Search PubMed⌕ Search

Biomedical subjects

W L Davis

Publications and source records attributed to W L Davis.

At least 91 records · Page 5Linked to original sources

Cytochemical localization of malate synthase in amphibian fat body adipocytes: possible glyoxylate cycle in a vertebrate.

The adipocytes of amphibian abdominal fat bodies contain typical microperoxisomes, as indicated by their fine structure. Electron microscopic cytochemistry showed that these organelles contain the enzymes catalase, typical for peroxisomes, and malate synthase. The latter is an enzymatic component characteristic of the glyoxylate cycle, a biochemical pathway known to exist in plant glyoxysomes (peroxisomes). This metabolic pathway makes possible the net conversion of lipid to carbohydrate. Toad adipocytes may represent yet another example of vertebrate peroxisomes which contain one of the marker enzymes (malate synthase) characteristic of the glyoxylate shunt.

Adipose Tissue↗

Activation of actin-containing microfilaments by vasopressin in the amphibian urinary bladder epithelium: a fluorescent study using NBD-phallacidin.

The effect of an antidiuretic hormone (ADH, vasopressin) on the microfilament system of the toad urinary bladder lumenal epithelium was investigated using NBD-phallacidin (NBD-ph). The latter material is a specific fluorescent label for F actin. In the presence of an osmotic gradient, both ADH and cyclic adenosine monophosphate (cAMP) appear to induce the polymerization of monomeric actin into F actin-containing microfilaments. The latter may then be involved in the morphological changes, including the formation of lateral intercellular lakes, associated with the typical hydroosmotic response.

Actins↗

Muscle cell membranes from early degeneration muscle cell fibers in Solenopsis are leaky to lanthanum: electron microscopy and X-ray analysis.

Lanthanum infusion techniques, transmission electron microscopy, and X-ray microanalysis were utilized to compare the permeability of muscle cell membranes from normal and degenerating muscle fibers of Solenopsis spp. In normal fibers, the electron-dense tracer was limited to components of the sarcotubular system. However, the insemination-induced degeneration of muscle fibers was characterized by the presence of an electron-dense precipitate within the myofibrils and mitochondria as well as in the extramyofibrillar spaces. The electron-dense material was subsequently identified by elemental analysis to be lanthanum. Such data indicate that one of the earliest stages of muscle degeneration involves an alteration in cell membrane permeability.

Animals↗

Leupeptin, a protease inhibitor, blocks insemination-induced flight muscle histolysis in the fire ant Solenopsis.

The effect of the protease inhibitor leupeptin on flight muscle histolysis in queen fire ants was studied by electron microscopy. In untreated animals artificially inseminated, muscle involution was apparent at 6 hr post-insemination and complete by 24 hr post-insemination. However, in animals pre-treated with leupeptin and subsequently artificially inseminated, no morphologic evidence of flight muscle breakdown was seen at any interval between 6 and 24 hr post-insemination. Such information appears to indicate that one or more proteases are involved in the process of insemination-induced muscle atrophy in fire ants. The most likely candidate is a soluble, calcium-activated myofilament-associated protease.

Animals↗

Peroxisomes in the absorptive cells of normal, rachitic, and vitamin-D replete chick intestine: ultrastructure and histochemistry.

Using routine transmission electron microscopy and light and electron microscopic techniques for the histologic demonstration (localization) of catalase (a peroxisomal enzyme), peroxisomes in chick duodenal epithelial cells were identified and studied. In these cells, peroxisomes were seen to be small, ovoid structures, delimited by a single unit membrane. They were concentrated in the supranuclear cytoplasm in initimate association with the smooth endoplasmic reticulum. As demonstrated histochemically, the heterogeneous matrix of these organelles was catalase positive. In addition, most of the larger peroxisomes revealed central nucleoids; however, the smaller peroxisomes were generally anucleoid. It thus appears that two classes of peroxisomes exist in chick intestinal absorptive cells: (1) small, anucleoid microperoxisomes, and (2) larger, nucleoid-containing peroxisomes. In addition to the above morphological characteristics, both peroxisome types were numerous in normal and vitamin-D-replete tissues, but were conspicuously decreased or absent from the apical cytoplasm of rachitic epithelial cells. From these observations it is hypothesized that these organelles may be involved in the overall vitamin-D response of the small intestine.

3,3'-Diaminobenzidine↗

Comparison of ocular disposition of free pivalic acid and pivalic acid esterified in dipivefrin.

Dipivefrin, a diester of pivalic acid and epinephrine that is used in the treatment of glaucoma, has been shown to be hydrolyzed in the cornea, and the ocular pharmacokinetics of the active moiety, epinephrine, have been determined. A comparison of the ocular disposition of the pivalic acid released by hydrolysis of dipivefrin and that of topically applied free pivalic acid showed that the released pivalic acid is not metabolized in the eye and is rapidly eliminated from the eye with no apparent sequestration in ocular tissues. In addition, results of two methods for preparing ocular tissues for analysis of drug distribution indicated that freezing and thawing processes can lead to redistribution of materials in ocular tissues.

Animals↗

Phenylbutazone kinetics and metabolite concentrations in the horse after five days of administration.

Phenylbutazone (PBZ) was administered (8.8 mg/kg of body weight) every 24 hours for 5 consecutive days, orally for the first 4 days and IV on day 5. The half-life (t 1/2) after this daily administration was 6.2 hours and the volume of distribution was 0.152 +/- 0.014 L/kg; the bioavailability after oral administration was 91.8 +/- 2.5%. The plasma concentration of PBZ at experimental hour (EH) 24 (24 hours after the 1st oral dose) was 1.7 +/- 0.39 micrograms/ml and increased to 4.2 +/- 0.29 micrograms/ml at EH 48 (24 hours after the 2nd oral dose). Values at EH 72, 96, and 120 (24 hours after administration of oral doses 3 and 4, and IV dose 5, respectively) were 4.8 +/- 0.62 micrograms/ml, 5.3 +/- 0.84 micrograms/ml, and 4.3 +/- 1.1 micrograms/ml, respectively. Significant increases (P less than 0.05) were measured between EH 24 and 48 with no changes during the subsequent 3 days. Changes in the urinary concentrations of PBZ were not seen over the 5-day dosing period. There were increases in the metabolites oxyphenbutazone and the alpha-alcohol during the 5-day dosing period. There was a delay in the appearance of alpha-alcohol in urine after oral and IV administration.

Administration, Oral↗

Calcium-containing lysosomes in the outer mantle epithelial cells of Amblema, a fresh-water mollusc.

The cells of the outer mantle epithelium contain numerous large pleomorphic electron dense bodies. In their fine structure they resemble lysosomes. Positive acid phosphatase histochemistry confirms that these supranuclear and subnuclear structures are lysosomes. A major portion of the intralysosomal material is resistant to high-temperature microincineration, indicative of an inorganic component. Subsequent microprobe analyses identified considerable calcium within these organelles. Such entities are similar in structure and ionic content to the lysosomes of avian intestinal absorbing cells, another calcium-transporting epithelium. These mantle lysosomes may function in transcellular calcium transport during shell formation, growth, and repair, especially since the mantle is the shell-forming organ in molluscs.

Animals↗

Electron-microscopic and morphometric study of vesiculation in the epithelial cell layer of the toad urinary bladder. Effect of antidiuretic hormone.

Basolateral vesicles are apparent in mitochondria-rich, granular and basal cells of the epithelium of unstimulated toad urinary bladder. Such vesicles are smooth-walled rather than coated. Because of the number of these vesicles, we decided to investigate the effect of arginine vasopressin (AVP) on this vesicle population during the hydroosmotic response. Both glutaraldehyde and glutaraldehyde-osmium tetroxide fixation procedures, as well as electron-microscopic morphometric techniques were utilized. Fifteen min after the administration of AVP in the presence of an osmotic gradient, the intracellular area occupied by basolateral vesicles was increased approximately threefold when compared to unstimulated control hemibladders (0.92 +/- 0.15% vs: 2.67 +/- 0.39%). The intracellular nature of the majority of these vesicles was confirmed by experiments utilizing lanthanum as an electron-dense tracer. It is hypothesized that this vesicular activity may be involved in water movement across the tissue after hormone treatment.

Animals↗

An electron microscopic histochemical and X-ray microprobe study of spherites in a mussel.

Transmission electron microscopic, histochemical and X-ray analytical microprobe techniques were used to study the inorganic-organic relationship in the spherites (calcospherules) from the mantle, i.e. subadjacent to the outer mantle epithelium, of the fresh-water mussel Amblema. These structures were shown to contain calcium which could be chelated by the flotation of sections on solutions of either formic acid or ethylene glycol bis-(beta-amino ethyl ether)-N, N1-tetra-acetic acid (EGTA), Analysis of both non-chelated and sections revealed a significant sulfur peak. Chelated spherites were also intensely stained with acid phosphotungstic acid (PTA), Such data is indicative of the presence of an organic glycoprotein (proteoglycan) matrix which could serve to bind mineral ions, thus forming organic-inorganic aggregates for calcium transport and homeostasis. In this regard, the spherites are analogous to both calcium phosphate containing mitochondrial granules and the initial calcification sites in vertebrate mineralizing tissues.

Animals↗

Lysosomal proliferation in rachitic avian intestinal absorptive cells following 1,25-dihydroxycholecalciferol.

Lysosomes in chick intestinal absorptive cells from rachitic (vitamin D-deficient) and vitamin D-replete animals were studied utilizing transmission electron microscopic histochemistry and ultrastructural morphometry. Absorptive cells from rachitic animals, serum calcium = 7.3 +/- 0.3 mg%, contained an average of 4.0 +/- 0.3 supranuclear lysosomes. In rachitic chicks sacrificed 9 hr post-injection of 1,25-dihydroxycholecalciferol, the active metabolite of vitamin D, the values for both serum calcium, 9.8 +/- 0.2 mg%, and the number of apical absorptive cell lysosomes, 12.9 +/- 0.6, were increased over non-injected or vehicle-only injected animals. Lysosomes in vitamin D-replete absorptive cells were characterized by their intense staining with pyroantimonate, indicative of their high calcium content. The same organelles also produced a positive reaction for acid phosphatase. Rachitic lysosomes, also acid phosphatase positive, were only lightly stained with pyroantimonate. The lysosomal proliferation apparently induced by 1,25-dihydroxycholecalciferol may be a further indication that these organelles play a role in intestinal calcium transport and/or intracellular calcium homeostasis within the absorptive cell.

Acid Phosphatase↗

Cartilage calcification: an ultrastructural, histochemical, and analytical x-ray microprobe study of the zone of calcification in the normal avian epiphyseal growth plate.

Sections from the zone of calcification of ruthenium red-fixed normal avian epiphyseal growth plates were analyzed by various morphological (histochemical) and analytical techniques. Calcium and phosphorus were identified in the chondrocyte pericellular rim, the uncalcified extracellular (territorial) matrix, and in both the peripheral and central aspects of the calcified accumulations within extracellular matrix. Cartilage proteoglycan, as determined by ruthenium binding, positive staining with acidic phosphotungstic acid, and the X-ray spectroscopic detection of sulfur, was identified in the same four zones. Thus, it appears that proteoglycans, in some form, are indeed retained at sites of biological calcification. Additionally, these macromolecules, synthesized in chondrocytes, may be involved in extracellular calcium translocation.

Animals↗

A histochemical and X-ray microanalysis study of calcium changes in insect flight muscle degeneration in Solenopsis, the queen fire ant.

Potassium pyroantimonate histochemistry, coupled with ethyleneglycoltetraacetic acid (EGTA)-chelation and X-ray microprobe analysis, was employed to localize intracellular calcium binding sites in the normal and degenerating flight musculature in queens of Solenopsis, the fire ant. In normal animals, calcium distribution was light to moderate within myofibrils and mitochondria. In the early contracture stages of the insemination-induced degeneration, both myofilament and mitochondrial calcium loading was markedly increased. In the terminal stages of myofibril breakdown, only Z-lines (isolated or in clusters) with an associated filamentous residue persisted. These complexes were also intensely calcium positive. This study further documents the presence of increased sarcoplasmic calcium during muscle necrosis. Surface membrane defects, mitochondrial calcium overload, and calcium-activated proteases may all be involved in this "normal" breakdown process.

Animals↗

Calcium lysosomes in rachitic and vitamin D3 replete chick duodenal absorptive cells.

Calcium-containing lysomes were described in a previous communication in this series (Davis et al., 1979). Their potential role in intestinal calcium uptake and transcellular transports was hypothesized. To further this notion, the effects of a rachitogenic diet and vitamin D3 repletion were investigated. Intestinal absorptive cells from chicks maintained on a vitamin D deficient diet were characterized by decreased numbers of supranuclear calcium lysosomes. In contrast, intestinal cells from chicks given vitamin D3 (cholecalciferol) subsequent to the rachitogenic diet showed numerous large compound supranuclear calcium lysosomes. Since other steroid hormones are known to effect lysosomes, it is tempting to speculate that vitamin D, itself a steroid hormone, may activate lysosomes which themselves might be involved in calcium homeostatic mechanisms.

Animals↗

Ultrastructural changes in the lysosomes of rachitic intestinal absorptive cells.

In chicks maintained on a rachitogenic (vitamin D deficient) diet, the number of intestinal absorptive cell calcium-lysosomes is markedly decreased in comparison to normal animals. In addition, the majority (better than 50%) of these rachitic calcium-lysosomes are atypical in their fine structure resembling the lamellar bodies seen in certain diseases (Tay-Sachs disease, Duchenne's muscular dystrophy). Such atypical organelles are characterized by their internal membranous swirls reminiscent of myelin figures. This information appears to be a further indication that lysosomes are normally involved in calcium homeostatic mechanisms and therefore sensitive to circulating vitamin D levels.

Animals↗

Microperoxisomes in the epithelial cells of the amphibian urinary bladder: an electron microscopic demonstration of catalase and malate synthase.

All cells that comprise the epithelium of the toad urinary bladder were found to contain small ovoid to tubular membrane-bound bodies with a finely granular matrix. Such organelles were devoid of dense cores (nucleoids). These microperoxisomes reacted positively when incubated for the demonstration of catalase or malate synthase activity. In the toad liver, peroxisomes as well as microperoxisomes were seen. Histochemically, both demonstrated catalase activity; neither showed malate synthase activity. The presence of malate synthase, a glyoxylate cycle enzyme, in toad urinary bladder microperoxisomes may make these latter organelles unique among vertebrates, since malate synthase has been thought to be absent in higher animals.

Animals↗