The mechanisms of reparative regeneration of venous endothelium

The reaction of endothelial cells of the inferior vena cava in response to freezing-induced lesions has been analysed in the experiments on 34 young adult Kyoto-Wistar normotensive rats. First the de-endothelialized surface is covered with flattened platelets and then, three days after surgery, the endothelium is restored as a result of migration and proliferation of endotheliocytes. The migrating endothelial cells removed the adhered platelets from de-endothelialized surface. The young endothelium was presented by a single layer of strongly elongated endothelial cells whose axis was parallel to the flow of blood. An immature endothelium is characterized by an increased number of endotheliocytes. No essential differences in the reaction of venous and aortic endothelium have been revealed in response to freezing-induced lesions.     

The morphofunctional characteristics of the thymus in rat fetuses and progeny in venous occlusion]

Using histological and morphometrical methods, morphofunctional state of the thymus was studied in fetuses and offspring of rats in physiological pregnancy and under conditions of phlebo-occlusion. Phlebo-occlusion resulted in hypertrophy of the fetus, thymus and offspring, changes in structural zones and cellular reactions.     

Protective effects of luteolin on the venous endothelium

Luteolin is a flavonoid with antioxidant properties already demonstrated in studies related to inflammation, tumor, and cardiovascular processes; however, there are no available information regarding its antioxidant effects at the venous endothelial site. We investigated the effects of luteolin (10, 20, and 50 μmol/L) in cultures of rat venous endothelial cells. Nitric oxide (NO) and reactive oxygen species (ROS) were analyzed by fluorimetry; 3-nitrotyrosine (3-NT) residues were evaluated by immunofluorescence, and prostacyclin (PGI₂) release was investigated by colorimetry. Intracellular NO levels were significantly enhanced after 10 min of luteolin incubation, with a parallel decrease in ROS generation. These results were accompanied by a significant reduction in the expression of 3-NT residues and enhanced PGI₂ rates. Therefore, luteolin is effective in reducing ROS thereby improving NO availability in venous endothelial cells. Besides, luteolin-induced decrease in 3-NT residues may correlate with the enhancement in endothelial PGI₂ bioavailability. These findings suggest the future application of this flavonoid as a protective agent by improving endothelial function in several circulatory disorders related to venous insufficiency.

     

The endothelium of the high endothelial venule: a specialized endothelium with unique properties

High endothelial venules (HEV) in lymphoid organs are specialized to facilitate the passage of lymphocytes into lymphoid parenchyma. This is accomplished by a ligand-receptor system on the endothelium and lymphocytes, which differs between lymph nodes and Peyer's patch. Experiments discussed in this paper show that other non-HEV-derived endothelial cells can acquire the characteristics of HEV and that HEV may differentiate under influences from their local micro-environments.     

The anticoagulant property of a lipoid anticoagulant and the mechanism of its influence on hemocoagulation]

A lipoid anticoagulant (LA) from human brain tissue was shown to possess a higher anticoagulant activity than previously obtained preparations. It has been established that out of four phospholipids present in LA, only phosphatidyl serine inhibits the coagulant activity of plasma. In isolated hemocoagulating systems LA and phosphatidyl serine were shown to inhibit prothrombin conversion catalyzed by thrombokinase and to exert the antithrombic action in the system thrombin--fibrinogen. Kinetic study of phosphatidyl serine- and LA-induced inhibition of thrombine formation and the thrombine--fibrinogen reaction by conjugated inhibition. Similar kinetic behaviour, observed upon inhibition of both processes by phosphatidyl serine and LA suggest that phosphatidyl serine is a main anticoagulant agent of LA.     

Arterial and venous circulation of the leg after intermittent venous occlusion

We study the arterial and venous circulation of the normal leg by strain gauge plethysmography and venous occlusion (thigh tourniquet). We propose the application of a simplified linear physical model of the venous circulation. It helps to analyse the plethysmographic data recorded during and after the congestion. It ignores the arterial inflow and consider the post-occlusive venous volume decay in function of time as being monoexponential. The venous compliance (C) is measured when the volume has reached a steady-state level during the congestion (known pressure). The time-constant (T) characterizes the volume decay in function of time when the occlusion is released. The tourniquet is successively inflated with two levels of pressure (30 and 60 mm Hg) in order to check if the system is actually linear as predicted by the model. The venous outflow is not strictly monoexponential and the model is only suitable to describe the beginning of the curve. The compliance does not behave linearly, the values measured at 30 mm Hg, being higher than at 60 mm Hg ($ 26%). The time-constant T is slightly influenced by the level of pressures. The calculated resistance is therefore lower at low pressure. We also study the arterial inflow before and after the venous congestion (3 min, 60 mm Hg). We observe a post-venous occlusion hyperaemia (mean rest flow: 5.2%/min, mean hyperemic flow: 12.1%/min) followed by a drop of the inflow (mean minimal flow: 3.4%/min). We evaluate the quantitative influence of neglecting the arterial inflow on the computing of the venous properties. The simplification appears acceptable.     

Insulin uptake by rat liver endothelium studied in fractionated liver cell suspensions

Summary Cellular distribution of insulin receptors was studied in fractionated rat liver cell suspensions using ¹²⁵1-insulin and a visual probe consisting of latex beads covalently linked to insulin (minibeads). Fractionation was done on metrizamide gradients which yielded two cellular fractions. The large cell fraction consisted mostly of hepatocytes and the small cell fraction consisted of 37% endothelial cells as well as Kupffer cells. The magnitude of insulin uptake by the endothelium-rich small cell fraction was at least double that of the uptake by the hepatocyte-rich fraction. The minibead technique demonstrated that in the small cell fraction only endothelial cells, and not Kupffer cells, were responsible for the insulin uptake. Our findings suggest that liver endothelium may be responsible for the uptake of circulating insulin and its transport to hepatocyte. This emphasizes the presence of a tissue-blood barrier in the liver.Abbreviations PRS phosphate-buffered saline - SEM scanning electron microscopy - TEM transmission electron microscopy     

The heat denaturation of bacterial ribonuclease studied by infrared spectroscopy]

The thermal denaturation of bacterial ribonuclease in the interval of pH 2.5-7.0 has been investigated by means of infra-red spectroscopy method. The protein melting for pH 2.5 begins at the temperature 25 degrees C and is accompanied by secondary protein structure reconstruction, partially destroying native beta-structure and leading to new denatured conformation appearance of different types of beta-turns. Spectral changes for pH 3.5 and 7.0 are significantly less in the same frequency areas. At the temperature more than 50 degrees C protein aggregation takes place with inter-molecule-beta-form formation.     

The role of the endothelium in the vascular effects of vintoperol]

The effects of vintoperol have been studied in the experiments on the vascular bed of femoral artery of 15 mongrel dogs under chlorazol-urethane narcosis and on isolated vascular preparations of rats. It is shown that ++de-endothelization of vascular bed using saponin and mechanical removal of endothelial stripes decreased the vasodilatation reaction and relaxation of stripes by 50-60% of the initial values. While infusing ventoperol (0.3 mg.kg-1 min-1) to ++de-endothelized vascular bed, the blood flow increased by 18 +/- 5% as against 47 +/- 3.9% of the initial value under intact endothelium. Blockade of guanylate cyclase by methylene blue decreased blood flow under vintoperol action to 24 +/- 3.5%. The similar results are obtained in the experiments in vitro. After ++de-endothelization of pulmonary artery++ the amplitude of relaxation of preactivated smooth muscles decreased vs. initial tone (21 +/- 3.7% vs. 56% +/- 5.3%). Inhibition of relaxation reaction of vascular stripe under vintoperol effect is also observed after treatment with gossypol, or methylene blue. Thus, endothelium is involved in the realization of vasodilating effect of vintoperol, its action being mediated by endothelium--derived relaxing factor.     

The HKA test revisited: a maximum-likelihood-ratio test of the standard neutral model

We present a maximum-likelihood-ratio test of the standard neutral model, using multilocus data on polymorphism within species and divergence between species. The model is based on the Hudson-Kreitman-Aguade (HKA) test, but allows for an explicit test of selection at individual loci in a multilocus framework. We use coalescent simulations to show that the likelihood-ratio test statistic is conservative, particularly when the assumption of no recombination is violated. Application of the method to polymorphism data from 18 loci from a population of Arabidopsis lyrata provides significant evidence for a balanced polymorphism at a candidate locus thought to be linked to the centromere. The method is also applied to polymorphism data in maize, providing support for the hypothesis of directional selection on genes in the starch pathway.     

Determination of pulmonary microvascular reflection coefficient in sheep by venous occlusion

We devised a technique that permitted elevation of pulmonary pressures in unanesthetized sheep by occluding their pulmonary veins. Using this technique, we raised pulmonary capillary pressure from a baseline of 13.2 +/- 2.2 to 35.3 +/- 5.1 mmHg. This increased lung lymph flow (from 8.8 +/- 2.7 to 53.1 +/- 13.9 ml/h). We estimated the pulmonary microvascular oncotic reflection coefficient and found it to be 0.82 +/- 0.05 (SD). The filtration coefficient was 0.019 +/- 0.005 ml.mmHg-1.min-1. During the period of increased pressure, the animals had stable arterial pressures and cardiac outputs. None of the animals developed blood coagulation problems. These data illustrate the usefulness of pulmonary venous occlusion to elevate pulmonary microvascular pressure to obtain plasma-to-lymph protein concentration ratios independent of flow, allowing for the calculation of the oncotic reflection coefficient.     

The fibronectin binding of salmonellae studied by a passive hemagglutination method]

The study has shown that the fibronectin-binding properties of different Salmonella species can be tested by means of passive hemagglutination with fibronectin-sensitized formulated sheep red blood cells (SRBC). S. enteritidis and S. typhimurium isolated from foodstuffs, as well as the representatives of the species S. kottbus, S. anatum, S. tshiongwe and S. newlands, are capable of binding fibronectin. Fibronectin-binding properties have also been detected in Shigella sonnei, as well as in eubiotic microorganisms Escherichia coli M 17 and Lactobacillus plantarum. In pH ranging from 6.5 to 7.4 salmonellae, shigellae and E. coli agglutinate fibronectin-sensitized SRBC much more effectively in acidic pH values, while changes in pH have been found to produce no essential influence on the fibronectin-binding capacity of Staphylococcus aureus and lactobacilli.