Cytotoxicity of iodine-131 radiopharmaceutical in tumor and non-tumor human cells and radioprotection by integral juices of Vitis labrusca L / Citotoxicidade do radiofarmacêutico iodo-131 em células humanas tumorais e não tumorais e radioprotecção por sucos integrais de Vitis labrusca L

Iodine-131 (I-131) radioisotope it causes the formation of free radicals, which lead to the formation of cell lesions and the reduction of cell viability. Thus, the use of radioprotectors, especially those from natural sources, which reduce the effects of radiation to healthy tissues, while maintaining the sensitivity of tumor cells, stands out. The objective of the present study was to evaluate the cytoprotective/radioprotective effects of whole grape juices manufactured from the conventional or organic production systems, whether or not exposed to ultraviolet (UV-C) light irradiation. The results showed that I-131 presented a cytotoxic effect on human hepatocellular cells (HepG2/C3A) at concentrations above 1.85 MBq/mL, after 24 and 48 hours of treatment, though all concentrations (0.0037 to 7.40 MBq/mL) were cytotoxic to non-tumor human lung fibroblast (MCR-5) cells, after 48 hours. However, grape juices (10 and 20 µL/mL) did not interfere with the cytotoxic effect of the therapeutic dose of I-131 on tumor cells within 48 hours of treatment, while protecting the non-tumor cells, probably due to its high antioxidant activity. In accordance with their nutraceutical potential, antioxidant and radioprotective activity, these data stimulate in vivo studies on the use of natural products as radioprotectants, such as grape juice, in order to confirm the positive beneficial potential in living organisms.

O radioisótopo iodo-131 (I-131) causa a formação de radicais livres, que levam à formação de lesões celulares e redução da viabilidade celular. Assim, destaca-se a utilização de radioprotetores, principalmente de origem natural, que reduzem os efeitos da radiação nos tecidos saudáveis, mantendo a sensibilidade das células tumorais. O objetivo do presente estudo foi avaliar os efeitos citoprotetores/radioprotetores de sucos de uva integral fabricados em sistemas de produção convencional ou orgânico, expostos ou não à radiação ultravioleta (UV-C). Os resultados mostraram que o I-131 apresentou efeito citotóxico nas células hepatocelulares humanas (HepG2/C3A) em concentrações acima de 1,85 MBq/mL, após 24 e 48 horas de tratamento, embora todas as concentrações (0,0037 a 7,40 MBq/mL) fossem citotóxicas para células de fibroblasto de pulmão humano não tumoral (MCR-5), após 48 horas. No entanto, os sucos de uva (10 e 20 µL/mL) não interferiram no efeito citotóxico da dose terapêutica de I-131 nas células tumorais em 48 horas de tratamento, protegendo as células não tumorais, provavelmente devido ao seu alto poder antioxidante. atividade. De acordo com seu potencial nutracêutico, atividade antioxidante e radioprotetora, esses dados estimulam estudos in vivo sobre o uso de produtos naturais como radioprotetores, como o suco de uva, a fim de confirmar o potencial benéfico positivo em organismos vivos.

Desafios para a reprodução assistida em animais de vida livre / Challenges for assisted reproduction in free-living animals

A reprodução assistida se faz necessária em programas de conservação de espécies ameaçadas de extinção, sendo um facilitador de transporte e troca de material genético. Neste contexto, o acesso ao material de animais de vida livre é essencial para incrementar o banco genético da espécie em questão, no entanto adaptar os métodos possíveis à realidade do campo torna essa área de pesquisa desafiadora. Ainda hoje os espermatozoides são os gametas mais acessados em animais de vida livre, porém com pouco uso efetivo para criopreservação e produção de filhotes. É pungente a necessidade de mais pesquisas nesta área, uma vez que há centenas de espécies brasileiras ameaçadas, com especificidades fisiológicas e que habitam habitats variados, o que demanda adaptações espécie-específicas e hábitat específicas.

Assisted reproduction is necessary for conservation programs for endangered species, facilitating transport and exchange of genetic material. In this context, access to material from free-living animals is essential to increase the genetic bank of the species in question. However, adapting the possible methods to the reality of the fieldwork makes this area of research a challenge. Even today, sperm are the most accessed gametes in free-living animals, but with little effective use for cryopreservation and production of offspring. The need for more research in this area is acute, as there are hundreds of Brazilian species under threat, with physiological specificities, and that inhabit varied habitats, which demand species-specific adaptations and specific habitats.

O que já sabemos sobre a reprodução de fêmeas de grandes felídeos? / What do we already know about the reproduction of big cats' females?

Os grandes felídeos são predadores de topo de cadeia com um papel essencial nos ecossistemas globais. O conceito de Conservação Única propõe a reprodução artificial como uma das ferramentas para reduzir a vulnerabilidade dessas espécies. Este manuscrito teve como objetivo avaliar o que há de novo na reprodução de grandes felídeos na última década. O conhecimento da fisiologia e do comportamento reprodutivo é o primeiro passo para o desenvolvimento de tecnologias reprodutivas em animais selvagens. Nos grandes felídeos, o comportamento copulatório é de fundamental importância, pois necessitam de mecanismos de indução da ovulação, que podem ser mecânicos, sensoriais ou via administração de hormônio luteinizante. O sucesso no cuidado neonatal representa o sucesso da tecnologia reprodutiva em fêmeas. Na última década, o sucesso da inseminação artificial foi relatado apenas em tigres-siberianos e leopardos da Anatólia, e a inseminação de onças-pintadas é foco de pesquisa do Instituto Reprocon, trocando material genético entre ambientes in situ e ex situ por meio de inseminação artificial. Para obter oócitos viáveis de alta qualidade, a técnica de escolha é a colheita de oócitos por laparoscopia. A produção de embriões in vitro enfrenta desafios para a maturação eficiente de oócitos e sua vitrificação eficiente. As técnicas reprodutivas precisam de estudos aprofundados em grandes felídeos para atingir a repetibilidade necessária para uma aplicação eficiente na conservação.

Big cats are apex predators with an essential role in global ecosystems. The One Conservation concept proposes artificial reproduction as one of the tools to reduce the vulnerability of these species. This manuscript aimed to assess what is new in big cat reproduction in the last decade. Knowledge of reproductive physiology and behavior is the first step towards developing reproductive technologies in wild animals. In big cats, copulatory behavior is of fundamental importance because they need ovulation induction mechanisms, which can be mechanical, sensory, or via the administration of the luteinizing hormone. The success in neonatal care represents the success of reproductive technology in females. In the last decade, successful artificial insemination was only reported in Siberian tigers and Anatolian leopards. Jaguar artificial insemination focuses on research at the Reprocon Institute, exchanging genetic material between in situ and ex situ environments thru artificial insemination. The technique of choice is laparoscopic ovum pick-up to obtain high-quality viable oocytes. The production of in vitro embryos faces challenges for the efficient maturation of oocytes and their efficient vitrification. Reproductive technologies need in-depth studies in big cats to achieve the repeatability necessary for efficient application in conservation.

Ointment of Ximenes americana promotes acceleration of wound healing in rats

Purpose:To evaluate the healing potential of the Ximenia americana hydroalcoholic extract in 10% cream in excisional wound models in rats.Methods:Sixty male adults Wistar rats were submitted to skin and subcutaneous tissue surgery in the right and left thoracic regions, divided into three experimental groups: Standard submitted to treatment with only the base vehicle, Treated wounds treated with hydroalcoholic extract of X. americana applied on 10%, Lanette base and Control, untreated wounds. The treatment was performed daily and the wounds evaluated microscopically by the quantification of fibroblasts, collagen fibers and blood vessels.Results:The histomorphometric analysis showed a significant increase in the number of fibroblasts, collagen fibers and blood vessels in the treated group.Conclusion:The topical action of the cream based on Ximenia americana shows angiogenic effects and improves the replacement of collagen, suggesting its use for the development of herbal remedy in the treatment of cutaneous wound healing.(AU)

The Effect of High Temperature on Viability, Proliferation, Apoptosis and Anti-oxidant Status of Chicken Embryonic Fibroblast Cells

ABSTRACT The effects of oxidative stress induced by high temperature on the cell viability, proliferation, apoptosis and oxidative status of chicken embryonic fibroblasts (CEF) were analyzed. The viability, proliferation, apoptotic and anti-oxidative status were measured after incubating CEF at the temperatures of 37ºC (control) and 40-44ºC (experimental groups) for 6,12 and 24 hours. The results showed that at high temperature (42-43ºC), the viability of CEF cells decreased after 6, 12 and 24 h of incubation, but the difference was significant only at 43ºC. Cell proliferation was significantly reduced at 44oC/6h. The apoptotic rate of CEF cells was increased following heat treatments in a time-dependent manner. ROS formation increased with increasing temperature, but the difference was only significant at 44ºC/6,12h. Heat stress did not significantly affect the superoxide dismutase (SOD) activity. CAT activity was significantly decreased at 43ºC/24h and 44ºC/12 and 24h. Malondialdehyde (MDA) formation was significantly increased at 43ºC/12h and 44ºC/12 and 24h. In conclusion, heat stress induced the oxidative stress, decreasing the viability, proliferation and anti-oxidative response of CEF cells.

The Effect of High Temperature on Viability, Proliferation, Apoptosis and Anti-oxidant Status of Chicken Embryonic Fibroblast Cells

The effects of oxidative stress induced by high temperature on the cell viability, proliferation, apoptosis and oxidative status of chicken embryonic fibroblasts (CEF) were analyzed. The viability, proliferation, apoptotic and anti-oxidative status were measured after incubating CEF at the temperatures of 37ºC (control) and 40-44ºC (experimental groups) for 6,12 and 24 hours. The results showed that at high temperature (42-43ºC), the viability of CEF cells decreased after 6, 12 and 24 h of incubation, but the difference was significant only at 43ºC. Cell proliferation was significantly reduced at 44oC/6h. The apoptotic rate of CEF cells was increased following heat treatments in a time-dependent manner. ROS formation increased with increasing temperature, but the difference was only significant at 44ºC/6,12h. Heat stress did not significantly affect the superoxide dismutase (SOD) activity. CAT activity was significantly decreased at 43ºC/24h and 44ºC/12 and 24h. Malondialdehyde (MDA) formation was significantly increased at 43ºC/12h and 44ºC/12 and 24h. In conclusion, heat stress induced the oxidative stress, decreasing the viability, proliferation and anti-oxidative response of CEF cells.(AU)

The Effect of High Temperature on Viability, Proliferation, Apoptosis and Anti-oxidant Status of Chicken Embryonic Fibroblast Cells

The effects of oxidative stress induced by high temperature on the cell viability, proliferation, apoptosis and oxidative status of chicken embryonic fibroblasts (CEF) were analyzed. The viability, proliferation, apoptotic and anti-oxidative status were measured after incubating CEF at the temperatures of 37ºC (control) and 40-44ºC (experimental groups) for 6,12 and 24 hours. The results showed that at high temperature (42-43ºC), the viability of CEF cells decreased after 6, 12 and 24 h of incubation, but the difference was significant only at 43ºC. Cell proliferation was significantly reduced at 44oC/6h. The apoptotic rate of CEF cells was increased following heat treatments in a time-dependent manner. ROS formation increased with increasing temperature, but the difference was only significant at 44ºC/6,12h. Heat stress did not significantly affect the superoxide dismutase (SOD) activity. CAT activity was significantly decreased at 43ºC/24h and 44ºC/12 and 24h. Malondialdehyde (MDA) formation was significantly increased at 43ºC/12h and 44ºC/12 and 24h. In conclusion, heat stress induced the oxidative stress, decreasing the viability, proliferation and anti-oxidative response of CEF cells.

Fator de crescimento fibroblástico 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro

Em 1995 a via Hippo foi inicialmente descoberta na Drosofila melanogaster, e desde então diversos pesquisadores vêm tentando entender melhor essa via. A via Hippo está relacionada a processos de proliferação celular, tanto em células foliculares ovarianas como no desenvolvimento embrionário. Os efetores dessa via, YAP e TAZ atuam como coativadores de transcrição de genes de proliferação e sobrevivência celular como CTGF e CYR61. Porém o fator de crescimento de fibroblasto também induz a expressão de CTGF. E segundo estudos recentes foi descoberto que o fator de crescimento fibroblastico 18 (FGF18) está presente na tuba uterina durante o desenvolvimento embrionário inicial. Isso nos levou a pensar que o FGF18 teria alguma função durante o desenvolvimento embrionário. Portanto, o objetivo deste estudo foi determinar se o FGF18 modula a via Hippo através da expressão de genes alvos de proliferação celular (CTGF e CYR61) durante a maturação oocitária e desenvolvimento embrionário inicial. Para isso três experimentos foram realizados. No primeiro experimento complexos cumulus oocito (CCOs) de bovinos foram maturados in vitro durante 6, 12 e 24 horas na presença de concentrações graduais de FGF18(0, 10 e 100ng/mL). No segundo experimento os CCOs foram maturados 3, 6 e 9 horas na presença de 0 ou 100ng/mL de FGF18. No experimento três CCOs foram colocados para maturação e foram divididos em três grupos. Um grupo considerado grupo controle no qual CCOs foram maturados, realizada a fertilização in vitro (FIV) e depois cultivados in vitro (CIV) sem a adição de FGF18. Um grpo no qual 100nf/mL de FGF18 foram adicionados durante as 24 horas da maturação in vitro e epós foi realizado a fertilização e o cultivo em meios sem adição de FGF18. E um terceiro grupos os CCOs foram cultivados (CIV) por sete dias na presença de 100ng/mL de FGF18. Neste grupo a MIV e FIV foram realizadas na ausência de FGF18. Os genes avaliados nos três experimentos foram os genes de proliferação celular, CTGF e CYR61. O experimento um que teve o objetivo de avaliar a adição de concentrações graduais durante a maturação houve um aumento (p<0,05) da expressão gênica para CTGF no grupo tratado com 100ng/mL as 12horas. No experimento dois em que o objetivo era avaliar a concentração de 100ng/mL houve uma diminuição (p<0,05) do grupo tratado em relação ao grupo controle as três horas. E no experimento três em que o objetivo era avaliar a adição de FGF18 durante o desenvolvimento embrionário, houve diferença estatística (p<0,05) do grupo tratado com FGF18 durante a CIV. Desde forma concluímos que FG18 modula a expressão de CTGF em período críticos da maturação nuclear do oócito, da expansão do cumulus e do desenvolvimento embrionário inicial

In 1995, the Hippo pathway was initially discovered in Drosophila melanogaster, and since then several researchers have been trying to better understand this pathway. The Hippo pathway is involved in cell proliferation processes, both in ovarian follicular cells in the early embryonic development. The effectors of this pathway YAP e TAZ act as transcription coactivators of cell proliferation and survival genes such as CTGF and CYR61. However, the fibroblast growth factor also induces CTGF expression. However, recent studies by the group found that fibroblast growth factor 18 (FGF18) is present in the fallopian tube during early embryonic development. This led us to think that FGF18 would have some role during embryonic development. Therefore, the aim of the following study was to determine whether FGF18 modulates the Hippo pathway through the expression of target genes for cell proliferation (CTGF and CYR61) during oocyte maturation and early embryonic development. To acquire the results of the objective, three experiments were carried out, with in vitro maturation (IVM) of cumulus oocyte complexes (COCs) in the first and second experiments, with the addition of FGF18 during IVM, and the structures were collected at different times. In the first experiment, gradual concentrations of FGF18 (0, 10 and 100ng / mL) were added and there was a control group without the addition of FGF18, and 6, 12 and 24 hours of maturation were collected. In the second experiment, a concentration of 100ng / mL of FGF18 was added during IVM, there was a control group and the structures were collected at 0, 3, 6 and 9 hours of maturation. In the experiment, three COCs were put to maturation and were divided into three groups. A group considered a control group in which COCs were matured, with in vitro fertilization (IVF) and then in vitro culture (IVC) without the addition of FGF18. A group in which, during maturation, FGF18 was added only during IVM and afterwards fertilization and culture were carried out in media without the addition of FGF18. In a third group, COCs were put to mature, fertilized and, when put to culture, FGF18 was added in the medium. The genes evaluated in the three experiments were the cell proliferation genes, CTGF and CYR61. In experiment one, which aimed to evaluate the addition of gradual concentrations during maturation, there was an increase (p <0.05) in CTGF gene expression in the group treated with 100ng / mL at 12 hours. In experiment two, in which the objective was to assess the concentration of 100ng / mL, there was a decrease (p <0.05) in the treated group compared to the control group at three hours. And in experiment three in which the objective was to evaluate the addition of FGF18 during embryonic development, there was a statistical difference (p <0.05) in the group treated with FGF18 during IVC. Therefore, we conclude that FGF18 modulates CTGF expression in critical periods of oocyte nuclear maturation, cumulus expansion and early embryonic development in cattle.

Pravastatin induces cell cycle arrest and decreased production of VEGF and bFGF in multiple myeloma cell line / Pravastatina induz parada no ciclo celular e diminuição na produção de VEGF e bFGF em linhagem de Mieloma Multiplo

Abstract Multiple myeloma (MM) is a B cell bone marrow neoplasia characterized by inflammation with an intense secretion of growth factors that promote tumor growth, cell survival, migration and invasion. The aim of this study was to evaluate the effects of pravastatin, a drug used to reduce cholesterol, in a MM cell line.Cell cycle and viability were determinate by Trypan Blue and Propidium Iodide. IL6, VEGF, bFGF and TGF were quantified by ELISA and qRT-PCR including here de HMG CoA reductase. It was observed reduction of cell viability, increase of cells in G0/G1 phase of the cell cycle and reducing the factors VEGF and bFGF without influence on 3-Methyl-Glutaryl Coenzyme A reductase expression.The results demonstrated that pravastatin induces cell cycle arrest in G0/G1 and decreased production of growth factors in Multiple Myeloma cell line.(AU)

Resumo O Mieloma Múltiplo é uma neoplasia de linfócitos B da medula óssea, caracterizada por inflamação com uma intensa secreção de fatores de crescimento que promovem o aumento do volume do tumor, sobrevivência celular, migração e invasão. O objetivo deste estudo foi avaliar os efeitos da pravastatina, uma droga usada para reduzir o colesterol, em um linhagem de MM. O ciclo celular e viabilidade foram determinadas por Trypan Blue e iodeto de propídio. IL6, VEGF, bFGF e TGF foram quantificadas por ELISA e qRT-PCR, incluindo aqui de HMG CoA redutase. Observou-se a redução da viabilidade das células, aumento de células na fase G0/G1 do ciclo celular e redução no VEGF e bFGF, sem influência na expressão da enzima 3-Metil-Glutaril Coenzima A redutase. Os resultados demonstraram que a pravastatina induz parada no ciclo celular em G0/G1 e diminuição da produção de fatores de crescimento em várias linhas de células de Mieloma.(AU)

Pravastatin induces cell cycle arrest and decreased production of VEGF and bFGF in multiple myeloma cell line

Abstract Multiple myeloma (MM) is a B cell bone marrow neoplasia characterized by inflammation with an intense secretion of growth factors that promote tumor growth, cell survival, migration and invasion. The aim of this study was to evaluate the effects of pravastatin, a drug used to reduce cholesterol, in a MM cell line.Cell cycle and viability were determinate by Trypan Blue and Propidium Iodide. IL6, VEGF, bFGF and TGF were quantified by ELISA and qRT-PCR including here de HMG CoA reductase. It was observed reduction of cell viability, increase of cells in G0/G1 phase of the cell cycle and reducing the factors VEGF and bFGF without influence on 3-Methyl-Glutaryl Coenzyme A reductase expression.The results demonstrated that pravastatin induces cell cycle arrest in G0/G1 and decreased production of growth factors in Multiple Myeloma cell line.

Resumo O Mieloma Múltiplo é uma neoplasia de linfócitos B da medula óssea, caracterizada por inflamação com uma intensa secreção de fatores de crescimento que promovem o aumento do volume do tumor, sobrevivência celular, migração e invasão. O objetivo deste estudo foi avaliar os efeitos da pravastatina, uma droga usada para reduzir o colesterol, em um linhagem de MM. O ciclo celular e viabilidade foram determinadas por Trypan Blue e iodeto de propídio. IL6, VEGF, bFGF e TGF foram quantificadas por ELISA e qRT-PCR, incluindo aqui de HMG CoA redutase. Observou-se a redução da viabilidade das células, aumento de células na fase G0/G1 do ciclo celular e redução no VEGF e bFGF, sem influência na expressão da enzima 3-Metil-Glutaril Coenzima A redutase. Os resultados demonstraram que a pravastatina induz parada no ciclo celular em G0/G1 e diminuição da produção de fatores de crescimento em várias linhas de células de Mieloma.

Assessment of the pathogenicity of cell-culture-adapted Newcastle disease virus strain Komarov

Newcastle disease vaccines hitherto in vogue are produced from embryonated chicken eggs. Egg-adapted mesogenic vaccines possess several drawbacks such as paralysis and mortality in 2-week-old chicks and reduced egg production in the egg-laying flock. Owing to these possible drawbacks, we attempted to reduce the vaccine virulence for safe vaccination by adapting the virus in a chicken embryo fibroblast cell culture (CEFCC) system. Eighteen passages were carried out by CEFCC, and the pathogenicity was assessed on the basis of the mean death time, intracerebral pathogenicity index, and intravenous pathogenicity index, at equal passage intervals. Although the reduction in virulence demonstrated with increasing passage levels in CEFCC was encouraging, 20% of the 2-week-old birds showed paralytic symptoms with the virus vaccine from the 18th(final) passage. Thus, a tissue-culture-adapted vaccine would demand a few more passages by CEFCC in order to achieve a complete reduction in virulence for use as a safe and effective vaccine, especially among younger chicks. Moreover, it can be safely administered even to unprimed 8-week-old birds.(AU)

EXPRESSÃO DE FIBROBLASTOS ASSOCIADOS AO CÂNCER EM MAMAS E LINFONODOS EM CADELAS COM NEOPLASMAS MAMÁRIOS E SEU PERFIL CLÍNICO.

Carcinomas mamários são neoplasmas de importância mundial, frequentes em cadelas e mulheres. Por se tratar de uma doença heterogênea, suas consequências variam de acordo com diferentes fatores prognósticos, como idade, comportamento biológico, presença de metástase e imunomarcação para determinados anticorpos. Sugere-se que os fibroblastos associados ao câncer (FACs) promovam a iniciação tumoral, progressão, e facilitam a formação de metástases; porém, seu papel no desenvolvimento tumoral ainda é desconhecido. Estas células expressam a marcação para - actina do músculo liso (-SMA), sendo que sua imunomarcação em diversos neoplasmas está associada ao pior prognóstico. Objetivou-se com este estudo avaliar se a intensidade de FACs em neoplasmas mamários e/ou linfonodos caninos tem relação com estadiamento mais avançados da doença e, consequentemente, pior prognóstico. Foram estudadas 18 cadelas com carcinoma mamários removidos por meio de mastectomia unilateral radical, sendo 8 com presença de metástase para linfonodos e 10 com linfonodos saudáveis. Foi realizada a imunohistoquímica dos neoplasmas mamários e dos linfonodos com o marcador -SMA. Na avaliação microscópica classificou-se as imunomarcações em negativa, fraca, moderada e forte. As associações entre os padrões de marcação e os fatores clínicos foram avaliadas através do Teste Exato de Fisher. Os resultados das imunomarcações moderadas e fortes de -SMA nos linfonodos foram associadas à presença de metástase (estadiamento IV), com valor de p = 0,03. A intensidade da marcação nas mamas estudadas não apresentou associação com a imunomarcação de seu linfonodo correspondente, com estadiamento IV ou com o tamanho do tumor. A idade das cadelas também não apresentou relação com estadiamentos III ou IV. A maior intensidade da imunomarcação de -SMA nos linfonodos metastáticos parece estar relacionada com a presença de células neoplásicas decorrentes do processo de transição epitélio-mesenquimal e consequente aumento na população de miofibroblastos. As imunomarcações moderadas/fortes nas mamas não foram estatisticamente relacionadas ao estadiamento IV da doença, embora alguns tumores conhecidos como altamente agressivos tiveram imunomarcação forte para o marcador -SMA. Conclui-se que o tamanho tumoral não influencia nas intensidades de imunomarcação de -SMA nas mamas. Além disso, não houve associação entre a intensidade de marcação nas mamas e seu linfonodo correspondente. Porém, linfonodos metastáticos apresentam maiores intensidade de FACs.

Breast carcinomas are neoplasms of worldwide importance, common in bitches and women. As it is a heterogeneous disease, its consequences vary according to different prognostic factors, such as age, biological behavior, presence of metastasis and immunostaining for certain antibodies. It is suggested that cancer-associated fibroblasts (CAFs) promote tumor initiation, progression, and facilitate the formation of metastases; however, its role in tumor development is still unknown. These cells express the -smooth muscle actin (-SMA) labeling, and their immunostaining in several neoplasms is associated with a worse prognosis. The objective of this study was to evaluate whether the intensity of CAFs in breast neoplasms and / or canine lymph nodes is related to more advanced stages of the disease and, consequently, a worse prognosis. Eighteen bitches with breast carcinoma removed by means of radical unilateral mastectomy were studied, 8 with metastasis to lymph nodes and 10 with healthy lymph nodes. Immunohistochemistry of breast neoplasms and lymph nodes was performed with the -SMA marker. In the microscopic evaluation, immunostaining was classified as negative, weak, moderate and strong. Associations between marking patterns and clinical factors were assessed using Fisher's exact test. The results of moderate and strong -SMA immunostaining in lymph nodes were associated with the presence of metastasis (stage IV), with a pvalue of 0.03. The intensity of the marking on the studied breasts was not associated with the immunostaining of its corresponding lymph node, with stage IV or with the size of the tumor. The age of the dogs was also not related to staging III or IV. The greater intensity of -SMA immunostaining in metastatic lymph nodes seems to be related to the presence of neoplastic cells resulting from the epithelial-mesenchymal transition process and a consequent increase in the population of myofibroblasts. Moderate / strong immunostaining in the breasts were not statistically related to stage IV of the disease, although some tumors known to be highly aggressive had strong immunostaining for the -SMA marker. It is concluded that the tumor size does not influence the immunostaining intensities of -SMA in the breasts. In addition, there was no association between the intensity of marking on the breasts and its corresponding lymph node. However, metastatic lymph nodes have greater intensity of CAFs.

Development of experimental in vitro burn model

To propose an experimental burn model in NIH-3T3 cell line. Induction of thermal injury in cultures of mouse fibroblast - NIH-3T3- cell line and determination of cell viability by MTT and imunofluorescence. The heating of the Petri dish increased proportionally to the temperature of the base and the time of exposure to microwave. In this in vitro burn model, using the cell line NIH-3T3 was observed drastic cellular injury with significant changes in cell viability and activity. It showed drastically modified cell morphology with altered membrane, cytoskeleton and nucleus, and low cellularity compared to the control group. The burn model in vitro using the cell line NIH-3T3 was reproductive and efficient. This burn model was possible to determine significant changes in cell activity and decreased viability, with drastic change in morphology, cell lysis and death.(AU)

Reconstruction of abdominal wall defects using small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor

To construct a new biomaterial-small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor, and to evaluate the new biomaterials for the reconstruction of abdominal wall defects. Thirty six Sprague-Dawley rats were used in the animal experiments and randomly divided into three groups. The new biomaterial was constructed by combining small intestinal submucosa with gelatin hydrogel for basic fibroblast growth factor release. Abdominal wall defects were created in rats, and repaired using the new biomaterials (group B), compared with small intestinal submucosa (group S) and ULTRAPROTM mesh (group P). Six rats in each group were sacrificed at three and eight weeks postoperatively to examine the gross effects, inflammatory responses, collagen deposition and neovascularization. After implantation, mild adhesion was caused in groups B and S. Group B promoted more neovascularization than group S at three weeks after implantation, and induced significantly more amount of collagen deposition and better collagen organization than groups S and P at eight weeks after implantation. Small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor could promote better regeneration and remodeling of host tissues for the reconstruction of abdominal wall defects.(AU)

Human beta defensin-4 and keratinocyte growth factor gene expression in cultured keratinocyte and fibroblasts of burned patients

To evaluate KGF and human beta defensin-4 (HBD-4) levels produced by dermic fibroblasts and keratinocytes cultivated from burned patients' skin samples. Keratinocytes and fibroblasts of 10 patients (four major burns, four minor burns and two controls) were primarily cultivated according to standard methods. HBD-4 and KGF genes were analyzed by quantitative PCR. In fibroblasts, KGF gene expression was 220±80 and 33.33±6.67 (M±SD; N=4), respectively for major and minor burn groups. In keratinocytes, KGF gene expression was 11.2±1.9 and 3.45±0.37 (M±SD; N=4), respectively for major and minor burn groups. In fibroblasts, HBD-4 gene expression was 15.0±4.0 and 11.5±0.5 (M±SD; N=4), respectively for major and minor burn. In keratinocyte, HBD-4 gene expression was 0.0±0.0 and 13.4±4.8 (M±SD; N=4), respectively for major and minor burn. KGF expression was increased in burn patient fibroblasts compared to control group. In keratinocytes culture, KGF suppression is inversely proportional to burn extension; it is active and increased in major burn but decreased in minor burn. HBD-4 expression was increased in fibroblasts and decreased in keratinocytes from all burned patients.(AU)

Keratinocyte growth factor, interleukins (1 beta, 6, 8, 10, 12), and tumor necrosis factor alpha in culture medium of dermal fibroblast of burned patients

PURPOSE: To evaluate the level of cytokines and keratinocyte growth factor (KGF) or Fibroblast Growth Factor 7 (FGF-7) in the culture medium of cultured human dermal fibroblasts from patients with large burn in comparison to small burn. METHODS: Fibroblasts of 10 patients (four large burns, four small burns and two controls) were initiated by the enzymatic method using collagenase. Cytokines and KGF in the supernatant of the culture medium was measured by, respectively, flow cytometry using Cytometric Bead Array Human Inflammation kit (CBA, BD Biosciences, USA) and the enzyme immunoassay method using the Quantikine (r) Human KGF. The experiments were performed in triplicate. RESULTS: The expression of IL-12 protein in patients with large burns showed a tendency to increase. IL- 6, IL- 10, and IL- 1beta were observed no difference. For IL - 8, TNF - alpha and KGF was observed a significant difference between the expression in large and small burned patient. CONCLUSION: That IL-8, TNF-alpha and KGF showed higher expression in cultured fibroblasts of large burned patients.(AU)

Alterações morfológicas que acometem o bulbo do olho de cães infectados naturalmente por Leishmania infantum/chagasi

A leishmaniose visceral é uma zoonose transmitida de forma vetorial, que sucede a picada de flebotomíneos infectados, e possui o agente etiológico protozoário do gênero Leishmania. O objetivo proposto é descrever as alterações oculares que podem afetar o bulbo do olho de cães infectados naturalmente pela leishmaniose e descrever as alterações morfológicas que ocorrem através da interação entre o parasito e as estruturas oculares. Foram realizadas duas coletas no município de São Luís MA do bulbo do olho de 12 cães ao todo. Os animais foram submetidos a uma avaliação clínica oftalmológica e individualizados quanto à presença de doenças oculares. Os cães foram doados com o consentimento do proprietário, de acordo com as regras do Conselho Nacional de Controle de Experimentação Animal (CONCEA). Todos os animais foram classificados como sintomáticos por avaliação clínica e as manifestações oculares observadas foram: uveíte, ceratoconjuntivite seca, úlcera de córnea, conjuntivite e secreção ocular purulenta, principalmente. De acordo com o tipo de infiltrado inflamatório, o mais prevalente foi o linfoplasmático. A conjuntiva bulbar, limbo e córnea foram a região do olho mais afetada por infiltrados. Pela IHC observamos a marcação de amastigotas pelo anticorpo anti-Leishmania nas túnicas bulbares e o anticorpo Vimentina para fibroblastos associados com as formas amastigotas. A marcação dos fibroblastos foi positiva na córnea e limbo do olho. Através da microscopia eletrônica de varredura (MEV), observamos a ultraestrutura do que apresentou hiperplasia do epitélio da córnea, desorganização do estroma e reação inflamatória exacerbada na túnica vascular e fibrosa. Diante os resultados obtidos, as manifestações são caracterizadas pela alta freqüência de infiltrados inflamatórios, principalmente do tipo linfoplasmático. Os fibroblastos presentes na córnea podem atuar como células hospedeiras importantes frente à doença sistêmica, o que traz uma nova perspectiva sobre o modo de ação do parasito nas túnicas oculares.

Visceral leishmaniasis is a vector-borne zoonosis that succeeds the bite of infected sandflies and has the protozoan etiological agent of the genus Leishmania. The main idea of this study is to describe the ocular changes that can affect the eye bulb of dogs naturally infected by leishmaniasis and to describe the morphological changes that occur through the interaction between the parasite and the ocular structures. Two collections were carried out in the city of São Luís - MA from the eye bulb of 12 animals. The animals were submitted to a clinical ophthalmologic evaluation and later individualized for the presence of ocular diseases. The animals were donated with consent of the owner, according to the rules of National Council for Control of Animal Experimentation (CONCEA). All animals were classified as symptomatic by clinical evaluation. Ocular manifestations were: uveitis, keratoconjunctivitis, corneal ulcer, conjunctivitis and purulent ocular secretion mainly. The bulbar conjunctiva, limbus and cornea were the most affected region by infiltrates. By IHC we observed amastigote labeling by anti- Leishmania antibody on bulbar tunics and Vimentin antibody for fibroblasts associated with amastigote forms. Fibroblast labeling was positive in the cornea and limbus of the eye. Through scanning electron microscopy (SEM), we observed the ultrastructure of corneal epithelium hyperplasia, stromal disorganization and exacerbated inflammatory reaction in fibrous tunic. Given the results obtained, the manifestations are characterized by the high frequency of inflammatory infiltrates, mainly lymphoplasmocitic type. Fibroblasts present in cornea may act as important host cells in face of systemic disease, which brings a new perspective on the action of the parasite in eye tunics.

Keratinocyte growth factor, tumor necrosis factor-alpha and interleukin-1 beta gene expression in cultured fibroblasts and keratinocytes from burned patients

PURPOSE: To evaluate the gene expression of KGF, TNF-alpha and IL-1 beta in skin fibroblasts and keratinocytes cultured from burned patients. METHODS: Three patients with large burns and three patients with small burns, as well as two controls, were included. The cell culture was initiated by the enzymatic method. After extraction and purification of mRNA, qPCR was used to assess the gene expression of KGF, TNF-alpha and IL-1 beta. RESULTS: The expression of KGF was increased on average 220-fold in large burns and 33.33-fold in small burns in fibroblasts, and 11.2-fold in large burns and 3.45-fold in small burns in keratinocytes compared to healthy patients (p<0.05). Expression of TNF-alpha was not observed. IL-1 beta is down-regulated in fibroblasts of burned patients, and much more repressed in small burns (687-fold, p<0.05). In keratinocytes, the repression of IL-1 beta expression occurs in patients with small burns (28-fold), while patients with large burns express this gene intensively (15-fold). CONCLUSIONS: The study showed a quantitative pattern in the expression of KGF gene, which is more expressed according to the size of the burn. TNF-alpha was not expressed. A qualitative pattern in the expression of IL-1 beta gene was demonstrated.(AU)

Envolvimento da subfamília do fator de crescimento fibroblástico 7 (FGF7) no controle da foliculogênese antral em bovinos / Involvement of fibroblast growth factor 7 (FGF7) subfamily in the control of antral folliculogenesis in cattle

Os mecanismos que coordenam o desenvolvimento folicular ainda não são completamente conhecidos e, portanto, constituem o alvo de numerosas investigações, seja por facilitarem a compreensão da fisiologia ou por serem promissoras ferramentas para a reprodução assistida. Recentemente, diversos peptídeos ovarianos de ação local têm sido descritos por participarem do controle de todas as fases do desenvolvimento folicular, bem como da modulação de hormônios esteróides ovarianos e gonadotrofinas; entre esses peptídeos estão os fatores de crescimento fibroblástico (FGFs). Os FGFs têm sido extensamente investigados em diversas fases do desenvolvimento folicular e parecem controlar processos como atresia folicular, esteroidogênese, bem como o desenvolvimento folicular pré-antral, sendo a subfamília do FGF7 uma das mais investigadas neste contexto. Assim, esta revisão tem como objetivo sumarizar a participação da subfamília do FGF7 no controle da foliculogênese antral de bovinos. (AU)

The mechanisms that coordinate follicular development are not well known and are, therefore, target of numerous investigations for facilitateing the understanding of physiology or as promising tools for assisted reproduction. Recently, several ovarian peptides with local action have been reported to participate in the control of follicular development in all stages and modulation of gonadotropins andovarian steroid hormones. In this context, fibroblast growth factors (FGFs) have been extensively investigated in different stages of follicular development and seem to control processes such as follicular atresia, steroidogenesis, and pre-antral follicle development, where FGF7 subfamily is one of the most investigated. Therefore, the objective of this review is to summarize the participation of the FGF7 subfamily in the control of bovine antral folliculogenesis. (AU)

Envolvimento da subfamília do fator de crescimento fibroblástico 7 (FGF7) no controle da foliculogênese antral em bovinos / Involvement of fibroblast growth factor 7 (FGF7) subfamily in the control of antral folliculogenesis in cattle

Os mecanismos que coordenam o desenvolvimento folicular ainda não são completamente conhecidos e, portanto, constituem o alvo de numerosas investigações, seja por facilitarem a compreensão da fisiologia ou por serem promissoras ferramentas para a reprodução assistida. Recentemente, diversos peptídeos ovarianos de ação local têm sido descritos por participarem do controle de todas as fases do desenvolvimento folicular, bem como da modulação de hormônios esteróides ovarianos e gonadotrofinas; entre esses peptídeos estão os fatores de crescimento fibroblástico (FGFs). Os FGFs têm sido extensamente investigados em diversas fases do desenvolvimento folicular e parecem controlar processos como atresia folicular, esteroidogênese, bem como o desenvolvimento folicular pré-antral, sendo a subfamília do FGF7 uma das mais investigadas neste contexto. Assim, esta revisão tem como objetivo sumarizar a participação da subfamília do FGF7 no controle da foliculogênese antral de bovinos.

The mechanisms that coordinate follicular development are not well known and are, therefore, target of numerous investigations for facilitateing the understanding of physiology or as promising tools for assisted reproduction. Recently, several ovarian peptides with local action have been reported to participate in the control of follicular development in all stages and modulation of gonadotropins andovarian steroid hormones. In this context, fibroblast growth factors (FGFs) have been extensively investigated in different stages of follicular development and seem to control processes such as follicular atresia, steroidogenesis, and pre-antral follicle development, where FGF7 subfamily is one of the most investigated. Therefore, the objective of this review is to summarize the participation of the FGF7 subfamily in the control of bovine antral folliculogenesis.