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Biomedical subjects

M Simionescu

Publications and source records attributed to M Simionescu.

At least 73 records · Page 4Linked to original sources

Organization of cell junctions in the peritoneal mesothelium.

Intercellular junctions in the mesothelium of the visceral (mesentery and omentum), and parietal (diaphragm, pre-aortic, and iliac region) peritoneum were examined in rats and mice by using freeze-cleaved preparations. In addition to usual intercellular junctions (cell body junctions), special junctions are found between cell processes and the surface of the neighboring cell (cell process junctions). Cell body junctions are provided with tight junctions and communicating (gap) junctions. The former consist of one to two junctional strands which show a characteristic staggered arrangement, and focal discontinuities. In cell process junctions, the strands form loops or appear as short, free-ending elements; their polymorphism suggests considerable lability, probably in connection with their assembly and disassembly. The existence of free-ending strands indicates that such structures can be used as attachment devices without being concomitantly involved in the formation of occluding zonules. In both types of junctions, the strands can be resolved into bars, approximately 80- 100nm long, frequently provided with terminal enlargements and intercalated particles which occur singly or in small clusters. These particles are morphologically similar to those present in communicating (gap) junctions. The mesothelium is also provided with isolate composite macular junctions. Throughout the mesothelium, the cleavage plane follows the outer contour of junctional strands and particles, suggesting that strand-to-strand interactions in the apposed membranes are weaker than interactions between each strand and underlying cytoplasmic structures. In their general geometry and cleavage characteristics, the mesothelial junctions resemble the junctions found in the venular endothelium.

Animals

Segmental differentiations of cell junctions in the vascular endothelium. Arteries and veins.

A systematic survey of endothelial junctions in elastic (aorta) and muscular (mesenteric) arteries and in medium (renal and mesenteric) and large (cava inferior) size veins has been carried out in the rat using freeze-cleaved preparations. The arterial endothelium is provided with a complex of occluding and communicating junctions (gap junctions) comparable to, though less elaborate than, that described in arterioles. The particles of the occluding junctions behave like "single unit" particles and have the tendency to remain on B faces upon membrane cleavage. In the venous endothelium the junctions take the form of long occluding junctions with few associated communicating junctions (maculae communicantes). As in arterial endothelium, the junctional particles appear preferentially on B faces in cleaved preparations. These structures, although continuous over long distances, are interrupted focally by areas in which the junctional elements are similar to those found in venules: the ridges and grooves are short, discontinuous, randomly distributed along the general line of cell contact, and often particle-free. In muscular arteries two unusual types of junctions are encountered. Both are disposed in loops over short distances along the perimeter of the cell. One type appears to be a strectched-out version of the usual combination of occluding and communcating junctions of the arterial endothelium (this type is also occasionally encountered in the venous endothelium). The other type is reminiscent of the septate junctions found in the epithelia of invertebrates but the apparent similarity remains to be checked by further work.

Animals

Galloylglucoses of low molecular weight as mordant in electron microscopy. I. Procedure, and evidence for mordanting effect.

Gallotannin, consisting mainly of low molecular weight esters such as penta- and hexagalloylglucoses (commercially available as tannic acid produced from Turkish nutgall), can be used for increasing and diversifying tissue contrast in electron microscopy. When applied on tissue specimens previously fixed by conventional methods (aldehydes and OsO4), the low molecular weight galloylglucoses (LMGG) penetrate satisfactorily the cells and induce general high contrast with fine delineation of extra- and intracellular structures, especially membranes. In some features, additional details of their intimate configuration are revealed. Various experimental conditions tested indicate that the LMGG display a complex effect on fixed tissues: they act primarily as a mordant between osmium-treated structures and lead, and concomitantly stabilize some tissue components against extraction incurred during dehydration and subsequent processing. Experiments with aldehyde blocking reagents (sodium borohydride and glycine) suggested that the LMGG mordanting effect is not dependent on residual aldehydes groups in tissues.

Connective Tissue

Galloylglucoses of low molecular weight as mordant in electron microscopy. II. The moiety and functional groups possibly involved in the mordanting effect.

Synthetic pentamonogalloylglucose applied to fixed tissues acts as a mordant, inducing high and diversified contrast similar to that obtained with natural gallotannins of low molecular weight (LMGG). By the separate use of each of the two moieties of the galloylglucose molecule, it was found that gallic acid is the mordanting agent. Glucose may contribute, however, to the effect by increasing the solubility and cross-linking potential of the compound, since the mordanting induced by gallic acid alone is weaker than that produced by its hexose esters. As suggested by results obtained with various phenolics and benzoic acid derivatives, the functional groups required for the mordanting effect of such agents are the carboxyl group, and at least one hydroxyl group concomitantly present on the benzene ring. In the case of galloylglucoses, it is assumed that the effect is due to hydrolysis products (gallic, digallic, or trigallic acids) or to the multiple hydroxyl groups of the intact molecule. Esters of gallic acid (propyl- and methylgallate), as well as pyrogallol, produce a "reversed staining" of all membranes, except for those of communicating (gap) junctions.

Cytological Techniques

Segmental differentiations of cell junctions in the vascular endothelium. The microvasculature.

Small vascular units consisting of an arteriole, its capillaries, and the emerging venule (ACV units) were identified in the rat omentum and mesentery. They were fixed in situ and processed for electron microscopy either as whole units or as dissected segments. Systematic examination of the latter (in thin sections, as well as in freeze-cleaved preparations) showed that the intercellular junctions of the vascular endothelium vary characteristically from one segment to another in the microvasculature. In arterioles, the endothelium has continuous and elaborate tight junctions with interpolated large gap junctions. The capillary endothelium is provided with tight junctions formed by either branching or staggered strands; gap junctions are absent at this level. The pericytic venules exhibit loosely organized endothelial junctions with discontinuous low-profile ridges and grooves, usually devoid of particles. No gap junctions were found in these vessels. The endothelium of muscular venules has the same type of junctions (discontinuous ridges and grooves of low profile); in addition, it displays isolated gap junctions of smaller size and lower frequency than in arterioles. The term communicating junction (macula communicans) is proposed as a substitute for gap junctions, since the latter is inappropriate, in general, and confusing in the special case of the vascular endothelium.

Animals

Decrease in ribosomal density of Proteus mirabilis exposed to subinhibitory concentrations of ampicillin or cephalothin.

The finding of reduced ribosomal densities at lower concentrations than those required to stop growth or cause visible defects in the cell wall is in contrast to the current view that the initial lesion produced by penicillins or cephalosporins is a defect in murein synthesis (11). This reduction in ribosomal density could be a primary or secondary effect and might be due to a decreased rate of growth, which has been shown to be associated with a lower ribosome frequency (12, 13) or to less stable ribosomes which disintegrated either spontaneously or as a result of the fixation procedures. The possibility that the decrease in ribosomal frequency was due to dilution (influx of liquid through a defective cell wall and/or cytoplasmic membrane, or efflux of ribosomes) remains, but no gross defect in cell wall was seen in more than 50 sections of cells showing reductions in ribosomal frequency. These findings suggest that an effect on ribosomes may antecede an effect on the cell wall.

Ampicillin

Morphometric data on the endothelium of blood capillaries.

Local differentiations within the endothelium of both muscular (diaphragm, myocardium) and visceral (pancreas, jejunal villi) capillaries have been studied in rats on sectioned and freeze-cleaved preparations. Four distinct parts have been recognized in the endothelial cells of all these vessels on the basis of subcellular components present in each part and on the basis of variations in the local frequency of plasmalemmal vesicles: (a) the parajunctional zone, (b) the peripheral zone, (c) the organelle region, and (d) the nuclear region. Our data indicate that approximately 16, approximately 7.0, and 8.5% of the endothelial cytoplasmic volume (in the peripheral zone) is accounted for by vesicles, their content, and their membranes, respectively. The average density of vesicular openings per microm(2) is 78 in diaphragm, 89 in myocardium, 25 in pancreas, and 10 in jejunal mucosa capillaries. The frequency of fenestrae is 1.7 times as high in jejunal (26/microm(2)) as in pancreatic capillaries (15/microm(2)), the corresponding fractional areas being approximately 9.5 and approximately 6%, respectively, of the endothelial surface. Intercellular spaces occupy a relatively small area ( approximately 0.08 to 0.2%) of the inner endothelial surface.

Animals

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

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