Contributo da exsudação de ácidos orgânicos para a tolerância ao alumínio em duas variedades de trigo da Madeira

O trigo encontra-se em terceiro lugar entre os cereais mais produzidos em todo o mundo. Um dos principais entraves ao seu cultivo e produção é a acidez dos solos, que proporciona a biodisponibilidade do alumínio, com a formação de catiões facilmente absorvidos que afetam o desenvolvimento radicular e podem levar à morte da planta. Algumas variedades de trigo desenvolveram a capacidade de tolerar a presença do alumínio. Esta tolerância pode resultar de diferentes estratégias, através da ação de diversos mecanismos, sendo que o presente trabalho pretende avaliar a importância da exsudação de ácidos orgânicos na tolerância ao alumínio por algumas variedades de trigo na Madeira. Ao longo deste trabalho foram analisados vários caracteres, cuja variação permite discriminar o comportamento de duas variedades regionais de trigo (Triticum aestivum erithrospermum Körn e Triticum aestivum var. milturum (Alef.) Velican) em condições de stress provocado pelo alumínio. As amostras de trigo foram colocadas em vasos herméticos na presença ou ausência da alumínio e o meio de crescimento final foi analisado para determinar a capacidade das plantas para exsudar malato e citrato. As plantas foram analisadas em relação a cinco marcadores moleculares para detetar a presença ou ausência do gene ALTM1 que codifica a proteína transmembranar de transporte do malato. Em resultado deste trabalho, conclui-se que uma das variedades regionais (T. aestivum erithrospermum) é tolerante ao alumínio e a outra (T. aestivum var. milturum) moderadamente tolerante. Ambas as variedades têm capacidade de exsudar ácidos orgânicos, ainda que a primeira tivesse uma exsudação mais proeminente. A variedade moderadamente tolerante apresentou uma taxa de alongamento radicular inferior e uma produção de calose superior devido à sua maior suscetibilidade ao alumínio. O gene ALMT1, responsável pelo transporte do malato do citoplasma para o exterior das células, foi detetado em ambas as variedades, levando a concluir que o que difere entre as variedades é a sua expressão.

Estudo comparativo de dois tipos de cultivo: monocultivo (camarões) versus cultivo integrado (algas/camarões)

The integrated culture of seaweed and aquatic animals is an ancient practice in Asian countries. The expansion of this practice to western countries is consequence of the recognition of this system as a sustainable alternative that allows economical diversification and mitigation of environmental impacts generated by effluents of aquaculture. This study evaluated the growth of the seaweed Gracilaria caudata and of the shrimp Litopenaeus vannamei in monoculture (shrimps) and integrated culture (shrimps and algae) systems, and accessed the effect of the seaweed in the water quality. There were two treatments in the experiment: monoculture (shrimps) and integrated culture (shrimps/ algae). The organisms were cultured in 6 aquaria (10L) filled with seawater (35.0±0.0 PSU and 28.1±0.4°C) for 28 days. The nutrients of water (PO43-, NH4+, NO2-, NO3- and DIN), the biomass and the relative growth rate (RGR, % day-1) of seaweed and shrimps were measured weekly. The parameters pH, temperature, salinity and dissolved oxygen were measured daily. The concentration of NH4+ in integrated culture (62.8±25.2µM) was lower (Mann-Whitney p<0.001) than in monoculture (85.6±24.3µM). The mean of PO4- in monoculture (10.4±4.6µM) was markedly higher (Mann-Whitney; p=0.024) than that in integrated culture (8.7±4.1µM). The level of dissolved oxygen in integrated culture (6.0±0.6mg/L) was higher (t-Student; P=0.014) than that in shrimp monoculture (5.8±0.6mg/L). The mean values of the parameters pH, NO2-, NO3- and DIN were 7.5±0.2, 10.1±12.2µM, 24.5±3.2µM and 120.17±30.76µM in monoculture, and 7.5±0.2, 10.5±13.2µM, 27.4±3.5µM and 100.76±49.59µM in integrated culture. There were not differences in these parameters between treatments. The biomass and RGR of seaweed reached 15.0±1.9g and 7.4±2.8% day-1 at the end of the experiment. The performance of shrimp was favorable in monoculture (1.5±0.8g; 5.7±1.6% dia-1) and in integrated culture (1.5±0.7g; 5.2±1.2% dia-1), and the rate of survival was 100% in both treatments. The tolerance and favorable performance of Gracilaria caudata suggest that this seaweed might be integrated into shrimp (Litopenaeus vannamei) culture systems

Caracterização limnológica e determinação da capacidade suporte do reservatório mendubim, (Rio Grande do Norte) para o cultivo de peixes em tanques-rede

The aims of this study were: i) assessing the trophic state of the Mendubim reservoir (semi-arid, Rio Grande do Norte, Brazil; 05° 38 99,0 S 36°55 98,0 W) based on chlorophyll-a, total phosphorus and nitrogen concentrations and water transparency; ii) relating the patterns of temporal variation of zooplankton and phytoplankton to the trophic state of the reservoir and iii) investigating the carrying capacity of the reservoir for cage fish farming. The samplingwas done monthly from July 2006 to July 2007 in three stations at the reservoir: next to the dam (barrage), in the central region and in the mouth of the main tributary. The abiotic and biotic variables analyzed were: Secchi depth, volatiles and fixed suspended solids, chlorophyll-a, total phosphorus and nitrogen, TN:TP ratio and mesozooplankton and phytoplankton composition and biomass. The results showed that the reservoir can be considered as mesotrophic with mean concentrations of total nitrogen, phosphorus and chlorophyll-a equal to 1711, 1 μg.L-1, 30,8 μg.L-1 and 5,62 μg.L-1 respectively. The Cyanophyceae class was the most representative in terms of density, with the presence of potentially toxic species such as Microcystis aeruginosa, Planktothrix planctonica, Cylindrospermopsis raciborskii, Aphanizomenon sp. ,Aphanocapsa delicatissima and Pseudanabaena acicularis. Among the zooplankton, the genus Notodiaptomus presented the largest biomass values. Overall, our results show that the light limitation should explain the weak relationship between chlorophyll-a and total phosphorus and nitrogen concentrations. We concluded that the water of Mendubim reservoir is suitable for intensive fish cage aquaculture. Based on the carrying capacity calculations for this reservoir, we found that the maximum sustainable yield of tilapias in cages in the reservoir is 126 ton per year assuming a factor of food conversion of 1.5: 1.0 and a phosphorus content in the fish food of 1%

Biodiversidade e abundância da ictiofauna associada ao cultivo orgânico de Litopenaeus vannamei (Boone, 1931)

The expansion of shrimp farming has caused a series of environmental impacts, often as a result of lack of planning and adequate management. Organic aquaculture has emerged as an alternative to conventional shrimp farming, and differently, aims at the economical, ecological and farming potential of other organisms, such as fishes, oysters and seaweeds. The present study aimed at evaluating the biological diversity and abundance of the ichthyofauna associated to Litopenaeus vannamei organic culture at PRIMAR farm (Rio Grande do Norte, Brazil). The sampling period consisted of four culture cycles (1, 2, 3, and 4) accomplished in four grow-out ponds (V1, V7, V2, and V4) in 2005. The first two culture cycles were carried out during the rainy season, and the last two cycles, during the dry season. Environmental (temperature, salinity, transparency, and rainfall), biodiversity (diversity and uniformity and abundance indexes) of the ichthyofauna, and shrimp productivity data were collected throughout these four culture cycles. The results obtained for the environmental variables showed that both salinity and transparency oscillated in a significant way (p < 0.001) between culture cycles. In relation to the abundance of the ichthyofauna, fifty four species associated to the organic culture of Litopenaeus vannamei were collected and identified at PRIMAR. For the biodiversity criterion, larger species richness (S = 46) and uniformity (E = 0.59 ± 0.16) were observed during cycles 3 and 4 than in cycles 1 and 2 (S = 41 and E = 0.15 ± 0.12). A positive and significant correlation was obtained for the transparency with diversity and uniformity of fish species between cycles 1 and 2 (wet season) and cycles 3 and 4 (dry season) (p < 0.017 - Shannon Index; p < 0.008 - Pielou Index, respectively). A correlation could not be established between shrimp productivity and fish biomass. However, fish biomass decreased from cycles 1 and 2 to cycles 3 and 4, concomitant with an increase in shrimp productivity, most probably due to a higher (from 58.0% to 71.0%) shrimp survival. Regarding the culture potential of the ichthyofauna, three species (Mugil curema, Mugil liza and Chaetodipterus faber) were identified as potential farming alternatives, either singly or in consortium with Litopenaeus vannamei. A fourth species (Centropomus undecimalis) was indicated as an alternative for single culture or in consortium with other fish species. In conclusion, the large diversity of estuarine fishes associated to Litopenaeus vannamei farming observed at PRIMAR clearly indicated the ecological feasibility for organic aquaculture in northeastern Brazil

Potencialidades de cultivo de macroalgas no litoral do Rio Grande do Norte (NE, Brasil): uma perspectiva de integração de dados ambientais em sistemas de informação geográfica (SIG)

Since centuries ago, the Asians use seaweed as an important source of feeding and are their greatest world-wide consumers. The migration of these peoples for other countries, made the demand for seaweed to increase. This increasing demand prompted an industry with annual values of around US$ 6 billion. The algal biomass used for the industry is collected in natural reservoirs or cultivated. The market necessity for products of the seaweed base promotes an unsustainable exploration of the natural banks, compromising its associated biological balance. In this context, seaweed culture appears as a viable alternative to prevent the depletion of these natural supplies. Geographic Information Systems (GIS) provide space and produce information that can facilitate the evaluation of important physical and socio-economic characteristics for the planning of seaweed culture. This objective of this study is to identify potential coastal areas for seaweed culture in the state of Rio Grande do Norte, from the integration of social-environmental data in the SIG. In order to achieve this objective, a geo-referred database composed of geographical maps, nautical maps and orbital digital images was assembled; and a bank of attributes including physical and oceanographical variables (winds, chains, bathymetry, operational distance from the culture) and social and environmental factors (main income, experience with seaweed harvesting, demographic density, proximity of the sheltered coast and distance of the banks) was produced. In the modeling of the data, the integration of the space database with the bank of attributes for the attainment of the map of potentiality of seaweed culture was carried out. Of a total of 2,011 ha analyzed by the GIS for the culture of seaweed, around 34% or 682 ha were indicated as high potential, 55% or 1,101 ha as medium potential, and 11% or 228 ha as low potential. The good indices of potentiality obtained in the localities studied demonstrate that there are adequate conditions for the installation of seaweed culture in the state of Rio Grande do Norte

Avaliação do efeito citotóxico da lectina da esponja marinha Cliona varians contra células de leucemia mielóide crônica

In this study, a BCR-ABL expressing human chronic myelogenous leukaemia cell line (K562) was used to investigate the antitumoral potential of a novel lectin (CvL) purified from the marine sponge Cliona varians. CvL inhibited the growth of K562 cells with an IC50 value of 70 g/ml, but was ineffective to normal human peripheral blood lymphocytes in the same range of concentrations tested (180 g/ml). Cell death occurred after 72 h of exposure to the lectin and with sign of apoptosis as analysed by DAPI staining. Investigation of the possible effectors of this process showed that cell death occurred in the presence of Bcl-2 and Bax expression, and involved a caspase-independent pathway. Confocal fluorescence microscopy indicated a major role for the lysosomal protease cathepsin B in mediating cell death. Accordingly, pre-incubation of K562 cells with the cathepsin inhibitor L-trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E-64) abolished the cytotoxic effect of CvL. Furthermore, we found upregulation of tumor necrosis factor receptor 1 (TNFR1) and down-modulation of p65 subunit of nuclear factor kappa B (NFB) expression in CvL-treated cells. These effects were accompanied by increased levels of p21 and downmodulation of pRb, suggesting that CvL is capable of cell cycle arrest. Collectively, these findings suggest that cathepsin B acts as death mediator in CvL-induced cytotoxicity possibly in a still uncharacterized connection with the membrane death receptor pathway

Bioprospecção de polissacarídeos sulfatados de macroalgas marinhas do litoral do Rio Grande do Norte: caracterização de uma heterofucana extraída da alga marron sargassum filipendula que induz apoptose em células hela

The coast of Rio Grande do Norte has more than 100 species of seaweed, mostly unexplored regarding their pharmacological potential. The sulfated polysaccharides (PS) are by far the more seaweed compounds studied, these present a range of biological properties, such as anticoagulant activity, anti-inflammatory, antitumor and antioxidant properties. In this study, we extract sulfated polysaccharide rich-extracts of eleven algae from the coast of Rio Grande do Norte (Dictyota cervicornis; Dictiopterys delicatula; Dictyota menstruallis; Dictyota mertensis; Sargassum filipendula; Spatoglossum schröederi; Gracilaria caudata; Caulerpa cupresoides; Caulerpa prolifera; Caulerpa sertularioides e Codim isthmocladum), and these were evaluated for the potential anticoagulant, antioxidant and antiproliferative. All polysaccharide extracts showed activity for anticoagulant, antioxidant and/or antiproliferative activity, especially D. delicatula and S. filipendula, which showed the most prominent pharmacological potential, thereby being chosen to have their sulfated polysaccharides extracted. By fractionating method were obtained six fractions rich in sulfated polysaccharides to the algae D. delicatula (DD-0,5V, DD-0, 7V, DD-1,0v, DD-1,3v, DD-1,5v and DD-2,0) and five fractions to the alga S. filipendula (SF-0,5V, SF-0,7V, SF-1,0v, SF-1,5v and SF-2,0v). For the anticoagulant assay only the fractions of D. delicatula showed activity, with emphasis on DD-1, 5v that presented the most prominent activity, with APTT ratio similar to clexane® at 0.1 mg/mL. When evaluated the antioxidant potential, all fractions showed potential in all tests (total antioxidant capacity, hydroxyl and superoxide radicals scavenging, ferrous chelation and reducing power), however, the ability to chelate iron ions appears as the main mechanism antioxidant of sulfated polysaccharides from seaweed. In antiproliferative assay, all heterofucanas showed dose-dependent activity for the inhibition of cell proliferation of HeLa, however, with the exception of SF-0,7V, SF- 1,0v and SF-1,5v, all fractions showed antiproliferative activity against MC3T3, a normal cell line. The heterofucana SF-1,5V had its antiproliferative mechanism of action evaluated. This heterofucan induces apoptosis in HeLa cells by a pathway caspase independent, promoting the release of apoptosis Inducing Factor (AIF) in the cytosol, which in turn induces chromatin condensation and DNA fragmentation into 50Kb fragments. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer.

Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos

studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- α are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteins

Metagenômica comparativa de solo de regiões de mata atlântica e caatinga do estado do Rio Grande do Norte - Brasil

The microorganisms play very important roles in maintaining ecosystems, which explains the enormous interest in understanding the relationship between these organisms as well as between them and the environment. It is estimated that the total number of prokaryotic cells on Earth is between 4 and 6 x 1030, constituting an enormous biological and genetic pool to be explored. Although currently only 1% of all this wealth can be cultivated by standard laboratory techniques, metagenomic tools allow access to the genomic potential of environmental samples in a independent culture manner, and in combination with third generation sequencing technologies, the samples coverage become even greater. Soils, in particular, are the major reservoirs of this diversity, and many important environments around us, as the Brazilian biomes Caatinga and Atlantic Forest, are poorly studied. Thus, the genetic material from environmental soil samples of Caatinga and Atlantic Forest biomes were extracted by direct techniques, pyrosequenced, and the sequences generated were analyzed by bioinformatics programs (MEGAN MG-RAST and WEBCarma). Taxonomic comparative profiles of the samples showed that the phyla Proteobacteria, Actinobacteria, Acidobacteria and Planctomycetes were the most representative. In addition, fungi of the phylum Ascomycota were identified predominantly in the soil sample from the Atlantic Forest. Metabolic profiles showed that despite the existence of environmental differences, sequences from both samples were similarly placed in the various functional subsystems, indicating no specific habitat functions. This work, a pioneer in taxonomic and metabolic comparative analysis of soil samples from Brazilian biomes, contributes to the knowledge of these complex environmental systems, so far little explored

Efeito de glucanas do fungo Caripia montagnei em modelo de inflamação intestinal induzida por tnbs em ratos Wistar e em células de carcinoma de cólon humano HT-29

Compounds derived from fungi has been the subject of many studies in order to broaden the knowledge of their bioactive potential. Polysaccharides from Caripia montagnei have been described to possess anti-inflammatory and antioxidant properties. In this study, glucans extracted from Caripia montagnei mushroom were chemically characterized and their effects evaluated at different doses and intervals of treatment. It was also described their action on colonic injury in the model of colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS), and its action on cells of the human colon carcinoma (HT-29). Compounds extracted of C. montagnei contain high level of carbohydrates (96%), low content of phenolic compounds (1.5%) and low contamination with proteins (2.5%). The (FT-IR) and (NMR) analysis showed that polysaccharides from this species of mushroom are composed of α- and β-glucans. The colonic damage was evaluated by macroscopic, histological, biochemical and immunologic analyses. The results showed a reduction of colonic lesions in all groups treated with the glucans of Caripia montagnei (GCM). GCM significantly reduced the levels of IL-6 (50 and 75 mg/kg, p < 0.05), a major inflammatory cytokine. Biochemical analyses showed that such glucans acted on reducing levels of alkaline phosphatase (75 mg/kg, p < 0.01), nitric oxide (p < 0.001), and myeloperoxidase (p < 0.001). These results were confirmed microscopically by the reduction of cellular infiltration. The increase of catalase activity suggest a protective effect of GCM on colonic tissue, confirming their anti-inflammatory potential. GCM displayed cytostatic activity against HT-29 cells, causing accumulation of cells in G1 phase, blocking the cycle cell progression. Those glucans also showed ability to modulate the adhesion of HT-29 cells to Matrigel® and reduced the oxidative stress. The antiproliferative activity against HT-29 cells displayed by GCM (p <0.001) can be attributed to its cytostatic activity and induction of apoptosis by GCM

Fucana ativa via das Map quinases e inibe a proliferação de células de ovário de hamster chinês (CHO)

Fucan is a term used to denominate L-fucose rich sulfated polysaccharides. The fucans have been studied due their pharmacological activities like antithrombotic, antiproliferative and antioxidant. We have extracted three fucan fractions from the brown seaweed Spatoglossum schröederi. These fucans were denominated Fuc B 1, Fuc B 1.5 and Fuc B 2. The chemical analyzes show that the fucans have very similar composition as demonstrated by agarose electrophoresis gel, sugar and sulfate content. The antiproliferative effect was determined by MTT and BrdU methodologies in CHO cells. The inhibition of proliferation effect of the three fractions was about 40%. Therefore this we proceed just with the Fuc B 2 due the higher yield. There is no apoptosis indication using the anexin V/propidium iodide test. We found a cell cycle phase G1 arrest. The western blotting show that the PKC; pFAK; pERK 1/2 are activated when the cells were treated with fucans. The treatement with inhibitor of MAPK PD98059 extinguished the fucan effect. These results indicates that fucan act by the ERK pathway inducing the cell death.

Indução de morte celular em células de adenocarcinoma cervical humano (HeLa) pela lectina da esponja Cinachyrella apion (CaL)

Cancer is a term used to represent a set of more than 100 diseases, including malignant tumors from different locations. The malignancies are the second leading cause of death in the population, representing approximately 17% of deaths of known cause. Strategies that induce differentiation have had limited success in the treatment of established cancers. In this work, a lectin purified from the marine sponge Cinachyrella apion (CaL) was evaluated due to its hemolytic, cytotoxic and antiproliferative properties, besides the ability to induce cell death via apoptosis in tumor cells. The antiproliferative activity of CaL was tested against cell lines, with the highest inhibition of tumor growth for HeLa, reducing cell growth at a dose dependent manner, with a concentration of 10 μg/mL. The hemolytic activity and toxicity against peripheral blood cells were tested using the concentration of IC50 for both trials and twice the IC50 for analysis in flow cytometry, indicating that CaL is not toxic to these cells. To assess the mechanism of cell death caused by CaL in HeLa cells, we performed flow cytometry and western blotting. The results showed the lectin probably induces cell death by apoptosis activation by pro-apoptotic protein Bax, promoting mitochondrial membrane permeabilization, cell cycle arrest in S phase, with accumulation of cells of approximately 57% in this phase, and acting as both dependent and/or independent of caspases pathway. These results suggest that CaL has the potential to be used as drug treatment against cancer.

Avaliação do estresse oxidativo induzido por superfícies de titânio

Commercially pure Titanium (cp Ti) is a material largely used in orthopedic and dental implants due to its biocompatibility properties. Changes in the surface of cp Ti can determine the functional response of the cells such as facilitating implant fixation and stabilization, and increased roughness of the surface has been shown to improve adhesion and cellular proliferation. Various surface modification methods have been developed to increase roughness, such as mechanical, chemical, electrochemical and plasma treatment. An argon plasma treatment generates a surface that has good mechanical proprieties without chemical composition modification. Besides the topography, biological responses to the implant contribute significantly to its success. Oxidative stress induced by the biomaterials is considered one of the major causes of implant failure. For this reason the oxidative potential of titanium surfaces subjected to plasma treatment was evaluated on this work. CHO-k1 cells were cultivated on smooth or roughed Ti disks, and after three days, the redox balance was investigated measuring reactive oxygen species (ROS) generation, total antioxidant capacity and biomarkers of ROS attack. The results showed cells grown on titanium surfaces are subjected to intracellular oxidative stress due to hydrogen peroxide generation. Titanium discs subjected to the plasma treatment induced less oxidative stress than the untreated ones, which resulted in improved cellular ability. Our data suggest that plasma treated titanium may be a more biocompatible biomaterial.

Produção e caracterização de quitooligossacarídeos produzidos pelo fungo Metarhizium anisopliae e avaliação da citotoxicidade em células tumorais

Chitosan is a natural polymer, biodegradable, nontoxic, high molecular weight derived from marine animals, insects and microorganisms. Oligomers of glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) have interesting biological activities, including antitumor effects, antimicrobial activity, antioxidant and others. The alternative proposed by this work was to study the viability of producing chitooligosaccharides using a crude enzymes extract produced by the fungus Metarhizium anisopliae. Hydrolysis of chitosan was carried out at different times, from 10 to 60 minutes to produce chitooligosaccharides with detection and quantification performed by High Performace Liquid Chromatography (HPLC). The evaluation of cytotoxicity of chitosan oligomers was carried out in tumor cells (HepG2 and HeLa) and non-tumor (3T3). The cells were treated for 72 hours with the oligomers and cell viability investigated using the method of MTT. The production of chitosan oligomers was higher for 10 minutes of hydrolysis, with pentamers concentration of 0.15 mg/mL, but the hexamers, the molecules showing greater interest in biological properties, were observed only with 30 minutes of hydrolysis with a concentration of 0.004 mg/mL. A study to evaluate the biological activities of COS including cytotoxicity in tumor and normal cells and various tests in vitro antioxidant activity of pure chitosan oligomers and the mixture of oligomers produced by the crude enzyme was performed. Moreover, the compound with the highest cytotoxicity among the oligomers was pure glucosamine, with IC50 values of 0.30; 0.49; 0.44 mg/mL for HepG2 cells, HeLa and 3T3, respectively. Superoxide anion scavenging was the mainly antioxidant activity showed by the COS and oligomers. This activity was also depending on the oligomer composition in the chitosan hydrolysates. The oligomers produced by hydrolysis for 20 minutes was analyzed for the ability to inhibit tumor cells showing inhibition of proliferation only in HeLa cells, did not show any effect in HepG2 cells and fibroblast cells (3T3)

Efeito do tratamento térmico do titânio sobre a proliferação de células pré-osteoblásticas

Titanium is a biomaterial widely employed in biomedical applications (implants, prostheses, valves, stents). Several heat treatments are usually used in order to obtain physical properties required to different applications. This work studied the influence of the heat treatment on microstructure of commercial pure titanium, and their consequences in growth and proliferation of MC3T3-E1 cells. Discs of titanium were treated in different temperatures, and characterized by optical microscopy, image analysis, wettabillity, roughness, hardness and X-ray diffraction. After the heat treatment, significant modifications in these properties were observed. Pattern images of titanium, before and after the cell culture, were compared by overlapping to analyze the influence of microstructure in microstructure and preferences guidance cells. However, in general, titanium discs that showed a higher residual strength also presented an increase of cells numbers on surface