These studies investigated the effects of retinoic Acid (RA) on angiogenesis. Three human, neoplastic cell lines were used to examine angiogenic promotion and/or inhibition. The cell lines, U-373MG glioblastoma, DU-145 prostate carcinoma, and TCCSUP bladder transitional cell carcinoma, were treated with the following: all-trans, 9-cis, or 13-cis RA, at doses from 0.0001 to 100 μM. Hpoxia was used to assist the expression of the angiogenic phenotype. Conditioned media (CM) were prepared by growing the tumor cells in the presence of RA and hypoxia for 24 hours, and then the CM was transferred to bovine, capillary endothelial cells (EC) for 48 hours. The EC were counted for each CM and compared to controls. Multiplex, reverse transcriptase-polymerase chain reaction (MRT-PCR) was used on tumor cell RNA to demonstrate that mRNA expression of several angiogenic growth factors was modified. Angiogenic proteins in the CM were identified by enzyme-linked immunosorbent assay (ELISA). The CM from the U-373 and DU-145 cells, but not the TCCSUP cells, treated with all-trans or 9-cis RA caused significant increases (P<0.05) in EC proliferation. RA in nonconditioned media inhibited EC proliferation. CM treated with 13-cis RA did not increase EC proliferation. MRT-PCR demonstrated that the tumor cells expressed vascular endothelial growth factor (VEGF), midkine, interleukin-8 (IL-8), thrombospondin (TSP), and leukemia inhibitory factor (LIF) mRNAs. Increases VEGF mRNA corresponded to increase EC proliferation. IL-8 mRNA increased at RA concentrations greater than 0.1 μM. Midkine mRNA concentrations were generally unaffected by RA treatments. ELISA demonstrated that VEGF was increased by all-trans RA, to a lesser degree by 9-cis, and even less by 13-cis. IL-8 protein secretion was inhibited by RA. The expression of TSP and LIF mRNAs was almost completely inhibited by hypoxia. EC were grown with sodium heparin at 0, 10, 100, 500, or 1000 μg/ml in CM produced with DU-145 cells and 0.0001, 0.001, or 1 μM all-trans RA. Heparin increased EC proliferation at 10 μg ml and inhibited proliferation at higher concentrations.

Glioblastoma multiforme (GBM) is an extremely aggressive and almost always fatal brain tumor. GBM literature indicates defective mismatch repair (MMR) mechanisms are not involved in GBM tumorigenesis as in other tumors, and instigating mechanisms of GBM tumorigenesis remain unclear. GBM and neural progenitor (NPR) cells were exposed to three concentrations of H2O2 (0, 0.5, and 1.0 μM), cultured, and then harvested 0, 2, 4, and 6 days post-exposure; DNA from cells was amplified with microsatellite primers, investigating whether or not H2O2 exposure affected microsatellite instability (MSI) in target sequences. Three out of six markers showed significant MSI in the H2O2-exposed NPR cells. Our results suggest H2O2, which generates reactive oxygen species (ROS), correlated with MSI accumulation that occurred in NPR cells in specific DNA regions. Thus, gene expression analysis to assess normal and abnormal gene expression of GBM and NPR cellss is warranted.

No description available.

No description available.

Traumatic injuries to the spinal cord (SCI) and peripheral nerve (PNI) affect several thousand people worldwide every year. At present, there is no effective treatment for SCI and despite continuous improvements in microsurgical reconstructive techniques for PNI, many patients are still left with permanent, devastating neurological dysfunction. This thesis investigates the effects of mesenchymal stem cells (MSC) derived from adipose (ASC) and dental (DSC) tissue and chitosan/microRNA-124 polyplex particles on regeneration after spinal cord and peripheral nerve injury in adult rats. Dental stem cells were obtained from apical papilla, dental pulp, and periodontal ligament. ASC and DSC expressed MSC surface markers (CD73, CD90, CD105 and CD146) and various neurotrophic molecules including BDNF, GDNF, NGF, VEGF-A and angiopoietin-1. Growth factor stimulation of the stem cells resulted in increased secretion of these proteins. Both ASC and DSC supported in vitro neurite outgrowth and in contrast to Schwann cells, ASC did not induce activation of astrocytes. Stimulated ASC also showed an enhanced ability to induce capillary-like tube formation in an in vitro angiogenesis assay. In a peripheral nerve injury model, ASC and DSC were seeded into a fibrin conduit, which was used to bridge a 10 mm rat sciatic nerve gap. After 2 weeks, both ASC and DSC promoted axonal regeneration in the conduit and reduced caspase-3 expression in the dorsal root ganglion (DRG). ASC also enhanced GAP-43 and ATF-3 expression in the spinal cord, reduced c-jun expression in the DRG and increased the vascularity of the implant. After transplantation into injured C3-C4 cervical spinal cord, ASC continued to express neurotrophic factors and laminin and stimulated extensive ingrowth of 5HT-positive raphaespinal axons into the trauma zone. In addition, ASC induced sprouting of raphaespinal terminals in C2 contralateral ventral horn and C6 ventral horn on both sides. Transplanted cells also changed the structure and the density of the astroglial scar. Although the transplanted cells had no effect on the density of capillaries around the lesion site, the reactivity of OX42-positive microglial cells was markedly reduced. However, ASC did not enhance recovery of forelimb function. In order to reduce activation of microglia/macrophages and the secondary tissue damage after SCI, the role of microRNA-124 was investigated. In vitro transfection of chitosan/microRNA-124 polyplex particles into rat microglia resulted in the reduction of reactive oxygen species and TNF-α levels and lowered expression of MHC-II. Upon microinjection into uninjured rat spinal cords, particles formed with Cy3-labeled control sequence RNA, were specifically internalized by OX42 positive macrophages and microglia. Alternatively, particles injected in the peritoneum were transported by macrophages to the site of spinal cord injury. Microinjections of chitosan/microRNA-124 particles significantly reduced the number of ED-1 positive macrophages after SCI. In summary, these results show that human MSC produce functional neurotrophic and angiogenic factors, creating a more desirable microenvironment for neural regeneration after spinal cord and peripheral nerve injury. The data also suggests that chitosan/microRNA-124 particles could be potential treatment technique to reduce neuroinflammation.

The chick chorio-allantoic membrane (CAM) contains a complex vascular network commonly used to study angiogenesis. The application of chemical factors and oxygen barrier films onto this tissue can easily influence the process of angiogenesis. In this study, oxygen barrier film patches (Krehalon, polyvinylidene chloride, 12 μm thick, O2 transmission rate = 2.19 cm3•ml/100 in2•day•atm) were applied to areas of the CAM. Holes were made in the film and alginate beads incubated in various chemical factors were placed in the holes. After 24 and 48 hours of exposure to the alginate beads, images were taken of the tissue using a stereomicroscope and then processed using ImageJ software (from the National Institue of Health (NIH)). The images were analyzed with the Fractal Analysis plugin of ImageJ (also from NIH) using four parameters. These parameters are the number of vessel segments, the number of vessel bifurcations, the total length of the vessels, and the complexity of the vascular network. From these parameters, the chemical factors can be identified as promoting angiogenesis (pro-angiogenic), inhibiting angiogenesis (anti-angiogenic), or having no effect on angiogenesis (not angiogenic). For the angiogenic beads, significant results were found in at least one of the four parameters. SNAP and H2O2 gave pro-angiogenic responses while Angiotensin II, Losartan, and Adenosine were anti-angiogenic. To test the effect of an oxygen barrier film patch on angiogenesis, images were taken of the tissue under the film patch (virtual holes) and holes exposed to atmospheric oxygen. Analysis of the virtual holes compared to the control holes gave significant results for several of the film patches. These film patches are distinguished by the chemical that was tested on each of the films. The virtual holes containing Angiotensin II, Losartan, Adenosine, and H2O2 gave pro-angiogenic results while SNAP and L-NAME virtual holes were anti-angiogenic. Thus, the chemical factors and the oxygen barrier film patches did have an effect on angiogenesis in the CAM.

Made available in DSpace on 2016-08-10T10:38:57Z (GMT). No. of bitstreams: 1 SUSY RICARDO LEMES PONTES.pdf: 2637281 bytes, checksum: e72835d2a5fe9725ba5481d3165a02bb (MD5) Previous issue date: 2014-12-19 / According to WHO, about 80% of world population, somehow, makes use of medicinal plants as medicine, and only 30% is done by medical indication. Currently the intense search for alternative therapies such as herbal medicine, is due to the inefficiency of some synthetic products, the high cost of allopathic and the search for less harmful to body treatments. The plant compounds responsible for serving in combat diseases are the secondary metabolites. The most outstanding terpenes; phenolic compounds and nitrogen compounds. In Brazil, the extracted oil of Carapa guianensis seed has been used as an herbal medicine, is applied in wound healing, treatment of febrile diseases, inflammation and pain. Its main constituents and possibly responsible for its biological activity are the fatty acids and limonoids compounds. In view of the wide and use and Andiroba oil plant characteristics, this study aimed to investigate the possible angiogenic, genotoxic and antigenotoxic activities. To assess angiogenesis was used the experimental model of MCA. Genotoxicity and antigenotoxicity tests were carried out in vivo, in mice through the micronucleus test. The test results on the MCA indicated that the oil caused a significant increase in vascular network (p<0.05) compared to neutral controls and inhibitor. In the micronucleus test, the effect of the oil antigenotoxic at doses of 250, 500 and 1000 mg kg b.w. Was observed in all treatment groups, since the number of micronuclei was lower (p<0.05) in the positive controls CP and MMC. With the methods and conditions employed, the results of this study suggest that the andiroba oil is not genotoxic and has important potential Physiotherapy, with angiogenic and antigenotoxic activity. / De acordo com a OMS, cerca de 80% da população mundial, de algum modo, faz uso de plantas medicinais como medicamento, e apenas 30% é feito por indicação médica. Atualmente, a intensa procura por terapias alternativas, como a fitoterapia, se deve à ineficiência de alguns produtos sintéticos, ao alto custo de medicamentos alopáticos e à busca por tratamentos menos agressivos ao organismo. Os compostos vegetais responsáveis por atuar no combate de doenças são os metabólitos secundários. Dentre eles se destacam os terpenos, compostos fenólicos e compostos nitrogenados. No Brasil, o óleo extraído da semente da Carapa guianensis tem sido usado como fitoterápico apesentando aplicabilidade na cicatrização de ferimentos, tratamento de doenças febris, inflamações e dores. Seus principais constituintes e possivelmente os responsáveis por sua atividade biológica são os ácidos graxos e compostos limonóides. Em virtude do amplo uso e das características do óleo da semente de andiroba, este estudo objetivou investigar suas possíveis atividades angiogênica, genotóxica e antigenotóxica. Para avaliar a angiogênese foi utilizado o modelo experimental da MCA. Na genotoxicidade e antigenotoxicidade foram realizados ensaios in vivo, em camundongo através do teste de micronúcleo. Os resultados do ensaio na MCA indicaram que o óleo provocou aumento significativo da rede vascular (p<0,05) em relação aos controles neutro e inibidor. No teste do micronúcleo, o efeito antigenotóxico do óleo nas doses de 250, 500 e 1000 mg Kg p.c., foi verificado em todos os grupos de tratamento, uma vez que o número de micronúcleos foi menor (p<0,05) em relação aos controles positivos CP e MMC. Através dos métodos e condições empregados, os resultados deste estudo sugerem que o óleo de andiroba não é genotóxico e apresenta importante potencial fitoterapêutico, com atividade angiogênica e antigenotóxica.

Submitted by admin tede (tede@pucgoias.edu.br) on 2016-11-22T18:16:35Z No. of bitstreams: 1 LUCIANA CARDOSO MARINHO.pdf: 2007688 bytes, checksum: 7b1d1ab427984f805bae729662ab0296 (MD5) / Made available in DSpace on 2016-11-22T18:16:35Z (GMT). No. of bitstreams: 1 LUCIANA CARDOSO MARINHO.pdf: 2007688 bytes, checksum: 7b1d1ab427984f805bae729662ab0296 (MD5) Previous issue date: 2016-08-10 / The latest research has shown that the angiogenic factors play an important role in the modulation of tumor angiogenesis process. Thus, this study aimed to analyze the influence of this process on the development of leukemias. a systematic review based on scientific articles that investigated the interference of angiogenic factors in cases of leukemia early and advanced stages was performed. The major evidence indicates that these factors, in special VEGF, are expressed intensely in malignant cells and may generate, for example, a hematologic malignancy. The regulation of expression of certain genes is also involved in abnormal proliferation, differentiation and apoptosis of these cells. The expression, function and regulation mechanism of angiogenic factors have been gradually elucidated in this review and in conclusion, it is noted that some of them have relation to the development and leukemias prognosis, and provide potential strategies for the treatment of patients with leukemia. / As últimas pesquisas demonstram que os fatores angiogênicos desempenham um importante papel na modulação do processo da angiogênese tumoral. Neste sentido, o presente estudo tem como objetivo a análise da influência deste processo no desenvolvimento de leucemias. Foi realizada uma revisão sistemática com base em artigos científicos que investigaram a interferência de fatores angiogênicos em casos de leucemias em estágios iniciais e avançados. As principais evidências indicam que estes fatores, em especial o VEGF, são expressos de forma intensa em células malignas e podem gerar, por exemplo, uma malignidade hematológica. A regulação de expressão de determinados genes também está envolvida na proliferação anormal, na diferenciação e apoptose destas células. A expressão, função e o mecanismo de regulação dos fatores angiogênicos foram elucidados gradualmente nesta revisão e como conclusão, salienta-se que alguns deles apresentam relação com o desenvolvimento e prognóstico de leucemias, além de fornecer possíveis estratégias para o tratamento de pacientes com leucemia.

Objetivos: Avaliar a influencia do estresse sobre os pesos maternos, placentários e fetais, as alterações histológicas placentárias e a expressão gênica dos fatores angiogênicos em ratas prenhes. Métodos: De setembro de 2007 a julho de 2008, foi realizado um estudo experimental do tipo casocontrole, em que 6 ratas prenhes foram submetidas a estímulos estressantes crônicos e agudos enquanto que 6 animais constituíram o grupo controle. No 20o dia de prenhez, todas as ratas foram sacrificadas. Os seguintes dados foram analisados e comparados entre os grupos: a. pesos maternos, placentários e fetais; b. alterações histológicas placentárias; e, c. expressão gênica dos fatores angiogênicos (VEGF-A e PlGF), dos seus receptores (VEGFR-1 e VEGFR-2) e do fator induzido por hipoxia (HIF-1). Resultados: Os pesos maternos, placentários e fetais foram significativamente menores no grupo de ratas prenhes-estressadas em relação ao controle. Histologicamente foram encontradas a presença de núcleos picnóticos no grupo prenhe-estressado. Observou-se expressão gênica significativamente maior de VEGF-A no grupo rata prenhe-estressada assim como redução significante da expressão gênica de PlGF, VEGFR-1, VEGFR-2, quando comparadas ao controle. Não houve alterações entre os grupos para o gene HIF-1. Conclusões: No modelo animal de estresse estudado, observou-se alterações significativas no peso placentário e da expressão gênica dos fatores angiogênicos placentários em ratas submetidas ao estresse. / Objectives: to evaluate the influence of the stress on the maternal, placental and fetal weights, on the histological findings and on the genetic expression of the angiogenic factors in pregnant rats. Methods: From September 2007 to Julie 2008, an experimental case-control study was conduced in which 6 pregnant rats were submitted to chronic and acute stress while six other were considered as controls. In the 20th day of gestation, all animals were sacrified. The following data were evaluated and compared between both groups: a. maternal, placental and fetal weights; b. histological findings; and c. genetic expression of angiogenic factors (VEGF-A and PIGF), receptors (VEGFR-1 and VEGFR-2) and hypoxic-induced factor (HIF- 1). Results: Maternal, placental and fetal weights were statistically smaller in the stressed animals comparing to controls. Histological analysis revealed picnotic nuclei in the placentas of stressed rats. A statistically significant increase in the genetic expression of VEGF-A as well as a reduction of the expression of the PlGF, VEGFR-1, VEGFR-2 were observed in the placentas of the stressed group in comparison to controls. There was no difference of expression of the gene HIF-1 between both groups. Conclusions: In the present animal model of stress, significant alterations in the placental weights, histology and genetic expression of the angiogenic factors among pregnant rats under stress.

Objetivos: Avaliar a influencia do estresse sobre os pesos maternos, placentários e fetais, as alterações histológicas placentárias e a expressão gênica dos fatores angiogênicos em ratas prenhes. Métodos: De setembro de 2007 a julho de 2008, foi realizado um estudo experimental do tipo casocontrole, em que 6 ratas prenhes foram submetidas a estímulos estressantes crônicos e agudos enquanto que 6 animais constituíram o grupo controle. No 20o dia de prenhez, todas as ratas foram sacrificadas. Os seguintes dados foram analisados e comparados entre os grupos: a. pesos maternos, placentários e fetais; b. alterações histológicas placentárias; e, c. expressão gênica dos fatores angiogênicos (VEGF-A e PlGF), dos seus receptores (VEGFR-1 e VEGFR-2) e do fator induzido por hipoxia (HIF-1). Resultados: Os pesos maternos, placentários e fetais foram significativamente menores no grupo de ratas prenhes-estressadas em relação ao controle. Histologicamente foram encontradas a presença de núcleos picnóticos no grupo prenhe-estressado. Observou-se expressão gênica significativamente maior de VEGF-A no grupo rata prenhe-estressada assim como redução significante da expressão gênica de PlGF, VEGFR-1, VEGFR-2, quando comparadas ao controle. Não houve alterações entre os grupos para o gene HIF-1. Conclusões: No modelo animal de estresse estudado, observou-se alterações significativas no peso placentário e da expressão gênica dos fatores angiogênicos placentários em ratas submetidas ao estresse. / Objectives: to evaluate the influence of the stress on the maternal, placental and fetal weights, on the histological findings and on the genetic expression of the angiogenic factors in pregnant rats. Methods: From September 2007 to Julie 2008, an experimental case-control study was conduced in which 6 pregnant rats were submitted to chronic and acute stress while six other were considered as controls. In the 20th day of gestation, all animals were sacrified. The following data were evaluated and compared between both groups: a. maternal, placental and fetal weights; b. histological findings; and c. genetic expression of angiogenic factors (VEGF-A and PIGF), receptors (VEGFR-1 and VEGFR-2) and hypoxic-induced factor (HIF- 1). Results: Maternal, placental and fetal weights were statistically smaller in the stressed animals comparing to controls. Histological analysis revealed picnotic nuclei in the placentas of stressed rats. A statistically significant increase in the genetic expression of VEGF-A as well as a reduction of the expression of the PlGF, VEGFR-1, VEGFR-2 were observed in the placentas of the stressed group in comparison to controls. There was no difference of expression of the gene HIF-1 between both groups. Conclusions: In the present animal model of stress, significant alterations in the placental weights, histology and genetic expression of the angiogenic factors among pregnant rats under stress.