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.

Lectina da esponja marinha Tedania ignis: purificação, caracterização e interação com leishmanias

Lectin obtained from the marine sponge Tedania ignis was purified and characterized by extraction of soluble proteins (crude extract) in 50mM Borax, pH 7.5. The purification procedure was carried out by crude extract precipitation with ammonium sulfate 30% (FI). The precipitated was resuspended in the same buffer and fractionated with acetone 1.0 volume (F1.0). A lectin was purified from this specific fraction by using an affinity chromatography Sepharose 6B. This lectin preferentially agglutinated human erythrocytes from B type previously treated with papain enzyme. The hemagglutinating activity lectin was dependent of divalent Mn2+ cation and was inhibited by the carbohydrates galactose, xylose and fructose. SDS-PAGE analysis indicated a molecular mass of the lectin around 45 kDa. This protein showed stability until 40°C for 1 h. Further, it showed activity between pH 2.5 and 11.5, with an enhanced activity at pH 7.5. Leishmania chagasi promastigotes stained with Coomassie brilliant blue R-250 were agglutinated by F1,0 and in the presence of galactose this interaction was abolished. These results show that this lectin could be implicated in defense procedures and it will can be used as biological tools in studies with this protozoon

Purificação e caracterização de uma ß-N-acetillhexosaminadase extraída do mamífero marinho Sotalia fluviatilis

This report shows 2232 times purification of a βNAcetylhexosaminidase from hepatic extracts from the sea mammal Sotalia fluviatilis homogenate with final recovery of 8,4%. Sequenced steps were utilized for enzyme purification: ammonium sulfate fractionation, Biogel A 1.5 m, chitin, DEAESepharose and hydroxyapatite chromatographies. The protein molecular mass was estimated in 10 kDa using SDSPAGE and confirmed by MALDITOF. It was found to have an optimal pH of 5.0 and a temperature of 60°C. Using pnitrophenylNAcetylβDglycosaminide apparent Km and Vmax values were of 2.72 mM and 0.572 nmol/mg/min, respectively. The enzyme was inhibited by mercury chloride (HgCl2) and sodium dodecil sulfate (SDS)

Caracterização estrutural e avaliação das atividades farmacológicas da fucana B extraída da alga Dictyota menstrualis

Seaweeds are a major source of biologically active compounds . In the extracellular matrix of these organisms are sulfated polysaccharides that functions as structural components preventing it against dehydration. The fraction 0.9 (FucB) rich in sulfated fucans obtained from brown seaweed Dictyota menstrualis was chemical characterized and evaluated for pharmacological activity by testing anticoagulant activity, stimulatory action on the synthesis of an antithrombotic heparan sulfate, antioxidant activity and its effects in cell proliferation. The main components were FucB carbohydrates (49.80 ± 0.10 %) and sulfate (42.30 ± 0.015 %), with phenolic compounds ( 3.86 ± 0.016 %) and low protein contamination ( 0.58 ± 0.001 % ) . FucB showed polydisperse profile and analysis of signals in the infrared at 1262, 1074 and 930 cm -1 and 840 assigned to S = O bonds sulfate esters , CO bond presence of 3,6- anhydrogalactose , β -D- galactose non- sulfated sulfate and the axial position of fucose C4 , respectively. FucB exhibited moderate anticoagulant activity , the polysaccharides prolonged time (aPTT ) 200 ug ( > 90s ) partial thromboplastin FucB no effect on prothrombin time (PT), which corresponds to the extrinsic pathway of coagulation was observed. This stimulation promoted fraction of about 3.6 times the synthesis of heparan sulfate (HS) by endothelial cells of the rabbit aorta ( RAEC ) in culture compared with cells not treated with FucB . This has also been shown to compete for the binding site with heparin. The rich fraction sulfated fucans exhibited strong antioxidant activity assays on total antioxidant (109.7 and 89.5 % compared with BHT and ascorbic acid standards ) , reducing power ( 71 % compared to ascorbic acid ) and ferric chelation ( 71 , comparing with 5 % ascorbic acid). The fraction of algae showed cytostatic activity on the RAEC cells revealed that the increase of the synthesis of heparan sulfate is not related to proliferation. FucB showed antiproliferative action on cell lines modified as Hela and Hep G2 by MTT assay . These results suggest that FucB Dictyota menstrualis have anticoagulant , antithrombotic , antioxidant potential as well as a possible antitumor action, promoting the stimulation of the synthesis of antithrombotic HS by endothelial cells and is useful in the prevention of thrombosis, also due to its inhibitory action on species reactive oxygen ( ROS ) in some in vitro systems , being involved in promoting a hypercoagulable state

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

Estudo do papel da proteína multifuncional APE1/Ref-1 sobre a resposta inflamatória na meningite bacteriana

Despite advances in antibiotic therapy, bacterial meningitis (BM) remains with high mortality and morbidity rates in worldwide. One important mechanism associated to sequels during disease is the intense inflammatory response which promotes an oxidative burst and release of reactive oxygen species, consequently leading to cell death. Activation of DNA repair enzymes during oxidative stress has been demonstrated in several neurological disorders. APE1/Ref-1 is a multifunctional protein involved in DNA repair and plays a redox function on transcription factors such as NFkB and AP-1.The aim of this study was assess the role of APE1/Ref-1 on inflammatory response and the possibility of its modulation to reduce the sequels of the disease. Firstly it was performed an assay to measure cytokine in cerebrospinal fluid of patients with BM due to Streptococcus pneumoniae and Neisseriae meningitides. Further, a cellular model of inflammation was used to observe the effect of the inhibition of the endonuclease and redox activity of APE1/Ref-1 on cytokine levels. Additionally, APE1/Ref-1 expression in cortex and hippocampus of rat with MB after vitamin B6 treatment was evaluated. Altogether, results showed a similar profile of cytokines in the cerebrospinal fluid of patients from both pathogens, although IFNy showed higher expression in patients with BM caused by S. pneumoniae. On the other hand, inhibitors of APE1/Ref-1 reduced cytokine levels, mainly TNF-α. Reduction of oxidative stress markers was also observed after introduction of inhibitors in the LPS-stimulated cell. In the animal model, BM increased the expression of the protein APE1/Ref-1, while vitamin B6 promoted reduction. Thereby, this data rise important factors to be considered in pathogenesis of BM, e.g., IFNy can be used as prognostic factor during corticosteroid therapy, APE1/Ref-1 can be an important target to modulate the level of inflammation and VIII oxidative stress, and vitamin B6 seems modulates several proteins related to cell death. So, this study highlights a new understanding on the role of APE1/Ref-1 on the inflammation and the oxidative stress during inflammation condition

Farelo de canola em dietas para o pacu Piaractus mesopotamicus (Holmberg 1987): efeitos sobre o crescimento e a composição corporal

Avaliaram-se os efeitos da inclusão de farelo de canola em dietas de juvenis de pacu (Piaractus mesopotamicus) sobre parâmetros de crescimento e composição corporal. Um total de 192 alevinos (9 a 15g) foram estocados em 24 tanques de cimento, de 100l de capacidade, durante 103 dias. O farelo de canola foi utilizado em quatro proporções: zero; 9,5%; 19% e 38% da dieta, com ou sem farinha de peixe (12%/dieta), totalizando oito tratamentos. A presença de farinha de peixe não afetou os parâmetros de crescimento avaliados. A inclusão de 38% de farelo de canola na dieta diminuiu o ganho de peso dos peixes, valores médios de 28,74g a 50,70g, e piorou a conversão alimentar aparente, de 1,66 para 2,85. A taxa de eficiência protéica também foi menor nos peixes alimentados com 38% de farelo de canola. As várias proporções de farelo de canola das dietas alteraram os teores de umidade, proteína bruta e lipídios dos peixes. A presença da farinha de peixe, nas dietas, somente influiu no teor de lipídios dos peixes alimentados com dietas contendo 9,5% de farelo de canola. Conclui-se que até 19% de farelo de canola pode ser adicionado às dietas de juvenis de pacu, sem que seu desenvolvimento seja prejudicado.

Farinha de resíduos da indústria de filetagem de tilápias como fonte de proteína e minerais para alevinos de tilápia-do-nilo (Oreochromis niloticus)

Neste trabalho, objetivou-se avaliar a utilização da farinha de resíduos da indústria de filetagem de tilápias (FR) como fonte de proteína e de minerais em rações práticas na alimentação de alevinos de tilápia do Nilo. Foram utilizados 120 alevinos de tilápias do Nilo com peso e comprimento iniciais de 0,58 ± 0,05 g e 3,49 ± 0,09 cm, respectivamente, distribuídos em 24 aquários com capacidade para 30 L, em um delineamento inteiramente casualisado, com quatro tratamentos e seis repetições. Os peixes foram alimentados com rações contendo 30% de proteína digestível e 3.000 kcal de energia digestível/kg, de acordo com os seguintes tratamentos: CO - ração à base de milho e farelo de soja, sem suplementação de fósforo; FB - ração à base de milho e farelo de soja, com fosfato bicálcico; FB + FR - ração à base de milho e farelo de soja, suplementada com fosfato bicálcico (50%) e farinha de resíduos (50%); FT - ração à base de milho e farelo de soja, suplementada com farinha de resíduos. Ao final do experimento, os melhores resultados de desempenho foram observados nos animais que receberam suplementação de P. Quanto às características de carcaça, os animais que receberam a ração CO apresentaram maior teor de gordura corporal e menores teores de cinzas, Ca e P. A FR pode ser utilizada em rações para alevinos de tilápia do Nilo como fonte de P, sem prejuízos no desempenho e na composição corporal.

Avaliação nutricional dos idosos institucionalizados no concelho de Viseu

O envelhecimento da população, bem como a alteração da dinâmica da família, têm contribuído diretamente para o aumento do número de idosos institucionalizados. A incidência da desnutrição alcança níveis elevados nestes idosos, sendo essencial o seu diagnóstico precoce para a melhoria da sua qualidade de vida. Constitui objetivo primordial do estudo a avaliação do estado nutricional de idosos institucionalizados no concelho de Viseu, Portugal. A amostra (n=120) é constituída por idosos de ambos os sexos, institucionalizados, com idade igual ou superior a 65 anos, com capacidade para se colocar de pé, sem demência e que não sejam portadores de patologia passível de afetar a digestão, absorção ou utilização dos nutrientes. A avaliação do estado nutricional é determinada tendo em conta parâmetros antropométricos, a aplicação do e Mini Nutricional Assessment (MNA) e dietéticos, através da avaliação da ingestão alimentar (registo de 3 dias). Os questionários aplicados estão validados para a população portuguesa idosa e foram aplicados após consentimento informado. Foram realizadas estatísticas descritivas e inferenciais às diferentes variáveis, de acordo com o género e estratos etários. Considerou-se um nível de confiança de 95%. Dos 120 indivíduos estudados 30% eram do sexo masculino e 70% eram do sexo feminino. As médias das idades eram de 82,36 ± 6,34 anos. A determinação do IMC mostrou que 12,5% dos idosos avaliados exibem baixo peso (IMC 19,6±2,1); 46,7% são eutróficos (IMC 25,7±1,3); 11,7% apresentam excesso de peso (IMC 29,0±0,6), sendo 29,2% classificados como obesos (IMC 33,8±2,9).A avaliação da população pelo MNA evidenciou uma percentagem de desnutrição de 1,7% com uma prevalência de risco de desnutrição (33,3%) mas identifica 65% da amostra como bem nutrida. Os 2 instrumentos utilizados para avaliação do estado nutricional, antropometria (IMC) e MNA mostraram uma correlação estatística entre si positiva. Independentemente do género e do grupo etário verificou-se que o Valor Calórico Total (VCT) ingerido diariamente apresentava valores abaixo do recomendado. Já a ingestão diária de hidratos de carbono e de proteínas apresentou valores médios superiores às recomendações nutricionais enquanto os teores de lípidos ingeridos se situava dentro dos limites recomendados estando porém os valores da ingesta média de fibra total (18g/dia) abaixo dos indicados para a população idosa (25g/dia). Quanto aos micronutrientes encontraram-se deficits de ingestão média diária de minerais e oligoelementos como o cálcio, ferro, magnésio selénio, zinco e iodo e também de algumas vitaminas (D, E, ácido fólico). Conclui-se então que no geral a população estudada apresenta um estado de nutrição normal avaliado através de IMC e do MNA dois instrumentos que estão correlacionados entre si e se complementam. Que a ingestão alimentar apresentada pelos idosos desta amostra é apenas ligeiramente inferior ao recomendado, não colocando em risco o aparecimento de desnutrição. Porém é importante a intervenção dietética no sentido de corrigir determinadas carências (vitaminas, minerais) e prevenir o risco de desnutrição detetado assim como a obesidade.