1. We have investigated the interaction of plasma proteins with the endothelial cell using cationized ferritin as a marker of the cell surface glycocalyx. 2. Single microvessels of the frog mesentery were sequentially perfused using glass micropipettes with solutions containing cationized ferritin (CF, 6.7 mg ml-1) in 0.10 M-NaCl and then with either frog plasma or bovine serum albumin (BSA; 50 or 10 mg ml-1), or protein-free Ringer solution, before suffusion fixation in 2.5% glutaraldehyde. 3. A layer of CF, usually two to four molecules thick, was associated with the luminal endothelial cell surface. In vessels post-flushed with protein-free Ringer solution the CF layer was closely adherent to all regions of the luminal endothelium, including the plasma membrane, vesicle diaphragms, coated pits and the entrances to clefts. However, when plasma was present during fixation the CF layer was separated from the cell surface by up to 100 nm over all regions. In vessels post-flushed with BSA the CF layer was also separated from the membrane but the effect was less striking. 4. The association of cationized ferritin with the endothelial cell surface was assessed quantitatively using electron micrographs of transverse sections (approximately 50 nm thick) of the perfused vessels, and expressed in terms of the depth of the layer of CF associated with the endothelial cell surface, its separation from the plasma membrane of the luminal endothelium, and the concentration of CF in the layer. The mean (+/- S.D.) separation in the presence of plasma, 32.3 +/- 10.5 nm (n = 12), was significantly greater (P less than 0.01) than that with either protein-free Ringer solution, 3.0 +/- 1.4 nm (n = 9), or BSA in Ringer solution, 8.3 +/- 3.0 nm (n = 8). The separation seen with BSA in Ringer was also significantly greater than that measured with a final Ringer solution perfusion (P less than 0.01). The effects of 10 and 50 mg ml-1 BSA were not different from one another. The total glycocalyx thickness, defined as the sum of the separation layer and depth of CF layer, with plasma present, 56.2 +/- 13.7 nm, was twice the value seen with Ringer solution, 28.0 +/- 9.1 nm (P less than 0.01), while the total thickness with BSA, 30.9 +/- 5.4 nm, was not different from the Ringer solution value.(ABSTRACT TRUNCATED AT 400 WORDS)

1. The proteolytic enzyme pronase, which degrades the endothelial cell glycocalyx, was perfused through single capillaries of frog mesentery. Hydraulic conductivity (Lp) of each vessel was determined before and after pronase perfusion. In three vessels in which Lp increased, the ultrastructure of interendothelial clefts was examined. In a separate group of frogs the effect of pronase on the endothelial glycocalyx was assessed by using cationized ferritin to label the capillary luminal surface. 2. Control Lp was 2.0 x 10(-7) cm s-1 cmH2O-1 (10 mg ml-1 bovine serum albumin, BSA, in frog Ringer solution). Vessels were then perfused with a solution containing 0.1 mg ml-1 pronase and 10 mg ml-1 BSA for 1 min. Lp measured in the same eleven vessels increased to 4.9 x 10(-7) cm s-1 cmH2O-1 (P less than 0.005). 3. Transverse sections of three of these vessels were examined by transmission electron microscopy at eight sites along each vessel. In these sections a total of 156 interendothelial cell clefts were found and photographed. No morphological features, such as fenestrations, transendothelial channels, or intercellular gaps associated with inflammation, were found which might account for the increases in Lp. 4. Measurement of cleft dimensions yielded a harmonic mean cleft depth (delta x) of 0.32 microns and an arithmetic mean cleft depth of 0.64 microns. Mean width (w) of the clefts outside the tight regions was 0.012 microns and the cleft length per unit area (L) was 1330 cm-1. The mean fractional pore area of vessel wall per unit cleft depth, Ap/delta x, calculated as Lw/delta x, was 48.7 cm-1. 5. There was less cationic ferritin (CF) labelling of the luminal glycocalyx in pronase-perfused than in control capillaries. On average, the proportion of the luminal surface covered by CF was 85% in controls and 42% in pronase-treated capillaries (P less than 0.01). In some vessels the CF pattern was greatly disrupted, indicating large changes in the glycocalyx structure. 6. It is concluded that the moderate increases in Lp induced by pronase perfusion are associated with partial digestion of the endothelial glycocalyx but are not accompanied by changes in the dimensions of the intercellular cleft. These observations support the fibre matrix hypothesis of capillary permeability and suggest that the endothelial glycocalyx contributes as much as 60% of the hydraulic resistance of the capillary wall.

The carcinogenic potential of 2‐amino‐3‐methylimidazo[4,5f]quinoline (IQ) was evaluated in cynomolgus monkeys. Monkeys received IQ, beginning at the age of one year, at doses of 10 or 20 mg/kg by gavage. Thus far, IQ has induced hepatocellular carcinoma in three monkeys with a latent period of 27 to 37 months. Metastases to the lung occurred in two of the three monkeys. Microscopically, the hepatocellular carcinoma in all three cases demonstrated a trabecular pattern. These data demonstrate that IQ is a potent carcinogen in nonhuman primates and support the idea that it is a potential carcinogen for humans.

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Lasko et al., 1991;Latif et al., 1992;Cunningham et al., 1993;Adamson et al., 1994). A number of oncogenic and tumoursuppressor gene functions have been demonstrated in squamous cell carcinoma of the head and neck (SCCHN) (Field et al., 1989(Field et al., , 1991Field, 1992) To date only two global analyses of the whole genome have been undertaken with the view to determine the fractional allele loss (FAL) of specific tumours and thus provide information concerning the 'genetic burden' of the disease during its progression as measured by clinicopathological parameters and survival data. This type of analysis has been undertaken in colorectal (Vogelstein et al., 1989) and bladder cancers (Knowles et al., 1994), and provides an indication of interacting genetic mechanisms in the development of these diseases. In addition, the results of such detailed allelotypes may aid the interpretation of carcinogenesis and the development of molecular progression models for specific tumours.We have undertaken a very comprehensive allelotype of SCCHN using 145 microsatellite markers in order to identify common regions of allelic imbalance and to analyse the interactions of these regions by calculating the fractional allele loss (FAL) in these tumours. Materias and methods Specimens