Immunohistochemical detection of receptors for oestrogen and progesterone in endometrial glands and stroma during the oestrous cycle in Nelore (Bos taurus indicus) cows

The aim of the present study was to monitor endometrial distribution and concentrations of oestrogen receptors alpha (ER alpha) and progesterone receptors (PR) by immunohistochemistry in Nelore cows (Bos taurus indicus) during the oestrous cycle. Blood samples were collected for progesterone measurement and endometrial samples were taken from the uterine horn contra lateral to the corpus luteum in 16 cows at days 0 (ovulation), 5, 9, 13 and 19 of the oestrous cycle. Immunostaining evaluation for ER alpha and PR in the glandular epithelium and uterine stroma was performed by two methods: positive nuclei counting and staining intensity of the nuclei. Specific positive staining reactions for both receptors were limited to cell nuclei and they were not identified in the cytoplasm. The proportion of ER alpha positive nuclei had a temporal variation throughout the oestrous cycle in both cell types evaluated and was higher in uterine stroma than the glandular epithelium (p < 0.05). The greatest proportion of ER alpha stained nuclei was observed at oestrus and during the initial and mid luteal phase (days 5, 9 and 13) (p < 0.05) in the glandular epithelium and at days 0, 5 and 9 in the uterine stroma (p < 0.01). The proportion of PR positive nuclei remained constant throughout the entire oestrous cycle for both cell types evaluated (p > 0.05). A higher proportion of PR positive nuclei was measured in the uterine stroma compared with the glandular epithelium (p < 0.05). Intensity of staining for ER alpha and PR varied throughout the oestrous cycle (p < 0.01). There was a higher staining intensity at days 0 and 5 in the stroma for ER alpha (p < 0.01) and PR (p < 0.01) and in the glandular epithelium at days 0, 5, 9 and 13 for ER alpha (p < 0.01) and at days 0, 5 and 9 for PR (p < 0.01) when compared with the other evaluated days. These data demonstrate that ER alpha and PR expression varied throughout the oestrous cycle in Nelore cows, in general with highest concentrations at oestrus and the lowest during the luteal phase. This is similar to patterns observed in Bos taurus taurus.

Diferenciação in vitro de células-tronco mesenquimais da medula óssea de cães em precursores osteogênicos

O objetivo principal da nossa pesquisa foi avaliar o potencial de diferenciação osteogênica de células-tronco mesenquimais (MSC) obtidas da medula óssea do cão. As MSC foram separadas pelo método Ficoll e cultivadas sob duas condições distintas: DMEM baixa glicose ou DMEM/F12, ambos contendo L-glutamina, 20% de SFB e antibióticos. Marcadores de MSC foram testados, confirmando células CD44+ e CD34- através da citometria de fluxo. Para a diferenciação osteogênica, as células foram submetidas a quatro diferentes condições: Grupo 1, as mesmas condições utilizadas para a cultura de células primárias com os meios DMEM baixa glicose suplementado; Grupo 2, as mesmas condições do Grupo 1, mais os indutores de diferenciação dexametasona, ácido ascórbico e b-glicerolfosfato; Grupo 3, células cultivadas com meios DMEM/F12 suplementado; e Grupo 4, nas mesmas condições que no Grupo 3, mais indutores de diferenciação de dexametasona, ácido ascórbico e b-glicerolfosfato. A diferenciação celular foi confirmada através da coloração com alizarin red e da imunomarcação com o anticorpo SP7/Osterix. Nós observamos através da coloração com alizarin red que o depósito de cálcio foi mais evidente nas células cultivadas em DMEM/F12. Além disso, usando a imunomarcação com o anticorpo SP/7Osterix obtivemos positividade em 1:6 células para o Meio DMEM/F12 comparada com 1:12 para o meio DMEM-baixa glicose. Com base nos nossos resultados concluímos que o meio DMEM/F12 é mais eficiente para a indução da diferenciação de células-tronco mesenquimais caninas em promotores osteogênicos. Este efeito provavelmente ocorre em decorrência da maior quantidade de glicose neste meio, bem como da presença de diversos aminoácidos.

Differential alterations in the activity of matrix metalloproteinases within the nervous tissue of dogs in distinct manifestations of visceral leishmaniasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Histochemical and morphological characteristics of the levator ani muscle in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Impact of proximal cytoplasmic droplets on quality traits and in-vitro embryo production efficiency of cryopreserved bull spermatozoa

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Distribution of NADPH-diaphorase-positive neurons in the prefrontal cortex of the Cebus monkey

We studied the distribution of NADPH-diaphorase (NADPH-d) activity in the prefrontal cortex of normal adult Cebus apella monkeys using NADPH-d histochemical protocols. The following regions were studied: granular areas 46 and 12, dysgranular areas 9 and 13, and agranular areas 32 and Oap. NADPH-d-positive neurons were divided into two distinct types, both non-pyramidal. Type I neurons had a large soma diameter (17.24 +/- 1.73 pm) and were densely stained. More than 90% of these neurons were located in the subcortical white matter and infragranular layers. The remaining type I neurons were distributed in the supragranular layers. Type II neurons had a small, round or oval soma (9.83 +/- 1.03 mu m), and their staining pattern varied markedly. Type II neurons were distributed throughout the cortex, with their greatest numerical density being observed in layers II and III. In granular areas, the number of type II neurons was up to 20 times that of type I neurons, but this proportion was smaller in agranular areas. Areal density of type II neurons was maximum in the supragranular layers of granular areas and minimum in agranular areas. Statistical analysis revealed that these areal differences were significant when comparing some specific areas. In conclusion, our results indicate a predominance of NADPH-d-positive cells in supragranular layers of granular areas in the Cebus prefrontal cortex. These findings support previous observations on the role of type II neurons as a new cortical nitric oxide source in supragranular cortical layers in primates, and their potential contribution to cortical neuronal activation in advanced mammals. (c) 2006 Elsevier B.V. All rights reserved.

Effects of cyclosporin, phenytoin, and nifedipine on the synthesis and degradation of gingival collagen in tufted capuchin monkeys (Cebus apella): Histochemical and MMP-1 and -2 and collagen I gene expression analyses

Background: The purpose of this experimental study was to evaluate the collagen fiber distribution histologically after phenytoin, cyclosporin, or nifedipine therapy and to correlate it with collagen I and matrix metalloproteinase (MMP)-1 and -2 gene expression levels.Methods: Gingival samples from the canine area were obtained from 12 male monkeys (Cebus apella). The mesial part of each sample was assessed by reverse transcription-polymerase chain reaction, whereas the distal part was processed histologically for picrosirius red and hematoxylin and eosin stainings, as well as for collagen IV immunostaining. One week after the first biopsy, the animals were assigned to three groups that received daily oral dosages of cyclosporin, phenytoin, or nifedipine for 120 days. Additional gingival samples were obtained on days 52 and 120 of treatment from two animals from each group on the opposite sides from the first biopsies.Results: Picrosirius red staining showed a predominance of mature collagen fibers in the control group. Conversely, there was an enlargement of areas occupied by immature collagen fibers in all groups at days 52 and 120, which was not uniform over each section. There was a general trend to lower levels of MMP-1 gene expression on day 52 and increased levels on day 120. Phenytoin led to increased levels of MMP-2 and collagen I gene expression on day 120, whereas the opposite was observed in the nifedipine group.Conclusion: Cyclosporin, phenytoin, and nifedipine led to phased and drug-related gene expression patterns, resulting in impaired collagen metabolism, despite the lack of prominent clinical signs.

Cyto-, myelo- and chemoarchitecture of the prefrontal cortex of the Cebus monkey

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Compromised Primary Implant Stability

The aim of this study was to evaluate implants installed with compromised primary stability associated or not with polylactide and polyglycolide copolymer (PLA/PGA). Two 0.4-mm overdrilled defects (3 mm in diameter and 6 mm long) were produced in the right tibia of 25 rabbits. Half of the defects were left untreated, and the other half was filled with PLA/PGA. Implants of 2.6 mm in diameter and 6 mm long were placed into all defects. Animals were killed at 5, 15, 40, and 60 postoperative days, and biomechanical analysis (torque-reverse), histomorphometry, and immunohistochemistry (osteoprotegerin, receptor activator of NFJB ligand [RANKL], osteocalcin, and collagen-1 [ COL- I] staining) were performed. All the implants achieved osseointegration. There were no statistically significant differences in the torque-reverse and in linear contact extension between bone tissue and implant surface and no statistically significant difference in osteoprotegerin, RANKL, osteocalcin, and collagen-1 expression between the studied groups in all studied periods (P > 0.05). We can conclude that osseointegration can occur in compromised primary implant stability situations, and the addition of PLA/PGA did not improve the osseointegration process in this experimental model.

Bone regeneration in surgically created defects filled with autogenous bone: an epifluorescence microscopy analysis in rats

Although the search for the ideal bone substitute has been the focus of a large number of studies, autogenous bone is still the gold standard for the filling of defects caused by pathologies and traumas, and mainly, for alveolar ridge reconstruction, allowing the titanium implants installation. Objectives: The aim of this study was to evaluate the dynamics of autogenous bone graft incorporation process to surgically created defects in rat calvaria, using epifluorescence microscopy. Material and methods: Five adult male rats weighing 200-300 g were used. The animals received two 5-mm-diameter bone defects bilaterally in each parietal bone with a trephine bur under general anesthesia. Two groups of defects were formed: a control group (n=5), in which the defects were filled with blood clot, and a graft group (n=5), in which the defects were filled with autogenous bone block, removed from the contralateral defect. The fluorochromes calcein and alizarin were applied at the 7th and 30th postoperative days, respectively. The animals were killed at 35 days. Results: The mineralization process was more intense in the graft group (32.09%) and occurred mainly between 7 and 30 days, the period labeled by calcein (24.66%). Conclusions: The fluorochromes showed to be appropriate to label mineralization areas. The interfacial areas between fluorochrome labels are important sources of information about the bone regeneration dynamics.

Influence of the proportion of particulate autogenous bone graft/platelet-rich plasma on bone healing in critical-size defects: An immunohistochemical analysis in rat calvaria

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of Artificial Aging on the Roughness and Microhardness of Sealed Composites

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Silver nanoparticles: influence of stabilizing agent and diameter on antifungal activity against Candida albicans and Candida glabrata biofilms

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Silver colloidal nanoparticles: antifungal effect against adhered cells and biofilms of Candida albicans and Candida glabrata

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Hybrid layer thickness and resin tag length of a self-etching adhesive bonded to sound dentin

Objectives: the purpose of this study is to employ optical microscopy to measure the thickness of the hybrid layer and the penetration (tags) of an aggressive self-etching adhesive system into sound dentin.Methods: occtusat cavities were prepared in 40 extracted human posterior teeth. The prepared teeth were randomly assigned to four experimental groups with 10 specimens each. The self-etching adhesive system Adper Prompt L-Pop was applied to the dentin surface as follows: Group 1: cavosurface enamel was etched for 60 s and dentin for 20 s with 35% phosphoric acid get, immediately followed by application of the self -etching adhesive with a brush to the entire cavity for 15 s; Groups 2, 3, and 4: no pre-etching was performed, and the self -etching adhesive was applied to both enamel and dentin for 15, 30 and 45 s, respectively. After curing, the cavities were fitted with composite resin Fittek Z250. Afterwards, the teeth were decalcified and the restorations were carefully removed for later embedding in paraffin. The specimens were serially sectioned at 6 mu m of thickness and sequentially mounted in glass slides. These sections were stained with Brown and Brenn staining for posterior analysis and measurement of the hybrid layer and resin tags on a tight microscope with a micrometric ocular 40/075. The results were submitted to analysis of variance at the 5% level.Results: whenever there was significance, the Tukey test was applied at the 5% level. The specimens receiving application of acid etching before the selfetching. adhesive displayed a larger thickness of the hybrid layer; on the other hand, specimens receiving only application of the self -etching adhesive on dentin for 15, 30 and 45 s exhibited similar thickness of the hybrid layer. As regards the resin tags, no statistically significant differences could be found between the study groups.Conclusions: it could be concluded that the increase in the time of application of the self-etching adhesive Adper Prompt L-Pop did not significantly influence the formation and thickness of hybrid layer, as well as its penetration into the sound dentin surface. (c) 2005 Elsevier Ltd. All rights reserved.