No abstract

Paclitaxel, an anti-mitotic anti-cancer agent, is active against solid tumors. The inhibition of depolymerization and promotion of microtubular assembly are essential for the anti-tumor activity of paclitaxel. Microtubule-associated proteins (MAPs) co-polymerize with tubulin and play some roles in microtubular dynamics. We examined the effect of paclitaxel on the interaction between tubulin and MAPs. Human lung-cancer cells, PC-14, were synchronized to G1/S border by the thymidine-double-block technique. After release from exposure to thymidine, the cells were treated briefly with 2 nM paclitaxel and the levels of alpha and beta tubulins and MAPs were examined after various times. Immunoblot analysis of paclitaxel-treated cells showed no changes in the overall expression of alpha and beta tubulins, microtubule-associated protein 2 (MAP2) or MAPs in comparison with controls. The samples were immunoprecipitated with anti-alpha- and anti-beta-tubulin antibodies and reblotted with an anti-MAP2 antibody, which showed that the amount of co-immuno-precipitated MAP2 in the synchronized cells, were increased by the brief paclitaxel treatment. These results suggest that paclitaxel treatment enhances the interaction between alpha and beta tubulins and MAP2. Since the phosphorylation state of MAP2 regulates the affinity of MAP2 for tubulins, and mitogen-activated protein (MAP) kinase is considered to be one of the kinases responsible for MAP2 phosphorylation, the effect of paclitaxel treatment on the MAP-kinase activity of synchronized PC-14 cells was examined. Two bands with molecular masses of 42 and 44 kDa were detected by an "intra-gel" MAP-kinase assay using myelin basic protein as the substrate. Paclitaxel treatment inhibited the MAP-kinase activity of PC-14 cells and inhibition was maximal at the G2/M phase of the cell cycle. Similar, concentration-dependent inhibition by paclitaxel of cellular MAP kinase of human synchronized small-cell lung carcinoma, H69, was observed. No inhibition of the MAP kinase of the paclitaxel-resistant sub-line H69/Txl by paclitaxel was observed, suggesting that some change of the MAP-kinase cascade had occurred in these cells. No direct inhibition of MAP-kinase activity by paclitaxel was observed in the cell-free assay (in vitro), suggesting that paclitaxel did not inhibit MAP kinase directly. Since it has been speculated that p34cdc2 kinase is also a kinase that phosphorylates MAP2, the effect of paclitaxel treatment on the p34cdc2-kinase activity of synchronized PC-14 and PC-9 cells was examined. Paclitaxel inhibited p34cdc2-kinase activation at the G2/M phase. These results suggest that paclitaxel inhibited MAP kinase and p34cdc2 kinase in vivo indirectly. These actions of paclitaxel may be responsible for the increased affinity between MAP2 and tubulins that it induces.

Time-dependent changes of bone mass in ambulant chronic respiratory failure patients 60 or more years of age were compared between those on home oxygen therapy (HOT) and those still free of HOT (non-HOT). HOT (n = 31) showed initial PaO2 of slightly greater than 60 Torr and non-HOT (n = 32) had PaO2 moderately greater than 60 Torr (64.4 Torr vs 75.1 Torr). PaCO2 in HOT was significantly higher than that of non-HOT (44.8 Torr vs 40.0 Torr). There was no difference in pulmonary function test results. The whole bone mineral density (BMD) as adjusted by age and sex was significantly lower in the HOT group than that in the non-HOT. At endpoints of the follow-up period over 2 years or more, daily bone losses in the whole BMD, whole bone mineral content, and lumber BMD were significantly more accelerated in HOT compared with non-HOT. When the Wistar rats were pair-fed and their locomotion was limited, the animal group placed for 4 weeks under hypoxic air showed a reduction in BMD as compared with the control. We suggest that hypoxemia contributes to bone mass loss.

In a review of cerebral metabolism and function in man, Kety (1950) reported that one subject, who demonstrated marked apprehension during a test situation, had an unusually high cerebral metabolic rate for oxygen (CMR02), 5.0 ml O2 100 g-l min-I, as compared to his normal range of 3.2-4.2 ml O2 100 g-I min-I. Kety suggested that anxiety, or psychic stress, can increase brain metabolism. A recent study in man also indicates that pain-induced stress significantly increases blood flow in gray matter by about 10%, particularly in the frontal lobes (lngvar et aI., 1976).These clinical observations appear to be sup ported by recent findings in rats. Immobilization for 5 -30 min (conscious rats were artificially ventilated while paralyzed by a muscle relaxant) increases both CMR02 and cerebral blood flow (CBF) up to twofold (Carlsson et aI., 1975(Carlsson et aI., , 1977. These cerebral effects in rats are ascribed to the f3-adrenergic re ceptor action of catecholamines, particularly epi-