suitable for nerve regeneration medium. In this perspective, this study aimed to evaluate the plasticity of mesenchymal stem cells from bone marrow of mice in the presence of culture medium conditioned with facial nerve explants (D-10) and fibroblast growth factor-2 (FGF-2). In this perspective, the cells were cultivated only with DMEM (group 1), only with D-10(group 2), only with FGF-2(group 3) or with D-10 and FGF-2(group 4). The growth and morphology were assessed over 72 hours. Quantitative phenotypic analysis was taken from the immunocytochemistry for GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200 on the fourth day of cultivation. Cells cultured with conditioned medium alone or combined with FGF-2 showed distinct morphological features similar apparent at certain times with neurons and glial cells and a significant proliferative activity in groups 2 and 4 throughout the days. Cells cultived only with conditioned medium acquired a glial phenotype. Cells cultured with FGF-2 and conditioned medium expressed GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200. On average, area and perimeter fo the group of cells positive for GFAP and the área of the cells immunostained for OX-42 were higher than those of the group 4. This study enabled the plasticity of mesenchymal cells (MCs) in neuronal and glial nineage and opened prospects for the search with cell therapy and transdifferentiation

Caracaterização do núcleo pré-geniculado do sagüi (Callithrix jacchus) :projeção retiniana, neuroquímica e atividade celular (expressão de FOS)

In rodents, the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) are the main components of the circadian system. The SCN is considerate the site of an endogenous biological clock because can to generate rhythm and to synchronize to the environmental cues (zeitgebers) and IGL has been related as one of the main areas that modulate the action of SCN. Both receive projections of ganglion cells of retina and this projection to SCN is called retinohypothalamic tract (RHT). Moreover, the IGL is connected with SCN through of geniculohypothalamic tract (GHT). In primates (include humans) was not still demonstrated the presence of a homologous structure to the IGL. It is believed that the pregeniculate nucleus (PGN) can be the answer, but nothing it was still proven. Trying to answer that question, the objective of our study is to do a comparative analysis among PGN and IGL through of techniques immunohystochemicals, neural tracers and FOS expression after dark pulses. For this, we used as experimental model a primate of the new world, the common marmoset (Callithrix jacchus). Ours results may contribute to the elucidation of this lacuna in the circadian system once that the IGL is responsible for the transmission of nonphotic information to SCN and participate in the integration between photic and nonphotic stimulus to adjust the function of the SCN. In this way to find a same structure in primates represent an important achieve in the understanding of the biological rhythms in those animals

Viabilidade econômica do cultivo de shiitake em diferentes escalas de produção

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

Influência da composição química do substrato no cultivo de Pleurotus florida

Com o objetivo de avaliar a influência da composição química do substrato no cultivo do cogumelo comestível Pleurotus florida, seis resíduos agrícolas foram testados: palha de arroz, palha de feijão, palha de trigo, folha de bananeira, palha de sorgo e sabugo de milho. O substrato palha de feijão apresentou resultados semelhantes para a produção (189,8 g kg-1), eficiência biológica (89,2%) e número de cogumelos (12) à palha de arroz, substrato utilizado tradicionalmente no cultivo de Pleurotus em escala comercial. Não foi possível atribuir apenas a um fator químico as altas produções e eficiências biológicas observadas em palha de arroz e palha de feijão e muito menos para a baixa produção em palha de sorgo (77,8 g kg-1) e sabugo de milho (53,2 g g-1). No geral, substratos com relação C/N em torno de 45 (N = 1,0%), maior conteúdo de cinzas, acrescido de altos teores de P, K, Ca, Mg and Mn foram os melhores para o cultivo de P. florida.

Extração e análise da fração lipídica da microalga monoraphidium sp., síntese e caracterização do seu biodiesel

The cultivation of microalgae biomass in order to produce biodiesel arises as an extremely promising aspect, in that the microalgae culture includes short cycle of reproduction, smaller areas for planting and residual biomass rich in protein content. The present dissertation evaluates the performance and features, through spectrometry in the region of infrared with transformed Fourier (FTIR) and spectrometry in the region of UVvisible (UV-Vis), of the extracted lipid material (LM) using different techniques of cell wall disruption (mechanical agitation at low and at high spin and agitation associated with cavitation). The technique of gas chromatography (GC) brought to light the success of alkaline transesterification in the conversion of oil into methyl monoesters (MME), which was also analyzed by spectroscopic techniques (FTIR, proton magnetic resonance (1H NMR) and carbon (13C NMR). Through thermogravimetric analysis (TGA) were analyzed the lipid material (LM), biodiesel and the microalgae biomass. The method which provided the best results concerning the efficiency in extraction of the LP of Monoraphidium sp. (12,51%) was by mechanical agitation at high spin (14 000 rpm), for 2 hours being the ideal time, as shown by the t test. The spectroscopic techniques (1H NMR, 13C NMR and FTIR) confirmed that the structure of methyl monoesters and the chromatographic data (CG) revealed a high content of saturated fatty acid esters (about 70%) being the major constituent eicosanoic acid (33,7%), which justifies the high thermal stability of microalgae biodiesel. The TGA also ratified the conversion rate (96%) of LM into MME, pointing out the quantitative results compatible with the values obtained through GC (about 98%) and confirmed the efficiency of the extraction methods used, showing that may be a good technique to confirm the extraction of these materials. The content of LM microalgae obtained (12,51%) indicates good potential for using such material as a raw material for biodiesel production, when compared to oil content which can be obtained from traditional oil for this use, since the productivity of microalgae per hectare is much larger and requires an extremely reduced period to renew its cultivation

Resíduos orgânicos como recondicionante de subsolo degradado e efeitos na atividade microbiana e fertilidade em cultivo de barbatimão

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Produção e composição bromatológica de forrageiras em sistema de integração lavoura-pecuária em diferentes épocas de semeadura

O objetivo deste trabalho foi avaliar a produção de matéria seca e a composição bromatológica de forrageiras em sistema de integração lavoura-pecuária, em diferentes datas de semeadura. Foi usado delineamento experimental de blocos ao acaso, em esquema fatorial 4x3, com quatro forrageiras - braquiária, Urochloa brizantha; capim-moa, Setaria italica; milheto, Pennisetum glaucum; e sorgo forrageiro, Sorghum bicolor - semeadas em três datas nas estações inverno/primavera e verão/outono, em sucessão à soja. O milheto e o sorgo forrageiro produziram maior quantidade de matéria seca em todas as datas de semeadura, nas duas estações de cultivo e, ao contrário da braquiária e do capim-moa, não diminuíram o teor de nutrientes digestíveis totais no inverno/primavera. Os teores de proteína bruta reduziram com o avanço das datas de semeadura no inverno/primavera e aumentaram no verão/outono, exceto para braquiária e milheto. O sorgo apresentou os menores teores de fibras em detergente neutro nas segunda e terceira datas de semeadura no inverno/primavera, e na primeira data de semeadura no verão/outono. Nas segunda e terceiras datas do inverno/primavera, o sorgo apresentou menor teor de fibras em detergente ácido. Quanto aos constituintes da parede celular, apenas os teores de hemicelulose não se alteram nas forrageiras em virtude das datas de semeadura, em ambas as estações.

Propriedades físicas de um Latossolo Vermelho sob cultivo de arroz de terras altas em diferentes manejos do solo e água

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

Manejo do solo, água e nitrogênio no cultivo de feijão

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

Avaliação de fontes de nitrogênio e épocas de aplicação em cobertura para o cultivo do trigo

The mineral nutrition, mainly to concern to nitrogen, is essential to obtain high productivity in the corn crop. This work was accomplished at experimental station of State University of São Paulo - UNESP - Ilha Solteira Campus, UNESP/FEIS, in Selviria-MS, in a Red dystrophic Latosol (Oxisol), in the agricultural year 2004/05. The objective of this study was to evaluate sources and times of nitrogen application on corn yield. A randomized blocks design was used with four replications, in a 3x5 factorial scheme. The treatments were three sources of nitrogen (Entec - Ammonium sulfonitrate with nitrification inhibitor, Ammonium sulfate and Urea) applied at sowing and/or at covering, stadium of 6-8 leaves completely unfolded, in 5 combinations (0 + 120; 30 + 90; 60 + 60; 90 + 30 and 120 + 0 kg of N ha(-1)). The nitrogen sources as Entec and the combination of 60 kg ha(-1) at sowing and at 6-8 leaf stadium provided the larger N leaf content. The higher grain yield was reached in the treatment that received the higher N doses at covering.

Estudo da descelularização tecidual na produção de arcabouços biológicos para enxertos

The regeneration of bone defects with loss of substance remains as a therapeutic challenge in the medical field. There are basically four types of grafts: autologous, allogenic, xenogenic and isogenic. It is a consensus that autologous bone is the most suitable material for this purpose, but there are limitations to its use, especially the insufficient amount in the donor. Surveys show that the components of the extracellular matrix (ECM) are generally conserved between different species and are well tolerated even in xenogenic recipient. Thus, several studies have been conducted in the search for a replacement for autogenous bone scaffold using the technique of decellularization. To obtain these scaffolds, tissue must undergo a process of cell removal that causes minimal adverse effects on the composition, biological activity and mechanical integrity of the remaining extracellular matrix. There is not, however, a conformity among researchers about the best protocol for decellularization, since each of these treatments interfere differently in biochemical composition, ultrastructure and mechanical properties of the extracellular matrix, affecting the type of immune response to the material. Further down the arsenal of research involving decellularization bone tissue represents another obstacle to the arrival of a consensus protocol. The present study aimed to evaluate the influence of decellularization methods in the production of biological scaffolds from skeletal organs of mice, for their use for grafting. This was a laboratory study, sequenced in two distinct stages. In the first phase 12 mice hemi-calvariae were evaluated, divided into three groups (n = 4) and submitted to three different decellularization protocols (SDS [group I], trypsin [Group II], Triton X-100 [Group III]). We tried to identify the one that promotes most efficient cell removal, simultaneously to the best structural preservation of the bone extracellular matrix. Therefore, we performed quantitative analysis of the number of remaining cells and descriptive analysis of the scaffolds, made possible by microscopy. In the second stage, a study was conducted to evaluate the in vitro adhesion of mice bone marrow mesenchymal cells, cultured on these scaffolds, previously decellularized. Through manual counting of cells on scaffolds there was a complete cell removal in Group II, Group I showed a practically complete cell removal, and Group III displayed cell remains. The findings allowed us to observe a significant difference only between Groups II and III (p = 0.042). Better maintenance of the collagen structure was obtained with Triton X-100, whereas the decellularization with Trypsin was responsible for the major structural changes in the scaffolds. After culture, the adhesion of mesenchymal cells was only observed in specimens deccelularized with Trypsin. Due to the potential for total removal of cells and the ability to allow adherence of these, the protocol based on the use of Trypsin (Group II) was considered the most suitable for use in future experiments involving bone grafting decellularized scaffolds

Fitomassa de plantas de cobertura em diferentes sucessões de culturas e sistemas de cultivo

Este trabalho foi realizado em Latossolo Vermelho Distrófico típico argiloso, na região de Selvíria (MS), com o objetivo de investigar a produção de massa verde e seca de plantas de cobertura sob três sucessões de culturas e dois preparos de solo. O delineamento experimental utilizado foi o de blocos ao acaso, com quatro repetições no esquema em faixas com parcelas subsubdivididas. As faixas foram constituídas por cinco coberturas: mucuna-preta, milheto, crotalária, guandu e vegetação espontânea (pousio) e subdivididas em dois sistemas de semeadura (direta e convencional), sendo essas últimas subsubdivididas com as culturas de milho, soja e algodão. As avaliações foram realizadas no ano agrícola de 2000/2001, e a área em estudo estava sob o efeito dos respectivos manejos desde o ano agrícola de 1997/1998. Verificou-se que entre as plantas de cobertura, as produções de massa verde e seca foram menores para o guandu e maiores para o milheto e a crotalária; a produção de fitomassa de plantas de cobertura é diferente entre os sistemas convencional e direto; as plantas de cobertura na área com algodão no primeiro e segundo ano agrícola produziram mais massa seca; as plantas de cobertura produziram menor quantidade de massa verde e seca sob a sucessão com a cultura da soja em semeadura direta.

Mecanismos de abertura do sulco e adubação nitrogenada no cultivo do feijoeiro em sistema plantio direto

Algumas culturas têm pouca adaptação ao sistema plantio direto, em vista da maior compactação da camada superficial do solo e, nesse caso, o mecanismo utilizado na semeadora para a abertura dos sulcos para deposição do fertilizante pode ter grande importância no sentido de facilitar a penetração das raízes. Este experimento foi desenvolvido em Selvíria (MS), com o objetivo de avaliar a produtividade do feijão de inverno cultivado em sistema plantio direto, em função da utilização de mecanismos de abertura para distribuição de fertilizantes na semeadura e da adubação nitrogenada em cobertura. O delineamento experimental utilizado foi em blocos casualizados, utilizando-se esquema fatorial 2 x 6, constituído por mecanismos de distribuição de fertilizante (haste escarificadora e disco duplo) e doses de N em cobertura (0, 25, 50, 75, 100 e 125 kg ha-1), com quatro repetições. Recomenda-se o uso da haste escarificadora como mecanismo de distribuição do fertilizante, para o cultivo do feijoeiro de inverno. A adubação nitrogenada em cobertura proporciona incrementos à produtividade do feijoeiro de inverno.