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Osmotic reflextion coefficients of capillary walls to low molecular weight hydrophilic solutes measured in single perfused capillaries of the frog mesentery.

1. Individual capillaries of the transilluminated frog mesentery have been perfused with suspensions of human red cells in frog Ringer solution containing 1-0 g albumin 100 ml.-1. The outer surface of the mesentery has been washed with normal frog Ringer solution and with frog Ringer solutions made hypertonic by addition of one of the following solutes: sodium chloride (100 m-mole. 1.-1); urea (100 m-mole.1.-1); sucrose (20-50 m-mole. 1.-1); cyanocobalamin (8-5 m-mole. 1.-1). The temperature of the mesentery was between 14 and 16 degrees C in all experiments. 2. Wtih the mesentery superfused with normal Ringer, the filtration coefficient was determined from measurements of the rate of fluid filtration across the capillary wall, at a series of known capillary pressures (Michel, Mason, Curry & Tooke, 1974). Filtration coefficient varied from 0-69 X 10(-3) to 4-45 X 10(-3) mum. sec-1 .cm H2O-1 with an average value of 1-87 X 10(-3) mum. sec-1. cm H2O-1. 3. When the superfusate was made hypertonic by the addition of a test solute, the osmotic reflextion coefficient (sigma) of the capillary wall to test solute was calculated from the additional rate of filtration, the concentration of test solute in the superfusate and the filtration coefficient. Average values for sigma were: sodium chloride, 0-068 +/- 0-03 (three capillaries); urea, 0-071 +/- 0.015 (four capillaries); sucrose, 0-115 +/- 0-023 (seven capillaries); cyanocobalamin, 0-100 +/- 0-03 (three capillaries). 4. In further experiments, the osmotic reflextion coefficients to sodium chloride, urea and sucrose were determined in the same capillary. Five technically acceptable experiments were carried out. Although there were differences in the value of sigma between different capillaries, in any one capillary values of sigma were of the same magnitude and there appeared to be no significant trend with the molecular size of the test solute. 5. Our findings are inconsistent with the hypothesis that there is a single pathway for water and small hydrophilic molecules across the capillary wall. 6. Our results may be interpreted in terms of an exclusive channel for water in parallel with a channel shared by both water and small hydrophilic molecules. It is suggested that the exclusive water channel may be the membranes and cytoplasm of the endothelial cells and the shared channel may be located in the intercellular junctions. 7. Our data suggest the exclusive water channel represents about 10% of the total filtration coefficient in frog mesenteric capillaries. The shared channel shows relatively little restriction to the molecules investigated. Estimates of the volume flow throught the two channels are made for conditions where hydrostatic pressure differences and osmotic pressure differences are the driving forces.

Animals

Capillary recruitment and heterogeneity of perfused capillary distribution in dog myocardium.

To estimate functional diffusion distances, the distribution of perfused capillaries was calculated in dog myocardium. A fluorescent dye was injected via a femoral vein in 6 anesthetized, open-chest dogs, and passed once through the coronary circulation (23 +/- 3 s). In 6 animals the dye circulated 4-20 min. In 6 animals the dye circulated for one pass following 2-3 min of asphyxia. The heart was then removed and frozen. Frozen sections from the left ventricule were cut, illuminated to excite the dye to fluoresce, and photographed. They were then stained by silver methenamine to mark all capillaries. The density of all capillaries was compared to that of capillaries containing fluorescent label. The distributions of capillaries were estimated by morphometry. In one pass of the normoxic coronary circulation, 66(+/- 4 SE)% of subepicardial and 60(+/- 4)% of subendocardial capillaries were detectably labeled. Their distribution approached a random pattern, and maximal distances to the nearest labeled capillary were lengthened by 50% compared to all capillaries. With multiple passes of the dye, or with asphyxia 76-79% of the capillaries were detectably labeled and their distribution approached the ordered pattern of the total capillary bed. We speculated that the unlabeled capillaries represented a spatially heterogeneous blood flow reserve.

Animals

Analysis of gas exchange between air capillaries and blood capillaries in avian lungs.

A number of models is analyzed to study gas exchange between blood capillaries and air capillaries in the avian parabronchial wall when diffusion is the only transport mechanism in the air capillaries. The existing anatomical arrangement of blood capillaries that traverse the periparabronchial tissue from peripherally located arterioles to draining venules at the luminal surface appears to provide a particularly high gas exchange efficiency. Application of the theory to measurements in the hen using histological estimates suggests that substantial concentration gradients exist inside the air capillary gas whose magnitude vary along the parabronchus. Thus at the gas inflow end of the parabronchus the partial pressure drop within the air capillaries could amount, for both O2 and CO2, to about 10--15 torr at rest and to 30--40 torr during exercise. Due to the peculiar arrangement of capillary blood flow to the air capillaries the effects of these gradients on gas exchange are very slight during rest. During exercise, however, the diffusional resistance inside the air capillaries may become limiting for the over-all gas exchange, and other mechanisms may be needed to secure respiratory gas transfer.

Animals

Maude Abbott Lecture--1991. The capillary then and now: an overview of capillary pathology.

The concept of capillary is traced through its historical development, beginning with the pathologic "synanastomoses" between arteries and veins as proposed by the ancient Greeks. Harvey assumed that arteriovenous connections existed but never saw them. Malpighi and van Leeuwenhoek were the first to observe the capillary circulation in vivo. In the 1800s, pathologists contributed two advances: a method for proving that capillaries have a structured wall (von Recklinghausen) and the notion of "capillary leakage" in inflammation (Cohnheim). Then physiologists took over for half a century (Starling, Krogh, Pappenheimer) until electron microscopy created a new viewpoint (Palade). Several topics of capillary pathophysiology are discussed: vascular labeling and the capillary as a privileged vessel; capillary versus venular leakage in inflammation; endothelial tone. The overview concludes with a summary of 15 pathologic events that may occur in capillaries.

Anatomy

Nailfold capillary density as a possible indicator of pulmonary capillary loss in systemic lupus erythematosus but not in mixed connective tissue disease.

Nailfold capillary density was measured in 24 patients with systemic lupus erythematosus (SLE), 14 with mixed connective tissue disease (MCTD) and 21 healthy subjects. Pulmonary function tests were performed on all subjects and needle muscle biopsies on 12 patients with SLE and 9 with MCTD. A significant correlation was documented between nailfold capillary density and pulmonary gas transfer (KCO) in patients with SLE (p less than 0.001) but not in patients with MCTD. This suggests that in SLE poor gas transfer may be dependent on alveolar capillary loss and that nailfold capillary density may be a good indicator of alveolar capillary density. There was no significant correlation between skeletal muscle fiber atrophy and nailfold capillary density in SLE or MCTD. Additional studies to optimize the nailfold capillary counting method are described.

Adolescent

Enzymophoresis of nucleic acids by tandem capillary enzyme reactor-capillary zone electrophoresis.

Enzymophoresis with coupled heterogeneous capillary enzyme reactor-capillary zone electrophoresis was developed and evaluated in the area of nucleic acids. Ribonuclease T1, hexokinase and adenosine deaminase were successfully immobilized on the inner walls of short fused-silica capillaries through glutaraldehyde attachment. These open-tubular capillary enzyme reactors were quite stable for a prolonged period of use under operation conditions normally used in capillary zone electrophoresis. The capillary enzyme reactors coupled in series with capillary zone electrophoresis served as peak locator on the electropherogram, improved the system selectivity, and facilitated the quantitative determination of the analytes with good accuracy. Also, they allowed the on-line digestion and mapping of minute amounts of transfer ribonucleic acids, and the simultaneous synthesis and separation of nanogram quantities of oligonucleotides.

Adenosine Deaminase

1-Deoxymannojirimycin inhibits capillary tube formation in vitro. Analysis of N-linked oligosaccharides in bovine capillary endothelial cells.

Capillary endothelial cells can be induced to form capillary-like structures in vitro by plating on fibronectin-coated dishes (Ingber, D. E., and Folkman, J. (1989) J. Cell Biol. 109, 317-330), thereby mimicking angiogenesis. To assess the role of glycoproteins bearing asparagine-linked oligosaccharides in this process, we tested the effect of oligosaccharide processing inhibitors on the formation of capillary tubes. Deoxymannojirimycin, a compound that prevents synthesis of hybrid and complex-type oligosaccharides, inhibited the formation of capillary tubes. In contrast, swainsonine, an inhibitor that blocks synthesis of complex- but not hybrid-type oligosaccharides, did not inhibit tube formation. Lectin affinity chromatography of 2-[3H] mannose-labeled glycopeptides from endothelial cells induced to form tubes did not reveal a striking difference in the spectrum of oligosaccharides compared to uninduced cells. Since endothelial cells formed tubes normally in the presence of swainsonine, we analyzed glycopeptides from swainsonine-treated induced and uninduced cells. Cells induced to form tubes were enriched in monosialylated hybrid-type oligosaccharides sensitive to alpha-fucosidase, beta-galactosidase, and beta-N-acetylhexosaminidase, suggestive of sialyl Lewis-X determinants. We used an enzyme-linked immunoassay to measure sialyl Lewis-X epitopes on capillary endothelial cells and found that both induced and uninduced cells expressed sialyl Lewis-X epitopes. Deoxymannojirimycin and, to a lesser extent, swainsonine reduced the level of sialyl Lewis-X epitopes in cells induced to form capillary tubes, but neither compound affected the level of epitopes in cell monolayers. We conclude that synthesis of at least hybrid-type oligosaccharides is required for capillary tube formation in vitro and that an increase in monosialylated, fucosylated glycans on asparagine-linked oligosaccharides occurs during this process.

1-Deoxynojirimycin

Capillary lengths, anastomoses, and estimated capillary transit times in skeletal muscle.

Total capillary length, capillary segment length, and number of anastomoses per capillary were measured in rat gracilis muscle at rest and after 2 min of phasic contraction. Mean values of the foregoing variables at rest (+/-SD) were, respectively, 1,012 micronm +/- 484, 409 micron +/- 274, and 0.83 +/- 1.09. Total capillary lengths are well described by the gamma distribution, number of anastomoses by the negative binomial distribution, and segment length by the Weibull distribution. Contraction has no significant effect on the means or the frequency distributions, indicating that: 1) pressure gradients between adjacent capillaries are small, and 2) intercapillary anastomoses do not improve flow distribution in exercise. Erythrocyte velocities observed in resting muscle (Burton, K. S., and P. C. Johnson. Am J. Physiol. 223: 517-524, 1972) were shown to be adequately characterized by the gamma distribution. From these velocities and the observed distribution of path lengths, we computed an estimated distribution of capillary transit times. Mean transit time was 4.29 s. The median was 2.45 s, and 11% of values exceeded 8 s. The range was 90 ms-43 s. This heterogeneity of transit times should profoundly affect calculations of O2 transport and the shape of indicator dilution curves.

Animals

Capillary diffusion capacity for I-131 and capillary filtration rate in female patients with idiopathic oedema.

In female patients with idiopathic oedema the capillary filtration rate and capillary diffusion capacity for I-131 were studied in lower-limb muscles and subcutaneous tissue. The capillary filtration rate averaged 0.276 ml/100 ml per minute, and the capillary diffusion capacity for I-131 averaged 9.8 mol/(mol/ml) per 100 g per minute in the gastroonemius muscle and 8.7 mol/(mol/ml) per 100 g per minute in the lower-limb subcutaneous tissue. All of these figures are statistically significantly higher than the corresponding control findings. The results are compatible with the concept of disturbed capillary permeability, and etiopathogenetic factor of idiopathic oedema.

Adolescent

Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries.

The abilities of several different capillary electrophoresis techniques to separate DNA restriction fragments up to 23,000 bp were investigated. Methods employing electroosmotic flow in an untreated silica capillary were found to provide, at best, only partial resolution of the 23 fragments in a 1-kbp DNA ladder. By coating the inner walls of a silica capillary with poly(acrylamide) and filling these capillaries with buffers containing methylcellulose as a sleving medium, all fragments in the 1-kbp DNA ladder were separated. In addition, this technique facilitated the separation of the very large fragments in a lambda DNA-HindIII digest. Optimum resolution was obtained at low separation potentials using buffers containing at least 0.5% methylcellulose. The performance of this technique, i.e., resolution and quantitation, make capillary electrophoresis a powerful complement to slab gel electrophoresis and may make it a preferred alternative to both agarose gel electrophoresis and HPLC for applications such as the confirmation of plasmid integrity.

DNA

Simultaneous measurements of capillary diffusion and filtration exchange during shifts in filtration-absorption and at graded alterations in the capillary permeability surface area products (PS).

The diffusion exchange of Cr-EDTA, using the single injection indicator diffusion method, was followed simultaneously with estimations of the capillary filtration capacity (CFC) in an "isogravimetric" rat hindquarter preparation during artificial perfusion and maximal dilatation. Measurements were performed at constant flow and during 1) shifts in filtration-absorbtion, 2) alterations of perfused capillary wall area (graded rarification of capillary network by microsphere injection) and 3) during alterations of permeability (i.a. infusion of histamine). At maximal vasodilatation CFC was 0.037 +/- 0.001 ml/min X mmHg X 100 g and PS for Cr-EDTA 5.67 +/- 0.13 ml/min X 100 g. During filtration or absorbtion, Cr-EDTA transfer from vessels to interstitium changed only slightly but the situation may well be different for solute transfer from interstitium to vessels. Alterations in capillary wall area resulted in proportional changes in PS for Cr-EDTA while the CFC changes were always relatively smaller. Histamine increased CFC some threefold with a marked increase in protein transfer, while PS for Cr-EDTA increased only marginally. This histamine effect could be ascribed mainly to an increase in the number of large pores which, because of their relative paucity, are of little importance for small molecular diffusion exchange but highly important for convective and macromolecular exchange.

Absorption

Fenestrated blood capillaries and lymphatic capillaries in rat skeletal muscle.

Capillary fenestrae occur in one of about 60 cross-sectioned blood capillaries in normal adult rat skeletal muscles. The fenestrae occur singly or in groups. Fenestrated capillaries are found close to muscle fibers as well as in the perimysial and perineurial connective tissue. Small numbers of lymphatic capillaries are also present, mostly in the perimysial connective tissue.

Animals

Flow patterns of blood cells in the retinal capillaries. Retinal capillary flow patterns.

We present a new technique for retinal blood cell angiography which was used to investigate flow patterns of blood cells in the retinal capillary net of normal cats. Capillary paths were defined to be the path taken by the labelled blood cells from the terminal arteriole to the entry point into a collecting venule. We were able to define two types of capillary paths. The 'simple' paths were minimally coiled and had a high blood cell flux. The 'complex' paths were coiled, structurally complex and had a low blood cell flux in comparison to the simple paths of the same eye. A simultaneous fluorescein angiogram was performed to reveal the complete anatomy of the capillary net in which the functional paths exist. The possibility that such flow patterns exist in the human retina should be considered.

Animals

Venous serum, capillary serum, and capillary plasma compared for use in determination of lactate dehydrogenase and aspartate aminotransferase activities.

Lactate dehydrogenase and aspartate aminotransferase activities in capillary serum or plasma were significantly greater than in simultaneously assayed venous serum, the greatest differences being between capillary and venous serum. Although some difference is attributable to tissue fluid contributions, platelets seem to account for most of it, with possible small contributions from leukocytes. Capillary tube size and type appear to be important factors. We recommend that when capillary blood is to be used for enzyme assay, it should be processed as plasma.

Aspartate Aminotransferases

[Capillary blood as diagnostic sample in pediatric laboratory diagnosis. Arterio-capillary-venous difference and blood collection technique].

It is preferable to use capillary blood as diagnostic material in paediatric laboratory tests rather than venous blood. The collection of capillary blood makes less demands on the child and is easy in principle. The objection to capillary blood primarily concerns differences in the analyte concentration in arterial, capillary and venous blood. Comparative studies in the same child under standardised conditions are not available for a number of parameters. However, according to the results available at present, for almost all routine clinical chemical and haematological examinations clinically usable results can be obtained. Using the example of total protein analysis for the same child we did not find any differences between arterial and venous blood. It is highly probably that the same would apply to many other components. Suspected differences are probably a result of short-comings in the collection technique.

Blood Chemical Analysis

Comparison of capillary ear blood and arterial blood to validate capillary sampling as an accurate assay of blood gas in swine.

Capillary sampling in swine can be performed as an accurate assay of arterial blood gases. Studies with swine provided results similar to, or slightly more favorable than, those reported for human beings, depending upon which cutaneous technique was used on human beings. On the basis of free flow or arterilization of the cutaneous sample and of the correlation between capillary and arterial pH, CO2 partial pressure (PCO2), and O2 partial pressure (PO2) values, the capillary sampling technique of complete incisement of a 2-mm section from the tip of the warmed porcine ear could be a substitution technique for arterial blood sampling. Free flow with this technique was maximized and high correlation coefficients (r) for pH (r = 0.96), PCO2 (r = 0.82), and PO2 (r = 0.90) capillary-arterial values (n = 37) were obtained.

Animals

Free solution capillary electrophoresis of proteins using untreated fused-silica capillaries.

Numerous efforts have been made to separate proteins by capillary zone electrophoresis (CZE). The most common optimization techniques are changing the pH of the running buffer, coating the capillary surface with a hydrophilic polymer, or using additives in the sample solution. Surface coatings and solution additives can reduce the adsorption of the protein onto the capillary surface, but they diminish the separation efficiency and the resolution of CZE. This paper reports the successful separation of proteins in a untreated fused-silica capillary by raising the pH of the running buffer and washing between runs with 1.0 M sodium hydroxide. Under these conditions, model proteins and proteins in human serum have been determined by CZE. It is shown that the results from CZE are compatible with those of sodium dodecyl sulphate-polyacrylamide gel electrophoresis.

Blood Proteins

[Characteristics of the vascular-capillary network and the relationship between capillaries and cells in the nucleus of Luys in the cat during postnatal ontogenesis].

The development of the Luy's nucleus of the cat is not accomplished by birth-time. Complex relationships between nerve cell and capillaries continue to change during postnatal ontogenesis. The function of the Luy's nucleus becomes perfect by the third month. It should be noted that maturation of nerve cells and the vascular network goes not in parallel: the enlargement of the nerve cell occurs smoothly while the increase of the vascular-capillary network density is undulatory. There are three peaks in the development of vascular-capillary network: the 20th day, one month and a half, adult cat.

Age Factors