Mast cell concentration and skin wound contraction in rats treated with Ximenia americana L

PURPOSE: To evaluate wound contraction and the concentration of mast cells in skin wounds treated with wild plum (Ximenia americana) essential oil-based ointment in rats. METHODS:Sixty rats were submitted to two cutaneous wounds in the thoracic region, on the right and left antimeres. Thereon, they were divided into three groups: GX (wounds treated once a day with hydro alcoholic branch extract of Ximenia americana), GP (wounds that received vehicle), and GC (wounds without product application). Wounds were measured immediately after the injury as well as 4, 7, 14 and 21 days post-topical application of the extract. At these days, five rats from each group were euthanatized. Thereafter, samples were fixed in 10% formaldehyde and processed for paraffin embedding. Sections were stained with H.E, Masson's Trichrome and toluidine blue for morphological, morphometrical and histopathological analysis, under light microscopy. The degree of epithelial contraction was measured and mast cell concentrations were also evaluated with an image analyzer (Image Pro-plus (R) software). RESULTS: The extract treated group showed lower mast cell concentrations in the 4th day of lesion, as compared to GP (GX GP = GCp<0.05). CONCLUSION: Ointment containing 10% X. americana induces a decrease in mast cell concentration, at the beginning of the healing process, and promotes early skin wound contraction in rats.FACEPEFAPEMIGCNPqUniv Fed Rio Grande do Norte UFRN, Dept Morphol & Anim Physiol, Postgrad Program Anim Biosci, Recife, PE, BrazilUniv Fed Rural Pernambuco UFRPE, Dept Morphol & Anim Physiol, Postgrad Program Anim Biosci, Recife, PE, BrazilUniv Fed Pernambuco UFPE, Dept Histol & Embryol, Histol Div, Recife, PE, BrazilUniv Fed Rural Pernambuco, Dept Morphol & Anim Physiol, Postgrad Program Anim Biosci, Recife, PE, BrazilUniv Sao Paulo FMUSP, Fac Med, Clin Hosp, Sao Paulo, BrazilUniv Fed Sao Paulo UNIFESP, Dept Morphol & Genet, Postgrad Program Morphol, Sao Paulo, BrazilUniv Fed Piaui, Dept Biol, Div Microbiol, Picos, PI, BrazilUniv Fed Rural Pernambuco, Dept Morphol & Physiol, Anat Div, Recife, PE, BrazilUniv Fed Sao Paulo UNIFESP, Dept Morphol & Genet, Postgrad Program Morphol, Sao Paulo, BrazilWeb of Scienc

Curvas de índice de sítio para leucena [Leucaena leucocephala (Lam.) De Wit] no agreste de Pernambuco.

The main goal of this work was to set and compare site index curves for leucena [Leucaena leucocephala (Lam.) de Wit] variety Hawaii (cv K8) in Agreste of Pernambuco. The data set came from Empresa Pernambucana de Pesquisa Agropecuária (IPA), Experimental Station of Caruaru, in which 544 trees cultivated with and without organic compound were measured 19 times during the period of 1990 – 2003. The models of Schumacher, Chapman-Richards, Silva-Bailey, Mitscherlich, Weibull and Clutter-Jones were tested and compared. To select the best equations, the Index of Fit (IF), standard error of estimate (), identity model test and the graphic of residuals distribution were used and results show that the model of Clutter-Jones provided a better IF for both treatments. The graphic of residual distribution did not show tendency among the models. The model identity test showed no differences among the models tested. As Schumacher’s model has been used frequently in forestry and presented the smallest number of parameters, it deserves special focus in studies of site index.O objetivo deste trabalho foi ajustar e comparar modelos de curvas de índice de sítio para Leucaena leucocephala (Lam.) de Wit., variedade Hawaii (cv. K8), no Agreste de Pernambuco. Os dados foram obtidos na Estação Experimental da Empresa Pernambucana de Pesquisa Agropecuária (IPA), no município de Caruaru - PE, considerando-se 544 árvores, cultivadas com e sem composto orgânico, com 19 mensurações ao longo do tempo (1990-2003). Foram ajustados e comparados os modelos de Schumacher, Chapman-Richards, Silva-Bailey, Mitscherlich, Weibull e Clutter-Jones. Para a seleção das equações, utilizaram-se os índices de ajuste (IA), o erro padrão da estimativa (), a distribuição gráfica dos resíduos e o teste de identidade dos modelos. O modelo de Clutter-Jones apresentou o maior valor de IA nos dois tratamentos. Nos gráficos das distribuições residuais, os modelos não apresentaram tendências. No teste de identidade dos modelos de regressão, verificou-se não haver diferença significativa ao nível de 5 % de probabilidade. Por ser um modelo com o menor número de parâmetros entre os testados e por ser freqüentemente utilizado na Ciência Florestal, o modelo de Schumacher merece especial destaque em trabalhos de índice de sítio

SITE INDEX CURVES FOR LEUCAENA [ Leucaena leucocephala (Lam.) de Wit] IN AGRESTE OF PERNAMBUCO

O objetivo deste trabalho foi ajustar e comparar modelos de curvas de \uedndice de s\uedtio para Leucaena leucocephala (Lam.) de Wit., variedade Hawaii (cv. K8), no Agreste de Pernambuco. Os dados foram obtidos na Esta\ue7\ue3o Experimental da Empresa Pernambucana de Pesquisa Agropecu\ue1ria (IPA), no munic\uedpio de Caruaru \u2013 PE, considerando-se 544 \ue1rvores, cultivadas com e sem composto org\ue2nico, com 19 mensura\ue7\uf5es ao longo do tempo (1990-2003). Foram ajustados e comparados os modelos de Schumacher, Chapman-Richards, Silva-Bailey, Mitscherlich, Weibull e Clutter-Jones. Para a sele\ue7\ue3o das equa\ue7\uf5es, utilizaram-se os \uedndices de ajuste (IA), o erro padr\ue3o da estimativa (Syx%), a distribui\ue7\ue3o gr\ue1fica dos res\uedduos e o teste de identidade dos modelos. O modelo de Clutter-Jones apresentou o maior valor de IA nos dois tratamentos. Nos gr\ue1ficos das distribui\ue7\uf5es residuais, os modelos n\ue3o apresentaram tend\ueancias. No teste de identidade dos modelos de regress\ue3o, verificou-se n\ue3o haver diferen\ue7a significativa ao n\uedvel de 5 % de probabilidade. Por ser um modelo com o menor n\ufamero de par\ue2metros entre os testados e por ser freq\ufcentemente utilizado na Ci\ueancia Florestal, o modelo de Schumacher merece especial destaque em trabalhos de \uedndice de s\uedtio.The main goal of this work was to set and compare site index curves for leucena [Leucaena leucocephala (Lam.) de Wit] variety Hawaii (cv K8) in Agreste of Pernambuco. The data set came from Empresa Pernambucana de Pesquisa Agropecu\ue1ria (IPA), Experimental Station of Caruaru, in which 544 trees cultivated with and without organic compound were measured 19 times during the period of 1990 \u2013 2003. The models of Schumacher, Chapman-Richards, Silva-Bailey, Mitscherlich, Weibull and Clutter-Jones were tested and compared. To select the best equations, the Index of Fit (IF), standard error of estimate (Syx%), identity model test and the graphic of residuals distribution were used and results show that the model of Clutter-Jones provided a better IF for both treatments. The graphic of residual distribution did not show tendency among the models. The model identity test showed no differences among the models tested. As Schumacher\u2019s model has been used frequently in forestry and presented the smallest number of parameters, it deserves special focus in studies of site index

DNA and bone structure preservation in medieval human skeletons

Morphological and ultrastructural data from archaeological human bones are scarce, particularly data that have been correlated with information on the preservation of molecules such as DNA. Here we examine the bone structure of macroscopically well-preserved medieval human skeletons by transmission electron microscopy and immunohistochemistry, and the quantity and quality of DNA extracted from these skeletons. DNA technology has been increasingly used for analyzing physical evidence in archaeological forensics; however, the isolation of ancient DNA is difficult since it is highly degraded, extraction yields are low and the co-extraction of PCR inhibitors is a problem. We adapted and optimized a method that is frequently used for isolating DNA from modern samples, Chelex® 100 (Bio-Rad) extraction, for isolating DNA from archaeological human bones and teeth. The isolated DNA was analysed by real-time PCR using primers targeting the sex determining region on the Y chromosome (SRY) and STR typing using the AmpFlSTR® Identifiler PCR Amplification kit. Our results clearly show the preservation of bone matrix in medieval bones and the presence of intact osteocytes with well preserved encapsulated nuclei. In addition, we show how effective Chelex® 100 is for isolating ancient DNA from archaeological bones and teeth. This optimized method is suitable for STR typing using kits aimed specifically at degraded and difficult DNA templates since amplicons of up to 250 bp were successfully amplified

Effects of soy isoflavones associated with low intensity mechanical vibration on bone tissue of ooforectomized rats

Introduction: It has been shown that both soy isoflavones (ISO) and low intensity high frequency mechanical vibration (VIB) have protective effects in bone tissue of ovariectomized (OVX) rats. Objective: We investigated whether combined treatments with ISO and VIB would exhibit cooperative effects in bone tissue of OVX rats. Methods: Fifty female Wistar rats at the age of 6 months were either sham-operated or OVX. After operation, the rats were divided in five groups: SHAM; OVX; OVX treated daily by gavage with 200mg/kg/body weight of ISO for 90 days (OVX-ISO); OVX treated with VIB, 20 min./day, at 60 Hz, 0.5g, 5 days/ week for 90 days (OVX-VIB) and OVX treated with ISO and VIB (OVX-ISO+VIB). After treatment, the femurs and tibias were collected and its length and thickness were measured. The femurs were then processed for histomorphometry, histochemistry and biochemical assay. The tibias were frozen for subsequent physical and mechanical tests. Results: Treatments did not have significant effects on body weight and in the bone measurements, except in the tibial thickness of the OVX-ISO and OVX-ISO+VIB groups that was significant higher than the SHAM group. Physical tests showed that the immersed weight, content of organic material and bone density were higher in the SHAM, OVX-VIB and OVX-ISO+VIB groups than the other groups, while the bone volume, bone mineral density, mineral material and the ash weight were higher in the OVX and OVX-ISO groups than the other groups and there were no significant differences in the mechanical tests among the groups. Treatment did not have significant effects in the trabecular bone volume, but it was not detected cortical bone loss in the group treated with the combined therapy. The treatments showed positive effects in the content of glycosaminoglycans, as well as in the organization of collagen fibers and these effects was higher in the OVX-ISO+VIB group. The TUNEL-positive cell profile was similar among the groups. Conclusion: These results demonstrate that the combined treatment (ISO+VIB) have better effects in the bone tissue constituents of ovariectomized osteopenic rats, when compared with the treatments isolated.Introdução: É sabido que tanto as isoflavonas da soja (ISO) e a vibração mecânica de baixa intensidade e alta freqüência (VIB) possuem efeitos protetores no tecido ósseo de ratas ooforectomizadas (OVX). Objetivos: estudar os efeitos dos tratamentos com ISO e VIB isolados e combinados no tecido ósseo de ratas OVX. Métodos: 50 ratas Wistar com 6 meses de idade foram SHAM operadas ou OVX e após três meses, divididas em 5 grupos: SHAM; OVX; OVX tratadas diariamente por gavage com 200mg/kg de ISO por 90 dias consecutivos (OVX-ISO); OVX tratadas com VIB, 20 min./dia, 30 Hz, 0.6g, 5 dias por semana por 90 dias (OVX-VIB) e OVX tratadas com ISO e VIB (OVX-ISO+VIB). Após o tratamento, os fêmures e tíbias foram coletados e seus comprimento e espessura foram medidos. Os fêmures foram processados para histomorfometria e aos métodos histoquímicos, bioquímico e do TUNEL. As tíbias foram congeladas e destinadas para testes físicos e biomecânicos. Resultados: Os tratamentos não exerceram efeitos significantes no peso corporal e nas medidas de comprimento e espessura, exceto na espessura das tíbias dos grupos OVX-ISO e OVX-ISO+VIB que foi significantemente maior comparado ao grupo SHAM. Os testes físicos mostraram que o peso imerso, a porcentagem de material orgânico e a densidade óssea foram maiores nos grupos SHAM, OVX-VIB e OVX-ISO+VIB quando comparados aos outros grupos, enquanto que o volume ósseo, a densidade mineral óssea, porc entagem de material mineral e o peso das cinzas foram maiores nos grupos OVX e OVX-ISO do que nos outros grupos e não houve diferenças significantes nos parâmetros biomecânicos analisados entre os grupos. Nos fêmures, os tratamentos não exerceram efeitos significantes no volume ósseo trabecular, porém, o tratamento combinado exerceu efeitos positivos no osso cortical. Os tratamentos tiveram efeitos positivos em relação à quantidade e na organização das fibras colágenas e esse efeito foi mais expressivo no grupo OVX-ISO+VIB. O método do TUNEL não detectou diferenças nas células TUNEL-positivas entre os grupos. Conclusão: Esses resultados demonstram que o tratamento combinado (ISO+VIB) exerce maior efeito nos constituintes do tecido ósseo de ratas ooforectomizadas osteopênicas, quando comparado aos tratamentos isolados.TEDEBV UNIFESP: Teses e dissertaçõe

Influência do estrógeno na regulação da autofagia e apoptose dos osteócitos no processo alveolar de ratas ovariectomizadas

Autophagy is a process that reduces cell stress and enhances cell survival. Estrogen is a hormone that inhibits bone resorption and osteocyte apoptosis, promoting bone homeostasis. However, it is not clear if estrogen interfere in the autophagy process to assure osteocyte survival. Objective: To evaluate the influence of estrogen in the incidence of autophagy and apoptosis in the osteocytes of alveolar process of ovariectomized rats. Material and methods: Sixty adult female rats were SHAM-operated (SHAM) or ovariectomized (OVX) and treated with estrogen (OVXE group) or received vehicle solution (SHAM and OVX groups) for 15, 30 and 45 days. Six SHAM and six OVX rats were euthanized before treatments and used as basal groups (SHAMBL and OVXBL). After treatments and euthanasia, the maxillae containing the first molars were processed for paraffin and Araldite embedding. HE stained sections were used for histomorphometry to obtain the interradicular bone area and the number of osteocyte and empty lacunae. Sections were also subjected to the silver impregnation method to analyze the osteocyte cytoplasmic processes, and to the TRAP method to obtain the osteoclast number. Sections were submitted to the TUNEL and immunohistochemical methods to analyze apoptosis (cleaved caspase-3, BAX, Bcl2 and HMGB1) and autophagy (beclin-1, LC3II, p62, or immunofluorescence for LC3II and p62 detection); other sections were subjected to the combined TUNEL/TRAP methods, whereas semi-thin sections were submitted to the Sudan Black method (lipid detection). Results: In comparison to the SHAMBL group, OVXBL group showed higher percentages of caspase-3, TUNEL, p62-positive osteocytes, empty lacunae and osteoclasts. On the other hand, this group showed lower percentages of beclin-1 and LC3II-positive osteocytes. The estrogen treated group for 15 days presented lower percentages of caspase-3 and TUNEL-positive osteocytes, and higher percentage of LC3II-positive osteocytes when compared to the OVX group. The group treated for 30 days showed lower number of empty lacunae and lower percentages of TUNEL and p62-positive osteocytes in comparison to the OVX group. The group treated for 45 days showed higher bone area and osteocyte number, lower percentages of caspase-3, BAX, TUNEL and p62-positive osteocytes; while higher percentages of beclin-1 and LC3II-positive osteocytes was observed when compared to OVX group. The treatment with estrogen for 15 and 30 days promoted significant reduction in osteoclast number, as compared to the OVX groups. A higher percentage of HMGB1-positive-citoplasm and a lower percentage of HMGB1-positive nuclei in osteocytes were seen in the OVX groups, when compared to the SHAM and estrogen treated groups. A high frequency of lipid droplets was observed in the osteocytes of the OVX groups. The ultraestrutural images showed large cytoplasmic regions of osteocytes occupied by lipid droplets, mainly in the OVX groups, and osteocytes with ultrastructural features of apoptosis. Conclusions: Our results indicate that estrogen deficiency reduces autophagy and increase apoptosis in osteocytes, whereas estrogen treatment maintains osteocyte viability by enhancing autophagy and inhibiting apoptosis. This inverse proportional association between autophagy and apoptosis supports the idea that estrogen may play a role in the autophagy control, reducing osteocytes apoptosis.A autofagia é um processo que reduz o estresse celular, aumentando a sobrevivência das células. O estrógeno é um hormônio que inibe a reabsorção óssea e a apoptose de osteócitos, promovendo a homeostase óssea. Porém, ainda não está claro se o estrógeno interfere no processo de autofagia para garantir a sobrevivência dos osteócitos. Objetivos: Avaliar a influência do estrógeno na incidência de autofagia e de apoptose nos osteócitos do processo alveolar de ratas ovariectomizadas. Material e métodos: Sessenta e seis ratas adultas foram SHAM-operadas (SHAM) ou ovariectomizadas (OVX) e tratadas com estrógeno (Grupo OVXE) ou receberam solução veículo (Grupos SHAM e OVX), por 15, 30 e 45 dias. Seis ratas SHAM e seis OVX foram eutanasiadas antes dos tratamentos para obtenção dos grupos basais (SHAMBL e OVXBL). Após os tratamentos e eutanásia, as maxilas contendo o primeiro molar foram processadas para inclusão em parafina e Araldite. Cortes corados com HE foram submetidos à histomorfometria para obtenção da área óssea interradicular, número de osteócitos e de lacunas vazias; outros foram submetidos aos métodos da prata para análise dos prolongamentos dos osteócitos e do TRAP para quantificação de osteoclastos. Cortes foram submetidos ao método do TUNEL e à imuno-histoquímica para análise de apoptose (caspase-3 clivada, BAX, Bcl2 e HMGB1) e autofagia (beclina-1, LC3II, p62, ou imunofluorescência para LC3II e p62), além da análise ultraestrutural. Cortes também foram submetidos aos métodos do TUNEL/TRAP combinados e cortes semifinos foram submetidos ao método de Sudan Black para detecção de lipídeo. Resultados: O grupo OVXBL mostrou maior porcentagem de osteócitos caspase-3, TUNEL e p62-positivos, menor porcentagem de osteócitos beclina-1 e LC3II-positivos, e maior número de lacunas vazias e de osteoclastos, em comparação ao grupo SHAMBL. O grupo tratado com estrógeno por 15 dias apresentou menor porcentagem de osteócitos caspase-3 e TUNEL-positivos, e maior porcentagem de osteócitos LC3II-positivos, em comparação ao grupo OVX. O grupo tratado por 30 dias mostrou menor número de lacunas vazias e de osteócitos TUNEL e p62-positivos em comparação ao OVX. O grupo tratado por 45 dias evidenciou maior área óssea e número de osteócitos, menor porcentagem de osteócitos caspase-3, BAX, TUNEL e p62-positivos, assim como maior porcentagem de osteócitos beclina-1 e LC3II-positivos, quando comparado ao grupo OVX. O tratamento com estrógeno por 15 e 30 dias promoveu diminuição significante no número de osteoclastos, em comparação aos grupos OVX. Notou-se maior porcentagem de osteócitos com citoplasma positivo ao HMGB1 e menor porcentagem de osteócitos com núcleo HMGB1- positivo nos grupos OVX. Gotículas de lipídios foram observadas com maior frequência nos osteócitos dos grupos OVX. A análise ultraestrutural evidenciou osteócitos com gotículas lipídicas ocupando amplas regiões citoplasmáticas, principalmente nos grupos OVX, e osteócitos com características ultraestruturais de apoptose. Conclusões: Nossos resultados indicam que a deficiência estrogênica diminui a autofagia e aumenta a apoptose dos osteócitos, enquanto o tratamento com estrógeno mantém a viabilidade dos osteócitos aumentando a autofagia e inibindo a apoptose, sugerindo uma associação inversamente proporcional entre esses dois processos nessas células. Esta associação inversamente proporcional entre autofagia e apoptose reforça a ideia de que o estrógeno deve participar no processo de controle da autofagia, além de reduzir a apoptose em osteócitos.Dados abertos - Sucupira - Teses e dissertações (2013 a 2016

Biology of Bone Tissue: Structure, Function, and Factors That Influence Bone Cells

Bone tissue is continuously remodeled through the concerted actions of bone cells, which include bone resorption by osteoclasts and bone formation by osteoblasts, whereas osteocytes act as mechanosensors and orchestrators of the bone remodeling process. This process is under the control of local (e.g., growth factors and cytokines) and systemic (e.g., calcitonin and estrogens) factors that all together contribute for bone homeostasis. An imbalance between bone resorption and formation can result in bone diseases including osteoporosis. Recently, it has been recognized that, during bone remodeling, there are an intricate communication among bone cells. For instance, the coupling from bone resorption to bone formation is achieved by interaction between osteoclasts and osteoblasts. Moreover, osteocytes produce factors that influence osteoblast and osteoclast activities, whereas osteocyte apoptosis is followed by osteoclastic bone resorption. The increasing knowledge about the structure and functions of bone cells contributed to a better understanding of bone biology. It has been suggested that there is a complex communication between bone cells and other organs, indicating the dynamic nature of bone tissue. In this review, we discuss the current data about the structure and functions of bone cells and the factors that influence bone remodeling