Apo B concentration in the normal human aorta.

The concentration of LDL-protein (apo B) was determined by electroimmunoassay in the grossly normal intima of 15 human aortas obtained at autopsy. The mean apo B concentration was: 1.02 ± 0.05 SEM (range 0.16 – 2.51) mg per cm³ tissue, or higher than literature values of plasma apo B. No detectable amounts of apo B were found in the neighboring tunica media in any of the cases. In one case apo B concentrations were also measured in tissues from liver, lung, spleen, kidney, brain, myocardium and skeletal muscle, but all had values at least ten-fold lower than aortic intima, except liver (one-fourth intima value). The mean concentrations of human serum albumin (HSA) in the intima was 13.52 mg/cm³, or only one-fourth plasma concentration. Thus the aortic intima not only has the highest apo B values of the tissues tested, but in the intima apo B is retained to a greater degree than another plasma macromolecule. These results may be relevant to the fact that arterial intima is the primary site of atherogenesis.     

Biaxial mechanical properties of the human thoracic and abdominal aorta,common carotid,subclavian, renal and common iliac arteries

The biomechanics of large- and medium-sized arteries influence the pathophysiology of arterial disease and the response to therapeutic interventions. However, a comprehensive comparative analysis of human arterial biaxial mechanical properties has not yet been reported. Planar biaxial extension was used to establish the passive mechanical properties of human thoracic (TA, \(n=8\) ) and abdominal (AA, \(n=7\) ) aorta, common carotid (CCA, \(n=21\) ), subclavian (SA, \(n=12\) ), renal (RA, \(n=13\) ) and common iliac (CIA, \(n=16\) ) arteries from 11 deceased subjects ( \(54\pm 21\)  years old). Histological evaluation determined the structure of each specimen. Experimental data were used to determine constitutive parameters for a structurally motivated nonlinear anisotropic constitutive model. All arteries demonstrated appreciable anisotropy and large nonlinear deformations. Most CCA, SA, TA, AA and CIA specimens were stiffer longitudinally, while most RAs were stiffer circumferentially. A switch in anisotropy was occasionally demonstrated for all arteries. The CCA was the most compliant, least anisotropic and least frequently diseased of all arteries, while the CIA and AA were the stiffest and the most diseased. The severity of atherosclerosis correlated with age, but was not affected by laterality. Elastin fibers in the aorta, SA and CCA were uniformly and mostly circumferentially distributed throughout the media, while in the RA and CIA, elastin was primarily axially aligned and concentrated in the external elastic lamina. Constitutive modeling provided good fits to the experimental data for most arteries. Biomechanical and architectural features of major arteries differ depending on location and functional environment. A better understanding of localized arterial mechanical properties may support the development of site-specific treatment modalities for arterial disease.     

Day-night variation in the in vitro contractility of aorta and mesenteric and renal arteries in transgenic hypertensive rats

TGR(mREN2)27 (TGR) rats develop severe hypertension and an inverted circadian blood pressure profile with peak blood pressure in the daytime rest phase. The present study investigated the in vitro responsiveness of different arteries of TGR rats during day and night. Twelve-week-old TGR rats and normotensive Sprague-Dawley (SPRD) controls, synchronized to 12h light, 12h dark (LD 12:12) (light 07:00 19:00), were killed at 09:00 (during rest) and 21:00 (during activity), and endothelium-dependent relaxation by acetylcholine and vascular contraction by angiotensin II were studied by measuring isometric force in ring segments of abdominal aorta and mesenteric and renal arteries. In SPRD rats, consistent day-night variation was found, with greater responses to angiotensin II during the daytime rest span. In TGR rats, biological time-dependent differences were found in the renal vasculature, but not in the aorta and mesenteric artery. Relaxation of SPRD rat aorta and mesenteric artery by acetylcholine was greater at 09:00, whereas in TGR rats, day-night variation was absent (mesenteric artery) or inverted (aorta). In conclusion, based on the study of two time points, daynight variation in vascular contractility of aorta and mesenteric artery is blunted in TGR rats, whereas renal artery segments showed an unchanged daynight pattern compared to SPRD controls. (Chronobiology International, 18(4), 665 681, 2001)     

The content of lipoperoxidation products in normal and atherosclerotic human aorta.

To evaluate the role of lipid oxidation in atherogenesis the levels of lipid- and protein-bound products of peroxidation in normal and atherosclerotic areas of human aorta were investigated. The level of fluorescent (360/430 nm) lipid products was measured in chloroform-methanol extracts of aortic tissue. Normal intima, initial lesions and fatty streaks had a similar content of fluorescent substances. On the other hand, high level of fluorescent products was found in atherosclerotic plaques. Cholesterol covalently bound to proteins, which serve as a marker of lipoperoxidation, was measured by high performance liquid chromatography after mild alkaline hydrolysis of delipidated tissue protein samples. The levels of protein-bound cholesterol in initial lesions and fatty streaks were close to its content in uninvolved intima (59 +/- 18 and 92 +/- 18 vs. 70 +/- 13 nmol/g protein). The content of covalently bound cholesterol in atherosclerotic plaques was dramatically higher (90-fold) than in the normal tissue. In addition to protein-bound cholesterol, considerable amount of lipofuscin was revealed in the cells of atherosclerotic plaques, but not in the cells of normal intima, initial lesions or fatty streaks. Thus, the contents of all investigated lipid- and protein-bound products of lipoperoxidation in earlier atherosclerotic lesions were similar to their levels in normal tissue. It can be due to a low rate of oxidized product formation and/or high rate of its degradation in or elimination from the vessel wall.     

Innervation of the human internal carotid arteries under normal conditions and in stenosis]

The neurohistological investigation of a portion of the internal carotid artery removed in operation on the occasion of occlusion revealed afferent, cholinergic and adrenergic nerve elements randomly located in the examined area. The histochemical and electronmicroscopic investigation of the superior cervical sympathetic ganglion removed from 42 patient operated on the occasion of occlusion of the carotid artery revealed a depletion in the ganglia of synaptic active zones, focal absence of catecholamines and neurohistological materials suggests that a substantial role in the process of stenosing of vascular walls is played by sophisticated effects of innervation connections upon the vessel sheaths.     

Primary cultures of enzyme-isolated cells from normal and atherosclerotic human aorta

A technique has been developed for isolating cells from the intimal and medial layers of the human aorta by enzymatic dispersion. After mechanical separation of intima, media and adventitia the intima and the media were dispersed by collagenase and elastase. Enzyme-isolated cells seeded in the culture with at a frequency of 30 to 50%. In the primary culture differentiated aortic cells were morphologically heterogenous. It was possible to define four main types of cells according to their shape: polygonal, elongated, asymmetrical and stellate. Polygonal and stellate cells are found only in cultures of grossly normal intima, whereas elongated and asymmetric cells are found in practically all cultures. The ratio of elongated to asymmetric cells in cultures obtained from healthy aorta and atherosclerotic plaque is more or less the same at approximately 3:1. In cultures of fatty streaks the proportion of asymmetric cells exceeds 50%. Using immunofluorescence, all four types of cell were identified as smooth muscle cells. The possible reasons for the cellular polymorphism in primary culture and the prospects of utilizing this culture for the study of cellular aspects of atherosclerosis' pathogenesis are discussed.     

Caliber and media thickness of intrahepatic arteries in a normal human liver. A morphometric study

Morphometrical data on the intrahepatic arterial wall were studied over a wide range of vascular diameters (100-1708 microns) in one human liver. The liver tissue containing Araldite-filled blood vessels was embedded in the water-soluble glycolmethacrylate (GMA). The calculation of the correction factor for the morphometric data obtained from 3 microns sections is described. The influence of formaldehyde fixation on the volume of the liver proves to be negligible. During dehydration a linear shrinkage of about 3% occurs. After infiltration and embedding in GMA only 1% of this shrinkage remains. During the drying phase about 4.3% further linear stretching occurs. No significant "residual compression factor" was found. Araldite plays a significant role in the retention of the dimensions of the liver specimen during histologic processing, while the dimensional changes of the Araldite itself are negligible. A positive linear correlation was found between the media thickness and the radius of the vessel. The physiological consequences are discussed. It is concluded that in the morphometric analysis of the arterial wall it is essential to apply a standardized procedure in histologic processing and in the measuring of the inner vascular diameter. The advantages of our method, in which blood vessels are perfused with Araldite under physiological pressure, are discussed.     

Flow in arteries in the presence of stenosis

Of concern in the paper is a study of blood flow in an arterial segment having a stenosis. The artery is modelled as an initially stressed orthotropic elastic tube filled with a viscous incompressible fluid. The analysis is based on the assumption of the presence of a mild stenosis in the artery. Blood is treated as a Newtonian fluid. The effect of the surrounding connective tissues on the motion of the wall has been incorporated. Propagation of small amplitude harmonic waves, generated due to the flow of blood where the wave length is large compared to the radius of the arterial segment, is is considered in detail.     

Specificity and inhibition studies of human renal dipeptidase

Purified human renal dipeptidase was shown to exhibit no detectable activity against substrates that are characteristic for other known mammalian peptidases. The enzymic activities that were assayed were: aminopeptidase A, aminopeptidase B, aminopeptidase M, aminopeptidase P, and tripeptidase. A quantitative assay for renal dipeptidase was developed which measures the rate of release of glycine from glycylpeptides by pre-column derivatization of the amino acid with phenylisothiocyanate followed by high-performance liquid chromatography. The ratio of Vmax/Km for a series of dipeptides was used as an index of the enzyme's preference for substrates. According to the data obtained, the enzyme prefers that a bulky, hydrophobic group of the dipeptide be located at the N-terminal position. This suggests that the substrate-binding site of the enzyme may provide a hydrophobic pocket to accommodate the hydrophobic moiety at the N-terminus of the dipeptide. The unsaturated dipeptide substrate, glycyldehydrophenylalanine, was employed in spectrophotometric assays to provide kinetic analyses of enzymic inhibition. The inhibitory effect of dithiothreitol was immediate, and the kinetic data indicated reversible, competitive inhibition. These results suggest that the inhibitor competes with substrate for a coordination site of zinc within the active site of the enzyme. The reaction of renal dipeptidase with the transition-state peptide analog, bestatin, was time dependent, and velocity measurements were made after the inhibitor had been incubated with the enzyme until constant rates were observed. These steady-state rate measurements, made following preincubation of enzyme with inhibitor, were employed to show that bestatin caused apparent non-competitive inhibition of the enzyme. The inhibitory effect of the beta-lactam inhibitor, cilastatin, upon the oligomeric dipeptidase was shown to be competitive. Graphical analysis of this inhibition indicated that the subunits of the enzyme react independently during enzymic catalysis and that the catalytic event is not influenced by cooperativity between sites on the subunits. The conversion of leukotriene D4 to leukotriene E4 in the presence of human renal dipeptidase was demonstrated by HPLC procedures. This bioconversion reaction was quantitated by derivatizing the glycine produced by cleavage of the cysteinylglycine bond and isolating this derivative as a function of time. The relationship between the purified enzyme concentration and enzyme activity against leukotriene D4 was shown to be linear over the enzyme concentration range of 1 ng through 69 ng in this assay.(ABSTRACT TRUNCATED AT 400 WORDS)