Different Types Of Atherosclerotic Lesions Research Articles

Local Accumulation of Lymphocytes in the Intima of Human Aorta Is Associated with Giant Multinucleated Endothelial Cells: Possible Explanation for Mosaicism of Atherosclerosis.

Distribution of different types of atherosclerotic lesions in the arterial wall is not diffuse, but is characterized by mosaicism. The causes of such distribution remain to be established. At the early stages of atherogenesis, low-density lipoprotein (LDL) particles and immune cells penetrate into the intimal layer of the arterial wall through the endothelium. In adult humans, the luminal surface of the arterial wall is a heterogeneous monolayer of cells with varying morphology including typical endothelial cells (ECs) and multinucleated variant endothelial cells (MVECs). We hypothesized that distribution of MVECs in the endothelial monolayer can be related to the distribution pattern of early atherosclerotic lesions. We obtained en face preparations of intact adult (22–59 years old) aortic wall sections that allowed us to study the endothelial monolayer and the subendothelial layer. We compared the distribution of MVECs in the endothelial monolayer with the localization of early atherosclerotic lesions in the subendothelial layer, which were characterized by lipid accumulation and immune cell recruitment. In primary culture, MVECs demonstrated increased phagocytic activity compared to mononuclear ECs. Moreover, we have shown that unaffected aortic intima contained associates formed as a result of aggregation and/or fusion of LDL particles that are non-randomly distributed. This indicated that MVECs may be involved in the accumulation of LDL in the subendothelial layer through increased transcytosis. Interaction of LDL with subendothelial cells of human aorta in primary culture increased their adhesive properties toward circulating immune cells. Study of unaffected aortic intima revealed non-random distribution of leukocytes in the subendothelial layer and increased localization of CD45+ leukocytes in the subendothelial layer adjacent to MVECs. Together, our observations indicate that MVECs may be responsible for the distribution of atherosclerotic lesions in the arterial wall by participating in LDL internalization and immune cell recruitment.

Heteroplasmic Variants and Copy Number of Mitochondrial DNA in Different Types of Atherosclerotic Lesions of Human Aorta

Heteroplasmic Variants and Copy Number of Mitochondrial DNA in Different Types of Atherosclerotic Lesions of Human Aorta

Use of Primary Macrophages for Searching Novel Immunocorrectors.

In this mini-review, the role of macrophage phenotypes in atherogenesis is considered. Recent studies on distribution of M1 and M2 macrophages in different types of atherosclerotic lesions indicate that macrophages exhibit a high degree of plasticity of phenotype in response to various conditions in microenvironment. The effect of the accumulation of cholesterol, a key event in atherogenesis, on the macrophage phenotype is also discussed. The article presents the results of transcriptome analysis of cholesterol-loaded macrophages revealing genes involved in immune response whose expression rate has changed the most. It turned out that the interaction of macrophages with modified LDL leads to higher expression levels of pro-inflammatory marker TNF-α and antiinflammatory marker CCL18. Phenotypic profile of macrophage activation could be a good target for testing of novel anti-atherogenic immunocorrectors. A number of anti-atherogenic drugs were tested as potential immunocorrectors using primary macrophage-based model.

Variability of heteroplasmic mitochondrial DNA mutations in different types of atherosclerotic lesions in human aorta: NGS data

Variability of heteroplasmic mitochondrial DNA mutations in different types of atherosclerotic lesions in human aorta: NGS data

Quantitative analysis of the expression of caspase 3 and caspase 9 in different types of atherosclerotic lesions in the human aorta

The existing data on apoptotic processes in human atherosclerotic lesions is insufficient and is often contradictory. This study was undertaken to evaluate the levels of the expression of key apoptosis-related genes, namely, caspase 3 (CASP3) and caspase 9 (CASP9) in the normal (non-atherosclerotic) intima of the human aorta in comparison with those in different types of atherosclerotic lesions. Twenty-five autopsy samples of thoracic aorta were examined by polymerase chain reaction (PCR) analysis. The study revealed that the expressions of CASP3 and CASP9 genes were changed in different types of atherosclerotic lesions in course of the progression of the disease, but not in a unanimous way. The mRNA expression of CASP3 was found to be steadily decreasing with the progression of atherosclerosis while the expression of CASP9 showed a pattern which can be described as a “bell-shaped” relationship between gene mRNA expression and the type of atherosclerotic lesion, with the maximum being observed in fatty streaks. The fall in CASP3 expression may be associated with cellular senescence as well as with the domination of necrotic processes in atherosclerotic lesions, as shown by electron microscopic analysis. Our study provides novel quantitative data on the expression of CASP3 and CASP9 genes in different atherosclerotic lesions in the human aorta and thus, might assist in better understanding of the processes occurring during the development of lesions in human atherogenesis.

Mosaicism of mitochondrial genetic variation in atherosclerotic lesions of the human aorta.

Objective. The aim of the present study was an analysis of heteroplasmy level in mitochondrial mutations 652delG, A1555G, C3256T, T3336C, 652insG, C5178A, G12315A, G13513A, G14459A, G14846A, and G15059A in normal and affected by atherosclerosis segments of morphologically mapped aortic walls. Methods. We investigated the 265 normal and atherosclerotic tissue sections of 5 human aortas. Intima of every aorta was divided according to morphological characteristics into segments with different types of atherosclerotic lesions: fibrous plaque, lipofibrous plaque, primary atherosclerotic lesion (fatty streak and fatty infiltration), and normal intima from human aorta. PCR-fragments were analyzed by a new original method developed in our laboratory on the basis of pyrosequence technology. Results. According to the obtained data, mutations G12315A and G14459A are significantly associated with total and primary atherosclerotic lesions of intimal segments and lipofibrous plaques (P ≤ 0.01 and P ≤ 0.05, accordingly). Mutation C5178A is significantly associated with fibrous plaques and total atherosclerotic lesions (P ≤ 0.01). A1555G mutation shows an antiatherosclerotic effect in primary lesion in lipofibrous plaques (P ≤ 0.05). Meanwhile, G14846A mutation is antiatherogenic for lipofibrous plaques (P ≤ 0.05). Conclusion. Therefore, mutations C5178A, G14459A, G12315A, A1555G, and G14846A were found to be associated with atherosclerotic lesions.

Changes of mitochondria in atherosclerosis: Possible determinant in the pathogenesis of the disease

Electron-microscopic analysis of atherosclerotic lesions demonstrated a high variability in the ultrastructural appearance of mitochondria in human aortic atherosclerotic lesions compared with the appearance of mitochondria in the normal parts of the aortic intima. This prompted us to suggest that the structural variations in the appearance of mitochondria might reflect the existence of somatic mutations in the human mitochondrial genome which could be a determinant of atherosclerosis. To test this hypothesis, we have compared the levels of heteroplasmy for several mitochondrial mutations previously proposed to be associated with different types of atherosclerotic lesions. The homogenates of unaffected aortic intimae and lipofibrous plaques of 12 male aortas were compared to reveal the average level of heteroplasmy for A1555G, C3256T, T3336T, G12315A, G14459A, and G15059A mutations of human mitochondrial genome. It has been shown at least four mutations of mitochondrial genome, namely, A1555G in MT-RNR1 gene, C3256T in MT-TL1 gene, G12315A in MT-TL2 gene, and G15059A in MT-CYB gene have significantly higher prevalence and mean value in lipofibrous plaques as compared to non-atherosclerotic intima, and therefore are associated with atherosclerosis. Somatic mutations in the human mitochondrial genome might play a role in the development of atherosclerosis. The mitochondrial mutations observed in our study should encourage further exploration of the concept that mitochondrial DNA heteroplasmy might be used as a biomarker of atherogenesis.

C-reactive protein levels are associated with the progression of atherosclerotic lesions in rabbits.

Elevated plasma levels of C-reactive protein (CRP) are associated with increased risk of cardiovascular disease. CRP immunoreactive protein is also detected in the lesions of atherosclerosis. However, it is not known whether the CRP contents of atherosclerotic lesions are associated with the initiation and progression of atherosclerosis. To examine this hypothesis, we investigated different types of atherosclerotic lesions of rabbits fed with a cholesterol-rich diet for 6, 12, 16, and 28 weeks and examined their relationship with CRP. We measured the aortic atherosclerotic area, macrophages, and smooth muscle cells along with CRP contents in the lesions. Atherosclerotic lesions of aortas began to form at 6 weeks and were characterized by accumulation of macrophages in the intima, and lesions became more fibrotic in the advanced stage. Both plasma CRP levels and the lesional CRP contents were associated with the lesion size. Our results suggest that plasma CRP, as well as lesional CRP, associated with the formation and progression of atherosclerotic lesions.

Peculiarities of cell composition and cell proliferation in different type atherosclerotic lesions in carotid and coronary arteries

Increased cell proliferation in early atherosclerotic lesions is recognized as an essential event in atherogenesis but the levels of cell proliferation in the different stages of atherosclerotic plague formation in different types of human large arteries are still insufficiently studied. In the present work, we studied intima thickness and proliferation of newly “infiltrated” hematogenous and resident cells in atherosclerotic lesions of the carotid and coronary arteries and compared these parameters with those in the aorta, which we reported in an earlier publication (Orekhov et al. [8]). Analysis of intima thickness and proliferation in grossly unaffected intima and in different types of atherosclerotic lesions (initial lesions, fatty streaks, lipofibrous plaques, and fibrous plaques) revealed that, although there were similar tendencies in the change of the infiltration levels of hematogenous cells and proliferation in different types of arteries, there were significant quantitative differences between different types of arteries. Hematogenous cells in lipofibrous plaques of the coronary and carotid arteries were found to account for a third and almost for a half of the total cell population, respectively, while atherosclerotic lesions in the aorta, as shown by us previously, contain no more than 15% of hematogenous cells. This suggests that the contribution of hematogenous cells to the development of atherosclerosis in the carotid and coronary arteries appears to be more significant than in the aorta. Despite the differences in the numbers of accumulating hematogenous cells in the intima, a similar “bell-shaped” dependence of cell numbers on the lesion type, involved the following sequence: unaffected intima – initial lesions – fatty streaks – lipofibrous plaques – fibrous plaques, was detected in the coronary and carotid arteries. The visualization of PCNA-positive cells in atherosclerotic and unaffected zones of the coronary and carotid arteries revealed similar patterns of the distribution of proliferating cells. The maximum numbers of PCNA-positive resident cells were identified in lipofibrous plaques. The changes in the total cell numbers were found to be accompanied by the changes in the numbers of both proliferating resident cells and proliferating hematogenous cells. According to our knowledge, this is the first report that provides factual data about the similarities and differences in cell composition and proliferation between different types of large arteries in which the development of atherosclerosis is of crucial importance.

Association Analysis of Allelic Variants of USF1 in Coronary Atherosclerosis

USF1 regulates the transcription of more than 40 cardiovascular related genes and is well established as a gene associated with familial combined hyperlipidemia, a condition increasing the risk for coronary heart disease. No detailed data, however, exists on the impact of this gene to the critical outcome at the tissue level: different types of atherosclerotic lesions. We analyzed the USF1 in 2 autopsy series of altogether 700 middle-aged men (the Helsinki Sudden Death Study) with quantitative morphometric measurements of coronary atherosclerosis. SNP rs2516839, tagging common USF1 haplotypes, associated with the presence of several types of atherosclerotic lesions, particularly with the proportion of advanced atherosclerotic plaques (P=0.02) and area of calcified lesions (P<0.001) of the coronary arteries. Importantly, carriers of risk alleles of rs2516839 also showed a 2-fold risk for sudden cardiac death (genotype TT versus CC; OR 2.10, 95% CI 1.17 to 3.75, P=0.04). The risk effect of rs2516839 was present also in aorta samples of the men. Our findings in this unique study sample suggest that USF1 contributes to atherosclerosis, the pathological arterial wall phenotype resulting in coronary heart disease and in its most dramatic consequence-sudden cardiac death.

Distribution and prevalence of inducible nitric oxide synthase in atherosclerotic vessels of long-term cholesterol-fed rabbits

The expression of inducible nitric oxide synthase (iNOS) as well as its functional activity has recently been reported in atherosclerotic lesions. The aim of the present study was to evaluate the expression of iNOS in various arteries of rabbits fed a long-term but low-level cholesterol-enriched diet which promotes different types of atherosclerotic lesions resembling human diseased vessels. No iNOS expression was revealed in arteries from control rabbits and in fatty streaks found in carotid and femoral arteries from hypercholesterolemic rabbits. In transitional lesions from the thoracic and abdominal aortas, the coronary and pulmonary arteries, a punctiform iNOS staining was detected in the intima. When lesions were more advanced, iNOS expression was found more intense and diffuse and localized in the subendothelial layer as well as in the media. Smooth muscle cell accumulation in intimal layers of the arteries is a marker of the degree of evolution of the atherosclerotic lesion; since we found a correlation between the smooth muscle cell infiltration in the intima and the iNOS expression in the intima and the subendothelial layer, our results suggest a link between the severity of the lesion and the iNOS expression.

Histopathology and pathogenesis of plaque instability and thrombus formation.

Our knowledge of the pathogenesis of plaque instability has undergone profound changes in recent years. Research in this field has been driven by the fact that atherosclerosis and its thrombotic complications continue to be the major cause of mortality and morbidity throughout the industrialized world. The different types of atherosclerotic lesions, mechanisms of atherosclerotic progression, plaque vulnerability and rupture are now better understood. This has led to evolution of therapeutic strategies designed to stabilize atherosclerotic plaque and to reduce progression. Further-more, knowledge of mechanisms leading to thrombosis after plaque rupture have led to the development of antithrombotic strategies to prevent and reduce complications arising from such an event. This review will describe the histopathology and pathogenesis leading to plaque instability, the factors associated with subsequent rupture and assess the role of thrombosis in the progression of atherosclerotic disease. We will focus on current therapeutic strategies to identify and reduce vulnerable plaques and speculate on future areas for research.

Effect of lercanidipine and its (R)-enantiomer on atherosclerotic lesions induced in hypercholesterolemic rabbits.

The in vivo antiatherogenic activity of the calcium antagonist lercanidipine and its (R)-enantiomer was investigated in two different types of atherosclerotic lesions (hyperplastic and fatty-streak lesions) in rabbits. Lercanidipine (0.3, 1, and 3 mg kg(-1) week(-1)) as well as its (R)-enantiomer at 3 mg kg(-1) week(-1) were given by subcutaneous injection for 10 weeks to White New Zealand rabbits, with cholesterol feeding beginning at week 2. The hyperplastic lesion was obtained by positioning a hollow silastic collar around one carotid artery, while aortic fatty streak lesions were induced by cholesterol feeding. In untreated animals (n=5), 14 days after collar positioning an intimal hyperplasia was clearly detectable: the arteries without collar showed a intima/media (I/M) ratio of 0.03+/-0.02, whereas in carotids with a collar the ratio was 2+/-0.42. In lercanidipine-treated animals a significant and dose-dependent effect on intimal hyperplasia was observed. I/M ratios were 0.73+/-0.4, 0.42+/-0.1, 0.32+/-0.1 for 0.3, 1, and 3 mg kg(-1) week(-1), respectively (P<0.05). The lercanidipine enantiomer (3 mg kg(-1) week(-1)) was as effective as the racemate (0.41+/-0.11). Proliferation of smooth muscle cells, assessed by incorporation of BrdU into DNA, was reduced by about 50%, 70%, 85%, and 80% by lercanidipine (0.3, 1, and 3 mg kg(-1) week(-1)) and its (R)-enantiomer, respectively. The area of fatty-streaks in the aorta (n = 11-15) was significantly reduced by lercanidipine (3 mg kg(-1) week(-1), 16% vs 27%, P<0.05), a trend was observed also with lower doses. When different segments of the aorta were considered (arch, thoracic, abdominal) a significant and dose-dependent effect in the thoracic and abdominal aorta was observed also at lower doses. The (R)-enantiomer was as effective as lercanidipine. These results suggest a direct antiatherosclerotic effect of lercanidipine, independent of modulation of risk factors such as hypercholesterolemia and/or hypertension as demonstrated by the absence of stereoselectivity.

Cell proliferation in normal and atherosclerotic human aorta: proliferative splash in lipid-rich lesions

Local accumulation of cells (hypercellularity) in the intima of the arterial wall as a result of cell proliferation is recognized as one of the major manifestations of human atherosclerosis. In the present study we have used a monoclonal antibody against PCNA to identify the proliferative activity, in uninvolved intima of human aorta classified as diffuse intimal thickening, and in different types of atherosclerotic lesions: specifically, initial lesion, fatty streak, fibroatheroma and fibrous plaque. As compared with a diffuse intimal thickening, the cell number in the initial lesions, fatty streaks and in a fibrolipid plaque (fibroatheroma) was 1.5–3-fold higher, while the cellularity in a fibrous plaque (fibrotic lesion) was lower than in a fibroatheroma and comparable with the cell number in the initial lesions. Using monoclonal antibodies, inflammatory cells (T- and B-lymphocytes as well as monocytes-macrophages) have been revealed in the intima. However, most (84–93%) of the intimal cells were noninflammatory cells classified as resident cells possessing the antigens of smooth muscle cells and pericytes as well as a small number of cells unidentifiable with the antibodies used. The highest number of proliferating cells was found in a fibroatheroma (11-fold higher as compared with a diffuse intimal thickening). A significant, but lesser increase of PCNA-positive cells was revealed in other types of lesions, too. The proliferative `splash' in lipid-rich lesions suggests a relationship between the lipid accumulation in atherosclerotic intima and the stimulation of proliferation. The highest proliferative index of resident cells (i.e. percentage of the PCNA-positive cells among the total number of resident cells) was revealed in fibrotic lesions. It was approximately eight-fold higher than in a diffuse intimal thickening. The proliferative index of inflammatory cells considerably exceeded that of resident cells. However, in all types of atherosclerotic lesions and in a diffuse intimal thickening it showed no significant differences and was similar to the proliferative index of inflammatory cells isolated from peripheral blood. This suggests that an increased number of resident cells in atherosclerotic lesions can be explained by stimulation of their proliferative activity, whereas an altered inflammatory cell number is rather a result of their penetration from the blood into the subendothelial intima with a constant proliferative index.

Comparative analysis of plasminogen activator inhibitor-1 expression in different types of atherosclerotic lesions in coronary arteries from human heart explants.

Clinical manifestations of coronary heart disease result primarily from the progressive development of atherosclerotic plaques and subsequent thrombus formation: processes which may be accelerated by an enhanced expression of plasminogen activator inhibitor (PAI-1) in the vessel wall. In the present study, content and expression of PAI-1 were comparatively analyzed in human coronary arteries in relation to the presence and severity of atherosclerotic lesions. Segments of coronary arteries obtained from heart explants (n = 15) were classified by the presence and types of atherosclerotic lesions. Antigen and activity levels of PAI-1 were determined in protein extracts of intimal and medial layers. In situ hybridization and immunohistochemical analyses were performed on serial sections of representative tissue specimens. Total PAI-1 antigen consistently increased from macroscopically normal areas (MNAs) to early lesions (ELs) and to maximal levels in fibrous (FPs) and calcified (CPs) plaques. No PAI activity was detected, although PAI-1 in its free form was present in all vascular specimens. Both free PAI-1 and PAI-1 complexed with plasminogen activators were significantly increased in extracts of advanced lesions. However, there was a 2-3 fold molar excess of free versus complexed PAI-1 in FPs and CPs. These findings suggest the presence of relevant amounts of PAI-1 in its substrate rather than in its inhibitor conformation in areas of advanced lesions. Compared with MNAs, PAI-1 mRNA was strongly expressed within the thickened intima of ELs. The highest PAI-1 expression was observed in FPs and CPs, being mainly localized in areas surrounding the necrotic cores in co-localization with infiltrating macrophages. PAI-1 content is consistently increased in relation to the severity of the lesions in atherosclerotic coronary arteries. The concomitant elevation of PAI-1 mRNA suggests that the PAI-1 increase in regulated by local synthesis in the areas of atherosclerotic lesions.

Effect of lacidipine on fatty and proliferative lesions induced in hypercholesterolaemic rabbits.

1. The in vivo antiatherogenic activity of the calcium antagonist, lacidipine, was investigated in two different types of atherosclerotic lesions (proliferative and fatty lesions) induced in rabbits. 2. The proliferative lesion was obtained by positioning a hollow silastic collar around one carotid artery, while aortic fatty lesions were induced by cholesterol feeding. Cholesterol (1%) and lacidipine (1, 3, and 10 mg kg-1) were given daily mixed with standard diet for 8 weeks to White New Zealand rabbits. The intimal hyperplasia (proliferative lesion) was induced 6 weeks after dietary and drug treatment started. 3. The neointimal formation was determined by measuring cross sectional thickness of intimal (I) and medial (M) tissue of fixed arteries. In untreated animals (n = 5), 14 days after collar positioning an intimal hyperplasia was clearly detectable: the arteries with no collar (sham) showed an I/M tissue ratio of 0.03 +/- 0.02, whereas in the carotid with collar the ratio was 0.62 +/- 0.12. In lacidipine-treated animals a significant and dose-dependent effect on proliferative lesions at all three doses tested, was observed. I/M ratios were 0.47 +/- 0.02, 0.40 +/- 0.09, 0.32 +/- 0.02 for doses 1, 3, and 10 mg kg-1 day-1, respectively (P < 0.05). 4. The fatty lesion extent was significantly reduced by lacidipine at the 10 mg kg-1 day-1 dose, although a trend was also observed with lower dosage. 5. These results suggest a direct antiatherosclerotic effect of lacidipine, independent of modulation of risk factors such as hypercholesterolaemia and/or hypertension. Furthermore, the proliferative lesions are apparently more sensitive to lacidipine than are lipid-rich lesions.

Fate of fibrinogen in human arterial intima.

Fibrinogen and fibrinogen/fibrin-related antigen (total FRA) was measured in human normal intima and different types of atherosclerotic lesions and mural thrombi. The amount showed marked variation between groups of tissue samples, but within each group there was a significant correlation between levels of total FRA and low density lipoprotein (LDL), suggesting that some common factor must influence their influx or retention. The total FRA were analyzed by gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting with antisera to whole fibrinogen and fragments D and E, and fibrinopeptide A (FPA). All intimal samples (but not thrombi) contained fragment X, the first product of plasmin digestion of fibrinogen, but fragment Y was present in only half the samples, and no core-fragment E containing FPA was detected in any sample, suggesting that fibrinogenolysis is limited. By contrast, all samples contained fragment E, which was negative for FPA, so presumably derived from fibrin; they also contained fragments D-dimer and DY, which are characteristic degradation products of cross-linked fibrin. There were no differences between samples obtained during reconstructive vascular surgery and samples obtained at autopsy, so the patterns appear to represent the steady state. This implies that within the intima there is continuous formation of cross-linked fibrin and continuous fibrinolysis, both processes generating fragments that may have atherogenic properties.

Fibronectin distribution in human aortic intima and atherosclerotic lesions: concentration of soluble and collagenase-releasable fractions

Fibronectin is associated with cell attachment and migration and interacts with fibrin, collagen and glycosaminoglycans; thus, it may be a factor in the focal proliferation of smooth muscle cells and collagen in atherosclerosis. We have measured, by rocket immunoelectrophoresis, the concentrations of soluble and collagenase-releasable fibronectin in normal human aortic intima and different types of atherosclerotic lesions. Soluble fibronectin concentration showed no significant difference between normal intima and lesions, but was 6–8-times higher than expected on the basis of plasma concentration and molecular mass. The concentration free in the interstitial fluid was about 3-times the expected level, suggesting that it originates from local synthesis as well as plasma insudation. In tissue, about half the fibronectin appeared to be reversibly associated with tissue components. Incubation with collagenase released fibronectin equal to twice the soluble fraction from normal intima and early proliferative lesions. In more advanced plaques that were accumulating lipid, the amount released was significantly higher ( P < 0.05) and more than 3-times the soluble fraction, suggesting that it might be involved in lipid accumulation. However, there was no correlation between release of fibronectin and bound low-density lipoprotein.

Heterogeneity of human aortic cells in regard to RNA content.

Cells of human aorta were isolated by dispersing the tissue with collagenase and elastase. The isolated cells were stained in suspension by the acridine orange fluorescent stain. The intensity of red fluorescence (greater than 580 nm) corresponding to the RNA content was measured in each individual cell and registered in a FACS II flow cytofluorometer. It was established that a cell population of human aorta is heterogenous with respect to RNA content. In a population of isolated cells, one can distinguish two subpopulations: (A) small cells with low RNA content, and (B) large cells with high RNA content. The ratio of both cell types varies in intima and media, and different types of atherosclerotic lesions. The share of cells belonging to subpopulation A is lower in media compared to intima. In intima, the number of these cells grows with the degree of atherosclerotic lesion. Possible reasons for the discovered metabolic heterogeneity of human aortic cells and prospects for the application of flow cytofluorometry to a research into cellular mechanisms of atherosclerosis is discussed.

Morphogenesis of aortic and coronary atherosclerosis in North India—a composite study

This paper deals with a composite gross, microscopic, and biochemical study of aortal and coronary arteries obtained from unselected medicolegal autopsies on subjects from 3 to 105 years of age in the Delhi area. Twenty-nine coronary arteries were studied grossly and microscopically; of these, biochemical analysis for total lipids and lipid fractions was made on 42 samples of normal intima and different types of atherosclerotic lesions. Eighty-eight aortas were studied microscopically in a similar manner. The study confirms previous observations that both in the aorta and coronary arteries, progression of early fatty streaks to more established lesions is distinctly slow in the local Indian population and that the frequency of fibrous plaques and complicated lesions is less than in the white population in the West. A diffuse sudanophilic thickening of the intima, as reported in the aorta, was observed in the coronary arteries also, starting as early as the second decade and increasing in incidence thereafter. This lesion is interpreted as a variant of the atherosclerotic lesion, representing a mode of evolution of the atherosclerotic process. The morphologic and biochemical studies show a sequential evolution of changes observed in the normal intima at one end of the spectrum, through diffuse intimal thickening, fatty streaks, and fibrous plaques to complicated lesions at the other end. A considerable overlap and histogenetic kinship of the lesions was obvious. The lipid pattern of the coronary intima showed interesting differences from that of the aorta, but in both vessels, it was basically similar to that reported in the Western white population. The observed geographic differences in the extent and severity of atherosclerosis cannot, therefore, be ascribed to differences in the quality and quantity of lipid deposition. The studies suggest two main bases for the observed differences. First, for reasons not fully understood, the intimal reaction to atherogenic stimuli among Indians is frequently of a diffuse nature, which does not compromise the lumen in an eccentric manner as the atheroma does. Second, fibrin deposits are rare in the early lesions of Indians. It is possible that these factors may account for tardy progression of the lesions to the occlusive variety. An unusual lesion of aortic medial calcification was found with high frequency. Its pathogenesis is obscure.