Free and polymerized tubulin in cultured bone cells and Chinese hamster ovary cells: the influence of cold and hormones

A low pH method of liposome-membrane fusion (Schneider et al., 1980, Proc. Natl. Acad. Sci. U. S. A. 77:442) was used to enrich the mitochondrial inner membrane lipid bilayer 30-700% with exogenous phospholipid and cholesterol. By varying the phospholipid-to- cholesterol ratio of the liposomes it was possible to incorporate specific amounts of cholesterol (up to 44 mol %) into the inner membrane bilayer in a controlled fashion. The membrane surface area increased proportionally to the increase in total membrane bilayer lipid. Inner membrane enriched with phospholipid only, or with phospholipid plus cholesterol up to 20 mol %, showed randomly distributed intramembrane particles (integral proteins) in the membrane plane, and the average distance between intramembrane particles increased proportionally to the amount of newly incorporated lipid. Membranes containing between 20 and 27 mol % cholesterol exhibited small clusters of intramembrane particles while cholesterol contents above 27 mol % resulted in larger aggregations of intramembrane particles. In phospholipid-enriched membranes with randomly dispersed intramembrane particles, electron transfer activities from NADH- and succinate-dehydrogenase to cytochrome c decreased proportionally to the increase in distance between the particles. In contrast, these electron- transfer activities increased with decreasing distances between intramembrane particles brought about by cholesterol incorporation. These results indicate that (a) catalytically interacting redox components in the mitochondrial inner membrane such as the dehydrogenase complexes, ubiquinone, and heme proteins are independent, laterally diffusible components; (b) the average distance between these redox components is effected by the available surface area of the membrane lipid bilayer; and (c) the distance over which redox components diffuse before collision and electron transfer mediates the rate of such transfer.     

Disparate effects of tumor necrosis factor-alpha/cachectin and tumor necrosis factor-beta/lymphotoxin on hematopoietic growth factor production and neutrophil adhesion molecule expression by cultured human endothelial cells

Tumor necrosis factor-alpha/cachectin (TNF-alpha) and tumor necrosis factor-beta/lymphotoxin (TNF-beta) are inflammatory mediators with similar spectrums of cytotoxic activity against tumors in vitro and in vivo. We compared the effect of purified recombinant human TNF-alpha and TNF-beta on neutrophil adhesion molecule expression and hematopoietic growth factor production by cultured human umbilical vein endothelial cells. Endothelial cells acquired adhesive properties for neutrophils after a 4-hr incubation with as little as 5 U/ml TNF-alpha. TNF-alpha stimulated a dose-dependent increase in endothelial cell adhesiveness for neutrophils, with a maximal effect at 250 U/ml. In contrast, TNF-beta did not enhance endothelial-dependent neutrophil adherence until a concentration of 600 to 1200 U/ml was reached. Endothelial cells cultured for 24 hr with TNF-alpha, 10 to 1,000 U/ml, released hematopoietic colony-stimulating activity. TNF-beta failed to augment growth factor production by endothelial cells at any concentration tested. Inhibitor assays showed that the absence of detectable colony-stimulating activity was not due to direct inhibition of colony growth by TNF-beta or to release of hematopoietic inhibitors by the TNF-beta-stimulated endothelial cells. Purified natural TNF-beta was similar to recombinant TNF-beta in its effect on neutrophil adhesion molecule expression and growth factor production by endothelial cells. These results indicate that the two immunomodulatory proteins TNF-alpha and TNF-beta differ in their effects on a common target tissue. TNF-beta, which retains tumoricidal properties, shows fewer proinflammatory activities on cultured endothelial cells than TNF-alpha in vitro.     

Tripsacum-maize interaction: a novel cytogenetic system

The genera Zea and Tripsacum cross readily when they are not isolated by gametophytic barriers, and it has been postulated that intergeneric introgression played a role in the evolution of maize. The basic x = 9 Tripsacum and x = 10 Zea genomes have little cytological affinity for each other in hybrids that combine 10 Zea with 18 Tripsacum chromosomes. However, one to four Tripsacum chromosomes sometimes associate with Zea chromosomes in hybrids between Z. mays (2n = 20) and T. dactyloides (2n = 72). These hybrids with 10 Zea and 36 Tripsacum chromosomes frequently produce functional female gametes with 36 Tripsacum chromosomes only. When they are pollinated with maize, their offspring again have 36 Tripsacum and 10 maize chromosomes, but the Tripsacum genome is contaminated with maize genetic material. In these individuals, intergenome pairing is the rule, and when they are pollinated with maize, their offspring have 36 Tripsacum and 10, 12, 14, 16, 18, or 20 Zea chromosomes. Plants with 36 Tripsacum and 20 Zea chromosomes behave cytologically as alloploids, although the Tripsacum genome is contimated with maize, and one basic maize genome is contaminated with with Tripsacum genetic material. When they are pollinated with maize, offspring with 18 Tripsacum and 20 Zea chromosome are obtained. Further successive backcrosses with maize selectively eliminate Tripsacum chromosomes, and eventually plants with 2n = 20 Zea chromosomes are recovered. Many of these maize plants are highly "tripsacoid." Strong gametophytic selection for essentially pure Zea gametes, however, eliminates all obvious traces of Tripsacum morphology within a relatively few generations.     

CALAMOPHYTON IN THE MIDDLE DEVONIAN OF NEW YORK STATE

Specimens of Calamophyton from the Middle Devonian Ashokan Sandstone near Kingston, Ulster County, N.Y., are shown to belong to C. bicephalum Leclercq and Andrews, a Belgian species. Steel needles and a stereoscopic binocular microscope were used to follow the path of the forking leaves and branching sporangiophores of the specimens through the matrix. The terete leaves dichotomized two to three times in more than one plane. Sporangiophores dichotomized once. Each branch bore three recurved lateral branches which in turn bore two sporangia. Each branch terminated in an elongate, filiform projection. Sporangia apparently dehisced longitudinally. Their walls were composed of elongate cells. Their spherical spores ranged from 86 to 166 μ in diameter and bore a trilete mark. Ornamentation consisted of coni and spinae up to 4.5 μ long. They resembled dispersed spores of Dibolisporites gibberosus var. major Richardson. This is the second occurrence of Calamophyton bicephalum and the first account of its spores. It is the second report of the genus in North America.     

CD18-dependent and -independent mechanisms of neutrophil emigration in the pulmonary and systemic microcirculation of rabbits

Neutrophil (PMN) migration in the systemic and pulmonary circulation of rabbits was compared by using different inflammatory stimuli to determine the role of the leukocyte adhesion complex, CD11/CD18, in each of these vascular beds. The adhesion complex was blocked by administering the anti-CD18 mAb 60.3. The data show that mAb 60.3 blocks PMN emigration into inflammatory foci in the abdominal wall produced by implanting sponges containing either hydrochloric acid, Streptococcus pneumoniae, Escherichia coli endotoxin, or PMA. mAb 60.3 also inhibited PMN emigration in response to peritoneal instillation of S. pneumoniae. The effect of mAb 60.3 on PMN emigration in the lungs varied depending upon the stimulus. PMN failed to migrate into the PMA-induced pneumonia; however, mAb 60.3 pretreatment only partially inhibited endotoxin-induced pneumonia and did not inhibit S. pneumoniae or hydrochloric acid-induced pneumonias. PMN lavaged from the alveolar spaces in the Streptococcal pneumonia had similar quantities of mAb 60.3 bound to their surfaces as the circulating PMN. We conclude that the CD11/CD18 complex mediates PMN adherence in the systemic circulation. However, PMN adherence in the pulmonary circulation may occur by either CD18-dependent or -independent mechanisms that are specific to the inciting stimulus.     

The effect of time lags on the stability of the equilibrium state of a population growth equation

Summary A scalar integrodifferential equation is considered which describes a single self-regulating species. Three results are presented towards showing that the carrying capacity equilibrium state becomes unstable as the self-regulating mechanism acts after a longer time lag.