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Biomedical subjects

W P Cunningham

Publications and source records attributed to W P Cunningham.

18 recordsLinked to original sources
PubMed1991

Improvement in RBC flux, acidosis and oxygenation in tumour microregions by Fluosol-DA 20%.

Using laser Doppler flowmetry, we investigated the effects of Fluosol-DA 20% on RBC flux in FSaII tumours and on RBC flux in normal skin of C3H mice. The RBC flux in tumours was significantly increased; however, the RBC flux in normal skin fluctuated only slightly after the treatment with Fluosol-DA 20%. Since an increase in RBC flux by Fluosol-DA 20% may facilitate the removal of acidic metabolites from the tumours, the effects of Fluosol-DA 20% on intratumour pH was also measured. We also measured intratumour pO2 after various dosages of Fluosol-DA 20% with carbogen inhalation. We concluded that the administration of Fluosol-DA 20% selectively increased the RBC flux in tumours compared to that in normal skin. Moreover, carbogen inhalation combined with increasing dosages up to 36 ml/kg of Fluosol-DA 20% effectively enhanced tumour oxygenation in FSaII tumours.

Animals↗
PubMed1982

Herpesviruses in metastatic Lucké renal adenocarcinoma.

The etiologic agent of the renal adenocarcinoma of leopard frogs, Rana pipiens, is the Lucké tumor herpesvirus (LTHV). The virus is easily detected with thin section electron microscopy in primary tumors of frogs which have been exposed to a cold environment. Several spontaneous metastatic nodules and a large primary tumor were detected at autopsy of a frog which had been maintained at 4 degrees C for 73 days. LTHV was found not only in the primary tumor, as previously reported, but also was present in metastatic tumor cells in the liver, fat body, and bladder. The presence of LTHV in metastatic cells demonstrates that the differentiated state of primary Lucké tumor cells is retained in its metastatic colonies even at the fine structure level revealed by electron microscopy.

Adenocarcinoma↗
PubMed1978

Surface topography of normal and neoplastic human anterior pituitary cells maintained in vitro.

Pituitary tissues were obtained from 25 patients who underwent surgery for excision of pituitary macroadenomas, selective excision of microadenomas, or removal of a normal gland for palliation of metastatic cancer. Cells thus obtained were maintained in vitro for varying intervals, fixed, and examined by light (phase contrast), microscopy, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Previous SEM reports indicate that surface topography of in vitro neoplastic cells displays features that may correlate with neoplastic behavior. Cultured normal and pituitary tumor cells did not display these surface differences, with one exception, a prolactin-secreting microadenoma. Characteristic patterns for the cell populations were identified. Certain cell types appeared in all the cultures: 1) large and small granule-containing cells; 2) flat and irregular cells; 31 spindle-shaped cells; and 4) spherical, irregularly surfaced cells. In one case of an endocrine-inactive juvenile pituitary chromophobe adenoma, unique cells were observed. Surface topography did not appear to be of predictive value in determining the neoplastic character of pituitary tumors.

Adenoma, Basophil↗
PubMed1977

Structure and organization of radial spokes in cilia of Tetrahymena pyriformis.

The structure and organization of radial spokes, the principal components between each of the peripheral doublet microtubules and the central sheath which surrounds the central pair of microtubules have been described in Tetrahymena pyriformis cilia. The radial spokes are grouped in triplets and are attached to the A-microtubule of each peripheral doublet at intervals of 200/280/360 A, the 200 A spacing being most distal to the base of the cilium. The radial spoke triplets are organized in the axoneme in a double helix with a pitch of 4,680 A. A method for determining the helical disposition by correcting for doublet sliding is presented.

Animals↗
PubMed1975

Effect of chloramphenicol on replication of mitochondria in Tetrahymena.

Tetrahymena pyriformis ST (3 X 10-4 cells/ml) was treated with 0.1 mg/ml chloramphenicol (CAP). Cell division ceased after 1.5 divisions with no decreased viability. Total mitochondrial volume and succinic dehydrogenase (SDH) activity/liter increased 1.7-fold and 3-fold, respectively. SDH activity/cell decreased whereas malate dehydrogenase activity/cell and respiratory control ratios and P:O ratios of isolated mitochondria were unchanged in treated cells. During 12 hours of growth in CAP the total surface area of mitochondrial inner and outer membrane was essentially unchanged or increased 4-fold, respectively. Mitochondria from cells treated with chloramphenicol had decreased size, buoyant density and protein:lipid ratio in the membranes. The membrane ubiquinone:protein ratio was unchanged. Tetrahymena cells contained 3.6 X 10-minus 12 g of mitochondrial DNA and 6,800 mitochondria in a volume of 41,000 mu-3. A 4-hour treatment with CAP caused a 4-fold increase in the number of mitochondria/cell and a 10-fold increase in mitochondria/liter in contrast to a 4-fold increase in number of mitochondria/liter in control cells. Thus CAP stimulated division of mitochondria. Individual mitochondria of treated cells had one-tenth the volume of control mitochondria. The rate of increase of mitochondrial DNA/liter was the same in control and CAP-treated cultures. The amount of DNA/mitochondrion decreased 75% in CAP-treated cells due to the rapid division of mitochondria. The cell volume, cell protein content and mitochondrial DNA content/cell decreased with growth of control cultures.

Animals↗
PubMed1974

Effects of phosphotungstate negative staining on the morphology of the isolated Golgi apparatus.

Isolated Golgi complexes can be recognized in phosphotungstate (PTA) negative stain as stacks of membranous plates surrounded by a complex anastomosing network of tubules and vesicles. The extent of this tubular network is, however, much greater than can be observed in thin sections of whole cells. To determine which of the steps leading to the final negatively stained image may produce the observed changes, we have monitored each of the steps by other electron microscope and biochemical methods. The first damage to the membranes seems to occur during the initial isolation procedure as judged by the appearance of smooth patches on the freeze-fractured membrane faces that are normally covered with particles. Subsequent suspension of the Golgi fraction in water, to dilute the sucrose for negative staining, leads to the disappearnce of the stacking, to some tubulation and some vesiculation of the membranes as judged by thin section and freeze-cleave microscopy. The latter technique also reveals an increase in smooth-cleaving membrane faces. Application of the negative stain to the water-washed Golgi fraction, finally, produces extensive tubular arrays and a simultaneous decrease in the remaining large membranous vesicles. The freeze-cleaved tubular membranes appear essentially smooth except for small patches of aggregated particles. Parallel gel electrophoresis studies of the membranes and of the water and negative stain wash extracts indicate that protein extraction is involved in these morphological changes. PTA seems to be a particularly effective solvent for certain membrane proteins that are not removed by the water wash. These observations suggest that removal of membrane proteins alters structural restraints on the membrane lipids so that they behave semiautonomously like myelinics and form new artificial structures. This does not eliminate the possibility, however, that some tubules also exist in the Golgi apparatus in vivo.

Animals↗
PubMed1973

Partial purification and phosphotungstate solubilization of basal bodies and kinetodesmal fibers from Tetrahymena pyriformis.

Previously devised methods for the isolation of basal bodies from ciliate protozoans were found to be inadequate for chemical analysis. We have modified and expanded these procedures and developed a method which gives preparations containing mainly basal bodies and kinetodesmal fibers. This procedure involved fixation of cells in 30% ETOH followed by digitonin or Triton X-100 solubilization and homogenization with a Brinkmann Polytron. This is followed by sucrose gradient centrifugation. Negative staining and thin sectioning revealed these preparations to be substantially more pure than those of previous workers. It was also found that neutralized phosphotungstate (PTA) solubilized many of the components present in fixed Tetrahymena. Neutralized 1.0% PTA solubilized axonemes, cortical, axonemal, and basal body microtubules as well as kinetodesmal fibers. These results have been confirmed by both electron microscope observations and gel electrophoresis of 100,000 g supernatants of the PTA extracts. A solution of 0.1% PTA did not affect the fibers but did solubilize basal bodies. Running 1.0% PTA extracts from our basal body fractions on sodium dodecyl sulfate (SDS) polyacrylamide gels allowed us to tentatively identify the peptides of basal bodies and kinetodesmal fibers. The latter structures appear to consist of a single 21,000 mol wt peptide. These results also suggest that great caution should be taken in interpreting PTA images, especially of microtubules and axonemes.

Animals↗
PubMed1972

Ultrastructural studies on a mutant of Bacillus subtilis whose growth is inhibited due to insufficient autolysin production.

The growth of Bacillus subtilis mutant betaA177 can be inhibited under special conditions in which not enough autolytic enzymes are produced for optimal growth. Electron microscopy studies show that during growth inhibition there is localized thickening of the cell wall at positions where cells bend. A model is proposed to explain this result. Rapid growth can be restored by adding lysozyme or a B. subtilis autolysin mixture to a growth-inhibited betaA177 culture. Such addition reduces the localized wall thickening and causes other changes in surface morphology which are described and discussed. Septum formation seems to be relatively less inhibited than cell elongation when lytic enzyme levels are reduced. Measurements were made demonstrating that walls at ends of cells are morphologically different from walls at sides of cells in cultures of betaA177 growing at 51 C.

Autolysis↗
PubMed1971

Isolation of germ cell Golgi apparatus from seminiferous tubules of rat testes.

Intact Golgi apparatus have been isolated with good purity from rat testis by a simplified sucrose gradient technique. The procedure is inherently selective in that most of the Golgi apparatus in the isolate are from germ cells in late spermatocyte or early spermatid development. No Sertoli cell Golgi apparatus are present in the fraction. Biochemical analyses showed that the enzyme N-acetylglucosamine galactosyltransferase is enhanced in the band containing Golgi apparatus. Because they are easy to isolate, and because they are involved with the formation of a specific internal cellular component (the acrosome), these Golgi apparatus will be useful objects for comparison with other kinds of Golgi apparatus and for other studies leading to a better understanding of the basic functioning of Golgi apparatus.

Animals↗
PubMed1970

Isolation of a Golgi apparatus-rich fraction from rat liver. I. Method and morphology.

Golgi apparatus were released without fixatives from rat hepatocytes by gentle homogenization, concentrated by differential centrifugation, and purified by sucrose gradient centrifugation. Examination of sections of purified fractions by electron microscopy showed fields of morphologically intact units of Golgi apparatus consisting of stacks of parallel flattened cisternae, secretory vesicles, and small vesicular profiles. Negative staining of unfixed pellets revealed a complex network of anastomotic tubules continuous with platelike structures and secretory vesicles. These structures corresponded, respectively, to the small vesicular profiles and parallel flattened cisternae with attached secretory vesicles of sectioned material. Small fragments of granular endoplasmic reticulum were often closely associated with the peripheral tubules, suggesting sites of continuity in intact hepatocytes.

Animals↗
PubMed1966

Structure of isolated plant Golgi apparatus revealed by negative staining.

Sucrose-gradient-purified dictyosomes of plant Golgi apparatus appear, after glutaraldehyde stabilization, as stacks of highly fenestrate and tubate cisternae when negatively stained with phosphotungstic acid, shadowed with heavy metal, or OsO(4)-stained in thin section. The tubular proliferations (diameter 200 to 400 A) extend for several microns from the central region and are united at intervals into an anastomosing network. Associated with the tubules are two kinds of vesicles which are distinguishable on the basis of texture, size, shape, and staining characteristics. One vesicle type is rough-surfaced, nearly spherical, and of uniform dimensions (diameter approximately 600 A). Metal shadowing shows that these vesicles remain spherical after drying. The other vesicle type is smooth-surfaced and varies in both size and shape. Intercisternal elements are revealed, by negative staining, on the surface of internal cisternae after fragmentation of the dictyosome. The progressive differentiation of cisternae from the forming face to the maturing face is observed in thin sections of these isolated preparations. The morphological characteristics observed in negatively stained dictyosomes indicate regions of functional specialization within the dictyosome cisternae and reveal a dictyosome structure more extensive than that envisioned from sections.

Golgi Apparatus↗