Supplementation with the histone deacetylase inhibitor trichostatin A during in vitro culture of bovine embryos

Summary Trichostatin A (TSA) is a histone deacetylase inhibitor that induces histone hyperacetylation and increases gene expression levels. The aim of the present study was to establish a suitable condition for the use of TSA in in vitro cultures of bovine embryos, and to determine whether TSA would increase blastocyst rates by improvement of chromatin remodelling during embryonic genome activation and by increasing the expression of crucial genes during early development. To test this hypothesis, 8-cell embryos were exposed to four concentrations of TSA for different periods of time to establish adequate protocols. In a second experiment, three experimental groups were selected for the evaluation of embryo quality based on the following parameters: apoptosis, total cell number and blastocyst hatching. TSA promoted embryonic arrest and degeneration at concentrations of 15, 25 and 50 nM. All treated groups presented lower blastocyst rates. Exposure of embryos to 5 nM for 144 h and to 15 nM for 48 h decreased blastocyst hatching. However, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay revealed similar apoptosis rates and total cell numbers in all groups studied. Although, in the present study, TSA treatment did not improve the parameters studied, the results provided background information on TSA supplementation during in vitro culture of bovine embryos and showed that embryo quality was apparently not affected, despite a decrease in blastocyst rate after exposure to TSA. © Cambridge University Press 2011.

Chromatin and morphometry alterations of bovine sperm head after Percoll (TM) density gradient centrifugation

The aim of this work was to assess the morphometry and chromatin integrity of bovine sperm head after a three layers discontinuous Percoll (TM) density gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Three ejaculates per bull were evaluated. The semen doses were thawed and two smears were made from each sample before (control) and after the Percoll (TM) centrifugation (Percoll (TM) group). The smears were stained with toluidine blue and grayscale digital images were captured and processed in Scilab environment software. It was observed that chromatin heterogeneity was reduced (P<0.05) and chromatin decondensation was increased (P<0.05) after the Percollm treatment utilized. In addition, it was observed that sperm head length was higher (P<0.05) and the side symmetry was lower (P<0.05) in centrifuged sperm cells. When analyzed separately by subspecies, it was observed that the decrease (P<0.05) in chromatin heterogeneity and the increase (P<0.05) in chromatin decondensation occurred in Zebu sperm heads. In addition, the length and the width:length ratio of sperm heads was affected by Percoll (TM) centrifugation in Zebu semen. In conclusion, the three layers discontinuous Percoll (TM) centrifugation increased the chromatin decondensation and the morphometric alterations of frozen-thawed bovine semen. However, the real implication of these findings in fertility rates of centrifuged sperm must be investigated. (C) 2011 Elsevier B.V. All rights reserved.

A three-dimensional network constructed from the assembly of 1,3-diaminopropane-copper(II) and tetracyanopalladate(II) moieties

The coordination polymer [Cu(Pd(CN)(4))(pn)](n) (pn = 1,3-diaminopropane) has been synthesized and characterized by elemental analysis, infrared spectroscopy and single-crystal X-ray diffraction. The crystal structure showed that three cyano groups of each [Pd(CN)(4)] unit bridge Cu(II) centers leading to the formation of a three-dimensional network. A series of bifurcated hydrogen bonds between the amino groups of the diamine and the nonbridging cyano groups of the cyanometallate result in the organization of suprarnolecular chains and rings along the polymer. (c) 2008 Elsevier B.V. All rights reserved.

Large nuclear vacuoles are indicative of abnormal chromatin packaging in human spermatozoa

The aim of this investigation was to determine the presence of abnormal sperm chromatin packaging in spermatozoa with large nuclear vacuoles (LNV) selected via high magnification by analysing the pattern of chromomycin A3 (CMA3) staining. A prospective observational study was designed to analyse semen samples obtained from 66 men undergoing infertility diagnosis and treatment. The numbers of cells with normal (dull yellow staining of the sperm head/CMA3-negative) and abnormal (bright yellow fluorescence of the sperm head/CMA3-positive) chromatin packaging were determined on slides with normal and LNV spermatozoa. The presence of bright yellow fluorescence (CMA3-positive) was significantly higher (p < 0.0001) in spermatozoa with LNV than in normal spermatozoa (719/1351; 53.2% vs. 337/835; 40.3%, respectively), reflecting a higher percentage of abnormal chromatin packaging in spermatozoa with large LNV. Our data support the hypothesis that the presence of LNV reflects the presence of abnormal chromatin packaging, which may facilitate sperm DNA damage. As sperm nuclear vacuoles are evaluated more precisely at high magnifications using motile sperm organelle morphology examination (MSOME), the present results support the use of high-magnification sperm selection for intracytoplasmic sperm injection (ICSI).

Significance of extruded nuclear chromatin (regional nuclear shape malformation) in human spermatozoa: implications for ICSI

The aim of this study was to determine the extent of DNA fragmentation and the presence of denatured single-strand or normal double-strand DNA in spermatozoa with extruded nuclear chromatin (ENC) selected by high magnification. Fresh semen samples from 55 patients were prepared by discontinuous isolate concentration gradient. Spermatozoa with normal nucleus (NN) and ENC were selected at 8400x magnification and placed on different slides. DNA fragmentation was determined by TUNEL assay. Denatured and double-stranded DNA was identified by the acridine orange fluorescence method. DNA fragmentation was not significantly different (p = 0.86) between spermatozoa with ENC (19.6%) and those with NN (20%). However, the percentage of spermatozoa with detectable denatured-stranded DNA in the ENC spermatozoon group (59.1%) was significantly higher (p < 0.0001) than in the NN group (44.9%). The high level of denatured DNA in spermatozoa with ENC suggests premature decondensation and disaggregation of sperm chromatin fibres. The results show an association between ENC and DNA damage in spermatozoa, and support the routine morphological selection and injection of motile spermatozoa at high-magnification intracytoplasmic sperm injection.

COX23, a homologue of COX17, is required for cytochrome oxidase assembly

Deletion of reading frame YHR116W of the Saccharomyces cerevisiae nuclear genome elicits a respiratory deficiency. The encoded product, here named Cox23p, is shown to be required for the expression of cytochrome oxidase. Cox23p is homologous to Cox17p, a water-soluble copper protein previously implicated in the maturation of the Cu-A center of cytochrome oxidase. The respiratory defect of a cox23 null mutant is rescued by high concentrations of copper in the medium but only when the mutant harbors COX17 on a high copy plasmid. Overexpression of Cox17p by itself is not a sufficient condition to rescue the mutant phenotype. Cox23p, like Cox17p, is detected in the intermembrane space of mitochondria and in the postmitochondrial supernatant fraction, the latter consisting predominantly of cytosolic proteins. Because Cox23p and Cox17p are not part of a complex, the requirement of both for cytochrome oxidase assembly suggests that they function in a common pathway with Cox17p acting downstream of Cox23p.

Exploring community assembly through an individual-based model for trophic interactions

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparative analysis of heteropyknotic chromatin and silver-stained material in Platycarenus umbractulatus (Pentatomidae: Discocephalinae)

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

Morphological patterns of the heteropycnotic chromatin and nucleolar material in meiosis and spermiogenesis of some Pentatomidae (Heteroptera)

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

Relics of eclogite facies assemblages in the Ceara Central Domain, NW Borborema Province, NE Brazil: Implications for the assembly of West Gondwana

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

Formation of TiO2 ceramic foams from the integration of the sol-gel method with surfactants assembly and emulsion

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efficient microwave-assisted hydrothermal synthesis of CuO sea urchin-like architectures via a mesoscale self-assembly

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

Self-assembly of cyclometallated Pd(II) compounds directed by C-H center dot center dot center dot pi interactions: A structural evidence for metalloaromaticity in a cyclopalladated ring

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)