Caracterização físico-química e efeitos de frações polissacarídicas da alga amansia multifida na coagulação, inflamação, radicais livres e viabilidade celular

Galactans are polysaccharides sulfated present in the cell wall of red algae. Carrageenans are galactans well known in the food industry as gelling polysaccharides and for induce inflammatory process in rodents as animal model. The extraction of polysaccharides from A. multifida has been carried out by proteolysis and precipitation in different volumes of acetone, which produced three fractions (F1, F2, and FT). Chemical and physical analyses revealed that these fractions are sulfated galactan predominantly. Results of the antioxidant activity assays showed that all of these fractions have antioxidant activity and that was associated with sulfate content of the analysis of reducing power and total antioxidant capacity. However, these fractions were not effective against lipid peroxidation. The fraction FT presented higher activity on the APTT test at 200 μg (> 240 s). The assessment of the hemolytic activity showed that the FT fraction has the best activity, increasing lyses by the complement system to 42.3% (50 μg) (p< 0,001). The fraction FT showed the best yield, anticoagulant and hemolytic activity between the three fractions and therefore it was choose for the in vivo studies. The Inflammation assessment using the FT fraction (50 mg / kg MB) showed that the cellular migration and the IL-6 production increased 670.1% (p< 0,001) and 531.8% (p< 0,001), respectively. These results confirmed its use as an inflammation inducer in animal model. Cytotoxicity assay results showed that all fractions have toxic effects on 3T3 and HeLa cells after exposition of 48 hours, except when 100 μg for both F1 and FT were used. These results arise the discussion whether these polysaccharides it should be used as additive in foods, cosmetics and medicines.

Avaliação do efeito do composto tipo heparina isolado do caranguejo Chaceon fenneri na hemostasia e na morte celular

Heparin is a pharmaceutical animal widely used in medicine due to its potent anticoagulant effect. Furthermore, it has the ability to inhibit the proliferation, invasion and adhesion of cancer cells to vascular endothelium. However, its clinical applicability can be compromised by side effects such as bleeding. Thus, the search for natural compounds with low bleeding risk and possible therapeutic applicability has been targeted by several research groups. From this perspective, this study aims to evaluate the hemorrhagic and anticoagulant activities and citotoxic effect for different tumor cell lines (HeLa, B16-F10, HepG2, HS-5,) and fibroblast cells (3T3) of the Heparin-like from the crab Chaceon fenneri (HEP-like). The HEP-like was purified after proteolysis, ion-exchange chromatography, fractionation with acetone and characterized by electrophoresis (agarose gel) and enzymatic degradation. Hep-like showed eletroforetic behavior similar to mammalian heparin, and high trisulfated /Nacetylated disaccharides ratio. In addition, HEP-like presented low in vitro anticoagulant activity using aPTT and a minor hemorrhagic effect when compared to mammalian heparin. Furthermore, the HEP-like showed significant cytotoxic effect (p<0.001) on HeLa, HepG2 and B16-F10 tumor cells with IC50 values of 1000 ug/mL, after incubation for 72 hours. To assess the influence of heparin-like on the cell cycle in HeLa cells, analysis was performed by flow cytometry. The results of this analysis showed that HEP-like influence on the cell cycle increasing S phase and decreasing phase G2. Thus, these properties of HEP-like make these compounds potential therapeutic agents

Influência da suplementação de retinol palmitato sobre os níveis de vitamina A no leite de puérperas saudáveis

The tendency towards reduction of serum retinol levels, an existing placental barrier and the increase of retinol demand, are factors that place puerperal and lactating women at risk for Vitamin A deficiency. This micronutrient is an essential component of vital processes such as differentiation, cellular proliferation, and apoptosis. The objective of this study is to evaluate the effect of palmitate retinol supplementation (100.000UI) upon the milk retinollevels in puerperal women at the Januário Cicco University Maternity Hospital. This intervention has been adopted by the Ministry of Health since 2002. The longitudinal experiment was conducted with 106 puerperal women (68 comprised the supplemented group and 38 the control group). The High Performance Liquid Chromatography (HPLC) method was used to dose the retinol of the milk and serum samples, and the creamtocrit method to determine the milk fat levels. The retinol means for the colostrums were 99.0 ± 64.4 ug/dL and 160.1 ± 94,4 ug/dl 6 hours afier supplementation; 68.9 ± 33.5 ug/dL for the transitional milk, and 30.6 ± 15.2 ug/dL for the mature milk of the supplemented group. Ali the difterences between means were statistically significant. The difterence between retinol means in the control group were also significant, with these being greater in the colostrum, 88.6 ± 62.1 ug/dL with 61.9 ± 30.1 ug/dl in the transition milk and 32.9 ±32.9 ± 17.6 ug/dL in the mature milk. No significant difference was observed in the retinol means of the three types ot milk in the supplemented group when compared to their respective means in the control group. The prevalence in serum (35.1 % and 81.1 % for the cutting point 20 ug/dL, respectively) and in milk (51.4%) revealed vitamin A deficiency as a public health problem. COlostrum, transition, and mature milk tats varied similarly in the supplemented group (1,92 ± 0,96; 3,25 ± 1,27 and 3,31 ± 1,36 grams) and in the control group (1,87 ± 1,14; 3,25 ± 1,31 and 3,36 ± 1,67 grams), with an observed difference between the colostrum/transition milk and the colostrum/mature milk fats. No difference was observed between the groups. The study showed that the 200.000UI supplementation was not sufficient to increase the milk retinol to the desired levels nor to meet the demands of the mothers with deprived hepatic reserves. It is suggested that another similar dose be offered within 30 days or less, and within 2 months post-partum, while continual/y monitoring for possible pregnancy

Análise da estabilidade genética de células-tronco mesenquimais humanas

Human multipotent mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, have become an important and attractive therapeutic tool since they are easily isolated and cultured, have in vitro expansion potential, substantial plasticity and secrete bioactive molecules that exert trophic effects. The human umbilical cord as a cell source for cell therapy will help to avoid several ethical, political, religious and technical issues. One of the main issues with SC lines from different sources, mainly those of embryonic origin, is the possibility of chromosomal alterations and genomic instability during in vitro expansion. Cells isolated from one umbilical cord exhibited a rare balanced paracentric inversion, likely a cytogenetic constitutional alteration, karyotype: 46,XY,inv(3)(p13p25~26). Important genes related to cancer predisposition and others involved in DNA repair are located in 3p25~26. Titanium is an excellent biomaterial for bone-implant integration; however, the use can result in the generation of particulate debris that can accumulate in the tissues adjacent to the prosthesis, in the local bone marrow, in the lymph nodes, liver and spleen. Subsequently may elicit important biological responses that aren´t well studied. In this work, we have studied the genetic stability of MSC isolated from the umbilical cord vein during in vitro expansion, after the cryopreservation, and under different concentrations and time of exposition to titanium microparticles. Cells were isolated, in vitro expanded, demonstrated capacity for osteogenic, adipogenic and chondrogenic differentiation and were evaluated using flow cytometry, so they met the minimum requirements for characterization as MSCs. The cells were expanded under different concentrations and time of exposition to titanium microparticles. The genetic stability of MSCs was assessed by cytogenetic analysis, fluorescence in situ hybridization (FISH) and analysis of micronucleus and other nuclear alterations (CBMN). The cells were able to internalize the titanium microparticles, but MSCs preserve their morphology, differentiation capacity and surface marker expression profiles. Furthermore, there was an increase in the genomic instability after long time of in vitro expansion, and this instability was greater when cells were exposed to high doses of titanium microparticles that induced oxidative stress. It is necessary always assess the risks/ benefits of using titanium in tissue therapy involving MSCs, considering the biosafety of the use of bone regeneration using titanium and MSCs. Even without using titanium, it is important that the therapeutic use of such cells is based on analyzes that ensure quality, security and cellular stability, with the standardization of quality control programs appropriate. In conclusion, it is suggested that cytogenetic analysis, FISH analysis and the micronucleus and other nuclear alterations are carried out in CTMH before implanting in a patient

Transcriptoma diferencial entre células-tronco mesenquimais humanas jovens e senescentes

Human mesenchymal stem cells (MSC) are powerful sources for cell therapy in regenerative medicine. The long time cultivation can result in replicative senescence or can be related to the emergence of chromosomal alterations responsible for the acquisition of tumorigenesis features in vitro. In this study, for the first time, the expression profile of MSC with a paracentric chromosomal inversion (MSC/inv) was compared to normal karyotype (MSC/n) in early and late passages. Furthermore, we compared the transcriptome of each MSC in early passages with late passages. MSC used in this study were obtained from the umbilical vein of three donors, two MSC/n and one MSC/inv. After their cryopreservation, they have been expanded in vitro until reached senescence. Total RNA was extracted using the RNeasy mini kit (Qiagen) and marked with the GeneChip ® 3 IVT Express Kit (Affymetrix Inc.). Subsequently, the fragmented aRNA was hybridized on the microarranjo Affymetrix Human Genome U133 Plus 2.0 arrays (Affymetrix Inc.). The statistical analysis of differential gene expression was performed between groups MSC by the Partek Genomic Suite software, version 6.4 (Partek Inc.). Was considered statistically significant differences in expression to p-value Bonferroni correction ˂.01. Only signals with fold change ˃ 3.0 were included in the list of differentially expressed. Differences in gene expression data obtained from microarrays were confirmed by Real Time RT-PCR. For the interpretation of biological expression data were used: IPA (Ingenuity Systems) for analysis enrichment functions, the STRING 9.0 for construction of network interactions; Cytoscape 2.8 to the network visualization and analysis bottlenecks with the aid of the GraphPad Prism 5.0 software. BiNGO Cytoscape pluggin was used to access overrepresentation of Gene Ontology categories in Biological Networks. The comparison between senescent and young at each group of MSC has shown that there is a difference in the expression parttern, being higher in the senescent MSC/inv group. The results also showed difference in expression profiles between the MSC/inv versus MSC/n, being greater when they are senescent. New networks were identified for genes related to the response of two of MSC over cultivation time. Were also identified genes that can coordinate functional categories over represented at networks, such as CXCL12, SFRP1, xvi EGF, SPP1, MMP1 e THBS1. The biological interpretation of these data suggests that the population of MSC/inv has different constitutional characteristics, related to their potential for differentiation, proliferation and response to stimuli, responsible for a distinct process of replicative senescence in MSC/inv compared to MSC/n. The genes identified in this study are candidates for biomarkers of cellular senescence in MSC, but their functional relevance in this process should be evaluated in additional in vitro and/or in vivo assays

Concreto celular polimérico: influência na adição de resíduo de poliéster insaturado termofixo

This work addresses the production of lightweight concrete building elements, such as plates, prefabricated slabs for pre-molded and panels of fencing, presenting a singular concrete: the Lightweight Concrete, with special properties such low density and good strength, by means of the joint use of industrial waste of thermosetting unsaturated polyesters and biodegradable foaming agent, named Polymeric Lightweight Concrete. This study covered various features of the materials used in the composition of the Polymeric Lightweight Concrete, using a planning of factorial design 23, aiming at studying of the strength, production, dosage processes, characterization of mechanical properties and microstructural analysis of the transition zone between the light artificial aggregate and the matrix of cement. The results of the mechanical strength tests were analyzed using a computational statistics tool (Statistica software) to understand the behavior and obtain the ideal quantity of each material used in the formula of the Polymeric Lightweight Concrete. The definition of the ideal formula has the purpose of obtaining a material with the lowest possible dry density and resistance to compression in accordance with NBR 12.646/92 (≥ 2.5 MPa after 28 days). In the microstructural characterization by scanning electron microscopy it was observed an influence of the materials in the process of cement hydration, showing good interaction between the wrinkled face of the residue of unsaturated polyesters thermosetting and putty and, consequently, the final strength. The attaining of an ideal formula, given the Brazilian standards, the experimental results obtained in the characterization and comparison of these results with conventional materials, confirmed that the developed Polymeric Lightweight Concrete is suitable for the production of building elements that are advantageous for construction