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S Seno

Publications and source records attributed to S Seno.

At least 37 records · Page 2Linked to original sources

Intestinal absorption of cationic and anionic ferric colloids and complexes: biochemical, histochemical and morphological observations in rats.

This experiment was undertaken to study the possible difference in the intestinal iron absorption efficiency among iron compounds with different electric charges. Observation of rats given oral administration of 59Fe-labeled cationic cacodylate ferric (59Fe-Cac) colloid, anionic citrate ferric (59Fe-Cit) colloid, cationic 59Fe-Cac complex and anionic 59Fe-Cit complex revealed that iron absorption was more efficient in the 59Fe-Cac colloid, moderate in the 59Fe-Cac complex, low in the 59Fe-Cit colloid, and lowest in the 59Fe- cases given 59Fe-Cac colloid and 59Fe-Cac complex, a very high ratio activity was found in the liver and in the erythrocyte or hemoglobin in circulating blood, while the blood plasma, bone marrow, and spleen were low in activity. Histochemical observations of rat jejunal mucosa exposed independently for 10 min to the Fe-Cac colloid, anionic Fe-Cit colloid, and Fe-Cac and Fe-Cit complexes revealed that the cationic Fe-Cac colloid and Fe-Cac complex adhered to the luminal surface of the mucosa covering the apical area of villi with some ferric iron in the capillaries, while the anionic Fe-Cit colloid and complex did not adhere to the epithelial cells and were found free in the jejunal lumen. Electron microscopy revealed that Fe-Cac colloid particles were taken into epithelial cells by pinocytosis at the webs of microvilli, moved to the Golgi area, exocytosed to the intercellular spaces, and then translocated into the basement membrane toward blood capillaries.

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A new method of the immunohistochemical detection of cellular antigens for light and electron microscopy.

A new immunohistochemical method for light and electron microscopy of tissue- and cell-specific antigens by using ferric colloid-labeled antibody is presented. The antibodies labeled with the cationic cacodylate ferric colloid are stable and bind specifically to the target antigens to show clearly the site of antigens in tissue sections and on free cells by Prussian blue reaction for light microscopy and by the specific figure of electron opaque ferric colloid particles for electron microscopy. The staining procedure is very simple and it gives clear picture. So the method will be of beneficial for general laboratory use in immunohistochemical researches.

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An experimental study on the pathway of iron transfer from macrophages to erythrocytes in rat liver.

In order to reveal the pathway of iron release from macrophages, a 59Fe-labelled ferric hydroxide-potassium polyvinyl sulfate complex (Fe-PVS) was injected intravenously into anemic rats and the level of radioactivity in the liver, spleen, bone marrow, blood plasma and red blood cells (RBC) was estimated at various time intervals after the injection. Histochemical observation of ferric iron and ferritin in the liver was also made on anemic rats treated using unlabelled Fe-PVS. Fe-PVS injection promoted the recovery of anemia causing a rapid increase in the RBC number, with activated erythropoiesis occurring in the spleen and bone marrow. Soon after the injection, most of the radio iron was found in the liver with a small amount in the circulating erythrocytes, bone marrow and spleen. The iron level in the liver decreased gradually with a rapid increase in the iron level of the erythrocytes which reached a very high level 6 days after the 59Fe-PVS injection. Histochemical observations showed a heavy deposition of ferritin in the Kupffer cells 3 days after Fe-PVS injection. This deposition was minimized after 6 days with an increase in the level of ferritin in the parenchymal cells in the central area of acini. The level of radioferritin estimated biochemically in the nonparenchymal cell fractions of the liver revealed that the level dropped by about one third approximately 3.5 days after the Fe-PVS injection, showing the stimulated ferritin release at this stage. Results indicate that Kupffer cells in the liver play an important role in ferritin synthesis from the phagocytized iron compounds and that the iron is supplied for erythroid cell proliferation.

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Adenine uptake cells in the rat kidney with remarks on the expression of Ia antigen.

The patterns of [14C] adenine ([14C]A) incorporation into DNA by proliferative cells in the kidney were studied by the autoradiographic technique. It was revealed that, after 3 daily injections of [14C]A (1 microCi/g body weight each), a portion of the glomerular cells and a few fibroblastoid cells in the cortical intexstitium incorporated [14C]A into DNA to a remarkable extent. Such cells also incorporated [3H]thymidine, but to a lesser extent. The cells which incorporate [14C]A to a particularly great extent (adenine uptake cells) also occur in other tissues. Such cells are confined to a few cell types of either the macrophage or fibroblast or reticulum cell lines. This fact suggests that the adenine uptake cells observed in the glomerulus are also of a similar cell line and most likely mesangial cells. By immunohistochemical examination for Ia antigen, adenine uptake cells are divided into Ia-positive and Ia-negative types. The present examination showed that the major portion of adenine uptake cells in the glomerulus are Ia-negative, and it is suggested that these cells are analogous to the Ia-negative macrophages in the lung. This suggestion is supported by the fact that, in the glomerulus, colloidal carbon uptake cells (macrophage-like cells) are present in fairly large numbers. The Ia-positive cells seem to be of the same cell line as the adenine uptake cells that express Ia antigen in other tissues, such as septal fibroblasts in the lung.

Adenine↗

Successful extended hypothermic cardiopulmonary preservation for heart-lung transplantation.

The inability to obtain sufficiently extended hypothermic organ preservation is a major restriction on clinical heart-lung transplantation. We used core cooling, nonrecirculating retrograde heart perfusion, and lung immersion with liposomal recombinant human superoxide dismutase in an attempt to provide effective 12-hour cardiopulmonary preservation. Donor dogs supported by cardiopulmonary bypass were rapidly cooled to 15 degrees C with cardioplegic arrest, and heterotopic heart and unilateral left lung transplantations were performed. In control dogs (n = 7), hearts and lungs, harvested after core cooling and cardioplegic arrest, were transplanted with a total mean ischemic time of 88 +/- 5 minutes. In group II (n = 7), heart-lung blocks were similarly excised but preserved at 4 degrees C for 12 hours (756 +/- 30 minutes) and then transplanted. During preservation, the lungs were immersed in hyperosmolar extracellular solution. For the heart, retrograde coronary sinus perfusion was performed with intracellular solution containing perfluorochemicals at a temperature of 4 degrees C and a rate of 30 ml/hr for 12 hours. In group III (n = 7), donor organs were similarly excised and preserved for 12 hours (726 +/- 39 minutes), except that liposomal recombinant human superoxide dismutase was administered during harvest, preservation, and reperfusion. Myocardial function, assessed by the ratio of end-systolic pressure to end-systolic dimension, after the 12-hour preservation period in both experimental groups was similar to that of the control group 4 and 6 hours after transplantation. The mean arterial oxygen capacity of the transplanted left lung during ventilation with an inspired oxygen concentration of 40% was also similar in each group. In contrast, the 12-hour preservation of pulmonary function assessed by pulmonary vascular resistance, the accumulation of extravascular lung water, and histologic evidence of alveolar wall injury, interstitial edema, and perivascular hemorrhage were significantly impaired in the absence of liposal recombinant human superoxide dismutase. These findings suggest that successful extended cardiopulmonary preservation for heart-lung transplantation is possible with core cooling, nonrecirculating retrograde heart perfusion, and hypothermic lung immersion incorporating liposomal recombinant human superoxide dismutase.

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[High energy phosphates and mitochondrial respiratory function after 24 hours cardiac preservation--comparison between simple hypothermic immersion and continuous hypothermic perfusion].

Cardiac preservation is one of the most important step during heart transplantation. Prolonged preservation period is necessary to get the greater donor pool. To get safe prolonged preservation two methods, simple hypothermic immersion method (group A) and continuous hypothermic perfusion method (group B), were compared in this report by evaluating the myocardiac high energy phosphate level and mitochondrial respiratory function. The values of ATP, TAN and Energy charge were significantly higher in group B than group A (p less than 0.001). Mitochondrial respiratory function such as state 3 QO2, state 4 QO2, RCI and ADP/O also showed significantly higher values in group B than group A (p less than 0.001). These date indicate that continuous hypothermic perfusion method provides better preservation than simple hypothermic immersion method for 24 hours cardiac preservation.

Adenosine Triphosphate↗

Core-cooling, heart-perfusion, lung-immersion technique provides successful cardiopulmonary preservation for heart-lung transplantation.

Mongrel dogs underwent heterotopic heart-orthotopic left lung transplantation. In Group I (N = 6), donor organs procured following core cooling to 15 degrees C on cardiopulmonary bypass (CPB) with cardioplegic arrest were immediately transplanted. In Group II (N = 6), following cardioplegic arrest without CPB core-cooling, the pulmonary artery was flushed with modified Collins' solution. Heart-lung blocks were immersed in extracellular solution for 6 hours and then transplanted. In Groups III and IV (N = 6 each), following CPB core-cooling to 15 degrees C and cardioplegic arrest, the organ blocks were immersed in extracellular solution (Group III) and the heart was perfused with oxygenated extracellular solution (Group IV). Evaluation of lung function using differences in arterial oxygen tension between the left and right atria demonstrated no differences between groups. However, extravascular lung water and pulmonary vascular resistance were significantly elevated in Group II. Cardiac function assessed by the ratio of end-systolic pressure to end-systolic dimension was significantly better in Group IV than in Groups II and III. Thus, adequate 6-hour hypothermic cardiopulmonary preservation with core cooling plus heart perfusion can be achieved for heart-lung transplantation.

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Oxygenated perfluorocarbon, recombinant human superoxide dismutase, and catalase ameliorate free radical induced myocardial injury during heart preservation and transplantation.

The effect of free radical scavengers on free radical-induced myocardial injury during heart preservation and transplantation was examined. Four groups of nine hearts each were harvested from mongrel dogs (12.5 to 16.5 kg) and orthotopically transplanted to size-matched recipients. All hearts received a continuous perfusion of oxygenated modified Collins' solution (group A). In addition, groups B, C, and D received Fluosol DA and albumin. Preservation perfusion was performed for 18 hours, at 4 degrees C, pH = 7.4, and 20 mm Hg. In group C, recombinant human superoxide dismutase (4,080 U/mg, 20 mg/kg) and bovine catalase (46,200 U/mg, 20 mg/kg) were administered only during preservation perfusion. In group D, these scavengers were administered just before and during reperfusion for 1 hour. Hemodynamic studies were performed before excision of the donor hearts and 1 hour after the termination of cardiopulmonary bypass. Creatinine kinase MB isoenzyme and thiobarbituric acid reactive substance levels in the coronary effluent were determined during preservation perfusion and reperfusion. Only group A showed a significant heart weight gain (p less than 0.05) and a decline in passive compliance (p less than 0.05) during preservation. Lactate release was higher in group A than in the groups receiving Fluosol DA. In contrast, pyruvate levels in group A were lower than in other groups. The generation of free radicals stayed at a low level during preservation, but significantly increased during reperfusion and was associated with a corresponding increase in creatinine kinase MB isoenzyme. Perfusion with a perfluorochemical solution (group B) inhibited the sharp rise in levels of thiobarbituric acid reactive substances and of creatinine kinase MB isoenzyme and improved cardiac function during reperfusion (versus group A). Exogeneous free radical scavengers administered just before and during reperfusion (group D) significantly ameliorated thiobarbituric acid reactive substances and creatinine kinase MB isoenzyme levels and also induced a significant hemodynamic improvement during reperfusion. However, administration of scavengers during preservation did not. This study demonstrates that the generation of free radicals is primarily significant during reperfusion and reoxygenation after ischemia. Thus the best time for administration of scavengers is just before and just after the onset of reperfusion. Furthermore, perfusion with perfluorochemicals effectively maintains aerobic metabolism and ameliorates free radical damage during this period.

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The circulating alpha 1-antitrypsin-elastase complex attacks the elastic lamina of blood vessels. An immunohistochemical study.

The aim of the study was to determine the destination of the alpha 1-antitrypsin-elastase complex, which is found in circulating blood after the peroral administration of elastase. The complex was made in vitro by mixing hog pancreatic elastase with human alpha 1-antitrypsin and then injected intravenously into rats and mice. Tissues taken at various times after injection were subjected to histochemical staining using an antibody against elastase. Light microscope observations revealed dense deposition of reaction products in the elastic lamina of the arterioles; moderate or slight deposits were seen in the tissues surrounding arteries, in the tubular epithelial cells of the proximal convoluted tubules in the kidney, and in the pancreatic ducts. Immunoelectron microscopy revealed heavy deposition of the reaction product in the elastic lamina of the small arteries and arterioles; some dissolution of the elastic fibers was also evident. Pinocytic uptake of the alpha 1-antitrypsin-elastase complex was observed on the abluminal surface of endothelial cells and in smooth-muscle cells bordering the elastic lamina of arterioles. The endothelial cells of the arteries and arterioles retained their normal morphological appearance, although local desquamation was observed in some animals. The results indicate that, when the alpha 1-antitrypsin-elastase complex is present in the circulating blood, it is incorporated into the elastic lamina through the endothelial layer. This results in liquefaction of the lamina, desquamation of endothelial cells and leakage of the complex into the perivascular tissues via the vascular walls. However, some of the complex seems to be excreted very quickly from the kidneys.

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Fine-granular cationic iron colloid. Its preparation, physicochemical characteristics and histochemical use for the detection of ionized anionic groups.

In order to obtain distinct and reliable information concerning the localization of ionized anionic groups in tissues, fine-granular cationic ferric hydroxide colloid solution (Fe-Cac-f) was newly devised. This can be obtained by boiling a mixture of ferric chloride and ammonium cacodylate solutions. The colloid particles of Fe-Cac-f are about 1.0 nm in size, i.e., one-fifth of the size of ferric cacodylate colloid (Fe-Cac; Seno et al. 1983a). As with Fe-Cac, Fe-Cac-f particles in the pH range of 1.6-7.6 carry a positive electric charge, but the latter show a better permeation of tissues. Using the Prussian blue reaction, Fe-Cac-f gives a distinct deep-blue color and can be used for the detection of anionic groups of acid mucopolysaccharides and proteins by light microscopy. It is also useful for detecting the exact sites of ionized anionic groups in deep tissue areas using electron microscopy.

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Isolation and characterization of nephritogenic antigen from bovine glomerular basement membrane.

A method for isolation of a potent nephritogenic antigen from bovine glomerular basement membrane has been established; the glomerular basement membrane was solubilized by trypsin digestion and fractionated successively by gel filtration on Ultrogel AcA-34, concanavalin A affinity chromatography and affinity chromatography on immobilized antibodies. The antigen thus prepared was found to be highly nephritogenic; it causes glomerulonephritis in rats by a single injection of 0.1 mg per individual. Amino acid and carbohydrate analyses revealed that the antigen is a glycoprotein which contains amino acids and sugars characteristic of collagen, namely, hydroxyproline, hydroxylysine, glycine, glucose and galactose, although the relative amounts of these amino acids and sugars are less than those found in Type IV collagen of glomerular basement membrane.

Amino Acids↗

The anionic barrier system on the mesonephric renal glomerulus of the brown hagfish, Paramyxine atami Dean (Cylostomi).

The distribution of anionic groups of acid mucopolysaccharides on the surface of glomerular constituents of a brown hagfish, Paramyxine atami Dean, has been studied morphologically. The ionized anionic groups of acid mucopolysaccharides were labeled on fixed tissues by staining with cationic cacodylate iron colloid (Fe-Cac) at pH 4.0. The glomerular permeability to cationic and anionic macromolecules was observed morphologically in the kidney of the animal injected with native anionic ferritin (NF) or cationized ferritin (CF) into the dorsal aorta. Histochemical staining of tissues with Fe-Cac (pH 4.0) revealed the ionized anionic groups of acid mucopolysaccharides on both luminal and abluminal surfaces of endothelial cells, within the glomerular basement membrane (GBM), and on the visceral epithelial cell surface facing the urinary space. The CF molecules introduced into the dorsal aorta easily passed through the fenestrae of the capillary endothelial cell layer and the thick fibrillar GBM, reaching the urinary space to be adsorbed to the visceral epithelial cell surface or taken up by these visceral epithelial cells. On the other hand, NF hardly passed through the capillary wall. These results show that the nonosmoregulating mesonephric glomerulus of the brown hagfish has a working anionic barrier system. The function of its glomerulus is compared to that of the mammalian metanephric glomerulus.

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The anionic barrier system in the mesonephric renal glomerulus of the arctic lamprey, Entosphenus japonicus (Martens) (Cyclostomata).

To examine the selective permeability of the nephrons of lower vertebrates, the permeability of the glomerulus in the kidney of an arctic lamprey, Entosphenus japonicus (Martens), to native anionic ferritin or cationized ferritin was studied by observing the distribution of ionized anionic groups in renal tissues. The cationized ferritin molecules injected into the dorsal aorta penetrated rapidly into the glomerular basement membrane layer through fenestrae present in the capillary endothelium and were subsequently excreted into the urinary spaces via the interstices between foot processes of the visceral epithelial cells. Native anionic ferritin, on the other hand, passed only minimally through the capillary wall. Cytochemical staining of fixed tissue or perfusion of the kidney in situ with cationic cacodylate-iron colloid revealed that the ionized anionic groups of acid mucopolysaccharides were distributed on both the luminal and abluminal surfaces of endothelial cells, and in the thick fibrous lamina rara interna of the glomerular basement membrane; they were especially dense on the surfaces of visceral epithelial cells and their foot processes. These results suggest that the mesonephric glomerulus of the arctic lamprey possesses a functionally well developed anionic barrier system comparable to that of the mammalian metanephric glomerulus.

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Absorption of elastase through the jejunal mucosa of the rat. An immunocytochemical study.

In order to reveal the absorption process of elastase from the intestine, hog pancreatic elastase was injected into the ligated jejunum lumen of the rat, and the tissues were cytochemically observed at various times after injection. The peroxidase anti-peroxidase (PAP) method using anti-hog-elastase rabbit antibody was used for light microscopy, and the anti-elastase Fab'-peroxidase conjugate was used for electron microscopy. The tissues stained by the PAP method exhibited a dense deposition of reaction products on the luminal surface of epithelial cells and a moderate deposition in the blood and lymph capillaries of the intestinal villi. Immunoelectron microscopy revealed that the reaction product was deposited on the surface of the microvilli and in their pocketing; some was found in the pinocytotic vesicles in the terminal-web area and on the inner surface of the enlarged smooth endoplasmic reticulum. Round droplets which gave a positive reaction were found in the widened intercellular cleft and the thick basement membrane lining the blood capillaries and lymphatics. The jejunum retained its normal ultrastructure. The results indicate that the elastase molecules, which were introduced into the rat jejunum lumen, were absorbed without being decomposed through healthy intestinal epithelial cells by pinocytosis and translocated into blood and lymph capillaries.

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Degranulation effect of ferric nitrilotriacetate (Fe3+-NTA) on the pancreatic islet beta-cells: its acute toxic effect on glucose metabolism.

A single injection of ferric nitrilotriacetate (Fe3+-NTA) caused a transitory increase in plasma immunoreactive insulin (IRI) and plasma immunoreactive glucagon (IRG) in rats. They reached maximum levels at 2 days after injection and returned to the normal range at 10 days. At 2 days after Fe3+-NTA injection, blood glucose level was normal but the glucose tolerance test (GTT) was impaired. There was a further increase in plasma IRI level and IRG level was suppressed after glucose loading. At 10 days after Fe3+-NTA injection, glucose tolerance was normal and IRI also returned to the normal range. No degenerative changes were found on H.E.-stained rat pancreatic tissue sections after Fe3+-NTA injection. Histochemical staining, however, showed a reduction in beta-granules and heavy metals (Timm's granules) from islet cells in the central area of the rat pancreatic islet 1 to 3 days after injection of Fe3+-NTA. The fading remained in some islets even at 10 days after injection, but by then the beta-granule distribution was restored in most islet cells. The results indicate a single Fe3+-NTA injection induced transitory instability of the pancreatic islet beta-cell granules and the glucose intolerance with a hyperresponse of IRI.

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