Dynamic Aspects of Glucocorticoid Receptors in the Spinal Cord of the Rat

In spite of biochemical and autoradiographic evidence for glucocorticoid binding sites in the spinal cord (SC), events occurring after the preliminary step of hormone binding were not studied. In this investigation, we have examined the transformation (activation) of the cytosolic receptor coupled to [3H]dexamethasone (DEX) and the in vivo interaction of adrenal hormone [corticosterone (CORT)] with purified nuclei from the SC, in addition to the CORT content of the SC before and after stress. Binding of [3H]DEX in the SC was 40% lower than in the hippocampus (HC), although the KD values were comparable. Transformation of [3H]DEX-receptor complexes in the cytosol was demonstrated by diethylaminoethane-cellulose chromatography, by DNA-cellulose binding, and by a combined minicolumn procedure including hydroxyapatite in addition to the last two techniques for separation of transformed, nontransformed, and meroreceptor complexes. In all these situations, SC glucocorticoid binding sites behaved similarly to those in the HC. Nuclear uptake of a tracer dose of [3H]CORT was much lower in the SC than in the HC; nuclear retention of CORT was more easily detected by radioimmunoassay after injection of 1 mg of CORT into adrenalectomized rats. Substantial amounts of CORT, which increased in level after stress, were measured in five regions in the SC, with higher concentrations in the cervical regions. These studies suggest that although SC and HC receptors show similar properties in vitro, differences emerged at the level of nuclear uptake in vivo, in that glucocorticoid action in the SC was similar to that in the optic nerve, where receptors seem to be localized mostly in glial cells.     

Uptake of immunisation in district health authorities in England

The uptakes of immunisation in the district health authorities in England were studied for the years 1983-5. Multiple regression analysis showed that the factors significantly associated with a low uptake of immunisation were mainly related to social conditions, particularly overcrowding of households and population density. Of the service factors, high proportions of elderly and singlehanded general practitioners and high average list sizes were also associated with a low uptake of immunisation in some of the analyses.The results suggest that the measures outlined in the government''s white paper on improving primary health care services are likely to lead to improved uptakes of immunisation. If, however, the uptakes of immunisation are used as a measure of standards of the services provided they should first be adjusted to control for variations in social conditions, and the quality of vaccination data would have to be improved.     

Control,Change, and Suffering: The Messages of Shan Buddhist Sermons

Shan villagers are Buddhists, but their main access to Buddhist ideology is through the sermons the local monks preach. We describe the content of a number of sermons. These outline a system where lack of control causes change which causes suffering. The sermons teach about the acquisition of knowledge where the objective is not to control but to escape from the system. Power, gained through withdrawal or relationships with powerful beings, gives more control, consequently less change, and less suffering. One can convert impermanent material wealth into changeless merit by offering it to the monks or temple. Offerings to non-reciprocating monks are competitive and define a hierarchy of both power and prestige. The ideology so outlined is one aspect of the Shan world view based on morally neutral power and is related to Shan political and economic forms. The monks serve state power not only by defining this ideology and receiving offerings but also by preaching about the benefits of making offerings and the importance of people behaving properly in their positions in the hierarchy.     

Purification of a murine leukemia inhibitory factor from Krebs ascites cells

A factor capable of inducing terminal differentiation in the murine myeloid leukemia cell line M1 has been purified to apparent homogeneity from the medium conditioned by Krebs II ascites tumor cells. The factor, termed leukemia inhibitory factor (LIF) is a single chain glycoprotein of apparent Mr 58,000 which induces differentiation and inhibits proliferation of the M1 cell line but not the WEHI-3B D+ murine myeloid leukemic cell line and has no detectable proliferative activity on normal myeloid progenitor cells. It was purified using four successive high-efficiency purification steps--anion-exchange chromatography on DEAE-Sepharose; cation-exchange chromatography on CM-Sepharose; affinity chromatography on lentil lectin-Sepharose; and reverse-phase high-performance liquid chromatography on a phenyl-silica matrix--to a specific biological activity of approximately 1.25 X 10(8) units/mg with an overall purification of 12,000-fold and a yield of 73% for the activity failing to bind to DEAE-Sepharose. Sufficient quantities of the factor (12 micrograms, 200 pmol) have been purified to allow structural and functional analysis of the molecule and comparison with other know differentiation inducers.     

Transformation of single myeloid precursor cells by the malignant histiocytosis sarcoma virus (MHSV): generation of growth-factor-independent myeloid colonies and permanent cell lines

Direct single-cell assays for oncogenic transformation are available for fibroblasts but not for other cell types. Using malignant histiocytosis sarcoma virus (MHSV), a member of the ras family of retroviruses, in vivo-infected granulocyte/macrophage and macrophage precursor cells lost the requirement for externally added hematopoietic growth factors. Factor-independent growth was demonstrated by colony-transfer experiments. More than 25% of the independent colonies were established as permanent macrophage cell lines following a phase of adaptation to tissue culture conditions. Factor-independent colony growth was also obtained by in vitro infection of single cells. As many as 50% of all myeloid precursor cells were target cells for MHSV as measured by this assay. About 2 x 10(-3) of these colony-forming cells acquired growth factor independence and immortality after in vitro infection. Cell lines derived from these colonies did not require adaptation to tissue culture conditions.     

Aberrant development of Trichophyton mentagrophytes hyphae cultured in the presence of Congo red

When Trichophyton mentagrophytes colonies were placed on a medium containing 150 micrograms/ml of Congo red, a dye which prevents chitin fibrillogenesis, their growth rate was reduced. The newly formed mycelium, examined under an ultraviolet microscope, consisted of thick, curled and branched hyphae endowed with swollen tips and subapical bulges. Short-time exposure revealed that the major sites of dye accumulation were the extension zones. Normally structured hyphae arose from aberrant mycelia when they were transferred onto a dye-free medium. The phenomena observed suggest that Congo red alters the wall properties of the extension zone, by inhibiting the gradual conversion of chitin chains, synthesized at the extreme tip, into microfibrils of increasing size and density.     

Compositional changes associated with plasma membrane thickening during floral induction of spinach

Floral induction in the long-day plant spinach ( Spinacia oleracea L. cv. Nobel) was accompanied by a thickening of the plasma membrane. Densitometry analyses showed that the light space of the dark-light-dark pattern of the membrane was not changed upon photoinduction. Rather, the increase was due to an enhancement of the dark layer adjacent to the cell wall. Parallel analyses of protein and phospholipid composition revealed no marked changes in protein composition or biosynthetic rate, protein phosphorylation, glycolipids and/or phospholipids as a result of the 24 h of continuous light sufficient to induce flowering. Photoinduction, however, was accompanied by an increase in the relative amount of plasma membrane sterols which may be related to the membrane thickening.     

Enzymatic recycling of oxidized ascorbate in pig heart: one-electron vs two-electron pathway

Enzymatic systems able to reduce either dehydroascorbate or ascorbyl radical back to ascorbate by "recycling" vitamin C may contribute to lowering the nutritional requirement of it and to increase tissue antioxidant capacity. The activities of two enzymatic activities, GSH-dehydroascorbate reductase (two-electron reduction pathway) and NADH-semidehydroascorbate reductase (one-electron reduction pathway) in pig tissues, have been investigated. The activity of glutathione-dependent reduction of dehydroascorbate, although measurable, appeared negligible taking into consideration the low physiological substrate concentration. On the other hand, the one-electron reduction of ascorbyl radical resulted fast enough to slow down the consumption of the antioxidant vitamin.     

Regulation of prostaglandin production by osteoblast-rich calvarial cells

The effect of various factors upon prostaglandin (PG) production by the osteoblast was examined using osteoblast-rich populations of cells prepared from newborn rat calvaria. Bradykinin and serum, and to a lesser extent, thrombin, were also shown to stimulate PGE₂ and 6-keto-PGF_1α (the hydration product of PGI₂) secretion by the osteoblastic cells. Several inhibitors of prostanoid synthesis, dexamethasone, indomethacin, dazoxiben and nafazatrom, were tested for their effects on the calvarial cells. All inhibited PGE₂ and PGI₂ (the major arachidonic acid metabolites of these cells) production with half-maximal inhibition by all four substances occuring at approximately 10⁻⁷ M. For dazoxiben and nafazatrom, this was in contrast to published results from experiments which have indicated that the compounds stimulated PGI₂ production. Finally, since the osteoblasts is responsive to bone-resorbing hormones, these were tested. Only epidermal growth factor (EGF) was shown to modify PG production. At early time EGF stimulated PGE₂ release, however, the predominant effect of the growth factor was an inhibition of both PGE₂ and PGI₂ production by the osteoblastic cells. The present results suggest that the bone-resorbing hormones do not act to cause an increase in PG by the esteoblast and that any increase in PG production by these cells may be in response to vascular agents     

Mechanics of the ligamentum nuchae of some artiodactyls

The ligamentum nuchae lies near the dorsal surface of the neck, connecting the occiput to the thoracic neural spines and helping to support the head. It consists largely of elastin, a protein with rubber-like elastic properties. Experiments have been performed on deer, a sheep and a camel to measure the elastic strain in the ligament, both when the head is raised and when it is lowered as in feeding from the ground. The elastic properties of the excised ligament were investigated so that the forces in it, in the intact animal, could be inferred from the observed strains. The forces required to support the head have been calculated, and are compared with the forces inferred from the strains.