Status of the capillary endothelium of the lung in normovolemic perfusion of the organ through the system of the pulmonary artery

Pulmonary perfusion for 30 min to the dog under conditions of normovolemia is not accompanied with any essential changes in parameters of alveolar capillaries endothelium. Just the opposite, transformation of endothelial lining of the peribronchial capillaries demonstrates possible disturbances of the liquor transport across the walls of these vessels. The volumetric part of the interstitial space near these capillaries increases, while in the alveolar septa it does not change. In lymph formation, flowing out of the lung, together with bronchial capillaries, blood capillaries of the alveoli must take part.     

Morphometric study of the blood-carrying capillaries during the administration of a liquid into the salivary gland]

The physiological saline was injected into the excretory duct of the cat parotid gland under the pressure of 30, 70 and 120 cm H2O. It was found with the aid of transmission electronic microscopy, morphometry and statistical analysis that the liquid injection into the gland produces compression of the blood capillaries weaving the acinus. The compression of the capillary tubes is shown by a significant reduction in space given to the lumen of the capillaries and their endothelial layer. The compression of the capillary tubes is combined with a two-fold lessening suggest that the additional blood volume entering the gland in response to its perfusion by the liquid does not reach the blood capillaries and is thrown off into the vein vessels through the shunt communications and that regulation of the blood volume getting into the cat parotid gland capillaries is likely to depend on the hydraulic and osmotic state in the interstitial space of glandular lobes.     

Stereologic analysis of the respiratory zone of the lungs of the laboratory rat and man during aging

By means of stereological analysis methods, lungs of mature and ageing laboratory rats and those of human beings at the age of 21-40, 41-60 and further have been studied. During the process of ageing the fraction of nonparenchymatous structures increases, while that of parenchyma decreases. Architectonics of acini is described: the volume of central passages increases and the volume of alveolar air decreases. The form of the alveoli approaches the spherical one. The area of the internal surface and the alveolar volume increase. The total number of the alveoli and the area of the respiratory lining decrease. The total number of the alveolar capillaries does not change, however, the density of their distribution grows small. The area and volume of a separate segment of the alveolar capillaries grow large. The age rearrangement of the respiratory zone morphologically differs from the changes that are observed at the obstructive emphysema.     

Alveolar vessel behavior in the zone 2 lung inferred from indicator-dilution data

To gain insight into the changes occurring in alveolar vessels when alveolar pressure exceeds venous pressure at the downstream end of the alveolar vessels (zone 2), we compared the uptake of serotonin and the extravascular volume accessible to 3HOH (Qev) under zone 2 and 3 conditions in isolated dog lung lobes. We also examined the influence of occluding some of the small pulmonary arteries with 58- to 548-micron-diam beads on the serotonin uptake and Qev. We found that, with the bead embolization, both the serotonin uptake and the Qev were reduced, whereas the change from zone 3 to 2 reduced serotonin uptake but did not change Qev. A plausible explanation for these observations is that the beads occluded vessels that were relatively large compared with those in which significant transvascular 3HOH exchange and serotonin uptake take place. Perfusion ceased in the collection of capillaries normally served by the obstructed arteries. Thus the extravascular water and the serotonin uptake sites downstream from the obstructions were not accessible to the indicators during the short time interval of the indicator passage through the lung. On the other hand, the change from zone 3 to zone 2 resulted in the collapse of small individual capillary segments within the alveolar vessel bed. Since the serotonin does not readily diffuse from the vessels through the tissue, it could not reach the endothelial cells of the collapsed capillaries. However, since the distances for diffusion between collapsed capillaries and neighboring perfused capillaries were small, the more highly diffusible 3HOH had access to the same Qev under both zone 2 and 3 conditions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Nucleolus degradation and growth induced by uv-microbeam irradiation of interphase cells grown in culture

In contrast to total cell irradiation, local UV-microbeam irradiation can stimulate a significant diminution in the irradiated mature nucleoli in interphase mammalian cells in culture. This diminution is accompanied by the concomitant expansion of the unirradiated nucleoli within the same nucleus, and the total nucleolar volume per nucleus does not change appreciably. It is suggested that these nucleolar volume changes are the result of the dispersion, migration, and redistribution of the nucleolar material between competitive nucleolar organizer regions of the interphase nucleus.     

Ultrastructure of tissue basophils and capillaries during different periods of pre- and postnatal development of the dermis in the rat

Dynamics of ultrastructure of mast cells (MC) and that of the capillaries of the derma have been studied by means of electron morphometry methods in rats during pre- and postnatal ontogenesis. The dynamics of the Golgi complex volume, that of the mitochondria and specific granules have been calculated in the total volume of the MC membrane organelles. Certain new data have been obtained on the process of the MC specific granules formation in the derma from progranules up to the stage of a mature granule. The process mentioned is most intensive during the first weeks of the postnatal ontogenesis. At the age of two weeks, signs of an active exocytosis of the granules are noted. Judging by certain morphological signs, development of transendothelial transport of substances in capillaries takes place in parallel with formation of the specific granules and corresponds to the beginning of exocytosis of substances in the MC granules. The correlative analysis proves that formation of the MC specific granules is connected with the number of microvesicles in cytoplasm of endotheliocytes.     

A study of the two-way transport of horseradish peroxidase across the visceral pleura

The authors report on a study of the transpleural transport of horseradish peroxidase after intrapleural and intracardiac application. Following intrapleural introduction, a retention of the marker on the apical membrane of mesothelial cells was observed, with subsequent transcellular transfer after incorporation into microvesicles. Following intracardiac injection, the marker moved out of the pulmonary capillaries across the endothelial vesicles and progressed to the pleural cavity across the intercellular spaces and mesothelial vesicles. With either route of injection, reaction product was noted in the basal lamina of the mesothelium, elastic membrane, alveolar macrophages and pneumocytes type II.     

A Novel Mechanism of Bacterial Toxin Transfer within Host Blood Cell-Derived Microvesicles

Shiga toxin (Stx) is the main virulence factor of enterohemorrhagic Escherichia coli, which are non-invasive strains that can lead to hemolytic uremic syndrome (HUS), associated with renal failure and death. Although bacteremia does not occur, bacterial virulence factors gain access to the circulation and are thereafter presumed to cause target organ damage. Stx was previously shown to circulate bound to blood cells but the mechanism by which it would potentially transfer to target organ cells has not been elucidated. Here we show that blood cell-derived microvesicles, shed during HUS, contain Stx and are found within patient renal cortical cells. The finding was reproduced in mice infected with Stx-producing Escherichia coli exhibiting Stx-containing blood cell-derived microvesicles in the circulation that reached the kidney where they were transferred into glomerular and peritubular capillary endothelial cells and further through their basement membranes followed by podocytes and tubular epithelial cells, respectively. In vitro studies demonstrated that blood cell-derived microvesicles containing Stx undergo endocytosis in glomerular endothelial cells leading to cell death secondary to inhibited protein synthesis. This study demonstrates a novel virulence mechanism whereby bacterial toxin is transferred within host blood cell-derived microvesicles in which it may evade the host immune system.     

The role of microvesicles containing microRNAs in vascular endothelial dysfunction

Many studies have shown that endothelial dysfunction is associated with a variety of cardiovascular diseases. The endothelium is one of the primary targets of circulating microvesicles. Besides, microRNAs emerge as important regulators of endothelial cell function. As a delivery system of microRNAs, microvesicles play an active and important role in regulating vascular endothelial function. In recent years, some studies have shown that microvesicles containing microRNAs regulate the pathophysiological changes in vascular endothelium, such as cell apoptosis, proliferation, migration and inflammation. These studies have provided some clues for the possible roles of microvesicles and microRNAs in vascular endothelial dysfunction‐associated diseases, and opened the door towards discovering potential novel therapeutic targets. In this review, we provide an overview of the main characteristics of microvesicles and microRNAs, summarizing their potential role and mechanism in endothelial dysfunction, and discussing the clinical application and existing problems of microvesicles for better translational applications.     

Myocardial capillaries in the fetal and the neonatal rat: a morphometric analysis of the maturing myocardial capillary bed

Developing myocardial capillaries from 16-day-gestation fetus to adult undergo several morphological changes including a thinning of the lateral extensions of the capillary endothelial cells, the formation of a basal lamina, and an increase in the number of plasmalemmal vesicles. A decrease in the extracellular space, an increase in the number of capillaries, and a decrease in the capillary diameter were also observed during the developmental period. In view of these ultrastructural changes, a morphometric analysis was made on the developing myocardial wall to demonstrate specific quantitative changes. The volumes which were occupied by capillary endothelial cells, capillary lumina, extracellular space, and myocardial myocytes within a reference volume of myocardium were measured; and we found that 8% of the reference myocardial volume was occupied by capillary endothelial cells, 85% was occupied by myocardial myocytes, 4% was occupied by capillary lumina, and, except for a significant change in extracellular space at 16 days gestation, 3% was occupied by extracellular space. Each volume ratio was found to be nearly constant throughout the studied period. In contrast to this constancy in the volume ratios, other parameters which were measured demonstrated significant changes during the developmental period studied. These overall changes include a 135% increase in capillary density, a 63% increase in luminal surface area of capillary endothelial cells, a 24% decrease in capillary diameter, a 12% decrease in diffusion distance, and a 35% decrease in the diameter of the erythrocyte population.(ABSTRACT TRUNCATED AT 250 WORDS)     

A study of the two-way transport of horseradish peroxidase across the visceral pleura

Summary The authors report on a study of the transpleural transport of horseradish peroxidase after intrapleural and intracardiac application. Following intrapleural introduction, a retention of the marker on the apical membrane of mesothelial cells was observed, with subsequent transcellular transfer after incorporation into microvesicles. Following intracardiac injection, the marker moved out of the pulmonary capillaries across the endothelial vesicles and progressed to the pleural cavity across the intercellular spaces and mesothelial vesicles. With either route of injection, reaction product was noted in the basal lamina of the mesothelium, elastic membrane, alveolar macrophages and pneumocytes type II.Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthday     

An analysis of the endothelial differentiation of the rat aorta during reparative regeneration

Following freeze-injury the arterial endothelium is able to restore completely its integrity without forming myointimal thickening. In the direction from the center of the former defect to its periphery the specific volume of biosynthetic and bioenergetic apparatus is reduced, specific volume of microvesicles rises, and parajunctional condensations of the microfilaments forms. Adhesion of monocytes to the endothelial surface is detected along with their migration into subendothelial space on all the stages of re-endothelialization.     

Permeability of regenerating and atrophic choriocapillaris in the rabbit

When rabbits receive intravenous injections of sodium iodate, large expanses of the retinal pigment epithelium are destroyed. The adjacent capillary bed, the choriocapillaris, atrophies in response to the loss of the pigment epithelium and then regenerates. This provides a model of the permeability of regenerating and atrophic choriocapillaris, which we studied using intravenously injected horseradish peroxidase and catalase. Regenerating capillaries were permeable to peroxidase but not catalase. The permeability to peroxidase was probably due to endothelial fenestrations, since catalase (which is larger than peroxidase and does not penetrate endothelial fenestrae) was retarded at interendothelial junctional complexes, indicating that they were intact. Atrophic choriocapillaries were impermeable to catalase but displayed a heterogeneous permeability to peroxidase. This was correlated with the presence or absence of fenestrae; capillary profiles lacking fenestrae retained peroxidase in their lumina, whereas if fenestrae were present the tracer penetrated into the pericapillary space. The observations indicate that: (1) the permeability of the regenerating choriocapillaris is qualitatively similar to the mature choriocapillaris, and (2) the atrophic choriocapillaris undergoes changes in permeability that are primarily correlated with the loss of endothelial fenestrae. The observations provide a functional correlate - change in permeability - for structural changes in choriocapillaris endothelium (thickening, loss of fenestrae) in response to destruction of the retinal pigment epithelium, which has been postulated to exert a trophic effect on these capillaries.     

Mixed meal and light exercise each recruit muscle capillaries in healthy humans

Intense exercise and insulin each increases total limb blood flow and recruits muscle capillaries, presumably to facilitate nutrient exchange. Whether mixed meals or light exercise likewise recruits capillaries is unknown. We fed 18 (9 M, 9 F) healthy volunteers a 480-kcal liquid mixed meal. Plasma glucose, insulin, brachial artery flow, and forearm muscle microvascular blood volume were measured before and after the meal. Brachial artery flow and microvascular volume were also examined with light (25% max), moderate (50%), and heavy (80%) forearm contraction every 20 s in 5 (4 M, 1 F) healthy adults. After the meal, glucose and insulin rose modestly (to approximately 7 mM and approximately 270 pM) and peaked by 30 min, whereas brachial artery blood flow (P < 0.05) and the microvascular volume (P < 0.01) each increased significantly by 60 min, and microvascular flow velocity did not change. For exercise, both 50 and 80%, but not 25% maximal handgrip, increased average forearm and brachial artery blood flow (P < 0.01). Flow increased immediately after each contraction and declined toward basal over 15 s. Exercise at 25% max increased microvascular volume threefold (P < 0.01) without affecting microvascular flow velocity or total forearm blood flow. Forearm exercise at 80% maximal grip increased both microvascular volume and microvascular flow velocity (P < 0.05 each). We conclude that light exercise and simple meals each markedly increases muscle microvascular volume, thereby expanding the endothelial surface for nutrient exchange, and that capillary recruitment is an important physiological response to facilitate nutrient/hormone delivery in healthy humans.     

The permeability of muscle capillaries to horseradish peroxidase

The main objective of this study was to determine the pathways by which horseradish peroxidase (HRP) can cross the endothelium of muscle capillaries. Specimens of mouse diaphragm were fixed for cytochemical analysis at various intervals after intervenous injection of 0.5 mg HRP, at 4 min after intervenous injection of varied amounts of HRP, and at 4 min after intervenous injections in various volumes of isotonic NaCl. Our findings indicate that endothelial junctions serve as a barrier which may allow passage of very limited amounts of HRP. They also suggest that endothelial vesicles transfer HRP from the capillary lumen to the pericapillary interstitium as well as in the reverse direction. Increasing the volume of solution injected to approximately 30% of total blood volume did not increase the amount of HRP that left the capillary lumen. Our results with HRP do not provide clearcut evidence that endothelial junctions are the site of the small pore.     

Structure of spinal cord capillaries in hypokinesia

Changes in spatial interrelations of the spinal cord capillaries and motoneurons and capillary ultrastructure were studied under hypokinesia. Spatial interrelations between the capillaries and neurons were not demonstrated to change under hypokinesia. They were estimated by the following parameters: area of neuronal profile field, number of capillaries, their length, distance from the nerve cell body, capillary bed area and index of capillary-neuronal interrelations. Quantitative investigation revealed capillary stenosis: their diameter was one and a half times less under hypokinesia. Morphologically, capillary stenosis was accompanied by the basal membrane thickening and endothelial cytoplasm vacuolization. There was a direct relation between endothelial villi and the places of the endothelial cells contacts, dilatation of the contact interstices and solidifying of their borders. Changes in the capillaries were followed by reactions in the pericapillary structures, such as: fibrillae were formed, mitochondrii accumulated in the perivascular glial projections, the membrane next to capillary astrocyte projections underwent desmosome-like condensation. Mitochondrial accumulations were also observed in the nerve cell projections and in their cytoplasm sites contacting with the blood vessels.     

A quantitative investigation of microvascular changes in the thyroid gland after infrared (IR) laser radiation.

We present an ultrastructural study of thyroid capillaries in which 50-day-old rats Wistar rats, were irradiated with an infrared (IR) laser, (total dose, 46.80 J/cm2), the tissue quantified 1 day after ending treatment and a quantitative capillary analysis carried out by light and electron microscopy. Light microscopy was used to calculate capillary volume density revealing a significant increase in the irradiated rats. The quantitative measurement of parameters by electron microscopy required a two stage analysis: Level I, Electron Microscopy (Magnification x5,000); and Level II, Electron Microscopy (Magnification x26,000). At Level I, the following parameters were measured in each capillary: capillary area, capillary diameter, luminal area, luminal diameter, endothelial area, nuclear area and mean endothelial thickness. At Level II, pinocytotic vesicle diameter and their numerical density in endothelial cells were evaluated. Electron microscopic analysis revealed an increased luminal area in the capillaries of the irradiated rats. They also presented a decrease in endothelial cell thickness and vesicular diameter and an increase in vesicle numerical density. This latter increase is indicative of presumptive changes in capillary permeability, but the possible functional significance of these morphological changes in the endothelial cells requires further investigation.     

Blood–Brain Barrier Pericytes Are the Main Source of γ-Glutamyltranspeptidase Activity in Brain Capillaries

Cerebral endothelial cells form the selective permeability barrier between brain and blood by virtue of their impermeable tight junctions and the presence of specific carrier systems. These specialized properties of brain capillaries are reflected in the presence of proteins that are not found in other capillaries of the body. gamma-Glutamyltranspeptidase (GGT) has been widely used as a marker for brain capillaries and differentiated properties of brain endothelial cells. By using histochemical and biochemical methods we have investigated the expression of GGT in isolated capillaries, cultured brain endothelial cells and pericytes, and cocultures of astrocytes and brain endothelial cells. It was surprising that the majority of GGT activity was associated with pericytes, but not endothelial cells, suggesting that GGT is a specific marker for brain pericytes. The remaining GGT activity that was associated with endothelial cells rapidly disappeared from cultured cells but was reinduced in cocultures with astrocytes. Our results emphasize the need for pure endothelial cells for the investigation of blood-brain barrier characteristics.     

Study of the rat parotid blood capillaries during a 3-hour feeding cycle]

Regularities attending the change in the lumen diameter and the area of the endothelial cells were studied in the capillaries of the rat parotid gland during the secretory cycle under conditions of circulatory ischemia. The mean lumen diameter and the area of endothelial cells failed to change during the secretory cycle and during the occlusion of the carotid artery. It is thus supposed that enhanced capillary blood flow during the discharge of saliva could be explained by the growing number of the functioning capillaries.     

Three-dimensional organization of the plasmalemmal vesicular system in directly frozen capillaries of the rete mirabile in the swim bladder of the eel

Summary Several recent studies comparing chemically fixed and cryofixed endothelium have indicated that glutaraldehyde fixation may result in increases in the population of vesicles in the cytoplasm. Other reports based on ultrathin serial-section reconstruction of chemically fixed endothelium have revealed that the vesicular system is comprised of interconnected membranous compartments, which are ultimately continuous with either cell surface but do not extend across the endothelial cell. In this study, we have investigated the three-dimensional organization of the vesicular system in directly frozen, freeze-substituted capillaries of the rete mirabile from the swim bladder of the eel, specifically using the same block of embedded capillaries in which frozen capillaries had previously been found to contain less vesicles than chemically fixed capillaries. The results show that essentially all vesicles remain inter-connected with each other and are part of two separate sets of invaginations from the luminal and abluminal cell surface like in chemically fixed tissue. Any increase in vesicle number resulting from glutaraldehyde fixation does not affect the overall three-dimensional organization of the vesicular system in these endothelial cells.