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.

Mass spectrometric characterization of two novel inflammatory peptides from the venom of the social wasp Polybia paulista

The social wasp P. paulista is relatively common in southeast Brazil causing many medically important stinging incidents. The seriousness of these incidents is dependent on the amount of venom inoculated by the wasps into the victims, and the characteristic envenomation symptoms are strongly dependent on the types of peptides present in the venom. In order to identify some of these naturally occurring peptides available in very low amounts, an analytical protocol was developed that uses a combination of reversed-phase and normal-phase high-performance liquid chromatography (HPLC) of wasp venom for peptide purification, with matrix-assisted laser desorption/ionization time-of-flight post-source decay mass spectrometry (MALDI-Tof-PSD-MS) and low-energy collision-induced dissociation (CID) in a quadrupole time-of-flight tandem mass spectrometry (QTof-MS/MS) instrument for peptide sequencing at the sub-picomole level. The distinction between Leu and Ile was achieved both by observing d-type fragment ions obtained under CID conditions and by comparison of retention times of the natural peptides and their synthetic counterparts (with different combinations of I and/or L at N- and C-terminal positions). To distinguish the isobaric residues K and Q, acetylation of peptides was followed by Q-Tof-MS analysis. The primary sequences obtained were INWLKLGKMVIDAL-NH2 (MW 1611.98Da) and IDWLKLGKMVMDVL-NH2 (MW 1658.98Da). Micro-scale bioassay protocols characterized both peptides as presenting potent hemolytic action, mast cell degranulation, and chemotaxis of poly-morphonucleated leukocyte (PMNL) cells. The primary sequences and the bioassay results suggest that these toxins constitute members of a new sub-class of mastoparan toxins, directly involved in the occurrence of inflammatory processes after wasp stinging. Copyright (C) 2004 John Wiley Sons, Ltd.

Jelleines: a family of antimicrobial peptides from the Royal Jelly of honeybees (Apis mellifera)

Four antimicrobial peptides were purified from Royal Jelly of honeybees, by using reverse phase-HPLC and sequenced by using Q-Tof-MS/MS: PFKLSLHL-NH2 (Jelleine-I), TPFKLSLHL-NH2 (Jelleine-II), EPFKLSLHL-NH2 (Jelleine-III), and TPFKLSLH-NH2 (Jelleine-IV). The peptides were synthesized on-solid phase, purified and submitted to different biological assays: antimicrobial activity, mast cell degranulating activity and hemolysis. The Jelleines-I-III presented exclusively antimicrobial activities against yeast, Gram+ and Gram- bacteria; meanwhile, Jelleine-IV was not active in none of the assays performed. These peptides do not present any similarity with the other antimicrobial peptides from the honeybees; they are produced constitutively by the workers and secreted into Royal Jelly. (C) 2004 Elsevier B.V. All rights reserved.

Análise comparativa de características seminais de Mazama cinzas brasileiros: morfometria espermática e proteômica do plasma seminal

Pós-graduação em Medicina Veterinária - FCAV

Estudo peptídico e determinação do perfil de metabólitos de venenos de escorpiões da família buthidae: Tityus serrulatus, Tityus bahiensis e Tityus obscurus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Structural and Biochemical Characterization of Peroxiredoxin Q beta from Xylella fastidiosa CATALYTIC MECHANISM and HIGH REACTIVITY

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

The tof-guard neuromuscular transmission monitor and its use in horses

Quando da utilização de bloqueadores neuromusculares, já foi enfatizado na literatura médica humana, que é de vital imporância a monitoração do bloqueio neuromuscular e que estes agentes nunca devem ser utilizados sem a mesma. O objetivo deste estudo foi o de avaliar o uso do monitor da transmissão neuromuscular TOF-Guard em eqüinos. Para tanto, doze eqüinos foram separados aleatoriamente para receberem como bloqueadores neuromusculares o pancurônio ou o atracúrio. Todos os eqüinos foram pré-medicados com romifidina, induzidos com diazepam e quetamina e mantidos com halotano. Foi administrado o atracúrio ou o pancurônio, seguindo-se a apnéia e início da ventilação mecânica controlada. O tempo entre a administração do relaxante muscular e a obtenção de um bloqueio máximo (T1=0), o retorno do T1 para 25% e da razão do TOF para 0,7 e o tempo do retorno do T1 de 25 para 75% foram mensurados. Concluiu-se que é de grande importância a monitoração do bloqueio neuromuscular quando da utilização de um bloqueador neuromuscular não-despolarizante, uma vez que ela torna o ato anestésico e cirúrgico mais seguro, com a utilização de doses adequadas que inviabilizam a superficialização do bloqueio durante o procedimento cirúrgico. O monitor TOF-Guard mostrou ser uma boa opção para a monitoração do bloqueio neuromuscular em eqüinos.

Coenzyme Q(10) and its effects in the treatment of neurodegenerative diseases

According to clinical and pre-clinical studies, oxidative stress and its consequences may be the cause or, at least, a contributing factor, to a large number of neurodegenerative diseases. These diseases include common and debilitating disorders, characterized by progressive and irreversible loss of neurons in specific regions of the brain. The most common neurodegenerative diseases are Parkinson's disease, Huntington's disease, Alzheimer's disease and amyotrophic lateral sclerosis. Coenzyme Q(10) (CoQ(10)) has been extensively studied since its discovery in 1957. It is a component of the electron transportation chain and participates in aerobic cellular respiration, generating energy in the form of adenosine triphosphate (ATP). The property of CoQ(10) to act as an antioxidant or a pro-oxidant, suggests that it also plays an important role in the modulation of redox cellular status under physiological and pathological conditions, also performing a role in the ageing process. In several animal models of neurodegenerative diseases, CoQ(10) has shown beneficial effects in reducing disease progression. However, further studies are needed to assess the outcome and effectiveness of CoQ(10) before exposing patients to unnecessary health risks at significant costs.

Collider aspects of flavor physics at high Q

This chapter of the "Flavor in the era of LHC" workshop report discusses flavor-related issues in the production and decays of heavy states at the LHC at high momentum transfer Q, both from the experimental and the theoretical perspective. We review top quark physics, and discuss the flavor aspects of several extensions of the standard model, such as supersymmetry, little Higgs models or models with extra dimensions. This includes discovery aspects, as well as the measurement of several properties of these heavy states. We also present publicly available computational tools related to this topic.

Expressão gênica dos REα, REβ e PR em tumores mamários de cadelas por meio do q-PCR

A expressão de receptores de estrógeno (ER) e progesterona (PR) por meio da técnica de q-PCR foi avaliada em 26 cadelas portadoras de neoplasias mamárias e cinco cadelas sem afecções mamárias (grupo controle). Os resultados mostraram que os três grupos de animais estudados - com tumor maligno ou benigno e controle - expressaram receptores de estrógeno alfa, beta e progesterona. A quantificação relativa mostrou tendência para uma expressão maior de receptores no grupo controle e menor no grupo de animais com neoplasias malignas. Além disso, observou-se expressão maior de ERα em relação ao ERβ, e as neoplasias malignas de origem mista apresentaram maiores concentrações dos receptores PR, ERα e ERβ que as neoplasias de origem epitelial.

The Saccharomyces cerevisiae COQ10 gene encodes a START domain protein required for function of coenzyme Q in respiration

Deletion of the Saccharomyces cerevisiae gene YOL008W, here referred to as COQ10, elicits a respiratory defect as a result of the inability of the mutant to oxidize NADH and succinate. Both activities are restored by exogenous coenzyme Q(2). Respiration is also partially rescued by COQ2, COQ7, or COQ8/ABC1, when these genes are present in high copy. Unlike other coq mutants, all of which lack Q(6), the coq10 mutant has near normal amounts of Q(6) in mitochondria. Coq10p is widely distributed in bacteria and eukaryotes and is homologous to proteins of the aromatic-rich protein family Pfam03654 and to members of the START domain superfamily that have a hydrophobic tunnel implicated in binding lipophilic molecules such as cholesterol and polyketides. Analysis of coenzyme Q in polyhistidine-tagged Coq10p purified from mitochondria indicates the presence 0.032-0.034 mol of Q(6)/mol of protein. We propose that Coq10p is a Q(6)-binding protein and that in the coq10 mutant Q(6) it is not able to act as an electron carrier, possibly because of improper localization.

Profiling the peptidome of the venom from the social wasp Agelaia pallipes pallipes

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