optimized-protocol-for-extraction-proteins-from-human-mitral

Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve

Analysis of the cellular proteome can help to elucidate the molecular mechanisms underlying diseases due to the development of technologies that permit ...

Centro Cardiologico Monzino IRCCS

No biological marker is currently available for evaluating pulmonary involvement and/or for monitoring the clinical course of sarcoidosis. The present pilot study focused on possible relationships between circulating plasma levels of surfactant protein type B (SP-B) and plasma receptor for advanced glycation end products (RAGE) and lung function abnormalities in patients with pulmonary sarcoidosis, since both SP-B and RAGE have been previously suggested as lung injury markers. The plasmatic levels of these two proteins were also investigated with respect to functional capacity, as assessed by a cardiopulmonary exercise test (CPET).

Opposite behavior of plasma levels surfactant protein type B and receptor for advanced glycation end products in pulmonary sarcoidosis.

In heart failure (HF) alveolar-capillary membrane is abnormal. Surfactant-derived proteins (SPs) and plasma receptor for advanced-glycation-end-products (RAGE) have been proposed as lung damage markers.

Surfactant-derived proteins as markers of alveolar membrane damage in heart failure.

It has long been known that Tissue Factor (TF) plays a role in blood coagulation and has a direct thrombotic action that is closely related to cardiovascular risk, but it is becoming increasingly clear that it has a much wider range of biological functions that range from inflammation to immunity. It is also involved in maintaining heart haemostasis and structure, and the observation that it is down-regulated in the myocardium of patients with dilated cardiomyopathy suggests that it influences cell-to-cell contact stability and contractility, and thus contributes to cardiac dysfunction. However, the molecular mechanisms underlying these coagulation-independent functions have not yet been fully elucidated. In order to analyse the influence of TF on the cardiomyocitic proteome, we used functional biochemical approaches incorporating label-free quantitative proteomics and gene silencing, and found that this provided a powerful means of identifying a new role for TF in regulating splicing machinery together with the expression of several proteins of the spliceosome, and mRNA metabolism with a considerable impact on cell viability.

Proteomics of tissue factor silencing in cardiomyocytic cells reveals a new role for this coagulation factor in splicing machinery control.

The long pentraxin PTX3 is an acute-phase multi-functional protein that might play both positive and detrimental effects under different pathophysiological conditions. We previously showed that statins down-regulate the release of PTX3 in human endothelial cells (ECs). The present study investigated the mechanism mediating this effect, its occurrence in other cells involved in atherogenesis, and whether it takes place in experimental atherosclerosis.

Atorvastatin reduces long pentraxin 3 expression in vascular cells by inhibiting protein geranylgeranylation.

This data article is related to the research article entitled Proteomics of Tissue Factor silencing in cardiomyocytic cells reveals a new role for this coagulation factor in splicing machinery control by Lento et al. [1]. Tissue Factor (TF) is a key player in the coagulation cascade, but it has additional functions ranging from angiogenesis, tumour invasion and, in the heart, the maintenance of the integrity of cardiac cells. This article reports the nano-LC-MS(E) analysis of the cardiomyocytic HL-1 cell line proteome and describes the results obtained from a Gene Ontology analysis of those proteins affected by TF-gene silencing.

Data for proteomic analysis of murine cardiomyocytic HL-1 cells treated with siRNA against tissue factor.

Gas exchange abnormalities are part of the heart failure (HF) syndrome and growing interest raised on possible biomarkers of alveolar-capillary unit damage. The present pilot single-center study sought to investigate the prognostic values of circulating surfactant protein type B (SP-B) in a cohort of systolic HF patients.

Plasma immature form of surfactant protein type B correlates with prognosis in patients with chronic heart failure. A pilot single-center prospective study.

[This corrects the article DOI: 10.1016/j.dib.2015.02.005.].

Corrigendum to "Data for proteomic analysis of murine cardiomyocytic HL-1 cells treated with siRNA against tissue factor" [Data Brief 3 (2015) 117-119].

The mitral valve is a highly complex structure which regulates blood flow from the left atrium to the left ventricle (LV) avoiding a significant forward gradient during diastole or regurgitation during systole. The integrity of the mitral valve is also essential for the maintenance of normal LV size, geometry, and function. Significant advances in the comprehension of the biological, functional, and mechanical behavior of the mitral valve have recently been made. However, current knowledge of protein components in the normal human mitral valve is still limited and complicated by the low cellularity of this tissue and the presence of high abundant proteins from the extracellular matrix. We employed here an integrated proteomic approach to analyse the protein composition of the normal human mitral valve and reported confident identification of 422 proteins, some of which have not been previously described in this tissue. In particular, we described the ability of pre-MS separation technique based on liquid-phase IEF and SDS-PAGE to identify the largest number of proteins. We also demonstrated that some of these proteins, e.g. αB-Crystallin, septin-11, four-and-a-half LIM domains protein 1, and dermatopontin, are synthesised by interstitial cells isolated from human mitral valves. These initial results provide a valuable basis for future studies aimed at analysing in depth the mitral valve protein composition and at investigating potential pathogenetic molecular mechanisms. Data are available via ProteomeXchange with identifier PXD004397.

Normal human mitral valve proteome: A preliminary investigation by gel-based and gel-free proteomic approaches.

Alteration of breathing pattern ranging from an increase of respiratory rate to overt hyperventilation during and after SCUBA diving is frequently reported and is associated with intrathoracic fluid overload. This study was undertaken to assess breathing efficiency after diving and the association with damage of alveolar cells. Ventilation efficiency (VE/VCO2) during maximal cardiopulmonary exercise test (CPET) before and 2h after a standard protocol dive has been analyzed in twelve professional males divers (39.5±10.5years). Furthermore, within 30min from surfacing, subjects underwent blood sample for surfactant derived proteins (SPs) determination, while thoracic ultrasound was performed at 30, 60, 90 and 120min. Dive consisted in a single quick descend to 18m of sea water, a 47min bottom stay and a direct ascent to the surface. CPET showed a preserved exercise performance with an increase of VE/VCO2 after diving (21.4±2.9 vs. 22.9±3.3, p<0.05). Mature SP-B increased while other SPs were unchanged. Ultrasound lung comets (ULC) were high in the first post-dive evaluation with a significant, but not complete, progressive reduction at 120min after surfacing. In conclusion we showed that, after a single dive, lung fluid increased with an increase of ventilation inefficiency and of the mature form of SP-B.

Article	Year
Opposite behavior of plasma levels surfactant protein type B and receptor for advanced glycation end products in pulmonary sarcoidosis. Respiratory medicine\| PubMed ID: 23948664	2013
Surfactant-derived proteins as markers of alveolar membrane damage in heart failure. PloS one\| PubMed ID: 25514679	2014
Proteomics of tissue factor silencing in cardiomyocytic cells reveals a new role for this coagulation factor in splicing machinery control. Journal of proteomics\| PubMed ID: 25668326	2015
Atorvastatin reduces long pentraxin 3 expression in vascular cells by inhibiting protein geranylgeranylation. Vascular pharmacology\| PubMed ID: 25849951	2015
Data for proteomic analysis of murine cardiomyocytic HL-1 cells treated with siRNA against tissue factor. Data in brief\| PubMed ID: 26217730	2015
Plasma immature form of surfactant protein type B correlates with prognosis in patients with chronic heart failure. A pilot single-center prospective study. International journal of cardiology\| PubMed ID: 26310985	2015
Corrigendum to "Data for proteomic analysis of murine cardiomyocytic HL-1 cells treated with siRNA against tissue factor" [Data Brief 3 (2015) 117-119]. Data in brief\| PubMed ID: 26977426	2015
Normal human mitral valve proteome: A preliminary investigation by gel-based and gel-free proteomic approaches. Electrophoresis\| PubMed ID: 27450324	2016
Diving and pulmonary physiology: Surfactant binding protein, lung fluid and cardiopulmonary test changes in professional divers. Respiratory physiology & neurobiology\| PubMed ID: 28467885	2017

Stefania Ghilardi

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