Some fibrinolytic properties of the human arteries and veins walls

The plasminogen activator of normal and atherosclerotic different arteries was studied with the histochemical method of Todd. An increase of plasminogen activator in atherosclerotic arteries of adventitia was found. The inhibition of plasmin fibrinolysis of intima-media and adventitia of normal and atherosclerotic different arteries was studied by means of the slide sandwich technique according to Noordhoek Hegt. In atherosclerotic arteries there was an increase of plasmin inhibitory activity of the intima-media layer in comparison with normal arteries. The mean plasmin inhibitory activity was higher in the vein wall of lower part of the body than in the higher one.     

Transport in rat vessel walls. I. Hydraulic conductivities of the aorta, pulmonary artery, and inferior vena cava with intact and denuded endothelia

In this study, filtration flows through the walls of the rat aorta, pulmonary artery (PA), and inferior vena cava (IVC), vessels with very different susceptibilities to atherosclerosis, were measured as a function of transmural pressure (DeltaP), with intact and denuded endothelium on the same vessel. Aortic hydraulic conductivity (L(p)) is high at 60 mmHg, drops approximately 40% by 100 mmHg, and is pressure independent to 140 mmHg. The trends are similar in the PA and IVC, dropping 42% from 10 to 40 mmHg and flat to 100 mmHg (PA) and dropping 33% from 10 to 20 mmHg and essentially flat to 60 mmHg (IVC). Removal of the endothelium renders L(p)(DeltaP) flat: it increases L(p) of the aorta by approximately 75%, doubles L(p) of the PA, and quadruples L(p) of the IVC. Specific resistance (1/L(p)) of the aortic endothelium is approximately 47% of total resistance; i.e., the endothelium accounts for approximately 47% of the DeltaP drop at 100 mmHg. The PA value is 55% at >40 mmHg, and the IVC value is 23% at 10 mmHg. L(p) of the intact aorta, PA, and IVC are order 10(-8), 10(-7), and 5 x 10(-7) cm.s(-1).mmHg(-1), and wall thicknesses are 145.8 microm (SD 9.3), 78.9 microm (SD 3.3), and 66.1 microm (SD 4.1), respectively. These data are consistent with the different wall structures of the three vessels. The rat aortic L(p) data are quantitatively consistent with rabbit L(p)(DeltaP) (Tedgui A and Lever MJ. Am J Physiol Heart Circ Physiol 247: H784-H791, 1984; Baldwin AL and Wilson LM. Am J Physiol Heart Circ Physiol 264: H26-H32, 1993), suggesting that intimal compression under pressure loading may also play a role in L(p)(DeltaP) in these other vessels. Despite very different driving DeltaP, nominal transmural water fluxes of these three vessels are very similar and, therefore, cannot alone account for their differences in disease susceptibility. The different fates of macromolecular tracers convected by these water fluxes into the walls of these vessels may account for this difference.     

Ca2+ removal mechanisms in rat cerebral resistance size arteries.

Tissue blood flow and blood pressure are each regulated by the contractile behavior of resistance artery smooth muscle. Vascular diseases such as hypertension have also been attributed to changes in vascular smooth muscle function as a consequence of altered Ca2+ removal. In the present study of Ca2+ removal mechanisms, in dissociated single cells from resistance arteries using fura-2 microfluorimetry and voltage clamp, Ca2+ uptake by the sarcoplasmic reticulum and extrusion by the Ca2+ pump in the cell membrane were demonstrably important in regulating Ca2+. In contrast, the Na+-Ca2+ exchanger played no detectable role in clearing Ca2+. Thus a voltage pulse to 0 mV, from a holding potential of -70 mV, triggered a Ca2+ influx and increased intracellular Ca2+ concentration ([Ca2+]i). On repolarization, [Ca2+]i returned to the resting level. The decline in [Ca2+]i consisted of three phases. Ca2+ removal was fast immediately after repolarization (first phase), then plateaued (second phase), and finally accelerated just before [Ca2+]i returned to resting levels (third phase). Thapsigargin or ryanodine, which each inhibit Ca2+ uptake into stores, did not affect the first but significantly inhibited the third phase. On the other hand, Na+ replacement with choline+ did not affect either the phasic features of Ca2+ removal or the absolute rate of its decline. Ca2+ removal was voltage-independent; holding the membrane potential at 120 mV, rather than at -70 mV, after the voltage pulse to 0 mV, did not attenuate Ca2+ removal rate. These results suggest that Ca2+ pumps in the sarcoplasmic reticulum and the plasma membrane, but not the Na+-Ca2+ exchanger, are important in Ca2+ removal in cerebral resistance artery cells.     

Exercise reduces angiotensin II responses in rat femoral veins

The control of blood flow during exercise involves different mechanisms, one of which is the activation of the renin-angiotensin system, which contributes to exercise-induced blood flow redistribution. Moreover, although angiotensin II (Ang II) is considered a potent venoconstrictor agonist, little is known about its effects on the venous bed during exercise. Therefore, the present study aimed to assess the Ang II responses in the femoral vein taken from sedentary and trained rats at rest or subjected to a single bout of exercise immediately before organ bath experiments. Isolated preparations of femoral veins taken from resting-sedentary, exercised-sedentary, resting-trained and exercised-trained animals were studied in an organ bath. In parallel, the mRNA expression of prepro-endothelin-1 (ppET-1), as well as the ET_A and ET_B receptors, was quantified by real-time PCR in this tissue. The results show that, in the presence of L-NAME, Ang II responses in resting-sedentary animals were higher compared to the other groups. However, this difference disappeared after co-treatment with indomethacin, BQ-123 or BQ-788. Moreover, exercise reduced ppET-1 mRNA expression. These reductions in mRNA expression were more evident in resting-trained animals. In conclusion, either acute or repeated exercise adapts the rat femoral veins, thereby reducing the Ang II responses. This adaptation is masked by the action of locally produced nitric oxide and involves, at least partially, the ET_B- mediated release of vasodilator prostanoids. Reductions in endothelin-1 production may also be involved in these exercise-induced modifications of Ang II responses in the femoral vein.     

Early changes in dissected small vessels: experimental study on rat arteries and veins

This study was undertaken to investigate the cause of the early endothelial damage that is seen at sites of microvascular anastomosis and in particular to study the possibility of a connection between damage to the vasa vasorum and subsequent endothelial denudation. Rat femoral vessels were subjected to a variety of experimental injuries, including simple dissection, clamping, and ligation. The vessels were examined in longitudinal section by light microscopy at intervals ranging from 5 minutes to 1 day. The endothelial cells were counted and the numbers were analyzed statistically. In addition, the anatomy of the vasa vasorum was studied using india ink perfusion. Simple dissection of the femoral vessels and excision of the vasa vasorum without interruption of blood flow were followed by ischemic lesions of the tunica media with subendothelial edema and ballooning and exfoliation of endothelial cells. Endothelial denudation reached a maximum level in 30 minutes. Adherence of leukocytes was found on damaged endothelial cells. Mural thrombi were seen in 13.6 percent of arteries and in 40 percent of veins following simple dissection.     

Transport of bacteria in porous media: II. A model for convective Transport and growth

A model is presented for the coupled processes of bacterial growth and convective transport of bacteria has been modeled using a fractional flow approach. The various mechanisms of bacteria retention can be incorporated into the model through selection of an appropriate shape of the fractional flow curve. Permeability reduction due to pore plugging by bacteria was simulated using the effective medium theory. In porous media, the rates of transport and growth of bacteria, the generation of metabolic products, and the consumption of nutrients are strongly coupled processes. Consequently, the set of governing conservation equations form a set of coupled, nonlinear partial differential equations that were solved numerically. Reasonably good agreement between the model and experimental data has been obtained indicating that the physical processes incorporated in the model are adequate. The model has been used to predict the in situ transport and growth of bacteria, nutrient consumption, and metabolite production. It can be particularly useful in simulating laboratory experiments and in scaling microbial-enhanced oil recovery or bioremediation processes to the field. (c) 1994 John Wiley & Sons, Inc.     

Differential expression of neuropilin-1 and neuropilin-2 in arteries and veins.

Neuropilin-1 (np1) and neuropilin-2 (np2) are receptors for class-3 semaphorins and for several isoforms of VEGF. We have cloned and characterized two chick isoforms of np2 cDNA. Expression patterns of np1, np2, and ephrin-B2 were compared in the developing vascular system of 24-72 h old chick embryos. We show for the first time that np2 is expressed in blood vessels in vivo from the earliest stages of their formation. In contrast to ephrin-B2, both np1 and np2 are expressed in blood islands of 24 h old chick embryos. At 48-72 h, np1 expression is localized preferentially in arteries with an expression pattern that resembles that of ephrin-B2. In contrast, np2 is expressed preferentially in veins. Thus, neuropilins may play a role in determining the arterial or venous identity of blood vessels.     

Length-tension relationships of small arteries, veins, and lymphatics from the rat mesenteric microcirculation

The passive and active length-tension relationships of isolated rat mesenteric lymphatics ( approximately 150 microm ID), and adjacent small arteries ( approximately 240 microm) and veins ( approximately 275 microm) were compared under isometric conditions using a wire myograph. About 60% of the lymphatic vessels developed spontaneous contractions in physiological saline solution at nominal preload. To maximally activate smooth muscle, 145 mM K(+) + 5 x 10(-5) M norepinephrine was used for arteries, and 145 mM K(+) + 1 x 10(-6) M substance P was used for lymphatics and veins. In response, arteries exhibited monotonic force development to a plateau level, whereas lymphatics and veins showed biphasic force development, consisting of a transient force peak followed by partial relaxation to a plateau over approximately 5 min. The passive and the active length-tension curves were similar in shape among all three vessels. However, the maximal active tension of arteries (3.4 +/- 0.42 mN/mm) was significantly greater than peak active tension (0.59 +/- 0.04 mN/mm) or plateau tension (0.20 +/- 0.04 mN/mm) in small veins and greater than peak active tension (0.34 +/- 0.02 mN/mm) or plateau tension (0.21 +/- 0.02 mN/mm) in lymphatics. Maximal active medial wall stress was similar between lymphatics and veins but was approximately fivefold higher in small arteries. For lymphatics, the pressure calculated from the optimal preload was significantly higher than that found previously in isobaric studies of isolated lymphatics, suggesting the capacity to operate at higher than normal pressures for increased responsiveness. Our results represent the first mechanical comparisons of arterial, venous, and lymphatic vessels in the same vasculature.     

Gamma-secretase inhibitor treatment promotes VEGF-A-driven blood vessel growth and vascular leakage but disrupts neovascular perfusion

The Notch signaling pathway is essential for normal development due to its role in control of cell differentiation, proliferation and survival. It is also critically involved in tumorigenesis and cancer progression. A key enzyme in the activation of Notch signaling is the gamma-secretase protein complex and therefore, gamma-secretase inhibitors (GSIs)--originally developed for Alzheimer's disease--are now being evaluated in clinical trials for human malignancies. It is also clear that Notch plays an important role in angiogenesis driven by Vascular Endothelial Growth Factor A (VEGF-A)--a process instrumental for tumor growth and metastasis. The effect of GSIs on tumor vasculature has not been conclusively determined. Here we report that Compound X (CX), a GSI previously reported to potently inhibit Notch signaling in vitro and in vivo, promotes angiogenic sprouting in vitro and during developmental angiogenesis in mice. Furthermore, CX treatment suppresses tumor growth in a mouse model of renal carcinoma, leads to the formation of abnormal vessels and an increased tumor vascular density. Using a rabbit model of VEGF-A-driven angiogenesis in skeletal muscle, we demonstrate that CX treatment promotes abnormal blood vessel growth characterized by vessel occlusion, disrupted blood flow, and increased vascular leakage. Based on these findings, we propose a model for how GSIs and other Notch inhibitors disrupt tumor blood vessel perfusion, which might be useful for understanding this new class of anti-cancer agents.     

Adrenergic innervation of the large arteries and veins of the semiarboreal rat snake Elaphe obsoleta

Fluorescence histochemistry was used to study the adrenergic innervation of the large arteries and veins at six points along the body of the semiarboreal rat snake Elaphe obsoleta. Apart from the vessels adjacent to the heart, there was a marked contrast in the density of adrenergic innervation of anterior and posterior systemic arteries and veins. The anterior arteries and veins have little adrenergic innervation in contrast to the extremely dense innervation of the arteries and veins posterior to the heart. The innervation pattern is consistent with known physiological adjustments to gravity and suggests a mechanism for regulating dependent blood flow via sympathetic nerves. In comparison to the posterior systemic arteries, parallel segments of pulmonary artery taken from the same body position of Elaphe contained a much sparser innervation by adrenergic nerves. The sparser innervation can be correlated with less gravitational disturbance in the pulmonary artery, which is relatively short in this and in other arboreal snakes.     

Shoot orientation affects vessel size, shoot hydraulic conductivity and shoot growth rate in Vitis vinifera L.

Vitis vinifera L. plants were grown in containers and each plant's single shoot was orientated upwards or downwards. Some plants were trained first upwards, then downwards, then again upwards (N-shaped plants). Vegetative growth was reduced in plants trained downwards compared to that in upward and N-shaped plants. Shoot growth rate slowed in downward shoot portions, but only after the apex had grown downwards for at least 10 internodes. Shoot hydraulic conductivity k _h, measured after elimination of xylem embolisms, was lower in downward than in upward plants. In N-shaped plants k _h was higher in the upward-growing shoot portions, and lower in the central, downward-growing portion. Shoot- and leaf-specific conductivities were also lower in downward than in upward shoot portions. Xylem cross-sectional area and xylem structure (number of wedges, number of vessels per unit xylem area) differed little in the three orientations. In contrast, vessel diameter and the sum of vessel cross-sectional areas were significantly smaller in downward than in upward shoot portions. These differences could explain the reduction in conductivity observed in the downward-orientated shoot portions. The measurements taken on N-shaped plants showed that the decreases in k _h and in vessel size were a result of shoot orientation, not shoot bending.     

Contractile responses to rat urotensin II in resting and depolarized basilar arteries

The effects of human urotensin II (hUII) on the vascular tone of different animal species has been studied extensively. However, little has been reported on the vasoactive effects of rat urotensin (rUII) in murine models. The aim of the present study was to investigate the effects of rUII on vasoreactivity in rat basilar arteries. Basilar arteries from adult male Wistar rats (300–350 g) were isolated, cut in rings, and mounted on a small vessel myograph to measure isometric tension. rUII concentrations were studied in both resting and depolarized state. To remove endothelial nitric oxide effects from the rUII response, we treated selected arterial rings with N_ω-nitro-L-arginine methyl ester (L-NAME). 10 μM rUII produced a potent vasoconstrictor response in rat basilar arteries with intact endothelium, while isometric forces remained unaffected in arterial rings treated with lower rUII concentrations. Although L-NAME did not have a significant effect on 10 μM rUII-evoked contraction, it slightly increased arterial ring contraction elicited by 1 μM rUII. In depolarized arteries, dose-dependent rUII increased depolarization-induced contractions. This effect was suppressed by L-NAME. Our results show that the rat basilar artery has a vasoconstrictor response to rUII. The most potent vasoconstrictor effect was produced by lower doses of rUII (0.1 and 1 μM) in depolarized arteries with intact endothelium. This effect could facilitate arterial vasospasm in vascular pathophysiological processes such as subarachnoid hemorrhage and hypertension, when sustained depolarization and L-type Ca²⁺ channel activation are present.     

Macromolecular organization and in vitro growth characteristics of scaffold-free neocartilage grafts.

Recent advances in tissue engineering offer considerable promise for the repair of focal lesions in articular cartilage. Here we describe (1) the macromolecular organization of tissue-engineered neocartilage grafts at light and electron microscopic levels, (2) their in vitro development, and (3) the effect of chondrocyte dedifferentiation, induced by monolayer expansion, on their resultant structure. We show that grafts produced from primary cultures of chondrocytes are hyaline in appearance with identifiable zonal strata as evidenced by cell morphology, matrix organization, and immunohistochemical composition. Like native articular cartilage, their surface zone contains type I collagen, surface zone proteoglycan, biglycan and decorin with type II collagen, aggrecan, chondroitin sulfate, chondroitin-4-sulfate, and keratan sulfate, becoming more prominent with depth. Assessment of cell viability by Live/Dead staining and cell-cycle analysis with BrDU suggest that the in vitro tissue has a high cellular turnover and develops through both appositional and interstitial growth mechanisms. Meanwhile, cell-tracker studies with CMFDA (5-chloromethyl-fluorescein diacetate) demonstrate that cell sorting in vitro is not involved in their zonal organization. Finally, passage expansion of chondrocytes in monolayer culture causes progressive reductions in graft thickness, loss of zonal architecture, and a more fibrocartilaginous tissue histology, consistent with a dedifferentiating chondrocyte phenotype.     

The source of NMR-detected motional anisotropy of water in blood vessel walls.

2H Double quantum-filtered (DQF) NMR spectroscopy of deuterated water is sensitive to the presence of order in biological systems. This is because the only nuclei that are detected are those with residual quadrupolar interactions due to their anisotropic motion. In the present study, samples of aorta, coronary and carotid arteries, and vena cava were studied in parallel by 2H DQF NMR and by light microscopy. The average quadrupolar splitting, calculated from the NMR data, varies considerably among the different blood vessels, with high reproducibility for each type of vessel. Polarization microscopy examinations using collagen-specific staining with picrosirius red, have shown a variety of color profiles for the different blood vessels. These reflect different physical modes of aggregation (packing and thickness) of collagen fibers. A correlation was found between the NMR parameters and the color profiles of the picrosirius red-stained sections. Treating the blood vessels with 90% formic acid resulted in the elimination of the 2H DQF NMR signal. Histological analysis demonstrated a complete degradation of collagen and muscle, whereas the elastin filaments were preserved. Evidence is given that the 2H DQF NMR signal is dominated by the contribution of water molecules interacting with the collagen fibers.