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Biomedical subjects

L V Leak

Publications and source records attributed to L V Leak.

At least 19 recordsLinked to original sources

Milky spots of the omentum: a source of peritoneal cells in the normal and stimulated animal.

The topography and ultrastructure of the omentum in normal and stimulated mice were investigated with combined transmission (TEM) and scanning electron microscopy (SEM). The present study demonstrated that lymphocytes and monocytes were the principle cell types in the non-stimulated milky spot. Following stimulation with bacterial toxin and adjuvant there was an increased microvascular permeability to fluid, neutrophils, monocytes and fibrin deposits within the connective tissue matrix of milky spots, and a subsequent increased cellular migration across the mesothelial lining into the peritoneal cavity. Cellular migration from the milky spot to the peritoneal cavity is facilitated by the absence of a basal lamina from the submesothelial connective tissue layer, therefore, cells can migrate from the interstitial spaces of the milky spot to intercellular gaps between mesothelial cells without having to penetrate a fibrous barrier.

Animals

Animal model of acute pericarditis and its progression to pericardial fibrosis and adhesions: ultrastructural studies.

To study the evolution of pericardial inflammation, we have developed a model of pericarditis in sheep by surgically injecting heat-killed staphylococci and Freund's adjuvant into the pericardial cavity under sterile conditions. The pericarditis evolved through the following phases: 1) inflammatory response, 2) mesothelial cell injury and desquamation, and 3) fibrotic phase. At 3-24 hr there was increased microvascular permeability, which resulted in the exudation of fluid, neutrophils, macrophages, and fibrin into the pericardial cavity and the pericardial interstitium. By 72 hr, large numbers of inflammatory cells were aggregated on the mesothelial surfaces and dispersed throughout the pericardial cavity, either as free-floating cells or located between strands of fibrin. At 6 days, fibrinolysis was apparent along the mesothelial surfaces; and newly formed collagen fibrils were deposited throughout the interstitial spaces and among the aggregated cells. These fibrils provided a matrix for the growth of new blood and lymphatic vessels into new connective tissue on both parietal and visceral pericardial surfaces. At 2 weeks, intrapericardial fibrosis had produced focal adhesions between the pericardial surfaces. At 1 month, extensive areas of the pericardial cavity were obliterated. By 9 months, there was a marked reduction in the numbers of cells and blood vessels and increased deposition of collagen and elastic fibers. The intrapericardial injection of heat-killed staphylococci and adjuvant provides a reproducible animal model to study the time course of pericardial inflammation.

Acute Disease

Distribution of cell surface charges on mesothelium and lymphatic endothelium.

The distribution of anionic sites on the luminal surfaces of the peritoneal mesothelium and lymphatic endothelium was investigated by injecting cationized ferritin (CF) intraperitoneally. After washing with phosphate-buffered saline, the diaphragm was fixed and processed for electron microscopy. CF label occurred in discontinuous patches along the mesothelial surface. Microvilli were heavily marked and often closely applied to the mesothelial surface. The intercellular cleft was also heavily labeled. The luminal aspect of the lymphatic endothelium was more extensively labeled, with the marker occurring in long discontinuous dense bands. The clefts of lymphatic endothelial intercellular junctions were extensively labeled especially along regions where cells were loosely apposed. The existence of a high density of anionic sites on membranes at the intercellular junctions of both mesothelial and lymphatic endothelial cells represent a salient feature which is very different from binding in blood capillary endothelium. The presence of a high density of anionic sites along the intercellular clefts of adjacent cells may play a role in the rapid movement of small solutes and molecules from interstitial spaces into the lymphatic lumen.

Animals

Concanavalin A receptor sites on lymph node cells in vivo and in vitro.

The distribution and density of receptors for concanavalin A (Con A) on the surfaces of cells of intact and isolated popliteal and axillary lymph nodes were investigated in the rabbit. Intact lymph nodes were perfused via the subcapsular (marginal) sinus with either Con A peroxidase or Con A ferritin, fixed with glutaraldehyde, and processed for electron microscopy. Both Con A peroxidase and Con A ferritin were distributed on the plasmalemma of lymphocytes, macrophages, neutrophils, plasma cells, reticular endothelial cells, and the vascular endothelium. Counts of Con A-conjugated ferritin particles indicated that the density of Con A receptors was generally similar for lymphocytes, macrophages, and neutrophils but lower on plasma cells. When lymph node cells were isolated by mechanical methods and exposed to Con A ferritin, the label was homogenously distributed on the cell surfaces of most cells. However, Con A binding was significantly higher on the surface of isolated cells than in the intact node. It is suggested that the increase in density of Con A binding sites on isolated cells may possibly be due to an unmasking of cell surface moieties in which additional Con A receptor sites become available as a result of the isolation procedure. The density of Con A ferritin binding sites was also significantly lower on the surface of isolated plasma cells than the lymphocyte and macrophage, suggesting that the density distribution of cell surface saccharides is different for various lymphoid cells.

Animals

Lymphatic removal of fluids and particles in the mammalian lung.

The structure and distribution of pulmonary lymphatics and their permeability to fluids and particulate materials have been investigated in the lungs of rats following fixation by combined intratracheal and vascular perfusion. In such preparations, the lymphatics remain in a distended state, and a close relationship to other structural components of the pulmonary interstitium is maintained. They were identified in regions with an abundant amount of connective tissue, forming an eleborate plexus within the pleura, the interlobular septum, peribronchial and perivascular areas. Recent data have shown that water-soluble molecules and particulate matter are removed from the interstitium along the lymphatic capillary (initial lymphatics) segment. It is distinguished by attenuated endothelial cells with extensively overlapping cell margins which are easily separated. We have studied this segment of the lymphatic vascular system following intratracheal injections of colloidal particles (ferritin and carbon) to determine the structural features responsible for the transport of large molecules and particulate materials across the lymphatic endothelial wall in the lung. The results showed that the tracer particles cross the lymphatic endothelial wall via the clefts of intercellular junctions. While the tracer particles were observed within vesicles, the question of transport across the lymphatic endothelium via plasmalemmal vesicles is still not settled since the number and size of vesicles containing tracer particles also increased with time. Intravascular injected dextran was also localized within the clefts of intercellular junctions and plasmalemmal vesicles. The results obtained with intratracheal and intravascular injected tracer substances are consistent with those observed in lymphatic capillaries for other tissues.

Animals

Lymphatic vessels of the mammalian heart.

An in situ heart lung preparation was developed to label lymphatics of the actively beating dog heart with subsequent fixation by vascular perfusion. Immediately after interstitial injections of trypan blue and colloidal carbon, a rich plexus of lymphatic vessels was visualized in the epicardium of the actively beating heart. With this method of fixation, tissue preservation is generally excellent and uniform throughout the heart. In thin sections examined with the electron microscope, lymphatic vessels are easily recognized by the content of plasma proteins which is preserved as an electron dense precipitate that is evenly dispersed throughout the lumen. An extensive plexus of thin walled lymphatic vessels is observed throughout the epicardial, myocardial and subendocardial regions. Numerous anchoring filaments are observed closely apposed to the abluminal endothelial surface which extend into the surrounding connective tissue. The distribution and ultrastructure of the cardiac lymphatic vessels are discussed in relation to their role in the removal of interstitial fluid from the heart.

Animals

The structure of lymphatic capillaries in lymph formation.

The lymphatic vascular system consists of endothelial lined vessels which begin as blind-end tubes or saccules that are located within the connective tissue areas. This system serves as a one-way drainage apparatus for the removal of diffusible substances as well as plasma proteins that escape the blood capillaries. If permitted to accumulate, these escaped components would deplete the circulatory system of its plasma colloids and disrupt the balance of forces responsible for the control of fluid movement and the exchange of gases and fluids across the blood vascular wall. The lymphatic capillaries are strategically placed and anatomically constructed to permit a continuous and rapid removal of the transient interstitial fluids, plasma proteins, and cells from the interstitium. Structurally the lymphatic capillaries consist of a continuous endothelium that is extremely attenuated over major aspects of its diameter, except in the perinuclear region which bulges into the lumen. These vessels lack a continuous basal lamina and maintain a close relationship with the adjoining interstitium by way of anchoring filaments. The adjacent cells are extensively overlapped and lack adhesion devices in many areas. When electron-opaque tracers are injected intravenously (i.e., horseradish peroxidase and ferritin), subsequent electron microscopic examination of tissues reveals the presence of tracer particles within the interstitium and the lymphatic capillary lumen. These particles gain access into the lymphatic capillaries via two major pathways: 1) the intercellular clefts of patent junctions and 2) plasmalemmal vesicles (pinocytotic vesicles). Another salient feature of the lymphatic endothelial cell includes the presence of numerous cytoplasmic filaments, which are similar in morphology to the actin filaments observed in a variety of cell types. The ultrastructural features of the lymphatic capillaries are discussed in relation to their role in the removal of interstitial fluids and particulate matter, and in the formation of lymph.

Animals

Studies on the permeability of lymphatic capillaries.

The passageway for interstitial fluids and large molecules across the connective tissue lymph interface has been investigated in dermal lymphatic capillaries in the ears of guinea pigs. Numerous endothelial cells overlap extensively at their margins and lack adhesion devices at many points. The observations suggest that these sites are free to move as a result of slight pressure changes. Immediately following interstitial injections of tracer particles (ferritin, thorium, carbon, and latex spheres), many of the overlapped endothelial cells are separated and thus passageways are provided between the interstitium and lymphatic lumen. Tracer particles also occur in plasmalemmal invaginations along both connective tissue and luminal fronts. All of the tracer particles accumulate within large autophagic-like vacuoles. Very few particles of ferritin are observed in the endothelium after 24 hr; however, the vesicles containing the nonprotein tracer particles (carbon, thorium, and latex) increase in size and content and remain within the lymphatic endothelial cells up to 6 months. The role of vesicles in the transport of large molecules and particles is discussed in relation to the accretion of tracer particles within large vesicles and autophagic-like vacuoles in the endothelial cytoplasm.

Animals