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

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

Evolução do reparo por excisão de nucleotídeo

The phylogeny is one of the main activities of the modern taxonomists and a way to reconstruct the history of the life through comparative analysis of these sequences stored in their genomes aimed find any justification for the origin or evolution of them. Among the sequences with a high level of conservation are the genes of repair because it is important for the conservation and maintenance of genetic stability. Hence, variations in repair genes, as the genes of the nucleotide excision repair (NER), may indicate a possible gene transfer between species. This study aimed to examine the evolutionary history of the components of the NER. For this, sequences of UVRA, UVRB, UVRC and XPB were obtained from GenBank by Blast-p, considering 10-15 as cutoff to create a database. Phylogenetic studies were done using algorithms in PAUP programs, BAYES and PHYLIP package. Phylogenetic trees were build with protein sequences and with sequences of 16S ribosomal RNA for comparative analysis by the methods of parsimony, likelihood and Bayesian. The XPB tree shows that archaeal´s XPB helicases are similar to eukaryotic helicases. According to this data, we infer that the eukaryote nucleotide excision repair system had appeared in Archaea. At UVRA, UVRB and UVRC trees was found a monophyletic group formed by three species of epsilonproteobacterias class, three species of mollicutes class and archaeabacterias of Methanobacteria and Methanococci classes. This information is supported by a tree obtained with the proteins, UVRA, UVRB and UVRC concatenated. Thus, although there are arguments in the literature defending the horizontal transfer of the system uvrABC of bacteria to archaeabacterias, the analysis made in this study suggests that occurred a vertical transfer, from archaeabacteria, of both the NER genes: uvrABC and XPs. According the parsimony, this is the best way because of the occurrence of monophyletic groups, the time of divergence of classes and number of archaeabacterias species with uvrABC system

Influencia da ativaçao acida na estabilidade termica de hibridos vermiculita/epoxido

ALVES, Ana Paula de M. ; GERMANO, A. F. S. ; ARAKAKI, L N H ; Ferreira, U. V. S. ; ARAUJO, A. S. Influencia da ativaçao acida na estabilidade termica de hibridos vermiculita/epoxido. In: CONGRESSO NACIONAL DE ANLISETERMICA E CALORIMETRIA, 7., 2010, São Pedro, SP. Anais... Sao Pedro, SP: ABRATEC, 2010

O estado como empregador de última instância: uma proposta de pleno emprego, estabilidade e condições dignas de trabalho para o Brasil

GOMES, Z. B. ; LOURENÇO, André Luís Cabral de . Atuação do Estado como empregador de última Instância: uma proposta para eliminar o desemprego estrutural do Brasil. In: Encontro Nacional de Economia Política, 13. 2008, João Pessoa/PB. Anais... João Pessoa: ENEP, 2008.

Avaliação demográfica, clinico-laboratorial e genética de indivíduos com lupus eritematoso sistêmico e artrite reumatóide residentes em região tropical

The aetiology of autoimmunes disease is multifactorial and involves interactions among environmental, hormonal and genetic factors. Many different genes may contribute to autoimmunes disease susceptibility. The major histocompatibility complex (MHC) genes have been extensively studied, however many non-polymorphic MHC genes have also been reported to contribute to autoimmune diseases susceptibility. The aim of the present study was to evaluate the influence of SLC11A1 gene in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Ninety-six patients with SLE, 37 with RA and 202 controls enrolled in this case-control study, were evaluated with regard to demographic, genetic, laboratorial and clinical data. SLE mainly affects females in the ratio of 18 women for each man, 88,3% of the patients aged from 15 to 45 years old and it occurs with similar frequency in whites and mulattos. The rate of RA between women and men was 11:1, with 77,1% of the cases occurring from 31 to 60 years. The genetic analysis of the point mutation -236 of the SLC11A1 gene by SSCP did not show significant differences between alleles/genotypes in patients with SLE or RA when compared to controls. The most frequent clinical manifestations in patients with SLE were cutaneous (87%) and joint (84.9%). In patients with RA, the most frequent out-joint clinical manifestation were rheumatoid nodules (13,5%). Antinuclear antibodies were present in 100% of the patients with SLE. There was no significant relation between activity of disease and presence of rheumatoid factor in patients with RA, however 55,6% of patients with active disease presented positive rheumatoid factor. Significant association between alleles/genotypes of point mutation -236 and clinical manifestations was not found

Estrutura genética populacional de Lutjanus analis cioba e Lutjanus jocu dentão (Lutjanidae) ao longo do litoral brasileiro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Efeito da incorporação de diatomita na estabilidade e permeabilidade de compósitos cimentíceos espumados aplicados a poços de petróleo

Lightweight oilwell cement slurries have been recently studied as a mean to improve zonal isolation and sheath-porous formation adherence. Foamed slurries consisting of Portland cement and air-entraining admixtures have become an interesting option for this application. The loss in hydrostatic pressure as a consequence of cement hydration results in the expansion of the air bubbles entrapped in the cement matrix, thus improving the sheath-porous formation contact. Consequently, slurries are able to better retain their water to complete the hydration process. The main objective of the present study was to evaluate the effect of the addition of an air-entraining admixture on the density, stability and permeability of composite slurries containing Portland cement and diatomite as light mineral load. Successful formulations are potential cementing materials for low fracture gradient oilwells. The experimental procedures used for slurry preparation and characterization were based on the American Petroleum Institute and ABNT guidelines Slurries containing a pre-established concentration of the air-entraining admixture and different contents of diatomite were prepared aiming at final densities of 13 to 15 lb/gal. The results revealed that the reduction of 15 to 25% of the density of the slurries did not significantly affect their strength. The addition of both diatomite and the air-entraining admixture increased the viscosity of the slurry providing better air-bubble retention in the volume of the slurry. Stable slurries depicted bottom to top density variation of less than 1.0 lb/gal and length reduction of the stability sample of 5.86 mm. Finally, permeability coefficient values between 0.617 and 0.406 mD were obtained. Therefore, lightweight oilwell cement slurries depicting a satisfactory set of physicochemical and mechanical properties can be formulated using a combination of diatomite and air-entraining admixtures for low fracture gradient oilwells

Resistotipagem e transferência genética de plasmídios de resistência entre coliformes e vibrios potencialmente patogênicos para o homem isolados de camarão (litopenaeus vannamei) comercializado em Natal-Rio Grande do Norte

Cinquenta amostras de camarão fresco e refrigerado (Litopenaeus vannamei) foram coletadas em diferentes pontos de comercialização na cidade de Natal RN. As amostras foram maceradas em um gral estéril e 25 gramas semeadas em 225mL de APA contendo 1% de NaCl e 25g em 225mL de CL incubadas a 35ºC - 24 horas. O crescimento em APA foi semeado em placas de Ágar TCBS, incubadas a 35ºC-24h para isolamento de Vibrio e Aeromonas. O crescimento do CL foi semeado em Agar EAM, para isolamento de coliformes. Dos 102 isolados, 91 (89,2%) pertenciam ao gênero Vibrio e 11 (10,8%) ao gênero Aeromonas, com predominância de V. cholerae não O1/não O139, V. alginolyticus, V. carchariae e V. parahaemolyticus K- e A. veronii biogrupo sobria , A. jandaei, A. schubertii, A. veronii biogrupo veronii e A. hydrophila. A menor eficiência entre os antimicrobianos foi da AMP (57,8% de resistência) seguida da AMK (29,4%) e TCY (21,6%). As 39 cepas de Vibrio e Aeromonas multirresistentes se distribuíram em 10 perfis distintos, sendo que um revelou cinco marcos (AMP, CHL, NIT, SXT e TCY) em um isolado de V. carchariae de camarão, adquirido em supermercados. O índice MAR, nas 39 cepas variou de 0,28 a 0,42, sugerindo que são de risco na transferência e difusão da resistência na cadeia alimentar. Após a cura plasmidial pelo tratamento com AO de 24 cepas multirresistentes e com resistência intermediária de víbrio e aeromonas escolhidas aleatoriamente, 13 perderam totalmente a resistência e 7 perderam parcialmente, sendo que o maior percentual de perda da resistência ocorreu nas cepas de V. cholerae não O1 e não O139 (6 cepas), se concentrando nos marcos de resistência a AMP (13), AMK (11), TCY(8) e CIP(3). Os resultados da conjugação realizada entre amostras de Vibrio xvi curadas e a E. coli K12C600 demonstraram que 78,5% das culturas de Vibrio testadas revelaram capacidade de transferência para o gene que confere resistência a AMP e 28,5% para a TCY. Dos coliformes, E. coli foi a mais frequente, seguida de Citrobacter spp, isoladas em 40,3% e 27,5% das amostras respectivamente. AMP foi o antimicrobiano menos eficaz, seguido de TCY. As 11 cepas multirresistentes se distribuíram em 9 perfis distintos, um deles constituído de cinco marcos (AMP, NIT, TCY, CHL, SXT), albergados em uma cepa de Klebsiella spp, oriunda de camarão adquirido em supermercado, similar ao resultado obtido em V. carchariae. Conclui-se que, os camarões marinhos frescos e refrigerados, comercializados em Natal-RN evidenciaram contaminação com coliformes, víbrios e aeromonas multirresistentes a antimicrobianos comumente utilizados na terapia médica e veterinária, e que, possivelmente, a transferência de genes de resistência entre bactérias se constitui um sério problema de saúde pública

O tratamento ortodôntico em uma perspectiva a longo prazo: avaliação da estabilidade oclusal e do grau de satisfação dos pacientes

This study aimed to evaluate factors associated to orthodontic treatment stability and patient satisfaction in the long-term. A total of 209 patients (88 class I and 121 class II) treated with straight wire fixed appliance were selected at least 5 years post treatment. Six hundred twenty seven dental casts were examined with the PAR Index at pretreatment (T1), end of treatment (T2), and at long-term follow up (T3, mean 8.5 years post treatment). At T3, a Dental Impact on Daily Living questionnaire was used to assess patient satisfaction with the dentition in the long-term. Friedman test and multiple regression analysis were used to evaluate changes among the time points and factors associated with stability and patient satisfaction. Predictive factors used to exam the occlusion were: PAR Index at T1 and T2, age at T1, the amount of time without retainer, length of Hawley retainer wear, length of follow-up, sex, extraction and third molar status. To assess patient satisfaction were considered: changes produced by the orthodontic treatment (PAR T2-T1), post treatment stability (PAR T3), age at the start of treatment (T1), length of treatment (T2-T1), gender, and extraction. Orthodontic treatment produced a significant improvement of 94.2% in the PAR Index (T2-T1), but this change was not associated with the level of satisfaction when the patient was questioned at T3. No significant change was observed between T2 and T3. However, when the sample was divided according to the level of finalization (PAR T2), it was observed that well-finished patients experienced some deterioration (P<.001), whereas the less well-finished ones showed some improvement (P<.05). Even with the deterioration, the well-finished patients still had a better PAR Index at T3 compared to the less well-finished ones (PAR T2- T3). Regression analysis showed that PAR Index at T1 and T2, age at T1, and length of retainer wear had a slight association with occlusal stability (R2 = 0.27). Patient satisfaction was significantly associated only with PAR Index at T3 (r2=0.125, P<.0001). We can conclude that, even thought orthodontic treatment is quite stable, not so well-finished treatments tend to show some improvement and well-finished ones deteriorate some in the long-term. Despite of that, well-finished patients still have better occlusal characteristics. Patient satisfaction is not related to the result of orthodontic treatment; nevertheless, there is a slight association with dentition in the long-term

Emulsões de óleo de copaíba: determinação do equilíbrio hidrófilo-lipófilo crítico (EHLC), propriedades e estabilidade físico-químicas

Durante as últimas décadas, as indústrias farmacêuticas têm despertado grande interesse em óleos vegetais e vários extratos de planta por causa da sua baixa toxicidade e alta biodegrabilidade. O óleo de copaíba (Cop) é usado in natura na medicina popular como anti-inflamatório e antimicrobiano para tratar várias doenças, tais como inflamação da garganta, úlceras e infecções urinárias e pulmonares. Emulsões são sistemas dispersos termodinamicamente instáveis que consistem em dispersões de gotículas microscópicas em outro líquido imiscível. O objetivo deste trabalho foi preparar diferentes emulsões de Cop, determinar o EHL crítico deste óleo e avaliar a sua estabilidade, além de realizar estudos comparativos entre diferentes métodos de construção de diagramas de fases. As emulsões foram preparadas pelo método de inversão de fases variando as proporções de EHL de 4,7 a 16,7. A estabilidade foi determinada por vários métodos e os diagramas de fases foram produzidos pelo método de titulação usando diferentes procedimentos de agitação. As emulsões a base de Cop com EHL entre 12,7 e 15,7 foram as mais estáveis. As emulsões apresentaram boa estabilidade em curto e longo prazo, aspecto leitoso e baixos valores de índice de cremagem. Diferentes sistemas coloidais foram produzidos a partir dos diagramas de fases dependendo do processo de agitação. Baseado nesses métodos, o valor determinado de EHL do Cop foi 14,8, as emulsões permaneceram estáveis por mais de um ano e estes resultados indicam que o estudo das emulsões de Cop pode ser um promissor veículo de liberação tópica de fármacos e ativos cosméticos

Influência de fármacos lipofílicos na estabilidade de emulsões de uso terapêutico

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Avaliação de um método de cromatografia em fase gasosa head space e estudo da estabilidade do etanol em amostras de sangue

The ethanol is the most overused psychoactive drug over the world; this fact makes it one of the main substances required in toxicological exams nowadays. The development of an analytical method, adaptation or implementation of a method known, involves a process of validation that estimates its efficiency in the laboratory routine and credibility of the method. The stability is defined as the ability of the sample of material to keep the initial value of a quantitative measure for a defined period within specific limits when stored under defined conditions. This study aimed to evaluate the method of Gas chromatography and study the stability of ethanol in blood samples, considering the variables time and temperature of storage, and the presence of preservative and, with that check if the conditions of conservation and storage used in this study maintain the quality of the sample and preserve the originally amount of analyte present. Blood samples were collected from 10 volunteers to evaluate the method and to study the stability of ethanol. For the evaluation of the method, part of the samples was added to known concentrations of ethanol. In the study of stability, the other side of the pool of blood was placed in two containers: one containing the preservative sodium fluoride 1% and the anticoagulant heparin and the other only heparin, was added ethanol at a concentration of 0.6 g/L, fractionated in two bottles, one being stored at 4ºC (refrigerator) and another at -20ºC (freezer), the tests were performed on the same day (time zero) and after 1, 3, 7, 14, 30 and 60 days of storage. The assessment found the difference in results during storage in relation to time zero. It used the technique of headspace associated with gas chromatography with the FID and capillary column with stationary phase of polyethylene. The best analysis of chromatographic conditions were: temperature of 50ºC (column), 150ºC (jet) and 250ºC (detector), with retention time for ethanol from 9.107 ± 0.026 and the tercbutanol (internal standard) of 8.170 ± 0.081 minutes, the ethanol being separated properly from acetaldehyde, acetone, methanol and 2-propanol, which are potential interfering in the determination of ethanol. The technique showed linearity in the concentration range of 0.01 and 3.2 g/L (0.8051 x + y = 0.6196; r2 = 0.999). The calibration curve showed the following equation of the line: y = x 0.7542 + 0.6545, with a linear correlation coefficient equal to 0.996. The average recovery was 100.2%, the coefficients of variation of accuracy and inter intra test showed values of up to 7.3%, the limit of detection and quantification was 0.01 g/L and showed coefficient of variation within the allowed. The analytical method evaluated in this study proved to be fast, efficient and practical, given the objective of this work satisfactorily. The study of stability has less than 20% difference in the response obtained under the conditions of storage and stipulated period, compared with the response obtained at time zero and at the significance level of 5%, no statistical difference in the concentration of ethanol was observed between analysis. The results reinforce the reliability of the method of gas chromatography and blood samples in search of ethanol, either in the toxicological, forensic, social or clinic

Soroprevalência da infecção pelo Trypanosoma cruzi no município de Caicó, RN, e caracterização genética por RAPD do parasito isolado de humanos e de triatomíneos no semiárido potiguar

Trypanosoma cruzi infection was evaluated in 390 resident individuals in different rural communities of Caicó municipality, State of Rio Grande do Norte (RN). Of 28 investigated communities the soroprevalence of T. cruzi infection was 2.8% in eight rural communities individuals. The epidemiological characteristics of seropositive shown that the age ranged from 22 to 64 years, being significantly raised from 31 years (90.9%). The female gender was predominant and low education degree. Those individuals reported that they never donated blood, but they had direct contact with triatomines bug. The isolation of the parasite was performed by blood culture and xenoculture methods to determine the genetic variability of the samples. Twenty seven T. cruzi isolates were analyzed by RAPD as genetic marker using three random primers (M13-40, gt11-F and L15996). The T. cruzi isolates showed 73.7% of shared bands considering the average obtained with the three primers, and were genetically well correlated. Using this marker it was possible to separate the populations of the parasite in three distinct groups. The first group composed by isolates obtained of triatomines and humans from four different districts (Caicó, Caraúbas, Serra Negra doNorte and Governador Dix-Sept Rosado); the second contained isolates obtained of triatomines of two different species (T. brasiliensis and P. lutzi) captured in Caraúbas and Serra Negra do Norte. The third grouped isolates obtained from humans of Angicos and Caicó municipalities. In different localities of distinct mesoregions, State of RN, a profile genetic well correlated was identified among all isolates and the presence of three distinct groups of the parasite circulating among vertebrate and invertebrate hosts

Estudo de compatibilidade e estabilidade térmica do ácido retinóico e hidroquinona por termogravimetria

Retinoic acid (RA) and hydroquinone (HQ) assets are widely used in pharmaceutical and cosmetic formulations, for having depigmenting properties and are largely produced in drugstores. To assist in the development of formulations containing the active RA and HQ National Forms of Brazilian Pharmacopoeia (2005 and 2012 ) proposes formulations with different excipients such as cetyl alcohol (AC), cetostearyl alcohol (ACT), methylparaben (MTP), propyl paraben ( PPB), glycerin (GLY), dipropylene glycol (DPG), imidazolidinil urea ( IMD ), cyclomethicone (CCM ), butylated hydroxytoluene (BHT), octyl stearate (ETO), EDTA, decil oleate (ODC) and hydroxipropymethyl celullose (HPMC). One of the difficulties found in most cosmetic formulations is the large number of incompatibilities between the components of the formulations, so the aim this study was to evaluate thermal stability and interactions between these active pharmaceutical ingredients and excipients. The depigmenting agents were analyzed by DSC and TG and excipients were analyzed by TG. The dynamic thermogravimetric curves were obtained on a SHIMADZU thermobalance, model DTG-60, using an alumina crucible, at the heating rate of 10ºC min-1, in the temperature range of 25-900 ºC, under an atmosphere of nitrogen at 50 mL min-1. The DSC curves were obtained using Shimadzu calorimeter, model DSC-60, using aluminum crucible, at the heating rate of 10ºC min-1, in the temperature range of 25-400ºC. The thermogravimetric and calorimetric curves were analyzed using TASYS software SHIMADZU. In this study no were found interactions between AR and the following excipients: MTP, PPB, IMD, ODC, EDTA, CCM, ETO, HPMC. However, were found interactions with the following excipients: AC, ACT, BHT, GLI and DPG. For HQ were found interactions with IMD and DPG. Interactions remained even changing proportions of the mixtures and the ternary. Thus, the studies conducted with excipients of National Formulary from 2005 and 2012 showed that these new excipients do not interact by thermogravimetry with the active pharmaceutical ingredients of this study