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PubMed1972

Permeability of intestinal capillaries. Pathway followed by dextrans and glycogens.

The pathway followed by macromolecules across the wall of visceral capillaries has been studied by using a set of tracers of graded sizes, ranging in diameter from 100 A (ferritin) to 300 A (glycogen). Polysaccharide particles, i.e. dextran 75 (mol wt approximately 75,000; diam approximately 125 A), dextran 250 (mol wt 250,000; diam approximately 225 A), shellfish glycogen (diam approximately 200 A) and rabbit liver glycogen (diam approximately 300 A), are well tolerated by Wistar-Furth rats and give no vascular reactions ascribable to histamine release. Good definition and high contrast of the tracer particles were obtained in a one-step fixation-in block staining of the tissues by a mixture containing aldehydes, OsO(4) and lead citrate in phosphate or arsenate buffer, pH 7.4, followed by lead staining of sections. The glycogens and dextrans used move out of the plasma through the fenestrae and channels of the endothelium relatively fast (3-7 min) and create in the pericapillary spaces transient (2-5 min) concentration gradients centered on the fenestrated sectors of the capillary walls. The tracers also gained access to the plasmalemmal vesicles, first on the blood front and subsequently on the tissue front of the endothelium. The particles are temporarily retained by the basement membrane. No probe moved through the intercellular junctions. It is concluded that, in visceral capillaries, the fenestrae, channels, and plasmalemmal vesicles, viewed as related parts in a system of dynamic structures, are the structural equivalent of the large pore system.

Animals↗
PubMed1973

Permeability of muscle capillaries to exogenous myoglobin.

Whale skeletal muscle myoglobin (mol wt 17,800; molecular dimensions 25 x 34 x 42 A) was used as a probe molecule for the pore systems of muscle capillaries. Diaphragms of Wistar-Furth rats were fixed in situ at intervals up to 4 h after the intravenous injection of the tracer, and myoglobin was localized in the tissue by a peroxidase reaction. Gel filtration of plasma samples proved that myoglobin molecules remained in circulation in native monomeric form. At 30-35 s postinjection, the tracer marked approximately 75% of the plasmalemmal vesicles on the blood front of the endothelium, 15% of those located inside and none of those on the tissue front. At 45 s, the labeling of vesicles in the inner group reached 60% but remained nil for those on the tissue front. Marked vesicles appeared on the latter past 45 s and their frequency increased to approximately 80% by 60-75 s, concomitantly with the appearance of myoglobin in the pericapillary spaces. Significant regional heterogeneity in initial labeling was found in the different segments of the endothelium (i.e., perinuclear cytoplasm, organelle region, cell periphery, and parajunctional zone). Up to 60 s, the intercellular junctions and spaces of the endothelium were free of myoglobin reaction product; thereafter, the latter was detected in the distal part of the intercellular spaces in concentration generally equal to or lower than that prevailing in the adjacent pericapillary space. The findings indicate that myoglobin molecules cross the endothelium of muscle capillaries primarily via plasmalemmal vesicles. Since a molecule of this size is supposed to exit through both pore systems, our results confirm the earlier conclusion that the plasmalemmal vesicles represent the large pore system; in addition, they suggest that the same structures are, at least in part, the structural equivalent of the small pore system of this type of capillaries.

Animals↗
PubMed1975

Permeability of muscle capillaries to small heme-peptides. Evidence for the existence of patent transendothelial channels.

Two heme-peptides (HP) of about 20-A diameter (heme-undecapeptide [H11P], mol wt approximately 1900 and heme-octapeptide [H8P], mol wt approximately 1550), obtained by enzymic hydrolysis of cytochrome c, were sued as probe molecules in muscle capillaries (rat diaphragm). They were localized in situ by a perixidase reaction, enhanced by the addition of imidazole to the incubation medium. Chromatography of plasma samples showed that HPs circulate predominantly as monomers for the duration of the experiments and are bound by aldehyde fixatives to plasma proteins to the extent of approximately 50% (H8P) to approximately 95% (H11P). Both tracers cross the endothelium primarily via plasmalemmal vesicles which become progressively labeled (by reaction product) from the blood front to the tissue front of the endothelium, in three successive resolvable phases. By the end of each phase the extent of labeling reaches greater than 90% of the corresponding vesicle population. Labeled vesicles appear as either isolated units or chains which form patent channels across the endothelium. The patency of these channels was checked by specimen tilting and graphic analysis of their images. No evidence was found for early or preferential marking of the intercellular junctions and spaces by reaction product. It is concluded that the channels are the most likely candidate for structural equivalents of the small pores of the capillary wall since they are continuous, water-filled passages, and are provided with one or more strictures of less than 100 A. Their frequency remains to be established by future work.

Animals↗
PubMed1975

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.

Animals↗
PubMed2005

MRI of perfluorocarbon emulsion kinetics in rodent mammary tumours.

Perfluorocarbon (PFC) emulsions can be imaged directly by fluorine-19 MRI. We developed an optimized protocol for preparing PFC droplets of uniform size, evaluated use of the resulting droplets as blood pool contrast agents, studied their uptake by tumours and determined the spatial resolution with which they can be imaged at 4.7 T. Perfluorocarbon droplets of three different average sizes (324, 293 and 225 nm) were prepared using a microemulsifier. Images of PFC droplets with good signal-to-noise ratio were acquired with 625 microm in-plane resolution, 3 mm slice thickness and acquisition time of approximately 4.5 min per image. Kinetics of washout were determined using a simple mathematical model. The maximum uptake of the PFC droplets was three times greater at the tumour rim than in muscle, but the washout rate was two to three times slower in the tumour. The results are consistent with leakage of the droplets into the tumour extravascular space due to the hyper-permeability of tumour capillaries. PFC droplets may allow practical and quantitative measurements of blood volume and capillary permeability in tumours with reasonable spatial resolution.

Animals↗
PubMed1982

Permeability of fenestrated capillaries in the isolated pig pancreas, with effects of bradykinin and histamine, as studied by simultaneous registration of filtration and diffusion capacities.

Combining an isogravimetric technique and a colorimetric 'on-line' method (Rippe & Stage 1978), filtration capacity (CFC) and diffusion capacity (PS) were simultaneously measured in the maximally vasodilated 'fenestrated' capillary bed of isolated, artificially perfused pancreatic glands in 12 juvenile pigs. Both CFC and PS for Cr-EDTA were about 20 times greater than in the 'continuous' capillary bed of skeletal muscle. With perfusate flow rates of 250 ml/min x 100 g during isogravimetry, PS-Cr-EDTA averaged 110 +/- 10.0 (S.E.) ml/min X 100 g, and diffusion limitation occurred first at flow rates above 300 ml/min X 100 g. CFC was independent of flow rate and averaged 0.641 +/- 0.027 ml/min X 100 g X mmHg. The parallel augmentation of PS-Cr-EDTA and CFC in the fenestrated capillary bed compared with continuous ones seems to reflect both a higher number of capillaries per unit tissue and an increased number of 'small pores' per unit capillary surface, whilst the 'large pore system' appears to be similar. Following bradykinin or histamine infusion, results were similar to those for continuous capillaries (e.g. Rippe, Kamiya & Folkow 1978). Thus, without further vasodilatation CFC increased 3-fold While PS-Cr-EDTA increased only some 25%, and subsequent isoprenaline infusion reversed these effects. Previous studies on continuous capillaries indicate that histamine-type agents act by opening additional 'large pores' in the venular exchange sections (cf. Rippe & Grega (1978, Svensjö 1978), while beta-adrenergic agonists block this effect. The results further suggest that the fenestrae are not involved in these bradykinin-histamine effects, but rather function as a high-density, small pore population.

Animals↗
PubMed1987

Permeability of renal capillaries. I. Preparation of neutral and charged protein probes.

This paper describes the preparation of charged and uncharged protein molecular probes for study of the permselectivity of renal capillaries. Horse heart myoglobin was used as a neutral myoglobin. Since it contained several fractions with different isoelectric points, it was purified by fast protein liquid chromatography (FPLC). To obtain a negatively charged myoglobin, the original horse heart myoglobin was treated with cyanate, resulting in net charge of -5.7 +/- 0.3 at physiological pH (mean +/- SEM). The charge was determined from the Donnan potential which develops over a semipermeable membrane separating the inside solution in which the protein was dissolved from a surrounding bath of equal ionic strength. Sperm whale myoglobin was similarly purified by FPLC and used as a positively (+1.7 +/- 0.2) charged isomer. Horseradish peroxidase (HRP) was purified by means of gel and ion-exchange chromatography and found to be neutral at physiological pH. Negatively charged (-14.0 +/- 0.5) HRP was obtained by succinylation. Two isomers of lactate dehyrogenase (LDH) were used, namely the slightly positive (+2) LDH-M4 and the strongly negative (-19) LDH-H4. These isomers, which occur naturally, did not require further purification. The Stokes-Einstein radii, as measured by gel chromatography, of inulin, myoglobin, HRP and LDH were 11, 17.5, 32 and 46 A, respectively. The chemical modifications did not alter the Stokes-Einstein radii. In biological studies on rat kidneys samples of both plasma and renal hilar lymph were found to contain radioactive low molecular weight degradation products in addition to the intact proteins. This necessitated separation of all individual samples on small Sephadex columns prior to analysis.

Animals↗
PubMed1987

Permeability of renal capillaries. II. Transport of neutral and charged protein molecular probes.

The permselectivity of the renal capillaries was investigated from the transport of a series of molecular probes: inulin, positive (+2, net charge at pH 7.4), neutral (0), and negative (-6) myoglobin, neutral (0) and negative (-14) horseradish peroxidase (HRP) and two isomers of lactate dehydrogenase (LDH), namely the positively charged (+2) LDH-M4 and the negatively charged (-19) LDH-H4. The determination of the concentration of tracer proteins necessitated gel separation of both plasma and renal hilar lymph. The reason for this is that the proteins, after filtration, will be reabsorbed and degraded by the proximal tubular cells into small molecular compounds (amino acids), which will return to both the renal interstitium and systemic plasma. Even if this degradation is of low degree, as for high-molecular-weight proteins, separation is still required, since the relative lymph concentration (plasma concentration put at 1) is also low, that is, even small amounts of low molecular compounds will distort the relative lymph concentration obtained. The transport from plasma to renal hilar lymph of the tracer molecules fell with increasing Stokes-Einstein radius. The relative lymph concentration of the 11 A inulin was 1.06 +/- 0.03 (mean +/- SEM), of the neutral 17.5 A myoglobin 0.76 +/- 0.05, of the neutral 32 A HRP 0.32 +/- 0.02 and of the neutral 46 A LDH 0.12 +/- 0.01. The data are compatible with a two-pore system. The negative tracer molecules were in general proportionally more restricted than the neutral (or positive) moieties (P less than 0.001) thus suggesting a negatively charged peritubular capillary membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Permeability of renal capillaries. III. Theoretical analysis of hydraulic conductivity, pore structure and electric properties.

The functional characteristics of the capillary membranes of rat kidneys were analysed in a computer-based model utilizing previous data on the hydraulic conductivity of the membranes and transport by diffusion, hydraulic flow and ion migration of: inulin, myoglobin (negative, neutral and positive), horseradish peroxidase (negative and neutral), lactate dehydrogenase (negative, neutral and positive) and albumin. The results showed that in the peritubular capillaries the main fluid reabsorption (26.8 x 10(-8) ml nephron-1 100 g body wt-1) occurs through a set of 20 A pores with a total pore area over pore length of 30 cm x (nephron 100 g body wt)-1, whereas the plasma proteins enter the renal interstitium through a few 140 A large pores with a total pore area over pore length of 7.2 x 10(-4) cm and a fluid reabsorption of 1.2 x 10(-10) ml nephron-1 100 g body wt-1. The intramembranous concentration of negative fixed charges in the large pore system was estimated at 20 mM (range 15-22 mM). Since this was accompanied by a potential difference of 0.3 mV, a net driving electro-osmotic force, favouring reabsorption, of 4 mmHg developed. The charges and the electro-osmotic force was found to be essential for protein transport, since if the membrane was uncharged, the transport and interstitial concentration of e.g. albumin was twice as high. The glomerular capillaries seem to have a more homogeneous structure, that is in essence a one-pore system; the pore radius was 44 A, the total pore area over pore length 2.8 cm and the concentration of negative fixed charges 40 mM (range 37-43 mM).

Animals↗
PubMed1994

Flavonoids. A review of the pharmacology and therapeutic efficacy of Daflon 500 mg in patients with chronic venous insufficiency and related disorders.

Daflon 500 mg consists of 450 mg diosmin and 50 mg flavonoid expressed as hesperidin. The authors describe its effects on the microcirculation in chronic venous insufficiency: improvement of venous tone, inhibition of prostaglandin synthesis, protection against free radicals, activation of the complement system, and increased lymphatic drainage. The authors summarize studies of its therapeutic efficacy in reducing venous capacitance, distensibility, and emptying time; reducing capillary permeability; and increasing capillary resistance.

Animals↗
PubMed1989

Permeability of continuous capillaries.

This review describes the theories of transport of solutes and solvent across the endothelial membrane of continuous capillaries which constitutes the microvascular exchange domain in peripheral tissues such as the heart, skeletal muscle, adipose tissue, and skin. The theories of equivalent endothelial pores and fiber matrix are considered together with discussions of the various methodological techniques for measurement of fluxes and flows, and the results and implications of anatomical and physiological examinations are considered.

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