Secretome of the preimplantation human embryo by bottom-up label-free proteomics

Abstract A bottom-up label-free mass spectrometric proteomic strategy was used to analyse the protein profiles of the human embryonic secretome. Culture media samples used for embryonic culture of patients undergoing intracytoplasmic sperm injection cycles were selected as a test case for this exploratory proof-of-principle study. The media were stored after embryo transfer and then pooled into positive (n = 8) and negative (n =8) implantation groups. The absolute quantitative bottom-up technique employed a multidimensional protein identification technology based on separation by nano-ultra-high pressure chromatography and identification via tandem nanoelectrospray ionization mass spectrometry with dataindependent scanning in a hydrid QqTOF mass spectrometer. By applying quantitative bottom-up proteomics, unique proteins were found exclusively in both the positive-and negative-implantation groups, which suggest that competent embryos express and secrete unique biomarker proteins into the surrounding culture medium. The selective monitoring of these possible secretome biomarkers could make viable procedures using singleembryo transfer

Micropropagation of Eucalyptus dunnii Maid

International audienc

Influência da ação antrópica sobre a fauna de macroinvertebrados aquáticos em riachos de uma região de cerrado do sudoeste do Estado de São Paulo

Foram estudadas a riqueza e a distribuição espacial da fauna de macroinvertebrados aquáticos em riachos da região de cerrado de Assis, SP, com a finalidade de verificar o efeito da ação antrópica sobre a fauna. As coletas foram realizadas em nascentes do Ribeirão Água do Cervo, principal fornecedor de água da cidade de Assis. Foram estabelecidos nove pontos de coleta nos quais foram colocados dez pacotes de seixos de tamanho padronizado (volume equivalente a 300 cm³ cada). Após 25 dias de exposição, os pacotes foram removidos do riacho. Os macroinvertebrados associados a cada pacote de seixos foram triados e identificados. Os resultados mostraram uma menor riqueza no local mais impactado. Não houve relação significativa entre os fatores físico-químicos e a fauna. No entanto, a ação antrópica foi significativamente relacionada com a fauna.The richness and spatial distribution of aquatic macroinvertebrates were studied in streams from a cerrado region, Assis, State of São Paulo, with the purpose of verifying the effect of the anthropic action on the fauna. The collections took place at the headwaters of the Ribeirão Água do Cervo (the main water supplier of the city of Assis). Nine collection stations were established in which ten standardized pebble packages were placed (equivalent volume, 300 cm³ each). After twenty-five days of exposure, the packages were removed from the stream. The macroinvertebrates associated to each of the pebble packages were removed and identified. The results showed a smaller richness in the most damaged station. There was not significant relation between the physicochemical factors and the fauna. The intensity of anthropic action was significantly related with the fauna.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Valor preditivo do potencial de implantação embrionário pela análise do perfil químico de meios de cultivo por espectrometria de massas (ESI-Q-ToF)

Objective: This case-control study analyzed mass spectrometry fingerprinting patterns of culture media samples used for embryo culture to predict embryo implantation. Methods: The culture medium harvested after embryo transfer of 22 embryos from 13 patients was used for the experiments. After embryo transfer, the remaining culture media were collected and samples were split in positive (n=8) and negative (n=14) implantation groups according to implantation outcomes (100% or 0% of implantation). Samples were individually diluted and injected directly to the Electrospray ionization (ESI) MS coupled to a Quadrupole Time-of-flight MS (Q-ToF-MS).Ions relative intensities of each spectrum were considered. Data analysis was conducted in MatLab 7.0 version using Partial Least Squares - Discriminant Analysis toolbox. Results: There were 3027 observed ions at 100% and 0% implantation groups by ESI-Q-ToF-MS. The statistical model could categorize the samples in two clusters, based on their positive and negative implantation outcomes. Less intense ions present in the mass spectra with statistical significance have contributed to the major differences to group distinction. Conclusions: Positive and negative implantation embryos showed a specific biochemical pattern present in culture media, which could be detected as a fast, simple and non-invasive way. This biochemical profile could help the selection of the most viable embryo, improving single embryo transfer and thus eliminating the risk and undesirable outcomes of multiple pregnancies. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida