Accumulation of Plasma-Derived Lipids in the Lipid Core and Necrotic Core of Human Atheroma: Imaging Mass Spectrometry and Histopathological Analyses

2021 ◽  
Author(s):  
Kazunori Nakagawa ◽  
Mitsuru Tanaka ◽  
Tae Hun Hahm ◽  
Huu-Nghi Nguyen ◽  
Toshiro Matsui ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Foam Cell ◽  
Collagen Fibers ◽  
Necrotic Core ◽  
Surrounding Tissue ◽  
Intimal Thickening ◽  
Lipid Core ◽  
Cholesteryl Linoleate ◽  
Lipid Pool ◽  
Human Atheroma

Objective: To clarify the pathogenesis of human atheroma, the origin of deposited lipids, the developmental mechanism of liponecrotic tissue, and the significance of the oxidation of phospholipids were investigated using mass spectrometry-aided imaging and immunohistochemistry. Approach and Results: Atherosclerotic lesions in human coronary arteries were divided into 3 groups: pathological intimal thickening with lipid pool, atheroma with lipid core, and atheroma with necrotic core. The lipid pool and lipid core were characterized by the deposition of extracellular lipids. The necrotic core comprised extracellular lipids and liponecrotic tissue. The proportion of cholesteryl linoleate in cholesteryl linoleate+cholesteryl oleate fraction in the extracellular lipid and liponecrotic regions differed significantly from that of the macrophage foam cell–dominant region, and the plasma-derived components (apoB and fibrinogen) were localized in the regions. The liponecrotic region was devoid of elastic and collagen fibers and accompanied by macrophage infiltration in the surrounding tissue. Non–oxidized phospholipid (Non-OxPL), OxPL, and Mox macrophages were detected in the three lesions. In the atheroma with lipid core and atheroma with necrotic core, non-OxPL tended to localize in the superficial layer, whereas OxPL was distributed evenly. Mox macrophages were colocalized with OxPL epitopes. Conclusions: In human atherosclerosis, plasma-derived lipids accumulate to form the lipid pool of pathological intimal thickening, lipid core of atheroma with lipid core, and necrotic core of atheroma with necrotic core. The liponecrotic tissue in the necrotic core appears to be developed by the loss of elastic and collagen fibers. Non-OxPL in the accumulated lipids is oxidized to form OxPL, which may contribute to the lesion development through Mox macrophages.

scholarly journals ULTRASTRUCTURAL STUDIES ON THE LYMPHATIC ANCHORING FILAMENTS

1968 ◽  
Vol 36 (1) ◽  
pp. 129-149 ◽  
Author(s):  
L. V. Leak ◽  
J. F. Burke
Keyword(s):  
Connective Tissue ◽  
Extracellular Space ◽  
Collagen Fibers ◽  
Capillary Wall ◽  
Surrounding Tissue ◽  
Elastic Fibers ◽  
Lymphatic Capillary ◽  
Ultrastructural Studies ◽  
Outer Leaflet ◽  
Tissue Area

The fine structure of the lymphatic capillary and the surrounding tissue areas was investigated. Instead of a continuous basal lamina (basement membrane) surrounding the capillary wall, these observations revealed the occurrence of numerous fine filaments that insert on the outer leaflet of the trilaminar unit membrane of the lymphatic endothelium. These filaments appear as individual units, or they are aggregated into bundles that are disposed parallel to the long axis of the lymphatic capillary wall and extend for long distances into the adjoining connective tissue area among the collagen fibers and connective tissue cells. The filaments measure about 100 A in width and have a hollow profile. They exhibit an irregular beaded pattern along their long axis and are densely stained with uranyl and lead. These filaments are similar to the microfibrils of the extracellular space and the filaments observed in the peripheral mantle of the elastic fibers. Infrequently, connections between these various elements are observed, suggesting that the lymphatic anchoring filaments may also contribute to the filamentous units of the extracellular space. It is suggested that these lymphatic anchoring filaments connect the small lymphatics to the surrounding tissues and represent the binding mechansim that is responsible for maintaining the firm attachment of the lymphatic capillary wall to the adjoining collagen fibers and cells of the connective tissue area.

scholarly journals Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes of Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea)

2020 ◽  
Author(s):  
Pavel Roudnický ◽  
David Potěšil ◽  
Zbyněk Zdráhal ◽  
Milan Gelnar ◽  
Martin Kašný
Keyword(s):  
Mass Spectrometry ◽  
Laser Capture Microdissection ◽  
Bacterial Infections ◽  
Protein Localization ◽  
Model Organism ◽  
Surrounding Tissue ◽  
Laboratory Animals ◽  
Tissue Specific ◽  
Laser Capture ◽  
Eudiplozoon Nipponicum

AbstractEudiplozoon nipponicum (Goto, 1891) is a hematophagous monogenean ectoparasite which inhabits the gills of the common carp (Cyprinus carpio). Heavy infestation can lead to anemia and in conjunction with secondary bacterial infections cause poor health and eventual death of the host.This study is based on an innovative approach to protein localization which has never been used in parasitology before. Using laser capture microdissection, we dissected particular areas of the parasite body without contaminating the samples by surrounding tissue, and in combination with analysis by mass spectrometry obtained tissue-specific proteomes of tegument, intestine, and parenchyma of our model organism, E. nipponicum. We successfully verified the presence of certain functional proteins (e.g. cathepsin L) in tissues where their presence was expected (intestine) and confirmed that there were no traces of these proteins in other tissues (tegument and parenchyma). Additionally, we identified a total of 2,059 proteins, including 72 peptidases and 33 peptidase inhibitors. As expected, the greatest variety was found in the intestine and the lowest variety in the parenchyma.Our results are significant on two levels. Firstly, we demonstrated how one can localize all proteins in one analysis and without using laboratory animals (antibodies for immunolocalization of single proteins). Secondly, this study offers the first complex proteomic data on not only the E. nipponicum but within the whole class of Monogenea, which was from this point of view until recently neglected.

scholarly journals Positive Expression of Human Cytomegalovirus Phosphoprotein 65 in Atherosclerosis

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Zhe Wang ◽  
Jun Cai ◽  
Mingming Zhang ◽  
Xiaojing Wang ◽  
Hongjie Chi ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Human Cytomegalovirus ◽  
Hcmv Infection ◽  
Foam Cell ◽  
Collagen Fibers ◽  
High Rate ◽  
Early Protein ◽  
Formalin Fixed Paraffin Embedded ◽  
Masson Trichrome Staining

Previous studies showed that human cytomegalovirus (HCMV) is associated with atherosclerosis. However, local vascular atherosclerosis related HCMV infection and protein expression remain unclear. This study aimed to assess the relationship between HCMV infection and atherosclerosis. Formalin-fixed, paraffin-embedded peripheral artery specimens were obtained from 15 patients with atherosclerosis undergoing vascular surgery from 2008 to 2010 at Zhongnan Hospital, Wuhan University. Pathological analyses were carried out after hematoxylin and eosin (H&E) and Masson trichrome staining. In situ hybridization and immunohistochemistry with two different monoclonal antibodies were employed to detect HCMV nucleic acids and proteins, respectively. H&E and Masson trichrome staining showed homogeneous extracellular matrix in femoral artery, while smooth muscle fibers were interlaced with collagen fibers; in carotid artery, inflammatory cell infiltration, foam cell vascular change, cholesterol crystals, and layered collagen fibers were observed. In situ hybridization showed no expression of HCMV nucleic acids in all 15 cases. Immunohistochemical staining for protein immediate-early protein (IE1 72) was negative in all cases, while phosphoprotein 65 (pp65) expression was detected in 14 cases. A high rate of positive pp65 signals was found in patients with atherosclerosis, suggesting that local HCMV infection may be associated with the pathogenesis of atherosclerosis. Further studies on this relationship are warranted.

Atomic Force Microscopy Study of Atherosclerosis Progression in Arterial Walls

2016 ◽  
Vol 22 (2) ◽  
pp. 311-325 ◽  
Author(s):  
Peter S. Timashev ◽  
Svetlana L. Kotova ◽  
Galina V. Belkova ◽  
Ekaterina V. Gubar’kova ◽  
Lidia B. Timofeeva ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Cross Sections ◽  
Microscopy Study ◽  
Collagen Fibers ◽  
Lipid Core ◽  
Force Microscopy ◽  
Atherosclerosis Progression ◽  
Plaque Type ◽  
Atomic Force ◽  
Thickened Intima

AbstractCardiovascular disease remains the leading cause of mortality worldwide. Here we suggest a novel approach for tracking atherosclerosis progression based on the use of atomic force microscopy (AFM). Using AFM, we studied cross-sections of coronary arteries with the following types of lesions: Type II—thickened intima; Type III—thickened intima with a lipid streak; Type IV—fibrotic layer over a lipid core; Type Va—unstable fibrotic layer over a lipid core; Type Vc—very thick fibrotic layer. AFM imaging revealed that the fibrotic layer of an atherosclerotic plaque is represented by a basket-weave network of collagen fibers and a subscale network of fibrils that become looser with atherosclerosis progression. In an unstable plaque (Type Va), packing of the collagen fibers and fibrils becomes even less uniform than that at the previous stages, while a stable fibrotic plaque (Vc) has significantly tighter packing. Such alterations of the collagen network morphology apparently, led to deterioration of the Type Va plaque mechanical properties, that, in turn, resulted in its instability and propensity to rupture. Thus, AFM may serve as a useful tool for tracking atherosclerosis progression in the arterial wall tissue.

scholarly journals Synthesis of (2β,3α,6-2H3)cholesteryl linoleate and cholesteryl oleate as internal standards for mass spectrometry

Steroids ◽  
2016 ◽  
Vol 107 ◽  
pp. 1-9 ◽  
Author(s):  
Yusuke Miura ◽  
Shu-Ping Hui ◽  
Rojeet Shrestha ◽  
Takahisa Hiruma ◽  
Seiji Takeda ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cholesteryl Oleate ◽  
Internal Standards ◽  
Cholesteryl Linoleate

scholarly journals Blood iron level and vulnerable coronary atherosclerotic plaques

2021 ◽  
Vol 10 (Supplement_1) ◽  
Author(s):  
A Oleksiak ◽  
M Kruk ◽  
KI Rucinska ◽  
K Marcinkiewicz ◽  
M Demkow ◽  
...  
Keyword(s):  
Regression Analysis ◽  
Vulnerable Plaque ◽  
Foam Cell ◽  
Necrotic Core ◽  
Atherosclerotic Plaques ◽  
Iron Level ◽  
Plaque Component ◽  
Science Centre ◽  
National Science ◽  
Low Attenuation Plaque

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Science Centre, Poland BACKGROUND Vulnerable plaque rupture is one of the causes of acute coronary syndromes. Preliminary research indicate that iron might accelerate the oxidation of low density lipoproteins (LDL) which can then be taken up by the LDL receptor on macrophages leading to their development into foam cells. Foam cell infiltration and necrotic core expansion are key events in atherogenesis and vulnerable plaque formation. However, the potential pathophysiological roles of iron in plaque development remain uncertain. PURPOSE The aim of the study was to investigate the relationship between iron and the type and composition of atherosclerotic plaques in the coronary arteries. METHODS In 200 patients with ≥1 stenosis ≥50% in computed tomography coronary angiography (CTCA) made for clinical indications we assessed: free iron level, the presence of high-risk plaque features: low-attenuation plaque (LAP), napkin-ring sign (NRS), positive remodeling (PR) and spotty calcium (SC) (CT Coronary, Syngo, Siemens), type of plaque (calcified, mixed, non-calcified) and their composition (calcified, fibrous, fibro-fatty, necrotic core) (QAngioCT, Medis). Fibro-fatty and necrotic core were analyzed together as vulnerable plaque component. The study was financed by the National Science Centre (2016/21/N/NZ5/01450). RESULTS Of 200 patients (125 men, 66 ± 10 years), the mean iron level (µg/dl) was 91 ±30 for women and 103 ±33 for men (p = 0.5). 3 patients had iron deficiency and 2 patients had iron overload. In CTCA analysis there were 815 calcified, 344 non-calcified and 438 mixed plaques. There was a trend in correlation between iron level and non-calcified plaque presence (p = 0.06). LAP was detected in 56 patients, NRS in 83, PR in 132, and SC in 125. Patients with LAP had higher iron levels (113 vs 93 µg/dl; p < 0.001). There was no association between iron and NRS, PR or SC (p > 0.05). In univariate regression analysis, the predictors of LAP were iron (p < 0.001) and male gender (p = 0.01). In multivariate regression analysis, iron was an independent predictor of LAP (p < 0.001; OR 1.02; 95%CI 1.01-1.03). Higher iron levels correlated with more fibro-fatty (p = 0.009) and necrotic core (p = 0.02); less calcified (p = 0.04); and with no relation to fibrous (p = 0.9), thus higher iron levels were associated with greater vulnerable plaque component (p = 0.003). CONCLUSIONS Higher iron levels are more likely to be associated with low-attenuation plaque and a greater vulnerable component of atherosclerotic plaques.

Pathology of stable coronary artery disease

ESC CardioMed ◽  
2018 ◽  
pp. 1315-1320
Author(s):  
Michael Joner ◽  
Maria Isabel Castellanos ◽  
Anna Bulin ◽  
Kristin Steigerwald
Keyword(s):  
Coronary Artery Disease ◽  
Smooth Muscle ◽  
Coronary Artery ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Necrotic Core ◽  
Intimal Thickening ◽  
Intraplaque Haemorrhage ◽  
Artery Disease ◽  
Plaque Destabilization

Coronary artery disease remains the major cause of morbidity and mortality on a global scale. Intimal thickening and fatty streaks represent early adaptive vascular changes, which are often regressive. Pathological intimal thickening represents the earliest progressive atherosclerotic lesion characterized by a focal accumulation of smooth muscle cells and acellular areas, often associated with lipid pools. Fibroatheroma is characterized by a necrotic core and can be split into early and late fibroatheroma, where macrophage apoptosis and defective efferocytosis play important roles in lesion progression. Intraplaque hypoxia is believed to result in neovascularization with subsequent intraplaque haemorrhage because of immature and leaky microvessels. Due to excessive cholesterol in remnants of erythrocyte membranes, intraplaque haemorrhage may result in rapid progression of necrotic core and plaque destabilization. Healed plaque rupture has been well delineated as an important mechanism of gradual luminal narrowing, where changes in collagen deposition allow recognition of rupture and healing sites. Calcification results from apoptosis of smooth muscle cells and macrophages or may also be caused by active release of cellular vesicles involved in calcium haemostasis. Extracellular matrix changes are associated with progression of atherosclerosis, while integrin signalling has been recognized as an important outside-in transcellular target of the inflammatory response.

scholarly journals Imaging Mass Spectrometry and Proteome Analysis of Marek’s Disease Virus-Induced Tumors

mSphere ◽  
2019 ◽  
Vol 4 (1) ◽  
Author(s):  
V. I. Pauker ◽  
L. D. Bertzbach ◽  
A. Hohmann ◽  
A. Kheimar ◽  
J. P. Teifke ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Disease Virus ◽  
Imaging Mass Spectrometry ◽  
Marek's Disease Virus ◽  
Marek's Disease ◽  
Surrounding Tissue ◽  
Marek’S Disease Virus ◽  
Marek’S Disease ◽  
Induced Tumors ◽  
Laser Capture

ABSTRACTThe highly oncogenic alphaherpesvirus Marek’s disease virus (MDV) causes immense economic losses in the poultry industry. MDV induces a variety of symptoms in infected chickens, including neurological disorders and immunosuppression. Most notably, MDV induces transformation of lymphocytes, leading to T cell lymphomas in visceral organs with a mortality of up to 100%. While several factors involved in MDV tumorigenesis have been identified, the transformation process and tumor composition remain poorly understood. Here we developed an imaging mass spectrometry (IMS) approach that allows sensitive visualization of MDV-induced lymphoma with a specific mass profile and precise differentiation from the surrounding tissue. To identify potential tumor markers in tumors derived from a very virulent wild-type virus and a telomerase RNA-deficient mutant, we performed laser capture microdissection (LCM) and thereby obtained tumor samples with no or minimal contamination from surrounding nontumor tissue. The proteomes of the LCM samples were subsequently analyzed by quantitative mass spectrometry based on stable isotope labeling. Several proteins, like interferon gamma-inducible protein 30 and a 70-kDa heat shock protein, were identified that are differentially expressed in tumor tissue compared to surrounding tissue and naive T cells. Taken together, our results demonstrate for the first time that MDV-induced tumors can be visualized using IMS, and we identified potential MDV tumor markers by analyzing the proteomes of virus-induced tumors.IMPORTANCEMarek’s disease virus (MDV) is an oncogenic alphaherpesvirus that infects chickens and causes the most frequent clinically diagnosed cancer in the animal kingdom. Not only is MDV an important pathogen that threatens the poultry industry but it is also used as a natural virus-host model for herpesvirus-induced tumor formation. In order to visualize MDV-induced lymphoma and to identify potential biomarkers in an unbiased approach, we performed imaging mass spectrometry (IMS) and noncontact laser capture microdissection. This study provides a first description of the visualization of MDV-induced tumors by IMS that could be applied also for diagnostic purposes. In addition, we identified and validated potential biomarkers for MDV-induced tumors that could provide the basis for future research on pathogenesis and tumorigenesis of this malignancy.

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