Characterization of monocarboxylate transporters (MCTs) expression in soft tissue sarcomas: distinct prognostic impact of MCT1 sub-cellular localization

Background: Soft tissue sarcomas (STSs) are a group of neoplasms, which, despite current therapeutic advances, still confer a poor outcome to half of the patients. As other solid tumors, STSs exhibit high glucose consumption rates, associated with worse prognosis and therapeutic response. As highly glycolytic tumors, we hypothesized that sarcomas should present an increased expression of lactate transporters (MCTs).Methods: Immunohistochemical expression of MCT1, MCT2, MCT4 and CD147 was assessed in a series of 86 STSs and the expression profiles were associated with patients' clinical-pathological parameters.Results: MCT1, MCT4 and CD147 were mainly observed in the plasma membrane of cancer cells (around 60% for MCTs and 40% for CD147), while MCT2 was conspicuously found in the cytoplasm (94.2%). Importantly, we observed MCT1 nuclear expression (32.6%). MCT1 and MCT4, alone or co-expressed with CD147 in the plasma membrane, were associated with poor prognostic variables including high tumor grade, disease progression and shorter overall survival. Conversely, we found MCT1 nuclear expression to be associated with low grade tumors and longer overall survival.Conclusions: The present work represents the first report of MCTs characterization in STSs. We showed the original finding of MCT1 expression in the nucleus. Importantly, opposite biological roles should be behind the dual sub-cellular localization of MCT1, as plasma membrane expression of MCT1 is associated with worse patients' prognosis, while nuclear expression is associated with better prognosis.

Programmed cell death-associated gene transcripts in bovine embryos exposed to bovine Herpesvirus type 5

In vitro-produced bovine embryos become infected after exposure to bovine Herpesvirus type 5 (BoHV-5), yet no changes in developmental rates, mitochondrial activity and inhibition of apoptosis are detected in comparison to unexposed embryos. Thus, the aim of the present study was to assess the transcription of mitochondria-mediated apoptosis genes using TaqMan real-time polymerase chain reaction. Transcripts of mcl-1, caspase-2, -3, Apaf-1 and Bax genes were measured after exposure to BoHV-5 in vitro. Mitochondrial dehydrogenase activity was evaluated by MIT test and compared between groups of exposed and unexposed embryos, at day 7 of development. The rate of oocyte maturation was assessed by the extrusion of the first polar body. In summary, BoHV-5 exposed embryos retained their viability, mitochondrial dehydrogenase activity and displayed up-regulation of transcription of survival mcl-1 gene and down-regulation of Bax transcription in relation to mitochondria-mediated pathway which might improve embryo viability. These findings demonstrate that BoHV-5 exposed embryos maintain their viability and mitochondrial dehydrogenase activity with no compromise of embryos produced in vitro. (c) 2013 Elsevier Ltd. All rights reserved.

Effects of the Temperature and Maturation Stages on the Germination of Roystonea regia Seeds

Roystonea regia (Kunth) O.F. Cook is largely used as ornamental. The propagation is done almost exclusively by seeds; however, there is a great variation in the germination process influenced by many factors. The objective of this work was to study the effects of the temperature and maturation stages on the germination of R. regia seeds. The experimental design was entirely randomized in a factorial arrangement 6x3 (six temperatures: constant at 20, 25, 30 and 35 degrees C and alternated at 20-30 and 25-35 degrees C, with a photoperiod of 12 hours; and three fruit maturation stages: brown, yellow and black), with four replications of 25 disseminules (seed with stucked endocarp) each. The disseminules had their mesocarp and exocarp were removed and shade dried. Their moisture content was determined, and then they were placed in plastic boxes (gerbox type) containing vermiculite. The disseminules, with the germinative intumescence, were daily noted until germination was steady. The germination rate and the germination speed index were calculated, and the data were submitted to the variance analysis. The means were compared by the Tukey test. It was concluded that the highest germination rate (99.7%) and germination speed were obtained by seeds from mature (black) fruits at the temperature of 35 degrees C.

Characterisation of the methylation pattern in the intragenic CpG island of the IGF2 gene in Bos taurus indicus cumulus cells during in vitro maturation

The aim of this study was to characterise the methylation pattern in a CpG island of the IGF2 gene in cumulus cells from 1-3 mm and a parts per thousand yenaEuro parts per thousand 8.0 mm follicles and to evaluate the effects of in vitro maturation on this pattern.Genomic DNA was treatment with sodium bisulphite. Nested PCR using bisulphite-treated DNA was performed, and DNA methylation patterns have been characterised.There were no differences in the methylation pattern among groups (P > 0.05). Cells of pre-IVM and post-IVM from small follicles showed methylation levels of 78.17 +/- 14.11 % and 82.93 +/- 5.86 %, respectively, and those from large follicles showed methylation levels of 81.81 +/- 10.40 % and 79.64 +/- 13.04 %, respectively. Evaluating only the effect of in vitro maturation, cells of pre-IVM and post-IVM COCs showed methylation levels of 80.17 +/- 12.01 % and 81.19 +/- 10.15 %.In conclusion, the methylation levels of the cumulus cells of all groups were higher than that expected from the imprinted pattern of somatic cells. As the cumulus cells from the pre-IVM follicles were not subjected to any in vitro manipulation, the hypermethylated pattern that was observed may be the actual physiological methylation pattern for this particular locus in these cells. Due the importance of DNA methylation in oogenesis, and to be a non-invasive method for determining oocyte quality, the identification of new epigenetic markers in cumulus cells has great potential to be used to support reproductive biotechniques in humans and other mammals.

Ultrastructural Morphology and Nuclear Maturation Rates of Immature Equine Oocytes Vitrified with Different Solutions and Exposure Times

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Midpalatal suture maturation: Classification method for individual assessment before rapid maxillary expansion

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

Cellular and Molecular Tissue Response to Triple Antibiotic Intracanal Dressing

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

Cellular and molecular studies of the effects of a selective COX-2 inhibitor celecoxib in the cardiac cell line H9c2 and their correlation with death mechanisms

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of follicular fluid supplementation during in vitro maturation on total cell number in bovine blastocysts produced in vitro

This study evaluated the influence of follicular fluid (FF) added to the maturation medium on the quality of bovine embryos produced in vitro. In the first experiment, oocytes were matured in media containing five different FF concentrations with different maturation times and classified according to meiotic progression and migration of cortical granules. In the second experiment, oocytes matured in the same media were fertilized at three different maturation times; thereafter, cleavage and blastocyst rates were evaluated. In the third experiment, oocytes were matured in media containing three different FF concentrations at two different maturation times, and embryo quality, inferred by the ratio of inner cell mass and trophectoderm cells compared with total cell number, was evaluated. Higher FF concentration (75 - 100% FF) slowed meiotic progression and CG migration (control - 78.13% vs. treated - 52.58% and control - 52.7% vs. treated - 11.59%, respectively, at 24 h of maturation). Also, FF at concentration of 75% or 100% had a negative influence on cleavage and blastocyst rates (control - 90.13% vs. treated - 82.64% and control - 35.73% vs. treated - 11.57%, respectively, at 24 h of maturation). The 50% FF resulted in embryos with increased inner cell mass numbers (control - 29.91 vs. treated - 35.49, at 24 h of maturation) and total cell numbers (control - 109.53 vs. treated - 120.67, at 26 h of maturation). Even though higher concentration of FF added to the maturation medium reduced embryonic development rates, in lower concentrations, FF slowed the meiotic progression and migration of CG and contributed to increases in inner cell mass number. Thus, FF added to the maturation medium enhances the number of cells in bovine embryos produced in vitro, especially for inner cell mass.

In vitro Maturation of Oocytes from Santa Ines Ewes Subjected to Consecutive Sessions of Follicular Aspiration by Laparoscopy

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

Dinâmica da vitelogênese durante a maturação ovariana em Artemesia longinaris

Pós-graduação em Aquicultura - FCAV

Oocyte maturation in the sloth's giant tick Amblyomma varium (Acari: Ixodidae) in an ecological context

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

Cellular behavior as a dynamic field for exploring bone bioengineering: A closer look at cell-biomaterial interface

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

DNA Damage and Its Cellular Response in Mother and Fetus Exposed to Hyperglycemic Environment

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

Control of oocyte maturation

Oocyte maturation is a complex process involving nuclear and cytoplasmic maturation. The nuclear maturation is a chromosomal segregation and the cytoplasmic maturation involves the reorganization of the cytoplasmic organelles, mRNA transcription and storage of proteins to be used during fertilization and early embryo development. The mechanism of oocyte maturation in vivo and in vitro still are not totally understood. However it is generally accepted that the second messenger cyclic adenosine monophosphate (cAMP) plays a critical role in the maintenance of meiotic blockage of mammalian oocytes. A relative increase in the level of cAMP within the oocyte is essential for maintaining meiosis block, while a decrease in cAMP oocyte concentration allows the resumption of meiosis. The oocyte cAMP concentration is regulated by a balance of two types of enzymes: adenylate cyclase (AC) and phosphodiesterases (PDEs), which are responsible for the synthesis and degradation of cAMP, respectively. After being synthesized by AC in cumulus cells, cAMP are transferred to the oocyte through gap junctions. Thus, specific subtypes PDEs are able to inhibit or attenuate the spontaneous meiotic maturation of oocytes with PDE4 primarily involved in the metabolism of cAMP in granulosa cells and PDE3 in the oocyte. Although the immature oocytes can resume meiosis in vitro, after being removed from antral follicles, cytoplasmic maturation seems to occur asynchronously with nuclear maturation. Therefore, knowledge of the oocyte maturation process is fundamental for the development of methodologies to increase the success of in vitro embryo production and to develop treatments for various forms of infertility. This review will present current knowledge about the maintenance of the oocyte in prophase arrest, and the resumption of meiosis during oocyte maturation, focusing mainly on the changes that take place in the oocyte.