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Biomedical subjects

D H Ausprunk

Publications and source records attributed to D H Ausprunk.

16 recordsLinked to original sources
PubMed1990

Human blood cells at microgravity: the NASA Initial Blood Storage Experiment.

The Initial Blood Storage Experiment (IBSE) probed the behavior of human red cells, white cells, and platelets during exposure to microgravity for 6 days and 2 hours on a National Aeronautics and Space Administration (NASA) shuttle mission, named STS 61-C, which was launched on January 12, 1986. IBSE involved carefully controlled comparisons between two identical sets of blood cells, one exposed to microgravity and the other held on the ground. Specially designed and fabricated, electrically powered environmental chambers provided appropriate environmental temperatures and air flow to support cell metabolism throughout the experiment. To circumvent the need for constant agitation of platelets during storage, a new thin-layer compression method for platelet preservation was developed. Blood cell samples were allocated to the two arms of the experiment, microgravity and earth gravity, by blind assignment. Moreover, to ensure unbiased assessment of the experiment's findings, postexperiment samples for measurement were identified by code. To optimize the chances of detecting possible gravitational effects, a wide array of measurements of cellular function, morphology, metabolism, and immunology were made. Analysis of variance was used in analyzing the data. The most striking finding was that platelets displayed markedly superior structural and functional integrity at microgravity. Granulocytes held on the ground were preserved slightly better than those that orbited in the shuttle, whereas red cells displayed few effects that were attributable to the gravitational variable. Polyvinylchloride-di-(2-ethylhexyl)phthalate (PVC-DEHP) was the plastic of choice for storage of red cells, while PVC-trioctyltrimellitate (TOTM) was superior to PVC-DEHP and polyolefin (PO) for platelets.

Aerospace Medicine↗
PubMed1986

Distribution of hyaluronic acid and sulfated glycosaminoglycans during blood-vessel development in the chick chorioallantoic membrane.

This study uses histochemical methods to determine the ultrastructural distribution of specific glycosaminoglycans (GAGs) during the development of blood vessels in the chick chorioallantoic membrane (CAM) and to correlate changes in GAG composition with the significant structural events in the development of these vessels. Tissues were stained with tannic acid, ruthenium red, and high iron diamine and digested in various GAG-degrading enzymes to identify specific GAGs. The results are consistent with a role for hyaluronic acid in the formation, alignment, or migration of the capillary plexus of the CAM and a role for sulfated GAGs (heparan sulfate, chondroitin sulfate, dermatan sulfate) in the differentiation and development of arterial and venous vessels of the chorioallantoic membrane.

Allantois↗
PubMed1986

Structural changes induced in capillary endothelial cells by growth factors: altered distribution of cytoskeletal elements and organelles in cells spreading in the presence of tumor conditioned medium and hypothalamus derived growth factor.

Using high-voltage and conventional electron microscopy of cell whole mounts, we have investigated the effects of tumor-conditioned medium and hypothalmus-derived growth factor on the structure of capillary endothelial cells during their attachment and spreading in tissue culture. Cells were cultured in A, Dulbecco's Modified Eagle's Medium (DMEM) and 10% calf serum; B, equal parts of A and 48 hr mouse sarcoma conditioned medium; and C, A containing 10 units of hypothalamus-derived growth factor. Cells cultured in all three media were fully spread, and to the same extent, by 4 hr after plating. While spreading, cells cultured in DMEM alone developed prominent stress fibers and contained numerous bundles of microtubules which formed radical tracts along which mitochondria and other organelles rapidly moved to the cell periphery. Stress fibers were thinner and microtubule tracts fewer in number in cells cultured in tumor-conditioned medium. In 4 hr, organelles moved only part of the distance to the cell margin. Stress fibers were rudimentary and microtubules randomly orientated in cells exposed to hypothalamus-derived growth factor. Most organelles remained near the cell nucleus. The dramatic decrease in stress fibers and microtubule tracts in cells grown in tumor-conditioned medium and hypothalamus-derived growth factor and the subsequent decreased capacity of the cells to move organelles toward their periphery could have some functional significance relative to the growth-promoting activity of these substances.

Adrenal Glands↗
PubMed1985

A morphologic and morphometric analysis of the aqueous outflow system of the developing cat eye.

The cellular and tissue changes accompanying the development and growth of the aqueous outflow system of the cat were investigated by quantitative light microscopy and by scanning and transmission electron microscopy. As in primates, the trabecular beams and sheets of the cat aqueous outflow system developed by reorganization of cells and extracellular matrix within the tissue filling the anterior chamber angle recess. Enlargement and coalescence of intercellular spaces gave rise to intertrabecular channels. From 3 to 9 days after birth, communications were established between the anterior chamber and intertrabecular spaces by perforation and resorption of tissue which initially covered the angle apex and appeared to be a peripheral extension of Descement's membrane and the corneal endothelium. Macrophage-like cells could be involved in this process. A rapid increase in the volume of the intertrabecular spaces and in the number of trabecular cells coincided with the opening of the trabecular meshwork to the anterior chamber. The trabecular meshwork grew 150-fold in volume from birth to adulthood, mainly as a result of a similar-fold expansion of its connective tissue components. The volume of the intertrabecular spaces increased 24-fold and trabecular cell number increased 14-fold during this same period. The disproportionate increase in volume of the various components of the trabecular meshwork was responsible for the decreased cell density and rarefaction displayed by this tissue as development progressed. Development of the aqueous outflow system of the cat is thus a complex, but highly co-ordinated, process, that depends on continued proliferation of cells and extracellular matrix, a progressive ordering of these components, and selective atrophy and removal of specific tissue components.

Aging↗
PubMed1981

Proteoglycans in the microvasculature. I. Histochemical localization in microvessels of the rabbit eye.

The ultrastructural organization of ruthenium red (RR) stainable material within small blood vessels located in the limbus of the rabbit eye was studied. Proteoglycans were identified in this material by digesting tissues with Streptomyces hyaluronidase, testicular hyaluronidase, chondroitinase ABC, or heparinase before ruthenium red staining. Neuraminidase digestion enabled separate identification of sialoglycoprotein. The luminal surface of endothelial cells demonstrates an RR-stained glycocalyx containing both sialoglycoprotein and proteoglycans, which are removed by testicular hyaluronidase and crude heparinase. The basal coat of endothelial cells and small granules (10-20 nm in diameter) located within the basal lamina stain with RR and are removed only by crude heparinase. The surface coat of smooth muscle cells and small granules (10-20 nm) within their basal laminas are also digested by crude heparinase. Large proteoglycan granules (20-50 nm), which are completely removed by testicular hyaluronidase and partially digested by Streptomyces hyaluronidase, are deposited between the connective tissue fibers of the media and adventitia. Other large granules that are attached to collagen fibers contain enzyme-resistant anionic materials. The surface coat of adventitial fibroblasts is removed only by crude heparinase. Thin filaments (3-5 nm in diameter) interconnect the cell coat material, basal lamina granules, and large connective tissue granules, to form a network of proteoglycans that traverses the intima, media, and adventitia. The highly ordered arrangement of proteoglycans in the microvascular wall suggests that these macromolecules play several roles in microvascular function.

Animals↗
PubMed1981

Proteoglycans in the microvascular. II. Histochemical localization in proliferating capillaries of the rabbit cornea.

The ultrastructural distribution of proteoglycans around capillaries growing in the cornea of the rabbit eye was determined after staining with ruthenium red (RR). Proteoglycans were identified by digesting tissues with glycosaminoglycan-degradative enzymes. Sialoglycoproteins were differentiated from proteoglycans by neuraminidase digestion. The capillary sprouts demonstrated a luminal glycocalyx containing testicular hyaluronidase-sensitive proteoglycans but little or no sialoglycoprotein. At the capillary tips, where mitosing and migrating endothelial cells are located, the basal cell surface displayed a network of small RR-stained granules (8 nm in diameter), which was partially removed by streptomyces hyaluronidase but not by testicular hyaluronidase. Thin filaments connected the granules to the endothelial cell plasmalemma and to a similar network of granules that is normally present in the corneal stroma. The stroma granules were partially digested by testicular hyaluronidase. In older capillary regions, where endothelial cells ceased proliferation, the basal network of proteoglycan granules was gradually infiltrated by fibrillar material until a basal lamina was formed. The proteoglycan granules were then arranged on both sides of the lamina densa, and a thin glycocalyx covered the basal endothelial cell surface. Thus, proteoglycans and anionic materials associated with growing capillaries serve to link proliferating and migrating endothelial cells to the extracellular matrix, help to organize the capillary basal lamina, form an anionic surface along the luminal front of capillaries, and probably help stabilize the structure of the capillary wall after proliferation ceases.

Animals↗
PubMed1978

Spreading of vascular endothelial cells in culture: spatial reorganization of cytoplasmic fibers and organelles.

The three-dimensional organization and fine structure of cytoplasmic components within whole non-embedded bovine aortic endothelial cells were examined during their attachment and spreading in tissue culture. Cells were cultured directly on Formvar-coated gold grids, fixed in glutaraldehyde and osmium tetroxide, critical point dried and examined by transmission electron microscopy (TEM) using stereoscopic methods, and by scanning electron microscopy (SEM). Reorganization of cytoplasmic structures during cell spreading occurred in four sequential stages: (1) spreading of the plasma membrane and unstructured cytoplasmic matrix; (2) spreading of cytoplasmic fiber systems (microtubules, microfilament bundles and microtrabecular system); (3) alignment of microfilament bundles and formation of radial tracts of microtubules; and (4) centripetal movement of organelles along radial tracts. These stages observed by TEM correlated with progressive degrees of cell flattening as visualized by SEM. These studies demonstrate that a characteristic reorganization of intracellular fiber systems and organelles accompanies the spreading of endothelial cells in culture.

Animals↗
PubMed1978

Endotoxin-induced changes in human platelet membranes: morphologic evidence.

Interaction between human platelets and bacterial endotoxin was studied in vitro with transmission and scanning electron microscopy. Washed human platelets, whose aggregation was blocked with apyrase, were incubated in a plasma-free medium containing crude endotoxin that had previously been complexed with copper. Thirty minutes of incubation resulted in adherence of endotoxin particles to the platelet surface, breaks in the platelet plasma membrane with apparent attempts at repair, pseudoped formation, and centralization of platelet organelles. Copper appeared to potentiate these phenomena, since neither Cu2+ at low concentrations nor endotoxin alone altered the morphology of the platelet membrane. This platelet-endotoxin interaction may be an intermediary step in the detoxification and clearance of endotoxin from the plasma.

Blood Platelets↗
PubMed1976

Vascular injury in transplanted tissues. Fine structural changes in tumor, adult, and embryonic blood vessels.

Adult, embryonic and tumor tissues from the rat were grafted to the chorioallantoic membrane (CAM) of the chick embryo. Fine structural changes in the microvasculature of the transplants were compared, with emphasis on the condition of endothelial cells. Endothelial cells in tumor grafts appeared irreversibly injured 2 h after grafting and most had degenerated by 8 h. In normal adult tissues, endothelial cells disintegrated more slowly, but lipid accumulation and blebbing leading to occlusion of vascular lumina, were prominent by 1 day. Cells were swollen and vacuolated 2 days after grafting and most were disrupted by 3 days. Embryonic endothelial cells were only mildly damaged and vessel lumina were patent for up to 3 days. Vessels were then reperfused with chick blood. It is concluded that the microvasculature in tumor tissue is extremely susceptible to ischemic damage and that this phenomenon may be involved in the central necrosis which develops in many animal tumors after they grow beyond a small size. Furthermore, only blood vessels and endothelium in embryonic tissues were sufficiently resistant to ischemic changes so that the graft microvasculature could be reperfused.

Animals↗
PubMed1975

Vascularization of normal and neoplastic tissues grafted to the chick chorioallantois. Role of host and preexisting graft blood vessels.

Adult, embryonic, and tumor tissues were grafted to the chorioallantoic membrane of the chick embryo to determine whether blood vessels originally within implants were reused in the establishment of a new blood supply. Grafts were examined daily by in vivo stereomicroscopy and in histologic sections. Colloidal carbon injections into the host vasculature served to confirm the precise onset of graft circulation; Preexisting tumor blood vessels disintegrated by 24 hours after implantation and revascularization occurred at 3 days by penetration of proliferating host vessels into the tumor tissue. Adult tissues did not revascularize, and the original graft vasculature progressively disintegrated during the 9 days of observation, Most embryonic tissues revascularized in 1 or 2 days by reperfusion of the existing graft vasculature. Anastomosis of host and graft blood vessels seemed to result from connections between newly formed vascular sprouts arising from both vasculatures. This study indicates that only tumor grafts acquire their blood supply solely by formation of new blood vessels from the host microvasculature. By contrast revascularization of normal tissues, when it does occur, is predominately the result of perfusion of the preexisting graft blood vessels.

Allantois↗