La proteómica y la metabolómica: los mecanismos de la enfermedad cardiovascular y el descubrimiento de biomarcadores

In the last decade, proteomics and metabolomics have contributed substantially to our understanding of cardiovascular diseases. The unbiased assessment of pathophysiological processes without a priori assumptions complements other molecular biology techniques that are currently used in a reductionist approach. In this review, we highlight some of the «omics» methods used to assess protein and metabolite changes in cardiovascular disease. A discrete biological function is very rarely attributed to a single molecule; more often it is the combined input of many proteins. In contrast to the reductionist approach, in which molecules are studied individually, «omics» platforms allow the study of more complex interactions in biological systems. Combining proteomics and metabolomics to quantify changes in metabolites and their corresponding enzymes will advance our understanding of pathophysiological mechanisms and aid the identification of novel biomarkers for cardiovascular disease.     

Abnormal ezrin localization is associated with clinicopathological features in invasive breast carcinomas

Summary The membrane-cytoskeleton crosslinker ezrin is associated with malignant progression and metastasis in human neoplasias. To study the role of ezrin in breast cancer, we first assessed ezrin expression in a panel of breast cancer cell lines by western blot and confocal microscopy. Western blot revealed no differences in total ezrin levels among these breast cell lines. However, immunofluorescence staining revealed that Estrogen receptor (ER)-positive, noninvasive and nontumorigenic cell lines concentrated ezrin at the apical surface, whereas invasive cell lines localized ezrin in motile structures (membrane ruffles and filopodia) but also had more diffuse cytoplasmic staining. We next studied ezrin expression in 509 breast carcinomas using tissue microarrays. Immunohistochemical staining for ezrin, p53, Ki-67, phospho-Akt, HER2, and hormonal receptors was performed. Ezrin staining in normal breast epithelium localized at the apical, but not lateral, cell surface, whereas, in most breast tumor cases (331, 70.3%), it localized in the cytoplasm. Complete membranous staining occurred in 89 (18.9%) samples, and apical staining was seen in 51 (10.8%) cases. There were significant positive associations between cytoplasmic ezrin localization and adverse tumor characteristics such as high grade, high level of Ki-67 expression, hormonal-receptor negativity, and lymph-node metastases. Apical ezrin staining was associated with favorable clinicopathological features and node-negative tumors. Membranous ezrin staining was associated with high grade, strong HER2 and p-Akt expression. In conclusion, the switch of ezrin localization from the apical membrane to either the complete membrane or to the cytoplasm is correlated with dedifferentiation and adverse features in invasive breast tumors and cancer cell lines.     

CPOE在韩国教学医院和普通医院使用情况的调查

本文介绍了调查CPOE在韩国的使用情况。原文曾在美国医学信息学协会杂志刊登。     

法舒地尔对脂多糖诱导的人脐静脉内皮细胞损伤的影响

目的:观察法舒地尔对脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVECs)损伤的影响,探讨其作用机制.方法:用0.1%的Ⅰ型胶原酶消化分离HUVECs,加入内皮细胞完全培养基中培养,采用胰蛋白酶进行消化传代培养,倒置显微镜下观察细胞形态并进行免疫组织化学鉴定.将HUVECs随机分为对照组(内皮细胞培养基培养)、LPS组(内皮细胞培养基+0.1 μg/mL LPS)和法舒地尔组(内皮细胞培养基+0.1 μg/mL LPS+3.275 μg/mL法舒地尔),Western blot法检测RhoA、ROCK2、p-ERM蛋白表达水平,实时荧光定量PCR检测RhoA、ROCK2和p-ERM mRNA表达量.结果:原代培养的内皮细胞在接种后24 h后完全贴壁生长,第3天融合呈铺路石样镶嵌排列,免疫组织化学检测可见第Ⅷ因子相关抗原阳性反应,证实为内皮细胞.LPS组RhoA、ROCK2和p-ERM蛋白及基因表达水平均显著高于对照组(P<0.01);而法舒地尔组ROCK2和p-ERM蛋白及基因表达均低于LPS组(P<0.05~P<0.01),但2组RhoA蛋白和基因表达水平差异均无统计学意义(P>0.05).结论:Rho/ROCK/ERM通路在LPS诱导的HUVECs细胞骨架改变中起重要作用,法舒地尔可拮抗LPS诱导的骨架蛋白重排等HUVECs损伤,其机制可能与抑制Rho/ROCK信号通路活化有关.     

The Preliminary Report of Pathological Changes of Epiretinal Membranes and Internal Limiting Membrane Removed during Idiopathic Macular Hole Surgery

Purpose:To investigate the pathological changes of epiretinal membranes(ERM)and internal limiting membrane(ILM)removed during idiopathie macular hole surgery.Methods:Ten consecutive patients with a unilateral idiopathic macular hole underwent pars plana vitrectomy(PPV)with the surgical removal of the ERMs overlying the hole and ILM surrounding the hole.The pathological features of the exciesed tissues were examined under the microscope.Results:According to the morphological changes,four ERMs showed cellular elements which looked like glia cells,macrophages,plasma cells,lymphocytes and fibroblast cells.Two of the ILM appeared as transparent membranes without cellular elements.The other eight ILM showed cellular elements on the transparent membranes.Conclusion:Our study supports the hypothesis that the tangential traction of vitreous and proliferative cellular elements on the inner surface of ILM causes indiopathic macular holes.Removal of the posterior cortical vitreous,ILM and proliferative cellular tissue is a valid treatment for IMH.     

Gene expression profiling in naïve and vaccinated rainbow trout after Yersinia ruckeri infection: insights into the mechanisms of protection seen in vaccinated fish

Despite the importance and success of vaccination against bacterial diseases in fish, little is known about the mechanisms of vaccine-induced disease resistance. In this study a known efficacious bacterial vaccine, to Enteric Redmouth Disease (ERM), was used to vaccinate rainbow trout, and sixty days later the fish were challenged with the causative agent of the disease, Yersinia ruckeri. The bacterial burden in the spleen, the spleen index, and the expression profiles of pro- and anti-inflammatory cytokines and marker genes for T helper (Th) cells in the spleen and gills were analyzed, in comparison to the profiles in naïve/challenged fish. As expected, the bacterial burden in the spleen of naïve fish increased over time and was correlated with the spleen index after Y. ruckeri challenge. The gene expression data showed that pro-inflammatory cytokines were upregulated post-infection in the spleen of both naïve and vaccinated fish after Y. ruckeri challenge although the pro-inflammatory cytokine expression was much lower in vaccinated fish compared to the naïve fish. A correlated expression between pro-inflammatory cytokines and anti-inflammatory cytokines was only seen in spleen of ERM vaccinated fish, where a Th1-like response was indicated by the correlated gene expression of IFN-γ, T-bet and IL-2. In contrast, in the gills, the inflammatory gene response was enhanced in vaccinated fish compared to naïve fish, but perhaps more importantly there was a strong upregulation of IL-22 which was negatively correlated with IFN-γ gene expression at this site. Thus, it is possible that different types of adaptive responses are on-going within the vaccinated fish during infection with Y. ruckeri, potentially affected by the site and stage of infection.     

Cocaine regulates ezrin–radixin–moesin proteins and RhoA signaling in the nucleus accumbens

The ezrin–radixin–moesin (ERM) proteins are a family of widely distributed membrane-associated proteins and have been implicated not only in cell-shape determination but also in signaling pathway. The nucleus accumbens (NAcc) is an important neuronal substrate mediating the effects of drugs of abuse. However, it has not been determined yet how ERM proteins are regulated in this site by drugs of abuse. Here we show in rat that the phosphorylation levels of ERM protein are dose- and time-dependently decreased in the NAcc by a single injection of cocaine (15 or 30 mg/kg i.p.). Further, we show that the amount of active RhoA, a small GTPase protein, is significantly reduced in the NAcc by cocaine, while the phosphorylation levels of ERM protein are also decreased by bilateral microinjections in this site of the Rho kinase inhibitors. Together, these results suggest that cocaine reduces phosphorylated ERM levels in the NAcc by making downregulation of RhoA–Rho kinase signaling, which may importantly contribute to initiate synaptic changes in this site leading to drug addiction.