Optimal single-embryo mass spectrometry fingerprinting

In pre-implantation embryos, lipids play key roles in determining viability, cryopreservation and implantation properties, but often their analysis is analytically challenging because of the few picograms of analytes present in each of them. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) allows obtaining individual phospholipid profiles of these microscopic organisms. This technique is sensitive enough to enable analysis of individual intact embryos and monitoring the changes in membrane lipid composition in the early stages of development serving as screening method for studies of biology and biotechnologies of reproduction. This article introduces an improved, more comprehensive MALDI-MS lipid fingerprinting approach that considerably increases the lipid information obtained from a single embryo. Using bovine embryos as a biological model, we have also tested optimal sample storage and handling conditions before the MALDI-MS analysis. Improved information at the molecular level is provided by the use of a binary matrix that enables phosphatidylcholines, sphingomyelins, phosphatidylserines, phosphatidylinositols and phosphoethanolamines to be detected via MALDI(±)-MS in both the positive and negative ion modes. An optimal MALDI-MS protocol for lipidomic monitoring of a single intact embryo is therefore reported with potential applications in human and animal reproduction, cell development and stem cell research. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2013 John Wiley & Sons, Ltd.

Effects of a single nucleotide polymorphism in the leptin gene on the productive traits of dairy buffaloes (Bubalus bubalis)

The gene responsible for coding the leptin hormone has been associated with productive and reproductive traits in cattle. In dairy cattle, different polymorphisms found in the leptin gene have been associated with several traits of economic interest, such as energy balance, milk yield and composition, live weight, fertility and dry matter consumption. The aim of this study was to detect genetic variability in the leptin gene of buffaloes and to test possible associations with milk yield, fat and protein percentages, age at first calving and first calving interval. Three genotypes (AA, AG and GG) were identified by polymerase chain reaction-restriction fragment length polymorphism, which presented genotypic frequencies of 0.30, 0.54 and 0.16, respectively. The allele frequencies were 0.57 for the A allele and 0.43 for the G allele. No significant effects were found in the present study, but there is an indicative that leptin gene affects lipid metabolism. © 2013 Springer Science+Business Media Dordrecht.

A single chart with supplementary runs rules for monitoring the mean vector and the covariance matrix of multivariate processes

The MRMAX chart is a single chart based on the standardized sample means and sample ranges for monitoring the mean vector and the covariance matrix of multivariate processes. User's familiarity with the computation of these statistics is a point in favor of the MRMAX chart. As a single chart, the recently proposed MRMAX chart is very appropriate for supplementary runs rules. In this article, we compare the supplemented MRMAX chart and the synthetic MRMAX chart with the standard MRMAX chart. The supplementary and the synthetic runs rules enhance the performance of the MRMAX chart. © 2013 Elsevier Ltd.

Use of platelet-rich plasma in the treatment of deep corneal ulcers induced in rabbits.clinical and histomorphometric evaluation

The repair process induced corneal ulcer in rabbits using platelet-rich plasma in the form of eyedrop or clot was clinically evaluated and compared. Sixty rabbits were divided into four groups of 15 animals, denominated platelet group (PG), clot group (CLG), control group (CG), and amniotic control group (AG). Experimental groups were then subdivided into three groups (M4, M7, M30), corresponding to the end of the evaluation period. There were no differences between treatments regarding ocular sensitivity, chemosis and ocular secretion. The groups treated with PRP either as eyedrop or a clot showed less opacity than the animals treated only with amniotic membrane at the moment of the final evaluation. The presence of corneal ulcers in the groups treated with PRP showed lower intensity than the other groups. Histomorphometric examination showed that corneal epithelization in the initial phase of the lesion was greater when using PRP. The use of amniotic membrane promoted corneal epithelial and stromal thickness, as well as synergism when associated to PRP.

Genetic diversity of the HLA-G coding region in Amerindian populations from the Brazilian Amazon: a possible role of natural selection

HLA-G has an important role in the modulation of the maternal immune system during pregnancy, and evidence that balancing selection acts in the promoter and 3′UTR regions has been previously reported. To determine whether selection acts on the HLA-G coding region in the Amazon Rainforest, exons 2, 3 and 4 were analyzed in a sample of 142 Amerindians from nine villages of five isolated tribes that inhabit the Central Amazon. Six previously described single-nucleotide polymorphisms (SNPs) were identified and the Expectation-Maximization (EM) and PHASE algorithms were used to computationally reconstruct SNP haplotypes (HLA-G alleles). A new HLA-G allele, which originated in Amerindian populations by a crossing-over event between two widespread HLA-G alleles, was identified in 18 individuals. Neutrality tests evidenced that natural selection has a complex part in the HLA-G coding region. Although balancing selection is the type of selection that shapes variability at a local level (Native American populations), we have also shown that purifying selection may occur on a worldwide scale. Moreover, the balancing selection does not seem to act on the coding region as strongly as it acts on the flanking regulatory regions, and such coding signature may actually reflect a hitchhiking effect.Genes and Immunity advance online publication, 3 October 2013; doi:10.1038/gene.2013.47.

Education and its impact on poverty: equity or exclusion?

Includes bibliography

Progress made in the implementation of the CARICOM single market and economy

Includes bibliography

Caracterização estrutural e das interações entre a Proteína G do hRSV e potenciais inibidores

Pós-graduação em Microbiologia - IBILCE

Estimativa do fluxo de nutrientes pelo trato digestivo de bovinos de corte alimentados com Brachiaria brizantha (Hochst A. Rich) Stapf. cv. Marandu em duas idades de rebrota

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

América Latina: población por años calendario y edades simples: período 1995-2005 = Latin America: population by calendar years and single ages, period 1995-2005

Publicación bilingüe (Español e inglés)

América Latina: población por años calendario y edades simples, 1995-2005 = Latin America: population by calendar years and single ages

Publicación bilingüe

Cytogenetic studies in fishes of the genera Hassar, Platydoras and Opsodoras (Doradidae, Siluriformes) from Jarí and Xingú Rivers, Brazil

We studied the karyotypes of Hassar cf. orestis and an undescribed Hassar species from the Jarí River and Opsodoras ternetzi, H. orestis and Platydoras cf. costatus from the Xingú River, all with 2n = 58. Constitutive heterochromatin is located in the centromere in most metacentric pairs; in some chromosomes this banding is not present, or it is located on the whole chromosome arm or in the distal regions. The NOR is located on a single biarmed pair at a distal region of the short arm in H. cf. orestis, H. orestis and P. cf. costatus at a distal region of the long arm in O. ternetzi and at a proximal region of the long arm in the Hassar species. In all species (except for Hassar sp.) the CMA3 analysis revealed a rich G-C region coincident with the NOR. Probably inversions occurred in the NOR chromosome during the chromosomal differentiation of the Doradidae species here described.

Análise de polimorfismo na região promotora do gene da Interleucina 18 (-137 G/C e -607 C/A) em pacientes portadores do vírus da hepatite C de Belém, Pará

Desde a sua descoberta, em 1989, o vírus da hepatite C (HCV) tem sido reconhecido como a maior causa de doença hepática crônica no mundo. Considerado um problema de saúde pública mundial que envolve entre 170 a 350 milhões de pessoas infectadas. Fatores genéticos do hospedeiro têm sido implicados na persistência da infecção pelo HCV. Estudos sugerem que dois polimorfismos de nucleotídeos únicos na posição -607 C/A (rs1946518) e -137 G/C (rs187238) na região promotora do gene da IL-18 têm sido encontrados e associados com a atividade de transcrição do promotor da IL-18 e, potencialmente, de IFN-γ, sendo associados ao atraso na depuração viral e na persistência da doença. Foi realizado um estudo do tipo transversal analítico no município de Belém-PA, em 152 amostras sanguíneas de pacientes infectados pelo HCV e 188 controles não infectados. As amostras foram submetidas à RT-PCR, para detecção do RNA viral e, posteriormente, à RFLP-PCR para avaliação do polimorfismo na região promotora do gene da IL-18, nas posições -137 G/C e -607 C/A. Os resultados não revelaram diferença significante para os polimorfismos da IL-18 entre os pacientes e grupo controle. Mas revelou diferença significante para os genótipos homozigotos G/G (39,1%), na posição -137 (OR = 3.00, IC [95%] = 1.24 – 7.22, p = 0.02), e A/A (21,7%), posição -607 (OR = 3.62, IC [95%] = 1.25 – 10.45, p = 0.03), entre as mulheres, em relação aos homens (22,6% e 7,6%). Os resultados demonstraram indícios que entre as mulheres, a presença do polimorfismo homozigoto A/A (-607) atue como fator protetor contra a infecção pelo HCV, já que o genótipo A/A (-607) tem sido relacionado em alguns estudos à doença hepática leve e à depuração viral.

Molecular analysis of three FUT3 gene single nucleotide polymorphisms and their relationship with the lewis erythrocytary phenotype in a human population of japanese-ancestry living in Tomé Açu, a town in the Brazilian Amazon

The Lewis blood group system involves two major antigens, Le^ª and Le^b. Their antigenic determinants are not primary gene products but are synthesized by the transfer of sugar subunits to a precursory chain by a specific enzyme which is the product of the FUT3 gene (Lewis gene). The presence of three FUT3 gene single nucleotide polymorphisms (SNPs) (59T > G; 508G > A and 1067T > A) was related to the Lewis phenotype of erythrocytes from 185 individuals of Japanese ancestry living in the town of Tomé-Açu in the Brazilian Amazon region. This relationship was detected using a serological hemagglutination test and the Dot-ELISA assay along with the molecular technique PCR-RFLP. We found that the three SNPs investigated in this study only accounted for a proportion of the Lewis-negative phenotype of the erythrocytes.

Improvement of friable callus production of Boerhaavia paniculata Rich and the investigation of its lipid profile by GC/MS

Neste estudo, um protocolo para induzir maiores quantidades de calos friáveis de Boerhaavia paniculata RICH e uma técnica de lipidômica foram aplicados para investigar o perfil dos lipídios e serem comparados com aqueles presentes nas raízes desta planta, os quais apresentaram atividade anti-inflamatória no extrato hexânico bruto. A cultura de calo foi induzida a partir de sementes em meio sólido contendo sais de Murashige e Skoog com diferentes quantidades de glicose e diferentes concentrações de ácido 2,4-diclorofenoxiacético. Os frascos foram mantidos em câmara de germinação a 30 ± 2°C, com fotoperíodo de 16h sob intensidade luminosa de 27 µmol m^–2 s^–1 durante 4 semanas. Os melhores resultados para a formação de calo friável e desenvolvimento da biomassa foram obtidos no tratamento contendo 2,26 µM de 2,4-D e de glicose (1,5 %; m/v). Técnica de lipidômica foi aplicada na fração hexânica mostrando as concentrações mais elevadas de esteróides β-sitosterol (3,53 mg/100 g de cs - células seca ), e ácidos graxos; especialmente o ácido 2-hidroxi-tetracosanóico (0,34 mg/100 g cs), ácido eicosanoico (86,25 mg/100 g cs), ácido esteárico (420,83 mg/100 g cs), ácido tetradecanóico (10,74 mg/100 g cs) e ácido linoléico (100,61 mg/100 g cs). O perfil lipídico do calo versus aquele encontrado nas raízes da planta silvestre é descrito neste trabalho.