In Situ DESI-MSI Lipidomic Profiles of Breast Cancer Molecular Subtypes and Precursor Lesions.

Clinically meaningful molecular subtypes for classification of breast cancers have been established, however, initiation and progression of these subtypes remain poorly understood. The recent development of desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) facilitates the convergence of analytical chemistry and traditional pathology, allowing chemical profiling with minimal tissue pretreatment in frozen samples. Here, we characterized the chemical composition of molecular subtypes of breast cancer with DESI-MSI. Regions of interest were identified, including invasive breast cancer (IBC), ductal carcinoma in situ (DCIS), and adjacent benign tissue (ABT), and metabolomic profiles at 200 µm elaborated using Biomap software and the Lasso method. Top ions identified in IBC regions included polyunsaturated fatty acids, deprotonated glycerophospholipids, and sphingolipids. Highly saturated lipids, as well as antioxidant molecules [taurine (m/z 124.0068), uric acid (m/z 167.0210), ascorbic acid (m/z 175.0241), and glutathione (m/z 306.0765)], were able to distinguish IBC from ABT. Moreover, luminal B and triple-negative subtypes showed more complex lipid profiles compared with luminal A and HER2 subtypes. DCIS and IBC were distinguished on the basis of cell signaling and apoptosis-related ions [fatty acids (341.2100 and 382.3736 m/z) and glycerophospholipids (PE (P-16:0/22:6, m/z 746.5099, and PS (38:3), m/z 812.5440)]. In summary, DESI-MSI identified distinct lipid composition between DCIS and IBC and across molecular subtypes of breast cancer, with potential implications for breast cancer pathogenesis. SIGNIFICANCE: These findings present the first in situ metabolomic findings of the four molecular subtypes of breast cancer, DCIS, and normal tissue, and add to the understanding of their pathogenesis.

Interactions of Lipid Polar Headgroups with Carbendazim Fungicide.

Carbendazim (MBC) is a fungicide widely used in agriculture which allows the high productivity of several cultures, a necessary condition considering the growing of the world population. Moreover, MBC has environmental impact mainly on the soil and water sources, and consequently, on animal and human lives. However, even though the toxicity of fungicides is well established, their action mechanism in cell membranes are not completely understood. Herein, we investigate the interaction of different polar headgroups: dimethyldioctadecylammonium bromide (DODAB), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DPPG); and different chain unsaturation degrees DPPC, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) with MBC. Lipid monolayers at the air/water interface were applied as mimetic systems of cell membranes to investigate the interaction with MBC dissolved in the ultrapure water subphase. It was found that the interaction is driven preferably by electrostatic forces of the headgroups, with higher affinity for DODAB (cationic), intermediate for DPPC (zwitterionic), and absent for DPPG (anionic), considering the monolayer in the condensed phase. DODAB-MBC electrostatic interaction was consistent with FTIR (cast films). We also investigated giant unilamellar vesicles (GUVs) of zwitterionic lipids (DPPC, POPC, and DOPC) with distinct chain unsaturations in the presence of MBC by confocal microscopy and molecular dynamic (MD) simulations. The results indicate that, unlike the chain unsaturation, the polar headgroups play key role on the lipid-MBC interaction.

Identificação dos perfis lipidômicos in situ dos tumores bifásicos da mama por DESI-MS (Desorption Electrospray Ionization/ Mass Spectrometry) / Identification of lipidomic profiles in situ of breast biphasic tumors by DESI-MS (Desorption Electrospray Ionization/ Mass Spectrometry)]

Os tumores bifásicos da mama formam um grupo heterogêneo de tumores que apresentam na histologia dois componentes, o epitelial e o estromal. Esses tumores podem ser benignos ou malignos, alguns podem recidivar ou dar metástase. O seu prognóstico varia conforme a sua classificação histológica, a presença de metástase e a adequabilidade do seu tratamento. Porém, por vezes, há ainda dificuldade em diferenciá-los quando há sobreposição das características histológicas que são utilizadas para realizar o diagnóstico no exame anatomopatológico. Visando um novo método para diferenciar esses tumores entre si, além do exame anatomopatológico habitual, buscou-se analisar esses tumores através de uma técnica da química analítica de ionização ambiente, utilizando espectrometria de massas, o DESI-MSI (Desorption Electrospray Ionization Mass Spectrometry Imaging). Foram utilizados cortes de amostras congeladas em lâminas para analisar no DESI-MSI e, através dessa análise, obteve-se os espectros de massas e as imagens químicas. A análise estatística foi realizada através do teste de Wilcoxon-Mann-Whitney. Nesse estudo encontramos maior intensidade dos ácidos graxos poli-insaturados (PUFAs); de fosfolipídios como as fosfatidilserina e fosfatidiletanolaminas; do ácido ascórbico e de peptídeos no sarcoma do estroma da mama e tumor Phyllodes maligno, comparados ao tumor Phyllodes borderline e, por sua vez, comparado ao tumor Phyllodes benigno, fibroadenomas e tecido normal. Alguns PUFAs, como o ácido araquidônico (m/z 303,2330) e o ácido adrênico (m/z 331,2643), são precursores dos eicosanóides e participam de vias pró-inflamatórias. Os PUFAs podem estar ligados a um fosfolipídio da membrana celular e, quanto mais estiver presente, menor a rigidez da membrana, o que diminui o gasto energético para a realização dos movimentos celulares, como a endocitose. O aumento da intensidade do ácido ascórbico nas células tumorais se deve provavelmente ao aumento da demanda metabólica de glicose pelas células tumorais; ocorrendo, portanto, aumento dos transportadores de glicose (GLUTS) e, como a molécula do ácido ascórbico e sua forma oxidada tem estrutura molecular semelhante à molécula da glicose, o ácido ascórbico seria carreado por esses transportadores para dentro da célula tumoral. A maior intensidade dos peptídeos no tecido maligno pode ser explicada pela maior demanda proteica da célula maligna; porém, além da importância dos peptídeos para construção das proteínas, sabe-se que alguns aminoácidos podem ser convertidos em metabólitos intermediários da glicólise aeróbia. Isso ocorre devido ao aumento da demanda de átomos de carbono para biossíntese de metabólitos. Essas diferenças lipidômicas e metabolômicas ocorrem pelo metabolismo diferenciado da célula maligna, a qual desenvolve mecanismos de adaptação para sobrevivência e proliferação. A análise das imagens químicas dos tumores bifásicos da mama obtidas por DESI-MSI mostrou que é possível diferenciar esses tumores. O uso futuro dessa técnica na rotina diagnóstica pode ser promissor devido a ser de execução rápida e de alta acurácia (AU)

Biphasic breast tumors form a heterogeneous group of tumors that present in histology two components, epithelial and stromal. These tumors may be benign or malignant, some may recur or metastasize. Its prognosis varies according to its histological classification, the presence of metastasis and the adequacy of its treatment. However, sometimes there is still difficulty in differentiating them when there is overlap of the histological features that are used to make the diagnosis in the pathological examination. Aiming at a new method to differentiate these tumors from each other, in addition to the usual pathological examination, we sought to analyze these tumors using an analytical chemistry technique of ambient ionization using mass spectrometry, the Desorption Electrospray Ionization Mass Spectrometry Imaging (DESI-MSI). Frozen specimen sections were used to analyze the DESI-MSI and, through this analysis, mass spectra and chemical images were obtained. Statistical analysis was performed using the Wilcoxon-Mann-Whitney test. In this study we found higher intensity of polyunsaturated fatty acids (PUFAs); phospholipids such as phosphatidylserine and phosphatidylethanolamines; ascorbic acid and peptides in breast sarcoma and malignant phyllodes tumor, compared to borderline phyllodes tumor and, in turn, compared to benign phyllodes tumor, fibroadenomas and breast normal tissue. Some PUFAs, such as arachidonic acid (m/z 303,2330) and adrenic acid (m/z 331,2643), are precursors of eicosanoids and participate in proinflammatory pathways. PUFAs may be linked to a cell membrane phospholipid and, the more it is present, the lower the membrane stiffness, which decreases the energy expenditure to perform cell movements, such as endocytosis. The increase in ascorbic acid intensity in tumor cells is probably due to the increased metabolic demand for glucose by tumor cells; there is therefore an increase in glucose transporters (GLUTS) and, since the ascorbic acid molecule and its oxidized form have a similar molecular structure to the glucose molecule, the ascorbic acid would be carried by these transporters into the tumor cell. The higher intensity of peptides in malignant tissue may be explained by the higher protein demand of the malignant cell; However, in addition to the importance of peptides for protein construction, it is known that some amino acids can be converted into intermediate metabolites of aerobic glycolysis. This is due to the increased demand for carbon atoms for metabolite biosynthesis. These lipidomic and metabolomic differences occur due to the differentiated metabolism of the malignant cell, which develops adaptation mechanisms for survival and proliferation. Analysis of chemical images of biphasic breast tumors obtained by DESI-MSI showed that it is possible to differentiate these tumors. The future use of this technique in the diagnostic routine may be promising due to its fast execution and high accuracy (AU)

A produção científica nacional sobre os direitos da criança hospitalizada / The national scientific production about on the rights of the hospitalized child

Este estudo objetivou identificar a produção científica nacional sobre os direitos da criança e do adolescente hospitalizados e analisar o papel do enfermeiro, enquanto profissional de saúde, na perspectiva desses direitos. Trata-se de uma revisão bibliográfica nas bases de dados LILACS e SciELO, no período de 1996 a 2008, em que foram analisados 13 artigos na íntegra. Os resultados revelam que, apesar de os artigos abordarem os direitos da criança hospitalizada, apenas um deles fazia referência à Resolução nº 41/95 que trata dos Direitos da Criança e do Adolescente. Conclui-se que é de suma importância que as enfermeiras pediátricas esclareçam e assegurem aos pais o direito de participação ativa na hospitalização de seu filho.

This study aim is to identify the national scientific production on the child?s rights and hospitalized adolescent and to analyze the paper of the nurse, as a health professional, in the perspective of these rights. One is about a bibliographical revision in the databases LILACS and SciELO, in the period of 1996 the 2008, where 13 articles in it complete form were analyzed. The results disclose that, although the articles approach the rights of hospitalized child, but 01 article make reference to the Resolution nº 41/95 wich deals with the Rights of the Child and Adolescent. The conclusion it is relevant that pediatric nurses clarify the parents about the right of active participation in the hospitalization of their kids or adolescents.

El objetivo de este estudio es identificar la producción científica nacional sobre los derechos del niño y adolescente hospitalizados y analizar el papel de la enfermera, como profesional de la salud, en la perspectiva de estos derechos. Uno de ellos es acerca de una revisión bibliográfica en las bases de datos LILACS y SciELO, en el período de 1996 hasta 2008, cuando 13 artículos en forma completa fueron analizados. Los resultados revelaron que, si bien los artículos enfoque de los derechos de los niños hospitalizados, sólo el artículo 01 hace referencia a la Resolución n º 41/95 que se ocupa de los Derechos del Niño y del Adolescente. Se concluye que es importante que las enfermeras pediátricas aclaren los padres sobre el derecho de participación activa en la hospitalización de sus hijos o adolescentes.