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

Atividades Biológicas De Xilana De Sabugo De Milho

The corn cob is an agricultural by-product still little used, this in part due to the low knowledge of the biotechnological potential of their molecules. Xylan from corn cobs (XSM) is a polysaccharide present in greater quantity in the structure of plant and its biotechnology potential is little known. This study aimed to the extraction, chemical characterization and evaluation of biological activities of xylan from corn cobs. To this end, corncobs were cleaned, cut, dried and crushed, resulting in flour. This was subjected to a methodology that combines the use of alkaline conditions with waves of ultrasound. After methanol precipitation, centrifugation and drying was obtained a yield of 40% (g/g flour). Chemical analysis indicated a high percentage of polysaccharides in the sample (60%) and low contamination by protein (0.4%) and phenolic compounds (> 0.01%). Analysis of monosaccharide composition indicated the presence of xylose:glucose:arabinose:galactose:mannose:glucuronic acid in a molar ratio 50:20:15:10:2.5:2.5. The presence of xylan in the sample was confirmed by nuclear magnetic resonance (¹H and ¹³C) and infrared spectroscopy (IR). Tests were conducted to evaluate the antioxidant potential of XSM. This showed a total antioxidant capacity of 48.45 EAA/g sample. However, did not show scavenging activity of superoxide and hydroxyl radical and also reducing power. But, showing a high capacity chelating iron ions with 70% with about 2 mg/mL. The ability to XSM to influence cell proliferation in culture was also evaluated. This polymer did not influence the proliferation of normal fibroblast cells (3T3), however, decreased the rate of proliferation of tumor cells (HeLa) in a dose-dependent, reaching an inhibition of about 50% with a concentration around 2 mg/mL. Analyzing proteins related to cell death, by immunoblotting, XSM increases the amount of Bax, Bcl-2 decrease, increase cytochrome c and AIF, and reduce pro-caspase-3, indicating the induction of cell death induced apoptosis dependent and independent of caspase. XSM did not show anticoagulant activity in the PT test. However, the test of activated partial thromboplastin time (aPTT), XSM increased clotting time at about 5 times with 600 μg of sample compared with the negative control. The presence of sulfate on the XSM was discarded by agarose gel electrophoresis and IR. After carboxyl-reduction of XSM the anticoagulant activity decreased dramatically. The data of this study demonstrate that XSM has potential as antioxidant, antiproliferative and anticoagulant compound. Future studies to characterize these activities of XSM will help to increase knowledge about this molecule extracted from corn and allow their use in functional foods, pharmaceuticals and chemical industries.

Influência das mudanças climáticas na estrutura funcional da comunidade fitoplanctônica em um reservatório da região do semiárido

The Brazilian Northeast is the most vulnerable region to climatic variability risks. For the Brazilian semi-arid is expected a reduction in the overall rates of precipitation and an increase in the number of dry days. These changes predicted by the IPCC (2007) will intensify the rainfall and droughts period that could promote the dominance of cyanobacteria, thus affecting the water quality of reservoirs, that are most used for water supply, in the semi-arid. The aim of this study was to evaluate the effects of increasing temperature combined with nutrient enrichment on the functional structure of the phytoplankton community of a mesotrophic reservoir in the semi-arid, in the worst case scenario of climate change predicted by the IPCC (2007). Two experiments were performed, one in a rainy season and another in the dry season. In the water sampled, nutrients (nitrate and orthophosphate) were added in different concentrations. The microcosms were submitted to two different temperatures, five-year average of air temperature in the reservoir (control) and 4°C above the control temperature (warming). The results of this study showed that warming and nutrient enrichment benefited mainly the functional groups of cyanobacteria. During the rainy season it was verified the increasing biomass of small functional groups of unicellular and opportunists algae such as F (colonial green algae with mucilage) and X1 (nanoplanktonic algae of eutrophic lake systems). It was also observed an increasing in total biomass, in the richness and diversity of the community. In the dry season experiment there was a greater contribution in the relative biomass of filamentous algae, with a replacement of the group S1 (non-filamentous cyanobacteria with heterocytes) for H1 (filamentous cyanobacteria with heterocytes) in nutrient- enriched treatments. Moreover, there was also loss in total biomass, species richness and diversity of the community. The effects of temperature and nutrients manipulation on phytoplankton community of reservoir Ministro João Alves provoked changes in species richness, the diversity of the community and its functional composition, being the dry period which showed the highest susceptibility to the increase in the contribution of potentially toxic cyanobacteria with heterocytes

Euler e os números pentagonais

The present investigation includes a study of Leonhard Euler and the pentagonal numbers is his article Mirabilibus Proprietatibus Numerorum Pentagonalium - E524. After a brief review of the life and work of Euler, we analyze the mathematical concepts covered in that article as well as its historical context. For this purpose, we explain the concept of figurate numbers, showing its mode of generation, as well as its geometric and algebraic representations. Then, we present a brief history of the search for the Eulerian pentagonal number theorem, based on his correspondence on the subject with Daniel Bernoulli, Nikolaus Bernoulli, Christian Goldbach and Jean Le Rond d'Alembert. At first, Euler states the theorem, but admits that he doesn t know to prove it. Finally, in a letter to Goldbach in 1750, he presents a demonstration, which is published in E541, along with an alternative proof. The expansion of the concept of pentagonal number is then explained and justified by compare the geometric and algebraic representations of the new pentagonal numbers pentagonal numbers with those of traditional pentagonal numbers. Then we explain to the pentagonal number theorem, that is, the fact that the infinite product(1 x)(1 xx)(1 x3)(1 x4)(1 x5)(1 x6)(1 x7)... is equal to the infinite series 1 x1 x2+x5+x7 x12 x15+x22+x26 ..., where the exponents are given by the pentagonal numbers (expanded) and the sign is determined by whether as more or less as the exponent is pentagonal number (traditional or expanded). We also mention that Euler relates the pentagonal number theorem to other parts of mathematics, such as the concept of partitions, generating functions, the theory of infinite products and the sum of divisors. We end with an explanation of Euler s demonstration pentagonal number theorem

Não vício assintótico, consistência forte e uniformemente forte de estimadores do tipo núcleo para dados direcionais sobre uma esfera unitária k-dimensional

In this work we studied the asymptotic unbiasedness, the strong and the uniform strong consistencies of a class of kernel estimators fn as an estimator of the density function f taking values on a k-dimensional sphere

Expressão imunoistoquímica de Glut-1 e marcadores de proliferação e apoptose em anomalias vasculares orais

Vascular anomalies constitute a distinct group of lesions, but they may present similar clinical and histopatological characteristics, which can lead to diagnostic mistakes. This study aimed by histopathology and immunohistochemical expression of human glucose transporter protein (GLUT-1), correctly identify and classify oral vascular anomalies, besides analyzing the immunoexpression of markers proliferation and apoptosis (Ki-67 and Bcl-2). All cases diagnosed as "oral hemangiomas" belonging to the archives of the Service of Pathological Anatomy from the subject of Oral Pathology of the Department of Dentistry (DOD), of the Federal University of Rio Grande do Norte (UFRN) were reviewed, totalizing 77 cases. Immunohistochemical analysis for GLUT-1 showed that only 26 (33.8%) of the specimens were true infantile hemangiomas (IHs). The 51 (66.2%%) GLUT-1 negative specimens were then reclassified as pyogenic granulomas (PGs) and vascular malformations (VMs) from their histopathologic characteristics,totalizing 26 (33.8%) cases of IHs, 20 (26.0%) of PGs and 31 (40.2) cases of oral VMs. The cases analyzed by the marker Ki-67 showed different median IH (13,85), PG (33,70) and VM (4.55) with statistically significant differences between them (p <0.001). In relation to the protein Bcl-2, the groups also showed different median of the established scores IH (1.00), PG (1.50), VMs (0.0) demonstrating statistically significant differences between them (p<0,001). No statistically significant correlation between the indexes of positivity for Ki-67 and the scores of immunoexpression of Bcl-2 were observed in any group. Thus, we can conclude that it is necessary a careful and parameterized review of cases of vascular anomalies making use of auxiliary tools such as GLUT-1, since the histopathological findings alone, sometimes, are not sufficient to differentiate some anomalies. Furthermore, analysis of the expressions of markers involved in the levels of proliferation of lesions is important for a better understanding of its biological behavior aspect

Biomonitoramento da qualidade da água e ações em educação ambiental em dois açudes da bacia hidrográfica Apodi-Mossoró, RN

Water scarcity is a reality for the inhabitants of the Brazilian semiarid region. The problem, in quantitative terms , is caused due to local climatic conditions due mostly to a water deficit. Qualitatively, results of multiple uses and resulting contamination by human activities. Eutrophication is presented as a threat to the sustainable use of water resources, in order to favor the predominance of dense populations of cyanobacteria, which can be potentially harmful to human health. The aim of this study is to understand the population dynamics of phytoplankton and cyanobacteria to assess water quality of reservoirs Santa Cruz do Apodi and Pau dos Ferros belonging to bacia Apodi Mossoró-RN . Water samples were collected monthly between October 2011 and May 2012 in reservoirs Santa Cruz do Apodi and Pau dos Ferros, in this latter , in Jan/12 we performed a diel profile . The abiotic variables obtained in the field and in the laboratory were: water temperature , dissolved oxygen , pH , turbidity , transparency , total nitrogen and total phosphorus . Phytoplankton samples were collected with a plankton net and bottle of Van dorn . Were performed : identification , quantification , calculation of biovolume , classification into functional groups and index calculation phytoplankton assembly ( Q ) , cyanotoxins were quantified by ELISA . Statistical analyzes supported the evaluation of the dynamics between biotic and abiotic factors. A questionnaire was conducted to examine the conceptions of the population, education professionals and students. The reservoir of Pau dos Ferros, shallow , turbid and eutrophic showed dominance of the functional group SN throughout the sample period . The phytoplankton biomass ranged from 20 to 70 mm ³ . L - 1, the lowest values coincided with the increase of the mixing zone and transparency, which contributed to the occurrence of a change in species composition phytoplankton . The application of the index Q proved relevant, the reservoir of Pau dos Ferros , depending on the species present , was classified ecologically as bad for almost the entire sample period . The reservoir Santa Cruz do Apodi showed low biomass ( 0.04 and 4.31 mm ³ . L - 1 ) and greater diversity in the phytoplankton composition . According to the index assembly (Q ), it showed moderate condition during most of the period influenced by different functional groups of typical meso- eutrophic environment (K S0, H1, C, F , J , E, D and N) . Associations of diatoms and green algae D and X1 succeeded populations of cyanobacteria in periods marked by greater instability in the system , caused by wind or rain. In summary , the occurrence of drought has a direct influence on the hydrological conditions of the reservoirs , in general, these events, reducing the reservoir level is directly related to decreased water quality and increased density of phytoplankton occurring predominance of cyanobacteria , the index Q reflected well to changes in phytoplankton composition , being a good indicator for biomonitoring of reservoirs in this study and survey of previous conceptions showed the need to work on environmental awareness for the preservation of water resources by conducting workshops for Environmental Education

Métodos estatisticos em cadeias de Markov

Este trabalho tem como objetivo o estudo do comportamento assintótico da estatística de Pearson (1900), que é o aparato teórico do conhecido teste qui-quadrado ou teste x2 como também é usualmente denotado. Inicialmente estudamos o comportamento da distribuição da estatística qui-quadrado de Pearson (1900) numa amostra {X1, X2,...,Xn} quando n → ∞ e pi = pi0 , 8n. Em seguida detalhamos os argumentos usados em Billingley (1960), os quais demonstram a convergência em distribuição de uma estatística, semelhante a de Pearson, baseada em uma amostra de uma cadeia de Markov, estacionária, ergódica e com espaço de estados finitos S

Expressão imuno-histoquímica das proteínas E-Caderina e BCL2 e nevos melanociticos orais e cutâneos

Melanocytic nevi (MNs) are benign melanocytic proliferations of cells, which can be found in the skin and mucous coat, including the oral mucosa. However, skin NMs are more common when compared to those that affect the oral mucosa. The molecular mechanisms involved in the development of nevi and the factors that can influence the migration pattern of the nevus cells are little explored. The aim of this study was to analyze the immunohistochemical expression of E-cadherin protein and Bcl-2 in oral / skin NMs and relate them to the clinical characteristics (gender, age, location, exposure to solar radiation) and histopathological types. 36 cases of oral NMs and 34 Skin NMs were analyzed. The immunohistochemistry was used of the protein E-cadherin and bcl-2, which were analyzed the intensity (weak, moderate and strong) and distribution marking (diffuse and focal). The immunoreactivity also analyzed as to the types of nevus cells (epithelioid cells -A, -B lymphocyte and fibroblast-like -C). Statistical analysis was performed using the chi-square tests of Pearson and Spearman correlation with significance level set at 5%. Of the 70 cases of NMs, 82.9% were female, 48.6% aged 26-50 years, 51.4% were diagnosed histologically as intradermal / intramucosal nevi and 80% were NMs acquired. Immunohistochemical expression of BCL2 and E-cadherin were variables in the sample and showed no association with clinical parameters. The expression of bcl-2 and E-cadherin were variable according to the types of nevus cells (A, B and C) (P = 0.001). The expression of bcl-2 was more diffuse in congenital MNs (p = 0.002). E-cadherin was positive in 83.3% of MNs <1cm (p = 0.001) and exhibited weak staining in 73.9% of MNs that were in exposed areas (p = 0.010). Based on these results, it is suggested that the E-cadherin has a modulating effect on the migratory properties of NMs, and bcl-2 is a marker of MNs with increased proliferative capacity.

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

Atividades biológicas de xilana de sabugo de milho

The corn cob is an agricultural by-product still little used, this in part due to the low knowledge of the biotechnological potential of their molecules. Xylan from corn cobs (XSM) is a polysaccharide present in greater quantity in the structure of plant and its biotechnology potential is little known. This study aimed to the extraction, chemical characterization and evaluation of biological activities of xylan from corn cobs. To this end, corncobs were cleaned, cut, dried and crushed, resulting in flour. This was subjected to a methodology that combines the use of alkaline conditions with waves of ultrasound. After methanol precipitation, centrifugation and drying was obtained a yield of 40% (g/g flour). Chemical analysis indicated a high percentage of polysaccharides in the sample (60%) and low contamination by protein (0.4%) and phenolic compounds (> 0.01%). Analysis of monosaccharide composition indicated the presence of xylose:glucose:arabinose:galactose:mannose:glucuronic acid in a molar ratio 50:20:15:10:2.5:2.5. The presence of xylan in the sample was confirmed by nuclear magnetic resonance (¹H and ¹³C) and infrared spectroscopy (IR). Tests were conducted to evaluate the antioxidant potential of XSM. This showed a total antioxidant capacity of 48.45 EAA/g sample. However, did not show scavenging activity of superoxide and hydroxyl radical and also reducing power. But, showing a high capacity chelating iron ions with 70% with about 2 mg/mL. The ability to XSM to influence cell proliferation in culture was also evaluated. This polymer did not influence the proliferation of normal fibroblast cells (3T3), however, decreased the rate of proliferation of tumor cells (HeLa) in a dose-dependent, reaching an inhibition of about 50% with a concentration around 2 mg/mL. Analyzing proteins related to cell death, by immunoblotting, XSM increases the amount of Bax, Bcl-2 decrease, increase cytochrome c and AIF, and reduce pro-caspase-3, indicating the induction of cell death induced apoptosis dependent and independent of caspase. XSM did not show anticoagulant activity in the PT test. However, the test of activated partial thromboplastin time (aPTT), XSM increased clotting time at about 5 times with 600 μg of sample compared with the negative control. The presence of sulfate on the XSM was discarded by agarose gel electrophoresis and IR. After carboxyl-reduction of XSM the anticoagulant activity decreased dramatically. The data of this study demonstrate that XSM has potential as antioxidant, antiproliferative and anticoagulant compound. Future studies to characterize these activities of XSM will help to increase knowledge about this molecule extracted from corn and allow their use in functional foods, pharmaceuticals and chemical industries.