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W L Davis

Publications and source records attributed to W L Davis.

At least 73 records · Page 4Linked to original sources

Planetary protection issues in advance of human exploration of Mars.

Current planetary quarantine considerations focus on robotic missions and attempt a policy of no biological contamination. The presence of humans on Mars, however, will inevitably result in biological contamination and physical alteration of the local environment. The focus of planetary quarantine must therefore shift toward defining and minimizing the inevitable contamination associated with humans. This will involve first determining those areas that will be affected by the presence of a human base, then verifying that these environments do not harbor indigenous life nor provide sites for Earth bacteria to grow. Precursor missions can provide salient information that can make more efficient the planning and design of human exploration missions. In particular, a robotic sample return mission can help to eliminate the concern about returning samples with humans or the return of humans themselves from a planetary quarantine perspective. Without a robotic return the cost of quarantine that would have to be added to a human mission may well exceed the cost of a robotic return mission. Even if the preponderance of scientific evidence argues against the presence of indigenous life, it must be considered as part of any serious planetary quarantine analysis for missions to Mars. If there is life on Mars, the question of human exploration assumes an ethical dimension.

Bacteria↗

The glyoxylate cycle in rat epiphyseal cartilage: the effect of vitamin-D3 on the activity of the enzymes isocitrate lyase and malate synthase.

The effect of vitamin-D deficiency and subsequent vitamin-D replacement on the metabolism of rat epiphyseal growth plate cartilage was studied. Biochemical analyses showed the presence of the two unique glyoxylate cycle enzymes isocitrate lyase and malate synthase in cartilage. The activity of these enzymes was markedly increased after treatment with the vitamin. Additionally, rat cartilage showed the capacity to oxidize fatty acid in the presence of cyanide. This cyanide-insensitive fatty acid oxidation is characteristic of peroxisomal B-oxidation rather than mitochondrial B-oxidation. Vitamin-D treatment also increased fatty acid oxidation. Lastly, incubation of rat cartilage in the presence of a fatty acid substrate such as palmitate, resulted in a higher tissue glycogen content. Tissue glycogen was further elevated by vitamin-D. Such data indicate the presence of glyoxylate cycle enzymes in a vertebrate tissue and raise the possibility that mammalian cartilage has the capacity to convert lipid to carbohydrate.

Animals↗

Glyoxylate cycle in the rat liver: effect of vitamin D3 treatment.

Evidence for the glyoxylate cycle in the mammalian rat liver was sought. Activity of two unique glyoxylate cycle enzymes, isocitrate lyase and malate synthase, was found in rat liver homogenates. Vitamin D3 treatment of rachitic animals produced a five- and fourfold increase, respectively, in the activity of these enzymes. Vitamin D3 also increased the peroxisomal fatty acid oxidation and the accumulation of glycogen in liver slices in the presence of palmitate. These data suggest that the mammalian rat liver can convert fatty acid carbon to carbohydrate carbon directly.

Animals↗

Electron microscopic cytochemical localization of Ca-ATPase in the rod outer segments of the toad Bufo marinus.

Transmission electron microscopy and ultracytochemistry were employed in an attempt to localize the enzyme calcium adenosine triphosphatase (Ca-ATPase) in the rod outer segments (ROS) of the toad retina. Utilizing a one-step incubation procedure, Ca-ATPase was identified as an electron-dense precipitate in the intradiskal spaces of the rod disks (vesicles) of the ROS. Analytical microscopy identified the reaction product as lead phosphate. The formation of the reaction product was dependent on the presence of ATP (the substrate) and calcium ions. However, calcium ions could be substituted for by magnesium ions. In addition, the reaction was vanadate sensitive. The latter is known to inhibit Ca-ATPase activity. Such data appear to indicate the presence of a Ca-Mg-ATPase in association with the rod disks. Since cyclic guanosine monophosphate (cyclic GMP), rather than calcium ions, is currently believed to be the primary intracellular messenger associated with phototransduction, the presence of an ROS Ca-ATPase may indicate other functions for this cation in the physiology and biochemistry of the visual process. Ca-ATPase might play a role in directional calcium fluxes between intracellular compartments.

Animals↗

Effects of contrast media on calcium transients and motion in cultured ventricular cells.

To investigate the mechanisms of the negative inotropic effects of contrast media, we superfused spontaneously contracting cultured chick embryo ventricular cells with Renografin-76 and iohexol (12% solutions), and hypertonic sucrose during simultaneous measurement of [Ca2+]i transients (indo-1) and motion (video-motion detector system). Exposure to contrast agents caused a significant reduction of contractility, with Renografin-76 having a much greater effect on amplitude of motion than iohexol. Renografin-76 significantly depressed [Ca2+]i transient amplitude, whereas iohexol had no effect. Addition of Ca2+ to correct for calcium binding by Renografin-76 completely reversed its depression of [Ca2+]i transients but only partially reversed the negative inotropic effects. Hypertonic sucrose caused a significant decrease in contraction amplitude, with no significant effects on [Ca2+]i transient amplitude. We conclude that the marked negative inotropic effect of Renografin-76 is caused by both calcium binding and hypertonicity. The less marked depression of contractility produced by iohexol likely is a result of hypertonicity and not caused by alteration of [Ca2+]i.

Animals↗

Quantitation of phagocytic cells in phenytoin-induced connective tissue proliferation in the rat.

This study demonstrates that, in rats, systemically administered phenytoin produces a statistically significant increase in accumulation of phagocytic and esterase positive cells in areas of phenytoin-induced connective tissue proliferation. Many of these cells are perceived to be macrophages. Since macrophages have been shown to produce chemoattractant and mitogens for fibroblasts, they may very well play a major role in the gingival hyperplasia seen in patients taking phenytoin. The results of the study show that the effects of phenytoin on fibroblasts may be more complex than previously thought.

Animals↗

Isthmus and ligament interrelationships: a computerized analysis of the spatial relationships of the hinge structures in unionid mussels.

A study of the isthmus and ligament in unionid mussels was undertaken employing methods specifically designed to preserve the in vivo relationships of these interrelated structures. Serial sections of the hard and soft tissues were used for two-dimensional analysis. From these, a tridimensional computerized reconstruction was developed. Special dissections of the undisturbed isthmus were also utilized. By using such methodologies, a new description of the ligament has been developed employing such terminology as the foliated ligament and the posterior folding laminae. Similarly, for the isthmus, an anterior lyre, a pallial crest, a pallial peduncle, and a posterior lyre are described. Such entities are both morphologically and physiologically related to the shell and ligament.

Animals↗

Histochemical and elemental localization of calcium in the granular cell subapical granules of the amphibian urinary bladder epithelium.

The ultrahistochemical analysis of apical granules in the epithelial cells, i.e., granular cells, of the amphibian urinary bladder using the N,N-naphthaloylhydroxylamine procedure identified the presence of calcium in these structures. Subsequent analytical microscopy employing fresh-frozen ultrathin cryosections for X-ray microanalysis of the granules further confirmed the above histochemical findings. In addition to calcium, elemental analysis indicated the presence of magnesium, phosphorus, sulfur, silicon, potassium, and chlorine either within or in close proximity to the granules. The possibility that these granules function as subcellular compartments for the uptake and storage of calcium ions, in a way similar to mitochondria, and thus function in intracellular calcium homeostasis, is discussed. Additionally, a role for this cation in the secretion of granular glycoproteins, i.e., stimulus-secretion coupling, is hypothesized.

Animals↗

Electron microscopic cytochemical localization of a basolateral calcium adenosine triphosphatase in vitamin D replete chick enterocytes.

A cytochemical technique for the electron microscopic localization of calcium adenosine triphosphatase (Ca-ATPase) was utilized to localize this enzyme in the enterocytes of rachitic and vitamin D-replete chicks. In animals treated with cholecalciferol (CC, vitamin D3), an electron-dense reaction product was located along the basolateral membranes of the absorptive cells within 72 hr after injection. Similarly, a reaction product was identified in association with the basolateral membranes within 24 hr after injection of 1,25-dihydroxycholecalciferol, the active metabolite of vitamin D. A microvillar reaction product was not seen in either of these two groups. Electron-dense reaction products were also seen in association with mitochondria and scattered throughout the cytoplasm of these enterocytes. The Ca-ATPase reaction product was dependent upon the presence of medium calcium and substrate (ATP), was inhibited by vanadate, and was heat labile. In the rachitic animals, a reaction product indicative of Ca-ATPase activity was not seen in association with either the basolateral membranes or the mitochondria. These data appear to indicate that an energy-requiring calcium-activated membrane pump plays a role in the flux of calcium across the enterocytes of the small intestine.

Animals↗

Electron microscopic cytochemical localization of Ca-ATPase in toad urinary bladder.

The calcium-regulating enzyme calcium adenosine triphosphatase (Ca-ATPase) was localized in the epithelium of amphibian urinary bladder by the one-step electron microscopic cytochemical procedure. The enzyme was identified along the basolateral border of the epithelial cells that comprise the bladder mucosa. The electron-dense precipitate indicating Ca-ATPase activity was seen in association with the outer leaflet of the basolateral plasmalemmae. Intracellularly, Ca-ATPase activity was seen in association with the mitochondrial matrix of the mitochondria-rich cells. Ca-ATPase was not seen along the apical microvillated border. Enzyme activity was also not seen after incubation in substrate-free media, calcium-free media, or incubation in the presence of vanadate. However, Ca-ATPase activity was evident when the calcium in the standard reaction medium was deleted in favor of magnesium. Addition of antidiuretic hormone (ADH; vasopressin) increased both the basolateral Ca-ATPase reaction and the mitochondrial reaction. Such data appear to indicate further that changes in cytosolic calcium ion concentration take place during the response of amphibian urinary bladder to the polypeptide hormone vasopressin.

Animals↗

Electron microscopic cytochemical localization of adenylate cyclase in amphibian urinary bladder epithelium: effects of antidiuretic hormone.

A cytochemical technique for electron microscopic localization of adenylate cyclase was used to identify this enzyme in quiescent and hormone-stimulated toad urinary bladder epithelium. In the absence of vasopressin (antidiuretic hormone), adenylate cyclase was detected along the outer surface of the basolateral plasma membranes of granular cells, mitochondria-rich cells, and basal cells, the major cell types comprising the hormone-sensitive urinary epithelium. In the presence of antidiuretic hormone, the basolateral precipitates were markedly increased. The latter was true for both tissues incubated in the presence of an osmotic gradient and those stimulated in the absence of such a gradient. A significant mucosal reaction was never seen. Such data indicate that the hormone receptors for vasopressin are located along the basolateral membranes of all epithelial cells comprising the mucosal hormone-sensitive epithelium. All cells of the epithelium also demonstrate a vasopressin-sensitive adenylate cyclase. We discuss possible mechanisms that attempt to integrate the cytochemical data into an overall scheme for the physiological action of this hormone on amphibian urinary bladder.

Adenylyl Cyclases↗

Bovine bone morphogenetic protein (bBMP) fraction-induced repair of craniotomy defects in the rhesus monkey (Macaca speciosa).

Large cranial defects do not always heal spontaneously, especially in humans; often they have to be obturated with metallic or acrylic fillers. Bilateral cranial trephine defects, measuring 14-20 mm in diameter, were created in three Rhesus monkeys, providing a typical primate model system for investigations in comparative physiology of bone regeneration. Each skull had one control and one experimental trephine defect. Control defects were implanted with bovine serum albumin (BSA). Experimental defects were implanted with 100-200 mg of a partially purified fraction of bovine bone morphogenetic protein (bBMP) in two monkeys. In one monkey, the BMP was incorporated in a 1:1 poly(lactic) poly(glycollic) acid copolymer, high-viscosity formula. The animals were killed at eight weeks, ten weeks, and 16 weeks after implantation. The tissue responses were analyzed by computed histomorphometry and routine histologic examination. At each time interval, the BMP implanted defects produced more complete regeneration than the control implants. The morphogenetic response occurred in the following sequences: mesenchymal cell proliferation, chondrogenesis, and increased bone formation. BMP-induced osteogenesis may initiate the regenerative process. The copolymer releases BMP but may constitute a barrier to the end stages of replacement by new bone, and would be more useful in a low-viscosity rapidly biodegradable form.

Animals↗

Phenytoin-induced connective tissue growth in the rat.

Rats were treated daily for 9 days with 100, 50, or 25 mg/kg phenytoin i.p. This treatment resulted in a significant increase in the thickness of the connective tissue capsules of the liver, spleen, and pancreas, and of the subepithelial connective tissue of the mesentery but not the epicardium or visceral pleura of the lung where exposure to the drug was via the vascular route. Many areas of connective tissue growth exhibited obvious proliferation of fibroblasts and in some areas contained seemingly large numbers of macrophages and an increase in vascularity. It was demonstrated by electron microscopy that the macrophages occasionally were seen in intimate contact with the fibroblasts. Our observations clearly showed that intraperitoneal exposure of visceral connective tissues of the rat to phenytoin rapidly resulted in a dose-related proliferation of that tissue. The presence of numerous macrophages leads to the suggestion that macrophage-derived growth factor could be responsible for the increased growth.

Animals↗

Fine structure and immunofluorescent studies of the WISH cell line.

Morphological studies of the WISH cell line reveal an epithelioid cell type with some characteristics of both the original human amnion epithelium and a transformed state. WISH cells have a cytoplasm filled with microtubules; however, actin filament bundles are few, with actin localized at areas of cell contact and arranged diffusely through the cytoplasm, as viewed by indirect immunofluorescence. Fingerlike projections or short filopodia are observed connecting cells that grow in a closely apposed monolayer. Other surface features, as viewed by scanning electron microscopy, include microvilli and blebs. Transmission electron microscopy shows that WISH cultures consist of light or dark cells with organelles that include lipid droplets, abundant free ribosomes, tubular mitochondria, lysosomes, annulate lamellae, rough endoplasmic reticulum, 6-nm microfilaments, 10-nm intermediate filaments, and microtubules. Pleomorphic nuclei with multiple nucleoli and fibrillar nuclear bodies are common. Desmosomes and subsurface confronting cisternae connect cells. To our knowledge, these structural studies are the first to describe WISH and lead to subsequent investigation of the cell surface phenomenon of blebbing and surface charge in WISH and another human cell line.

Actin Cytoskeleton↗

Cytochalasin D and cationized ferritin as probes for the morphological investigation of blebbing in two human cell lines.

The potent fungal metabolite cytochalasin D (CD) and cationized ferritin (CF) are used in combination to test for negative charge distribution on blebs (knobs). Two established human epithelial cell lines, WISH and HeLa, that display blebs in various phases of the cell cycle or under certain culture conditions are investigated. CD alone, applied at a low concentration (1.0 micrograms/ml) and for a short time period (3 min), causes blebs to appear as the prevalent surface feature. These are filled mainly with free ribosomes. Additionally, feltlike mats, presumed to be disorganized, compacted microfilaments, are formed directly beneath the cell membrane. These are especially evident in the cortical cytoplasm below the blebs or bleb clusters. CF (0.345 mg/ml), applied for a 5-min period after CD administration (1.0 microgram/ml) for 3 min, appears along the surface of microvilli, at the base of blebs, and in vesicles beneath the bleb clusters. In some cases, microfilaments (6 nm in diameter) are closely related to the vesicles. CF does not preferentially bind to the apical cell membrane of blebs. Above areas of the subplasmalemmal microfilaments, CF membrane binding is apparent, even under circumstances where the filaments are disorganized by cytochalasin treatment. These results seem to show the following: bleb membranes are different from the remainder of the cell and do exhibit a loss of negative charge and surface charge may be dependent on the presence or structural integrity of membrane-related 6-nm microfilaments.

Actin Cytoskeleton↗

Antidiuretic hormone response in the amphibian urinary bladder: time course of cytochalasin-induced vacuole formation, an ultrastructural study employing ruthenium red.

Cytochalasin is known to inhibit the antidiuretic hormone-induced hydro-osmotic response (bulk water flow) in the amphibian urinary bladder without altering hormone-stimulated diffusional water permeability or short-circuit current. In addition, histological studies have shown that the mold metabolite induces the formation of large intracellular vacuoles or lakes in the epithelial cells. We report here a transmission electron microscopic time-course study which indicates that during the early phases of the ADH response cytochalasin causes the formation of numerous multivesicular bodies or aggregates derived from individual basolateral pinocytotic vesicles. Because of their apparent hypertonic nature, the vesicles, as well as the vesicular aggregates, accumulate water during hormone-stimulated hydro-osmotic flow. As a result, the multivesicular bodies dilate and fuse to form the large intracellular lakes characteristic of cytochalasin treatment in the presence of both an applied osmotic gradient and vasopressin. In the presence of mucosal ruthenium red, the luminal glycocalyx was heavily stained with this tracer. At no time, however, even in the presence of hormone, was there any evidence for the uptake of this dye at the apical epithelial border. In the presence of serosal ruthenium red, the lateral intercellular spaces, basolateral pinocytotic vesicles, basal lamina, and collagen, as well as other subepithelial structures, were ruthenium positive. With cytochalasin D, vasopressin, and serosal ruthenium red, both the pinocytotic vesicles and the multivesicular bodies demonstrated an apparent membrane associated ruthenium positive coat. The tracer data indicates that the basolateral pinocytotic vesicles, increased by the presence of hormone, are indeed endocytotic in nature. The mucopolysaccharide coat associated with these structures may be involved in ionic and/or fluid transport.

Animals↗

Effects of ocular inflammation on tear proteins. I. Thiol protease inhibitors.

Concentrations of protease inhibitors found in tears have been observed to vary during ocular inflammation. Possible stimuli for these variations were tested for by studying thiol protease inhibitor concentrations in models of ocular inflammation. Inflammatory responses were elicited either by topical application of arachidonic acid (AA), or compound 48/80. For each model, four rabbits were treated in one eye with the inflammatory stimulant and tear samples were collected before and up to five hours following treatment. Control animals were treated with buffer. When compared to pretreatment values, tears from AA-treated eyes showed decreased inhibitor at one and two hours and an increased inhibitor concentration at three hours. In the 48/80 model, inhibitory activity and protein levels were elevated when compared over time to the control group (p greater than or equal to 0.02, p greater than or equal to 0.0001 respectively). When compared to pretreatment values, inhibitor values were elevated at all times after treatment and protein values were elevated at one and three hours in this model. Serum proteins were also increased in the tears of rabbits treated with 48/80. The results suggest that one or more of the mediators released by basophil or mast cell degranulation stimulate increased tear protease inhibitory activity and tear serum proteins.

Animals↗

Effects of contrast media on cultured myocardial cells.

To quantify and characterize mechanisms of negative inotropic effects of ionic and nonionic angiographic contrast media, spontaneously contracting monolayers of cultured chick ventricular cells were studied using the Renografin-76 and iohexol as characteristic ionic and nonionic contrast agents. Utilizing microscopic video-motion detector techniques the effect of each contrast agent on contraction amplitude was measured. Also, the effect of changes in the ionized calcium concentration and osmolality induced by the contrast agents was evaluated. The study demonstrated a linear depression of contractility by increasing contrast concentrations. The negative inotropic effect of Renografin-76 was significantly greater than that of iohexol. Renografin-76 demonstrated significant calcium binding, whereas iohexol had none. Correction for calcium binding by Renografin-76 partially reversed the negative inotropic effect of this agent. Hypertonic sugar solutions of equivalent osmolality to the contrast agents demonstrated dose-related negative inotropic effects such that, in this preparation, calcium binding by Renografin-76 and its high osmolality accounted for all its negative inotropic effect. In contrast, iohexol demonstrated an intrinsic negative inotropic effect not completely explained by its osmolality.

Animals↗