Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2015-01-30T10:44:15Z No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertacao - Priscilla Regina Nasciutti - 2014.pdf: 4368236 bytes, checksum: 303bde7df2549243e3355b74c882e0ee (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-01-30T12:50:15Z (GMT) No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertacao - Priscilla Regina Nasciutti - 2014.pdf: 4368236 bytes, checksum: 303bde7df2549243e3355b74c882e0ee (MD5) / Made available in DSpace on 2015-01-30T12:50:15Z (GMT). No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertacao - Priscilla Regina Nasciutti - 2014.pdf: 4368236 bytes, checksum: 303bde7df2549243e3355b74c882e0ee (MD5) Previous issue date: 2014-02-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / In dogs, as in humans, arterial hypertension is characterized by an excessive increase of systemic blood pressure which can cause renal, ocular, neurological and cardiac lesions. Currently, five classes of antihypertensive agents are commonly used. A drug prototype (LASSBio 897) capable of promoting vasodilation effects was developed and became a possible alternative for new therapeutic strategy. The main goal of this study was to investigate its vasodilatory effect, as well as to identify possible changes in clinical, cardiovascular, renal and hepatic functions. Six adult and healthy beagle dog were used. They received oral doses of 0.5mg/kg, 1.0mg/kg and 2.0mg/kg of LASSBio 897 and 0.5mg/kg of benazepril, as a positive control. Assessments of liver and kidney function, doppler echocardiographic and electrocardiographic examinations and blood pressure measurements, were performed. No clinical changes suggesting acute toxicity were observed and all measured parameters were within the normal values for canine species. Blood pressure values decreased gradually until time of 120min, in which its lowest value was observed (120.58mmHg), among the three studied doses, similar to that observed with benazepril. No hypotension or arrhythmias were observed. In echocardiography, values of stroke volume and cardiac output were lower than those described as normal for the species, with averages that decreased until time of 240min. In conclusion the oral administration of the prototype LASSBio 897, in single doses, causes vasodilation and is safe since no significant changes in haematological, renal and hepatic biochemical evaluations were observed, as well as in urinary review. The prototype does not cause side effects, hypotension or abnormal heart rhythms. / Em cães, assim como nos humanos, a hipertensão arterial é caracterizada por uma elevação excessiva da pressão arterial sistêmica e pode produzir lesões renais, oculares, neurológicas e cardíacas. Atualmente, existem cinco classes de agentes anti-hipertensivos que são utilizados para o tratamento. Um protótipo a fármaco (LASSBio 897) capaz de promover efeitos vasodilatadores foi desenvolvido, tornando-se um possível candidato a uma nova estratégia terapêutica. O objetivo principal foi verificar o efeito vasodilatador, bem como identificar possíveis alterações clínicas, cardiovasculares, renais e hepáticas. Foram utilizados seis cães da raça beagle, adultos, clinicamente saudáveis, que receberam, por via oral, doses de 0,5mg/kg, 1,0mg/kg e 2,0mg/kg do LASSBio 897 e 0,5mg/kg de benazepril, como controle positivo. Foram submetidos à avaliações da função hepática e renal, exames ecodopplercardiográficos, eletrocardiográficos e mensuração da pressão arterial. Não foram observadas alterações clínicas que sugerissem toxicidade aguda e todos os parâmetros mensurados contemplavam valores normais para a espécie canina. Os valores de pressão arterial foram diminuindo gradativamente até o tempo de 120 minutos, quando se observou seu menor valor (120,58mmHg), nas três doses estudadas, semelhante ao observado com o benazepril. Não foram observadas hipotensão ou arritmias. No ecodopplercardiograma os valores de volume sistólico e débito cardíaco foram menores do que os descritos como normais para a espécie, com médias que foram diminuindo até o tempo de 240 minutos. Concluindo, a administração oral do protótipo a fármaco LASSBio 897 nas doses únicas utilizadas produz vasodilatação e é segura, pois não foram observadas alterações importantes nas avaliações hematológicas, bioquímicas renais e hepáticas, bem como da avaliação urinária. O protótipo não causou efeitos adversos, hipotensão ou anomalias do ritmo cardíaco

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-26T14:49:08Z No. of bitstreams: 2 Dissertação - Jakeline Ferreira de Araújo Lôbo - 2016.pdf: 4418775 bytes, checksum: bf4e8486761257498acba551292fe110 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-26T14:51:49Z (GMT) No. of bitstreams: 2 Dissertação - Jakeline Ferreira de Araújo Lôbo - 2016.pdf: 4418775 bytes, checksum: bf4e8486761257498acba551292fe110 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-26T14:51:49Z (GMT). No. of bitstreams: 2 Dissertação - Jakeline Ferreira de Araújo Lôbo - 2016.pdf: 4418775 bytes, checksum: bf4e8486761257498acba551292fe110 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-06-10 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The equine presents itself as an important kinesiotherapeutic tool, this occurs because when walking, it carries out tridimensional movements that cause coordinated and synchronized muscle actions in the rider, what can justify its therapeutic application in disabled human persons. It was established that the frontal plan up and down movements performed by the equine has impact on the rider’s pelvis in movements of lateral inclination. The back and forth movements in sagittal plan generate ante and retroversion movements. On the other hand, the right and left movements in transverse plan result in pelvic rotations. Such movements are similarly executed by the human being pelvis, this way, the rider physically prevented from walking, passively experiences while riding, the same amount of pelvis displacement and rotation during the biped march. Due to the importance of this movement quality, this work has as objective analyze the equine conformation of the equine center of the Rehabilitation and Readaptation center Dr. Henrique Santillo (CRER). It is a transverse study, which used 180 cm tape measure, hipometer, reflexive markers, cameras and kinematical analysis software Kinovea, to evaluate the linear and angular parameters, together with five zootechnical indexes. Eight half-breed equines, aged between 3 to 17 years, were evaluated, they were destinated exclusively to the equine therapy practice. The statistical analysis was descriptive and analytic. The corporeal index average was 0, 85, the relation between the back height and the substernal fail height was 1, the conformation index 2, 1125, indicating the suitable animals for march and saddle. In the actual study, the animals presented smaller neck and head scapular length in relation to other equine conformation studies. The thoracic cavity depth determines cardiopulmonary performance and provide to the equine therapy, long working hours. On the CRER equines, the average obtained was high. On the forearm, knee and shank regions, it is ideal to be hypertrophied, to protect from possible injuries, reflex of good management practices and then, it was observed in the animals. There should be correspondence between the angles of the thoracic, or accommodation members and the pelvic or propulsive ones, and some angles of this study had correspondence, as the humerus-radius and the coxofemoral and in relation to the solo phalanges. As a conclusion, The CRER equines kept proportionality regarding to some variables as the thorax perimeter, the head length and width and the scapula angulation and the solo phalanges. The zootechnical indexes determine that the animals are suitable for the equine therapy practice. Regarding to the other parameters, they didn’t obtain proportionality, which can interfere in the pace progress, this justifies the using of specific tools as the ones used in this study for the adequate choice of equines for equine therapy and the need for more studies in other centers for comparison. / O equino apresenta-se como uma importante ferramenta cinesioterapêutica, isto ocorre porque ao andar realiza movimentos tridimensionais que se desdobram em ações musculares coordenadas e sincronizadas no cavaleiro, o que pode justificar sua aplicação terapêutica em humanos pessoas com deficiências. Constatou-se que os movimentos para cima e para baixo no plano frontal executado pelo equino, repercutem na pelve do cavaleiro em movimentos de inclinação lateral. Os movimentos para frente e para trás no plano sagital geram movimentos de antero e retroversão. Por sua vez, os movimentos para a direita e esquerda no plano transversal resultam em rotações pélvicas. Tais movimentos são similarmente executados pela pelve do ser humano, e desse modo, o cavaleiro fisicamente impedido de andar, experimenta passivamente ao cavalgar a mesma quantidade de deslocamento e rotação da pelve durante a marcha bípede. Devido a importância da qualidade deste movimento este trabalho tem por objetivo analisar a conformação de equinos do Centro de Equoterapia do Centro de Reabilitação e Readaptação Doutor Henrique Santillo (CRER). Trata-se de um estudo transversal que utilizou fita métrica de 180 cm, hipômetro, marcadores reflexivos, máquina fotográfica e software de análise cinemática Kinovea®, para avaliação dos parâmetros lineares e angulares, juntamente com cinco índices zootécnicos. Foram avaliados oito equinos mestiços, com idades de 3 à 17 anos, destinados exclusivamente á prática da Equoterapia. A análise estatística foi descritiva e analítica. A média do índice corporal foi de 0,85, da relação entre a altura do costado e a altura do vazio subesternal foi 1, do índice de conformação 2,1125, indicando os animais aptos para marcha e sela. No presente estudo, os animais apresentaram menor comprimento escapular, de pescoço e cabeça em relação a outros estudos de conformação com equinos. A profundidade da caixa torácica determina o rendimento cardiorrespiratório e proporciona para equoterapia períodos longos de trabalhos. Nos equinos do CRER, a média obtida foi alta. Nas regiões do antebraço, joelho e canela o ideal é que sejam hipertrofiados, para proteção de possíveis lesões, reflexo de boas práticas de manejo e, assim, foi observado nos animais. Deve existir correspondência entre os ângulos dos membros torácicos, ou de acomodação, e pélvicos ou propulsores, e alguns ângulos deste estudo tiveram correspondência, como o úmero-radial e coxofemoral e em relação às falanges solo. Como conclusão, os equinos do CRER mantiveram proporcionalidade em relação a algumas variáveis como o perímetro do tórax, o comprimento e largura da cabeça e as angulações da escápula e as falanges solo. Os índices zootécnicos determinaram que os animais estão aptos para a prática de Equoterapia. Em relação aos demais parâmetros não obtiveram proporcionalidade, o que pode interferir no andamento do passo, o que justifica a utilização de ferramentas específicas como as utilizadas neste estudo para a escolha adequada de equinos para Equoterapiae a necessidade de mais estudos em outros centros para comparação.

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2016-12-06T16:54:56Z No. of bitstreams: 2 Dissertação - Bruno César Ferreira Gonzaga - 2016.pdf: 2851863 bytes, checksum: 3316d056c52b9a8ca6ed7608909e2b80 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2016-12-13T15:35:12Z (GMT) No. of bitstreams: 2 Dissertação - Bruno César Ferreira Gonzaga - 2016.pdf: 2851863 bytes, checksum: 3316d056c52b9a8ca6ed7608909e2b80 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-12-13T15:35:12Z (GMT). No. of bitstreams: 2 Dissertação - Bruno César Ferreira Gonzaga - 2016.pdf: 2851863 bytes, checksum: 3316d056c52b9a8ca6ed7608909e2b80 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-09-30 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The Brachiaria grass, with broad geographic distribution in Brazil, has been shown to cause photosensitization and appearance of foamy macrophages in the liver and liver and mesenteric lymph nodes of ruminants. Weight loss of 2.52% in cattle with subclinical hepatogenous photosensitization was documented and it is estimated that in one year in a single slaughterhouse more than R$ 100,000 in annual losses are caused by liver damage resulting from consumption of Brachiaria spp. The aim of this study was to corelate the extent of liver parenchyma containing foamy macrophages with hot carcass weight of cattle. Carcasses of cattle aged 20-36 months were weighed and divided into two groups: Group 1 (544.87-607.13 pounds) and Group 2 (689.42-757.24 pounds) The livers of bovines were weighed and samples of the right lobe were collected for histopathologic analysis. The areas of hepatic parenchyma containing foamy macrophages were measured using the ImageJ® program. Statistical analysis of the parameters (liver weight, hot carcass weights and affected area of liver tissue) was performed quantitatively, by calculating the mean, standard deviation and coefficient of variation. These data were analyzed for normality and homogeneity of variances, through the Kolmogorov-Smirnov and Bartlett tests. As the variables were homogeneous and had a normal distribution were evaluated by parametric test analysis of variance. The correlation of variables was performed by Spearman test. The significance level was set at 5% and all descriptive analyzes were performed by EXCEL program and statistical tests by the software R. Cattle grazing Brachiaria spp. presented hepatic lesions characterized by degeneration, necrosis, presence of mild inflammatory infiltrates and small area of hepatic tissue containing foamy macrophages. There was no influence of the affected area of the hepatic parenchyma containing foamy macrophages with hot carcass weight of cattle. / O capim braquiária, com ampla distribuição geográfica no Brasil, vem sendo apontado como causa de fotossensibilização e do aparecimento de macrófagos espumosos no fígado e linfonodos hepáticos e mesentéricos de ruminantes. Foram relatadas perdas de peso na ordem de 2,52% em bovinos com fotossensibilização hepatógena subclínica e estimou-se que mais de R$ 100.000 em perdas anuais por lesões hepáticas em apenas um frigorífico fosse decorrente do consumo de Brachiaria spp. O objetivo deste estudo foi correlacionar a extensão do parênquima hepático contendo macrófagos espumosos com o peso de carcaça quente dos bovinos. As carcaças dos bovinos, com idade entre 20 a 36 meses, foram pesadas e distribuídas em dois grupos: Grupo 1 (247,15 a 275,39 kg) e Grupo 2 (312,72 a 343,48 kg). Os fígados dos bovinos foram pesados e amostras do lobo direito coletadas para análise histopatológica. As áreas do parênquima hepático contendo macrófagos espumosos hepáticos foram mensuradas utilizando-se o programa ImageJ®. A análise estatística dos parâmetros (peso do fígado, peso da carcaça quente e área comprometida do tecido hepático) foi realizada de forma quantitativa, calculando a média, desvio padrão e coeficiente de variação. Esses dados foram analisados quanto à normalidade e homogeneidade das variâncias, por meio dos testes de Kolmogorov-Smirnov e Bartlett. Como as variáveis foram homogêneas e tiveram uma distribuição normal, foram avaliadas pelo teste paramétrico de análise de variâncias. A correlação das variáveis foi realizada pela correlação de Spearman. O grau de significância adotado foi de 5% e todas as análises descritivas foram realizadas pelo programa EXCEL e os testes estatísticos pelo programa R. Os bovinos sob pastejo de Brachiaria spp. apresentaram lesões hepáticas caracterizadas por degeneração, necrose, presença de infiltrados inflamatórios de intensidade leve e pequena área do tecido hepático contendo macrófagos espumosos. Não houve influência da área comprometida do parênquima hepático contendo macrófagos espumosos com o peso de carcaça quente dos bovinos.

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2017-03-31T17:23:22Z No. of bitstreams: 2 Dissertação - Thamiza Carla Costa dos Santos - 2016.pdf: 1439728 bytes, checksum: 19a51f9a39cdb7d376515edd24484335 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-04-03T12:02:47Z (GMT) No. of bitstreams: 2 Dissertação - Thamiza Carla Costa dos Santos - 2016.pdf: 1439728 bytes, checksum: 19a51f9a39cdb7d376515edd24484335 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-03T12:02:47Z (GMT). No. of bitstreams: 2 Dissertação - Thamiza Carla Costa dos Santos - 2016.pdf: 1439728 bytes, checksum: 19a51f9a39cdb7d376515edd24484335 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-08-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The healing is a complex and dynamic event aimed at the restoration of tissue integrity. However, some factors, such as diabetes, can harm this physiological event. Chitosan, a derivative of chitin, has been considered as a versatile and effective option for the treatment of wounds. This study aimed to evaluate and compare the effect of chitosan 2% gel and chitosan spray based on secondary intention healing of skin wounds in diabetic rats. 54 rats were used, divided randomly into three groups: control group (CG), chitosan 2% gel group (GG), and Chitosan spray group (GS) with 18 animals each. The rats in GC, GG, and GS had their wounds treated daily with saline solution 0.9%, chitosan 2% gel, and chitosan spray, respectively. The groups were divided into three subgroups (n = 6) according to the times of histological analysis of wound (third, seventh, and 14th day). The induction of diabetes in the rats took place by intraperitoneal application of alloxan (120 mg/kg). After confirming the state of diabetes, the animals were anesthetized and the wounds made with the aid of a punch 9.0mm in the dorsal region. Macroscopic and microscopic analyzes were performed. Macroscopic analysis showed that after 14 days, the chitosan 2% gel and chitosan spray gave a contraction rate of 100% and 99%, respectively. At the histological level, there was the anticipated recruitment of mononuclear cells, fibroblasia, and deposition of dense fibrovascular tissue and marked fibrosis in the wounds of GG and GS, emphasizing the ability of this biopolymer to optimize the healing of diabetics. It can therefore be a therapeutic option for the development of other formulations that provide wound repair in diabetic patients. / A cicatrização é um evento complexo e dinâmico que visa a restauração da integridade tecidual. No entanto, alguns fatores, como o diabetes, podem prejudicar este evento fisiológico. A quitosana, um derivado da quitina, tem sido cogitada como uma opção versátil e eficaz para o tratamento de feridas. O presente estudo teve como objetivo avaliar e comparar o efeito do gel de quitosana a 2% e spray à base de quitosana na cicatrização por segunda intenção de feridas cutâneas em ratas diabéticas. Foram utilizadas 54 ratas, divididas, aleatoriamente, em três grupos: grupo controle (GC) grupo gel de quitosana a 2% (GG) e grupo spray de quitosan (GS) com 18 animais, cada. As ratas dos grupos GC, GG e GS tiveram suas feridas tratadas diariamente com solução fisiológica 0,9%, gel de quitosana a 2% e spray à base de quitosana, respectivamente. Os grupos foram subdivididos em três subgrupos (n=6) conforme os tempos de análise histológica da ferida (terceiro, sétimo e 14º dia). A indução do diabetes mellitus, nas ratas, ocorreu por meio da aplicação intraperitoneal da aloxana (120mg/kg). Após a confirmação do estado de diabetes, os animais foram anestesiados e as feridas confeccionadas com auxílio de um punch de 9,0mm na região dorsal. Foram realizadas análises macroscópicas e microscópicas. As análises macroscópicas mostraram que ao 14º dia, o gel de quitosana 2% e o spray de quitosana proporcionaram uma taxa de contração de 100% e 99%, respectivamente. A nível histológico, observou-se o recrutamento antecipado de células mononucleares, fibroblasia e deposição de tecido fibrovascular denso acentuados e fibrose nas feridas dos GG e GS, enfatizando a capacidade deste biopolímero em otimizar a cicatrização de diabéticos. Podendo, portanto, ser uma opção terapêutica como também a base para o desenvolvimento de outras formulações que proporcionem a reparação de feridas em pacientes diabéticos.

Stroke and traumatic brain injury are among the leading causes of death in the world. Until now, there are no effective treatments available. Current pharmaceutical treatments have limited benefits to repair the damaged tissue. Brain tissue engineering is a promising strategy to help brain regeneration after the damage induced by stroke or traumatic brain injury. In this thesis, our work focused on designing and evaluating appropriate silk fibroin-based hydrogels combined with stem cells therapy for brain tissue regeneration. The work initially started from looking for appropriate silk fibroin-based hydrogel substrates which can support the viability and neural differentiation of pluripotent cells. Mouse embryonic stem cells (mESC) were used as a model. Different processing procedures of silk fibroin-based hydrogel substrates were prepared by chemical genipin crosslinking and physical sonication crosslinking. The viability and neural differentiation of pluripotent cells on these hydrogel substrates were evaluated, using tissue culture plates (TCP) as control. Different crosslinking processes were found to modulate the neural differentiation of pluripotent cells. Chemical genipin crosslinked hydrogel substrates could inhibit the neural differentiation of mESC compared to control TCP, while the physical sonication crosslinked hydrogel substrates could support the neural differentiation as TCP. According to the results obtained in the first stage, the physically sonication-crosslinked 3D silk fibroin hydrogel was produced to encapsulate human neural stem cells (hNSC). In order to improve the hNSC attachment and neuronal differentiation, the isoleucine-lysine-valine-alanine-valine (IKVAV) peptide derived from laminin was covalently conjugated to the silk fibroin. The viability and neural differentiation of hNSC were evaluated in the unmodified and IKVAV peptide modified silk fibroin hydrogels. We found that the IKVAV peptide modified silk fibroin hydrogel could increase the viability, proliferation and neuronal differentiation of hNSC. Furthermore, the angiogenesis potential of sonication-induced 3D silk fibroin unmodified and modified with IKVAV and a scramble peptide VVIAK (as control) were evaluated in a human outgrowth endothelial cells (OEC) mono-culture system and a co-culture system in which OEC were cultured with human bone marrow mesenchymal stem cells (BM-MSC). Both the silk fibroin unmodified and modified with IKVAV peptide could not induce angiogenesis in the mono-culture system under the VEGF condition. However, in the co-culture system, we found that unmodified, IKVAV-modified and VVIAK-modified silk fibroin hydrogels all could support angiogenesis. Furthermore, there were no significant differences among unmodified, IKVAV modified and VVIAK modified silk fibroin hydrogels influencing on angiogenesis structure and gene expression related to angiogenesis. The thesis will introduce the detailed work in three different chapters (from chapter 3 to chapter 5) respectively.

Organ failure is one the biggest problems, doctors face every day. Many patients are not able to get a transplant, but even those who recieved it, may undergo painful process of organ rejection and be on the transplant waiting list again. Organ transplants shortage is severe problem in current medicine that has many ethical and medical issues. To solve this problem, the new direction in regenerative medicine was formed, organ prinitng. The main goal of organ printing is fabrication of organ replacements that would mimic the original ones in terms of complexity and functionality. By direct fabrication and maturation of organs in vitro, the problem of organ shortage can be solved, moreover, based on the advances in cell therapy, these organs can be printed with patients own cells, which will eliminte the problem of transplant rejection. Organ printing is multistep and complex process, composed of three main steps: tissue design, or theoretical modelling of replacement composition; tissue fabrication, or direct cell encapsulation and controlled assembly of building units; at last, tissue maturation to reach desirable functionality of the replacement. In the past decade, there was developed a variety of methods for the second step of organ printing, cell encapsulation, which is practicaly the main procedure for tissue fabrication. However, all these methods of cell encapsulation are complex and they might affect cells viability and functionality, which will result in changed tissue function. Thus, starting from the detailed analysis of the tissue fabrication process (encapsulation and assembly methods) the list of possible cell behavior affectors was composed. Based on this list, we designed a multistep protocol for coherent evaluation of cells behavior parameters, in terms of viability, functionality and activity during the tissue fabrication and its maturation steps. Three main materials were used for this study, two naturally (alginate and modified gelatin) and one synthetically (polyethilene glycol) derived polymers. The encapsulation step was performed with two different methods based on chemical or photo crosslinking of the material. Cell parameteres were evaluated on the molecular level for variety of parameters, including viability, activity, proliferation, stress markers expression, at last ability to adapt artificial environment to the cell functional niche with extracellular matrix markers expression, and proteoglycans. The innovation of the presented study consists in the developing a unique protocol for detailed cell functionality evaluation during the organ printing procedures. In fact, based on the conducted study, it was proved the safety of the encapsulation methods. Moreover, based on the cell parameters post-encapsulation, there was suggested the optimal time for tissue maturation for application of the fabricated structures in organ printing, but also in other fileds, like developmental and pathological biology, or drug screening. Eventully, a novel way of simple blocks assembly into 3D complex structures was developed and proved to be safe for cell parameters. At last, for the future research in organ printing, a detailed study over a cell behavior and functionality has to be performed for every fabrication method, what will improve the organ production process drastically.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-07-17T11:35:24Z No. of bitstreams: 2 Tese - Daniel Silva Goulart -2017.pdf: 3553497 bytes, checksum: 6ec13ae3cfbad63a4fdfdcf4e9b0dab1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-07-18T10:31:42Z (GMT) No. of bitstreams: 2 Tese - Daniel Silva Goulart -2017.pdf: 3553497 bytes, checksum: 6ec13ae3cfbad63a4fdfdcf4e9b0dab1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-07-18T10:31:42Z (GMT). No. of bitstreams: 2 Tese - Daniel Silva Goulart -2017.pdf: 3553497 bytes, checksum: 6ec13ae3cfbad63a4fdfdcf4e9b0dab1 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-03-02 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The genus Brachiaria has provided important forage species to the countries located in tropical regions. However, this genus of grass is associated with photosensitization and poisoning in animal production. Thus, the aim of this study was to establish the standard for collection, processing, and conservation of Brachiaria brizantha for protodioscin quantification and spore count from Pithomyces chartarum and the evaluation of guinea pig (Cavia porcellus) as an experimental model of poisoning by diosgenin. For Brachiaria brizantha evaluation, samples were collected from eight pastures, which were subdivided into young leaves, mature leaves, old leaves, and whole plant. After that, the samples were submitted to nine different drying and conservation treatments. After separation and treatment of the samples, saponins were quantified and spores were counted. In most of treatments, the young leaves presented larger amounts of protodioscin than the old leaves and the whole plant. The treatments that maintained the highest amounts of protodioscin were room temperature, oven, and oven with forced air circulation. No fungal spores were found in the pasture samples. Thus, we concluded that the concentration of protodioscin varies with the maturation stage of the leaves; drying at room temperature, in an oven, or in an oven with forced air circulation are the best methods, and freezing plants for preservation of protodioscin is not recommended. In the guinea pigs evaluation, 14 guinea pigs were divided into two groups, the treatment group (TG) and the control group (CG). The TG received 480 mg/kg of diosgenin and the CG received only vegetable oil. Both groups received the treatment for 30 days. Blood samples were collected on day 0 (M0) and day 30 (M30) for blood cell count and biochemical analyzes. In clinical biochemistry, the analyzes were performed for alanine aminotransferase, aspartate aminotransferase, gamma glutamyl trasferase, alkaline phosphatase, urea, creatinine, glucose, cholesterol, HDL, triglycerides, lactate, cholinesterase, and calcium. Hematology and biochemistry did not present alterations, which could not be related to hepatic disease caused by saponins. In histology, only one animal had bile duct hyperplasia, the other alterations observed were generally nonspecific. Therefore, we concluded that the administration of 480 mg/kg of diosgenin to guinea pigs did not produce significant hepatic lesions or hematological and biochemical alterations related to liver disease. / O gênero Brachiaria possui importantes espécies de forrageiras utilizadas em países localizados em regiões tropicais. No entanto, esse gênero de gramíneas está associado à fotossensibilização e intoxicações em animais de produção. Deste modo, objetivou-se com este estudo estabelecer o padrão para colheita, processamento e conservação de amostras de Brachiaria brizantha para quantificação de protodioscina e contagem de esporos de Pithomyces chartarum e a avaliação da Cavia porcellus (cobaias) como modelo experimental na intoxicação por diosgenina. Para a avaliação da Brachiaria brizantha, amostras foram colhidas de oito pastagens as quais foram subdivididas em folhas jovens, folhas maduras, folhas velhas e planta inteira e submetidas a nove diferentes métodos de processamento. Após separação e tratamento das amostras foi realizada a quantificação de saponinas e a contagem de esporos. Observou-se que na maioria dos tratamentos as folhas jovens apresentaram quantidades maiores de protodioscina que as folhas velhas e a planta inteira. Os tratamentos que mantiveram as maiores quantidades de protodioscina foram temperatura ambiente, estufa e estufa com ventilação forçada. Nas amostras das pastagens não foram encontrados esporos do fungo. Desta forma, conclui-se que a concentração de protodioscina é maior nas folhas jovens; a secagem em temperatura ambiente, em estufa ou em estufa de ventilação forçada são os melhores métodos para preservação de protodioscina e o congelamento de plantas para a quantificação de protodioscina não é recomendado. Para a avaliação das cobaias como modelos experimentais na intoxicação por diosgenina foram utilizadas 14 cobaias, divididas em dois grupos, o grupo tratado (GT) e o grupo controle (GC). O GT recebeu 480 mg/kg de diosgenina diluída em óleo vegetal e o GC recebeu somente o óleo vegetal, ambos por 30 dias. Para avaliação do hemograma e análises bioquímicas foram colhidas amostras sanguíneas no dia 0 (M0) e no dia 30 (M30). Na bioquímica plasmática foram realizadas análises para ALT, AST, GGT, ALP, ureia, creatinina, glicose, colesterol, HDL, triglicerídeos, lactato, colinesterase e cálcio. A hematologia e a bioquímica não apresentaram alterações relacionadas à doença hepática causada por saponinas. Na histologia apenas um animal apresentou hiperplasia de ductos biliares, as outras alterações observadas foram de forma geral inespecíficas. Conclui-se que, na dose utilizada, a administração da diosgenina a cobaias não produz lesões hepáticas expressivas nem alterações hematológicas e bioquímicas relacionadas a hepatopatia.

Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-01-06T16:16:41Z No. of bitstreams: 2 Dissertação - Ygor Roberto de Carvalho - 2009.pdf: 1167127 bytes, checksum: 7fe8cf1100e0875c875d048be01ffbbc (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-01-07T08:16:45Z (GMT) No. of bitstreams: 2 Dissertação - Ygor Roberto de Carvalho - 2009.pdf: 1167127 bytes, checksum: 7fe8cf1100e0875c875d048be01ffbbc (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-01-07T08:16:45Z (GMT). No. of bitstreams: 2 Dissertação - Ygor Roberto de Carvalho - 2009.pdf: 1167127 bytes, checksum: 7fe8cf1100e0875c875d048be01ffbbc (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2009-09-28 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Peritoneal adhesions are fibrous bands that form abnormal unions between two or more serosal coating surfaces, may cause obstructions of abdominal tubular organs and pelvic pain. Several studies have been performed using antiinflammatory, anticoagulant, proteolytic agents and biomaterials in the form of film or gel, with the aim of preventing or simply reduce the formation of peritoneal adhesions. The chitosan is a polymer obtained by deacetylation of chitin, a polysaccharide obtained from the exoskeleton of crustaceans and has been used in various medical specialties as a biomaterial. The aim of this study was to evaluate the film based on chitosan as a barrier in the formation of postoperative intestinal adhesions in rabbits. For this, 30 rabbits used were male adult, race New Zealand, with an average weight of three kilograms and clinically healthy, of which 15 were submitted to enterotomy (control group) and 15 to enterotomy with overlap of the chitosan-based film at the intestinal incision (treatment group). Five animals from each group were assessed at seven, 14 and 21 days postoperatively, when held to the macroscopic evaluation of adhesions with regard presence, extent and type, and also performed microscopic evaluation of the enterorrhaphy site and hematimetric exams. The adhesions occurred in 80% of subjects in the control group and 86.7% of treatment group. In the control group 50% of the animals developed adhesions between the site of enterorrhaphy and abdominal wall, while in the treatment group 76.9% of individuals had adhesions between the site of enterorrhaphy and adjacent bowel. The control group had formed adhesions extensions higher than in treatment group. We found statistical differences between subgroups T7 and C7, C14 and T14 on the extent of adhesions (p ≤ 0.05) but not between C21 and T21. Also no significant differences between subgroups on the type of adhesions were seen. As for the histopathological evaluation, there was significant difference (p ≤ 0.05) between the subgroup C7, where 80% of samples the score was considered strong, and T7 in that 80% of the samples was considered moderate score on the concentration of fibroblasts and angiogenesis. Regarding the variable necrosis, was observed in 40% of samples of T7 and 0% of C7 and therefore statistically different (p ≤ 0.05). Significant differences (p ≤ 0.05) were also found between the groups C14 and T14 on variables heterophils (C14, 80% score discreet, T14, 80% score marked), eosinophils (score of 60% in discrete T14, but absent in C14) and necrosis (mild in 60% of C14 and severe in 80% of T14). In subgroups C21 and T21 only the scores of mononuclear cells were statistically different (p ≤ 0.05). The hematimetric variables remained within the normal values for the species. Based on the results we can conclude that the chitosan-based film reduces the extent of intestinal adhesions in rabbits and is useful as a barrier system in this species and that this film, when overlaid with a wound of rabbits induces intestinal tissue response to formation of caseous material. / Aderências abdominais ou peritoniais são faixas fibrosas que formam uniões anormais entre duas ou mais superfícies serosas de revestimento, podendo causar obstruções de órgãos abdominais tubulares e dores pélvicas. Diversos estudos já foram realizados utilizando antiinflamatórios, anticoagulantes, agentes proteolíticos e ainda biomateriais na forma de filme ou gel, com a finalidade de prevenir ou simplesmente reduzir a formação de aderências peritoniais. A quitosana é um polímero obtido pela desacetilação da quitina, que é um polissacarídeo oriundo do exoesqueleto de crustáceos e vem sendo utilizado em diversas especialidades médicas como biomaterial. O objetivo deste estudo foi avaliar o filme à base quitosana como barreira na formação de aderências intestinais pós-operatórias em coelhos. Para isso, utilizou-se 30 leporinos machos, adultos, da raça Nova Zelândia, com peso médio de três quilos e clinicamente sadios, dos quais 15 foram submetidos à enterotomia (grupo controle) e 15 à enterotomia e sobreposição do filme à base de quitosana sobre a incisão cirúrgica intestinal (grupo tratamento). Cinco animais de cada grupo foram avaliados aos sete, 14 e 21 dias pós-operatórios, quando se realizou a avaliação macroscópica das aderências quanto à presença, extensão e o tipo, sendo também realizada avaliação microscópica do sítio da enterorrafia e hematimetria. As aderências ocorreram em 80% dos indivíduos do grupo controle e 86,7% do grupo tratamento. No grupo controle em 50% dos animais as aderências formaram-se entre o sítio da enterorrafia e parede abdominal, enquanto no grupo tratamento 76,9% dos indivíduos apresentaram aderências entre o sítio da enterorrafia e alças intestinais adjacentes. No grupo controle as aderências formadas possuíam extensões maiores que no grupo tratamento. Foram encontradas diferenças estatísticas entre os subgrupos C7 e T7; C14 e T14 quanto à extensão das aderências (p≤ 0,05), mas não entre C21 e T21. Também não foram observadas diferenças significativas entre os subgrupos quanto ao tipo de aderências. Quanto à avaliação histopatológica, houve diferença significativa (p≤ 0,05) entre o subgrupo C7, em que 80% das amostras o escore foi considerado acentuado, e T7 em que 80% das amostras o escore foi considerado moderado quanto à concentração de fibroblastos e angiogênese. Em relação à variável necrose, esta foi observada em 40% das amostras de T7 e 0% de C7 sendo, portanto estatisticamente diferente (p≤ 0,05). Diferenças significativas (p≤ 0,05) também foram encontradas entre os subgrupos C14 e T14 quanto às variáveis heterófilos (C14, 80% escore discreto; T14, 80% escore acentuado), eosinófilos (escore discreto em 60% de T14, porém ausentes em C14) e necrose (discreta em 60% de C14 e acentuada em 80% de T14). Já nos subgrupos C21 e T21 apenas os escores das células mononucleares foram estatisticamente diferentes (p≤ 0,05). As variáveis hematimétricas permaneceram dentro dos valores considerados normais para a espécie. Com base nos resultados pode-se concluir que o filme à base de quitosana reduz a extensão das aderências intestinais em coelhos, sendo útil como sistema de barreira nessa espécie e que este filme, quando sobreposto a uma ferida intestinal de coelhos induz a resposta tissular com formação de material caseoso.

In the last two decades several studies have been trying to explore a possible role for gut microbiota in Autism Spectrum Disorders (ASD), supported by the high incidence of gastrointestinal disorders among ASD children and by the now well recognized existence of the brain-gut-microbiota axis (the complex system of bidirectional interactions between central nervous system, gastrointestinal tract and microorganisms inhabiting the gut). Nevertheless, results about alterations in gut microbiota composition and/or activity in ASD are to date strongly contrasting. A possible explanation could be that these studies tend to treat ASD as a unique pathology, whereas it includes different cognitive-behavioural phenotypes. Moreover, they do not consider factors which are important for children’s gut flora development, such as type of delivery, nutritional history (e.g. formula milk during lactation) and medical history (e.g. antibiotics intake) as well as factors that may affect the present composition of microbiota, such as the current diet (e.g. the strong food selectivity that often occurs in ASD children) and the presence of gastrointestinal disorders. In this study, I developed an interview to parents to assess whether there are differences related to the above mentioned aspects between ASD children and typically developing children and among ASD themselves, considering differences in cognitive level and severity of autistic traits. I also explored the use of special diets such as gluten-, lactose and casein free diets, the reasons for their adoption and the possible benefits for the child. Moreover, I decided to include in this interview also a section dedicated to parental difficulties in managing mealtime in order to collect information useful to plan future interventions. I found differences between ASD- and typical children in the incidence of gastrointestinal disorders and food selectivity. Especially, some children initially eat everything and then switch to a more and more restricted diet. This could be considered an early symptom of the pathology. I also found an association between gastrointestinal disorders and severity of autistic traits. Furthermore, I collected faecal samples from ASD families (two parents, an ASD child and a typically developing sibling) and analysed them with metaproteomics and bioinformatics techniques in order to assess microbiota activity and evaluate it in light of ASD phenotype, nutritional habits, gastrointestinal disorders and genetic proximity. Demonstrating the existence of a different microbiota composition in ASD or at least in a subgroup could allow to identify a biomarker of a possible development of ASD and to design preventive interventions, even through probiotics intake. Moreover, it could help to better understand the molecular mechanism underlying this pathology.

Made available in DSpace on 2015-03-04T18:58:02Z (GMT). No. of bitstreams: 1 thesis (1).pdf: 2502829 bytes, checksum: 20035b341686418a94516986bc16b476 (MD5) Previous issue date: 2013-11-22 / In this dissertation, we present a detailed study of the model proposed by Hinow et al. 2009, in which, through a continuum model based on equations of reaction-diffusion type, the authors simulate the spatial dynamics of tumor growth. The dynamics of proliferating, hypoxic, necrotic and endothelial cells, extracellular matrix and nutrients are included in this model. An angiogenesis model is also included, allowing the evaluation of different treatment protocols for cancer through antiangiogenic and cytotoxic drugs. Numerical and computational models have been developed for this class of models. Further examination of the model from Hinow et al. 2009 indicated some inconsistencies in the model. By adding new hypotheses, a new model is developed, overcoming the observed non-biological behaviors. The new model aggregates new dynamics, including the interactions between healthy cells and other tissues. One of the major contribution of this model is the proposal of a haptotactic term responsible for the directional movement of healthy cells towards dead cells. The inclusion of such term provides a more realistic approach to the treatment of the disease. As an example, healthy cells are able to return to regions formerly occupied by the tumor after a successful cytotoxic treatment. Several numerical experiments are presented to justify the development of such model and to show its behavior when different treatment protocols for the disease are considered. / Nesta dissertacao de mestrado apresentamos um estudo detalhado do modelo proposto por Hinow et al. 2009, no qual os autores por meio de um modelo continuo baseado em equacoes do tipo reacao-difusao simulam a dinamica espacial do crescimento tumoral. Neste modelo sao incluidas as dinamicas das celulas proliferativas, hipoxicas, necroticas e endoteliais, alem da matriz extracelular e de nutrientes. Um modelo de angiogenese e tambem incluido, permitindo a avaliacao de diferentes protocolos de tratamento da doenca por meio de drogas antiangiogenicas e citotoxicas. Modelos numericos e computacionais foram desenvolvidos para esta classe de modelos tendo como base formulacoes consistentes de elementos finitos. O estudo aprofundado do modelo de Hinow et al. 2009 indicou algumas inconsist^encias. A partir da inclusao de novas hipoteses um novo modelo foi desenvolvido, eliminando as situacoes nao-biologicas observadas. O novo modelo agrega novas dinamicas, incluindo as interacoes das celulas saudaveis com as demais do tecido. Uma das maiores contribuicoes do novo modelo proposto e a consideracao de um termo haptotatico responsavel pelo movimento direcional das celulas saudaveis na direcao das celulas mortas. A inclusao de tal termo propiciou uma abordagem mais realista do tratamento da doenca, captando por exemplo, apos o tratamento citotoxico, a retomada pelas celulas saudaveis da regiao outrora ocupada pelo tumor. Diversos experimentos numericos sao apresentados para justificar o desenvolvimento de tal modelo, assim como a simulacao de diversos protocolos de tratamento da doenca.