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Biomedical subjects

D E Bockman

Publications and source records attributed to D E Bockman.

At least 19 recordsLinked to original sources

Inhibition of epibranchial placode-derived ganglia in the developing rat by bisdiamine.

Although bisdiamine has been shown to affect the development of mammals, its effect on the nervous system has gone largely unrecognized. In the present study, rats were given bisdiamine by gavage on days 9 and 10 of pregnancy. They were sacrificed at intervals and the fetuses were prepared for study of serial sections stained with hematoxylin and eosin, or by immunohistochemical reaction with HNK-1 monoclonal antibody. HNK-1 reacted strongly with the nervous system, allowing precise analysis of the components and their relationships. Controls receiving no bisdiamine were prepared and studied in parallel with the experimental fetuses. Administration of bisdiamine inhibited development of the petrosal and nodose ganglia, altered associations of the glossopharyngeal, vagus, and hypoglossal nerves, and inhibited contributions of vagal nerve fibers to the developing enteric system. The proximal ganglia of the glossopharyngeal and vagus nerves developed normally. It is concluded that bisdiamine affects, directly or indirectly, the differentiation of nervous components derived from the epibranchial placodes. It seems likely that these placode-derived components serve as pioneer neurons in establishing the pathway for the posteriorly extending trunks of the glossopharyngeal and vagus nerves. The early changes in congenital conditions such as the DiGeorge syndrome may not be limited to alterations in neural crest derivatives. It may be worthwhile to investigate more closely whether there are alterations in the nervous system associated with these syndromes.

Abnormalities, Drug-Induced

Microvasculature of the pancreas. Relation to pancreatitis.

Local or generalized alteration of microcirculation may be expected in diseases of the pancreas. Changes may range from increased permeability of capillaries to hemorrhage. Tissue necrosis may result from prolonged ischemia owing to intravascular coagulation and severely impaired blood flow. It is possible to observe early microvascular changes by intravital microscopy. Klar and coworkers have demonstrated by this method that isovolemic hemodilution improves blood flow under conditions that would otherwise lead to tissue damage. This paper presents the basic microcirculation of the pancreas and the changes that accompany pancreatic disease. It emphasizes that concentration on the changes in microcirculation that accompany the early manifestations of pancreatic diseases, particularly pancreatitis, may reveal important clues to their pathogenesis.

Acute Disease

Cytological changes in the pancreas of transgenic mice overexpressing transforming growth factor alpha.

Transgenic mice overexpressing human transforming growth factor alpha (TGF-alpha) predictably develop an enlarged, firm pancreas. The present study investigated the changes that occur in the different components of the pancreas in these animals. The increase in size of the pancreas may be accounted for by increased connective tissue. The added collagen is mainly type I. Thin, elongate fibroblasts are frequently bordered by a basal lamina, a relationship that is normally restricted to the perineurium. Collagen is intimately associated with epithelial cells. Fingers of connective tissue extend close to acinar lumina. Redifferentiation of acinar cells produces tubular complexes. In some cases, acinar cells take on the appearance of ductular cells. In some, there is a transition to mucin-producing cells. Intermediate forms between acinar and mucin-producing cells are present. The growth factor is localized in acinar cells and decreases with redifferentiation. The pancreas of these animals routinely displays characteristics that also are observed in diseases of the exocrine pancreas in humans, including fibrosis and redifferentiation. It is likely that the changes are the result of both direct and indirect effects of TGF-alpha, some of which may parallel altered control mechanisms in human pancreatic disease. Study of this model may provide clues to understanding the initiation of fibrosis and redifferentiation in human pancreas.

Animals

Capacity of neural crest cells from various axial levels to participate in thymic development.

In order to assess the capacity of neural crest from different sources to participate in thymic development, neural crest from selected axial levels was transplanted unilaterally from quail donors to the region in chick hosts from which neural crest cells normally migrate to interact with the primordial thymus. The greatest representation of donor cells was observed after isotopic transplantation and when donor tissue was taken from the hyoid and mesencephalic regions of the neural crest. The capacity for transplants to contribute cells decreased both anteriorly and posteriorly, so that neural crest close to the usual origin of mesenchyme-producing cells contributed a large number of donor cells around the developing thymus than neural crest from anterior and posterior regions. Cells from the transplant were inserted as an addition to the host chick cells. Thus, a special relationship and capacity for interaction in thymic development is expressed by neural crest at usual levels over a limited span of axial regions, but to some extent by all regions. This study has established that the capacity for neural crest cells from different axial levels to interact with developing organs is not uniform, but may vary, depending upon the nature of the interaction with a particular organ.

Animals

Impaired development of the thymic primordium after neural crest ablation.

Impaired thymic development as a result of ablation of neural crest has been observed in embryos late in development. The present study was initiated to determine what changes are effected early in thymic development by neural crest ablation. The epithelial primordia of the thymus were studied in chick embryos on the sixth day of incubation. Embryos with neural crest ablations were compared with sham-operated and untreated controls. Neural crest ablation inhibited formation of epithelial thymic primordia. Primordia in experimental embryos were fewer in number and were smaller than in shams and untreated controls. When primordia from shams and controls were transplanted to the chorioallantoic membrane of chick hosts, they were able to develop into organs with the typical features of embryonic thymus. Similar transplantation from neural crest-ablated animals, on the other hand, led to small, predominantly epithelial structures with meager lymphoid development. These findings are consistent with the hypothesis that mesenchyme derived from cranial neural crest is critical in initiating and sustaining the development from pharyngeal pouches of epithelial structures competent to attract and support the proliferation and differentiation of lymphoid stem cells.

Allantoin

The participation of neural crest derived mesenchymal cells in development of the epithelial primordium of the thymus.

The purpose of this study was to correlate the contributions by derivatives of the neural crest with the development of the epithelial primordium of the thymus. The monoclonal antibody E/C8 was used to localize derivatives of the neural crest in chick embryos. Neural crest was ablated by microcautery of neural folds. Evaluation of thymic development was carried out on serial sections of embryos sacrificed on the sixth day of incubation. The size of the epithelial thymic primordium was smaller in experimental animals than in shams. E/C8-immunoreactivity was concentrated around the periphery of the primordium. It was determined, by quantifying reaction product using the Core-SCAN computer color analysis program, that the amount of immunoreactivity was decreased after ablation of neural crest. Statistical analysis showed that the quantity of reaction product was positively and significantly correlated with the size of the thymic primordium. It is concluded that mesenchymal derivatives of the neural crest, through participation in the early development of the epithelial primordium, play an important role in thymic development, and therefore with development of the immune system.

Animals

Localization of fibronectin in megakaryocytes of fetal liver.

Antibodies specific for fibronectin were utilized to determine the sites of localization in the liver during development. The livers of fetal rats from each of gestation days 11-19, and from days 1 and 8 postpartum, were studied by fluorescence microscopy. Fibronectin was localized predominantly in megakaryocytes and megakaryocyte precursors, and to a lesser extent in the extracellular matrix surrounding blood vessels and between hepatocytes and sinusoids. The cytoplasm of megakaryocytes and their precursors displayed bright fluorescence but their nuclei were negative for fibronectin. Hepatocytes had negative or faint fluorescence. Megakaryocytes were present in the liver from day 12, and were numerous from day 13 through most of the rest of gestation. The relative numbers of megakaryocytes decreased in later gestation; at 8 days postpartum only a few were observed per section. Hepatic megakaryocytes appeared before megakaryocytes were established in spleen and bone marrow. The early and persistent high levels of fibronectin in hepatic megakaryocytes, in the absence of comparable localization within hepatocytes, leads us to the hypothesis that megakaryocytes are important in establishing circulating fibronectin levels in the fetus. Similarly, bone marrow megakaryocytes may contribute to circulating fibronectin in the adult.

Animals

Alteration of early vascular development after ablation of cranial neural crest.

A previous study has shown that, subsequent to ablation of cranial neural crest, heart morphology and pharyngeal arch vessels (aortic arches) are altered before septation of the outflow tract normally occurs. In the present study, we concentrated on very early development of the aortic arch apparatus in the chick (incubation days 3-5). The three-dimensional organization of the arch vessel apparatus was studied by scanning electron microscopy after intravascular injection of Mercox, and by serial sections of embryos embedded in plastic. Alterations in the arch vessel apparatus were already present by day three in embryos with neural crest ablation at stage 9-10. Bilateral symmetry frequently was lost. Arch vessels sometimes were enlarged and occupied most of the arch, with little surrounding mesenchyme. Some arch vessels were small or occluded. Mesenchyme was significantly reduced in quantity in the arches, and was not condensed and symmetrical as in controls. There was a significant increase in the proportion of direct apposition of vessel endothelium with epithelium, without the intervening mesenchyme typical of controls. The surgical manipulation used in this study leads to distinct alterations in the arches of components and relationships which are important in development. Altered blood flow likely affects the development of the heart.

Animals

Bypass circuits as the source of thromboemboli during extracorporeal membrane oxygenation.

To determine the presence and extent of thrombus formation in the apparatus used for extracorporeal membrane oxygenation we studied various portions of the polyvinylchloride circuit from five infants who received extracorporeal membrane oxygenation for 70 to 330 hours. All infants had right-sided cannulation. Sections were cut from the circuit at the time of decannulation and subjected to light and scanning electron microscopy. The site that contained the most thrombus formation was the membrane oxygenator bypass circuit, which is subjected to repeated periods of unclamping and clamping to direct blood flow through the membrane oxygenator. Autopsy results from nonsurvivors showed evidence of pulmonary and renal infarcts, a left frontal lobe infarct, a thromboembolus of the left external and internal carotid arteries, and thrombi in the lungs, kidney, brain, and coronary arteries. One survivor had computed tomographic evidence of infarction of the left middle cerebral artery distribution. We suggest that the areas of the extracorporeal membrane oxygenation circuit subjected to repeated changes in flow dynamics may be the source of microemboli.

Catheters, Indwelling

Early association of duodenal contents and blood with acini during experimental pancreatitis.

This study was directed toward understanding the mechanism by which the closed duodenal loop induces experimental pancreatitis. Carbon black was introduced into the duodenal lumen as a tracer before formation of the closed loop. Within four hours, the carbon, marking the location of duodenal contents, had passed retrogradely through pancreatic ducts and broken out to localize at the periphery of acini in various regions of the pancreas. In this periacinar location the tracer was mingled with erythrocytes, neutrophils, platelets, and fibrin. Acinar cells were damaged as the mixed duodenal and blood components passed along their base and between cells. It is concluded that acinar cells are involved very early and directly in the initiation of pancreatitis by this method, and not secondary to ductal inflammation.

Acute Disease

Analysis of nerves in chronic pancreatitis.

We sought to identify characteristics of pancreatic nerves that were altered in chronic pancreatitis. Pancreatic tissue removed from patients with chronic pancreatitis was analyzed for the number and size of nerves, their association with inflammatory infiltrates, and their fine structure. The mean diameter of nerves in these patients was significantly greater than in controls, whereas the mean area of tissue served per nerve was significantly less than in controls. Foci of inflammatory cells, prominent in some specimens, sometimes were associated with nerves and ganglia, but inflammatory foci and neural elements also existed separately. Invasion of nerve tissue by inflammatory cells was observed but was not massive. Ultrastructural changes were detected in nerves. Individual nerve fibers showed evidence of damage, and there was evidence of edema in the nerve bundle. The perineurial sheath was altered so that it no longer provided a barrier between the surrounding connective tissue and the internal neural components. The results of this study indicate that nerves are preferentially retained while parenchyma degenerates and is replaced by fibrosis during chronic pancreatitis, but that they are retained in an altered condition. Increased mean diameters of nerves in chronic pancreatitis argues against pain being caused by constriction due to fibrosis. It is likely that both sensory and motor nerve fibers are affected by this alteration.

Adult

Effect of neural crest ablation on development of the heart and arch arteries in the chick.

Mesenchymal derivatives of the neural crest contribute to the connective tissues and blood vessels of the pharyngeal arches, and participate in the septation of the outflow tract of the heart. The present study was designed to determine the nature and timing of alterations in the development of the heart and arch arteries subsequent to diminished neural crest contributions. The neural crest contributing to the three caudalmost pharyngeal arches was ablated bilaterally in chick embryos and compared with sham or unoperated controls. Heart development was studied by scanning electron microscopy. Arch artery development was studied microscopically after intravascular injection of India ink and clearing of the specimen. Neural crest ablation caused morphological changes in most hearts. Hearts in experimental animals commonly were elongate and were subject to inappropriate development of ventricular and atrial areas. A surgical effect delayed the disappearance of arch arteries one and two, and removal of neural crest produced an additional delay. Neural crest ablation caused failure of arch arteries three, four (right), and six to develop to the proper size in some animals. Survival of those whose sixth arch arteries achieved the proper size caused group measurements to reach normal values again by stage 32. Closure of arch arteries in some animals and maintenance in others produced greater variability in experimental animals than in controls. It is significant that heart morphology was altered before septation of the outflow tract normally occurs. This indicates at the least that another factor, such as altered blood flow, contributes to the abnormal development. Altered flow may result from changes in pharyngeal arch mesenchyme and arch artery endothelium.

Animals

Chronic obstructive pancreatitis, nesidioblastosis, and small endocrine pancreatic tumor.

A patient is described in whom a small 0.8-mm-diameter endocrine tumor was found in association with localized pancreatitis, stricture of the main pancreatic duct, and nesidioblastosis. The endocrine tumor, localized 7 mm from the main duct, obstructed a collateral pancreatic duct and four of its branches. Pancreatitis was localized into two areas. It was fairly severe in a small area of the body drained by the obstructed collateral duct and extended around it, reaching the main pancreatic duct which was partly occluded at this level by periductal fibrosis. Pancreatitis was mild in the left part of the pancreas distally from the incomplete stricture of the main duct. The head and body of the pancreas between the stricture and the duodenum were normal. Nesidioblastosis was present in the obstructed parts of the pancreas. It is suggested that the tumor could have caused pancreatitis, stenosis of the main duct, and nesidioblastosis.

Chronic Disease

Pancreatic extracellular matrix alterations in chronic pancreatitis.

The proliferation of pancreatic extracellular matrix, which characterizes chronic pancreatitis, has been analysed using immunohistochemistry. The relationship of matrix components to intraductal precipitates and the presence of serum proteins in precipitates were also studied to investigate the suggestion that ductal permeability increases in chronic pancreatitis. Pancreatic tissue from organ donors was compared with that from patients with chronic calcifying or chronic obstructive pancreatitis. Frozen sections were labeled with monospecific antibodies to collagen types I, III, pro-III and IV, laminin, fibronectin, IgG, IgA, and IgM and then visualized by indirect immunofluorescence. In chronic pancreatitis, interstitial collagens and fibronectin appeared increased and disorganized in both fibrous tissue and areas that appeared histologically normal. Type IV collagen distribution was abnormal and in some sites was present with interstitial collagen. In addition, intraductal precipitates were shown to contain immunoglobulins, and defects were identified in the duct basal lamina associated with precipitates. These results demonstrate that in chronic pancreatitis interstitial collagens are extensively disorganized, the fibrosis possibly being relatively labile. The presence of serum proteins in intraductal precipitates confirms an increase in ductal permeability, and associated defects in the basal lamina appear to define a route via which serum proteins may enter the intraluminal compartment.

Adult

Ultrastructure of human acute pancreatitis.

Few studies have been published on the ultrastructural changes which accompany human acute pancreatitis, and these have concentrated primarily on parenchyma. The present study concentrates on extraparenchymal changes, compares acute pancreatitis occurring alone with that on a background of chronic pancreatitis, and tests for similarity with observations made previously in an experimental model. Pancreatic tissue came from 16 patients undergoing surgery for pancreatic disease and five subjects without pancreatic disease. Regressive changes in parenchymal cells were consistent with ischemia, and with previously described studies. Polymorphonuclear leukocytes infiltrated into stroma and parenchyma. Platelets accumulated intra- and extravascularly. Fibrin deposits were common in the connective tissue, and could be observed in intercellular spaces at the base of acini, mingled with degenerating acinar cells and secretion product. Microthrombi occurred in blood vessels. These alterations were consistent with those in experimental acute pancreatitis. Similar changes were observed whether or not acute pancreatitis occurred on a background of chronic pancreatitis. The vascular component is important in acute pancreatitis, and altered epithelial barriers allow interaction between blood-borne material and pancreatic exocrine secretions.

Acute Disease

Structure and function of specialized cilia in the exocrine pancreas.

Cilia are found in acinar and ductular lumina. Most descriptions of human or dog pancreatic tissue have indicated that these are probably motile structures which move and/or mix pancreatic secretion. A recent study interpreted arrangements of microtubules within cilia which deviated from the classic 9 + 2 pattern as indicating pathological change. There have been suggestions, however, from studies on nonmammalian and human pancreas, that the structure of pancreatic cilia suggests a sensory function. The present paper reports studies on the ultrastructural organization of the specialized cilia in human and canine pancreatic tissue. The pattern of microtubular organization resembles that of cilia modified for chemoreception rather than that of classic kinocilia. It is concluded that pancreatic ductular cilia are sensory rather than motile, providing a mechanism for monitoring general or specific molecular concentrations.

Animals