Bark stripping of hinoki cypress by sika deer in relation to snow cover and food availability on Mt Takahara,central Japan

Bark stripping of hinoki cypress (Chamaecyparis obtusa (Sieb. et Zucc.) Endl.) by sika deer (Cervus nippon Temminck) was monitored for 4years on approximately 300 trees at three sites on Mt Takahara, central Japan. We investigated the effects of snow cover and food availability on seasonal and yearly changes in bark stripping. Bark stripping occurred during snowy periods (January and February) when less food was available because of the snow cover. The most serious bark stripping was observed in 1996 when the snow was the deepest and food resources were the poorest among the 4years of the study. In contrast, bark stripping did not occur in 1999 when the snow was thin and, consequently, more food was available. These results suggest that snow cover and food avail-ability influence the occurrence and intensity of bark stripping both seasonally and annually.     

Seasonal change in browsing by sika deer on hinoki cypress trees on Mount Takahara,central Japan

Browsing by sika deer (Cervus nippon Temminck) on hinoki cypress (Chamaecyparis obtusa (Sieb. et Zucc.) Endl.) was monitored for approximately 3years on 300 trees at three sites on Mount Takahara, central Japan. We investigated the seasonal change in browsing on hinoki cypress in relation to the availability of other food plants (dominated by Sasa nipponica Makino et Shibata) and preferred food plants (deciduous broad leaves). Browsing occurred mostly from November to April, while no browsing damage occurred from June to October. Browsing was observed in November, even when the amount of other food plants did not decrease. Therefore, the cause of browsing cannot be fully explained by a decreased availability of other food plants. Comparison of fecal composition and food availability showed that deer had a preference for deciduous broad leaves. The availability of these preferred plants decreased in November and increased in May. Chemical analysis of the hinoki cypress and deciduous broad leaves (crude protein and neutral detergent fiber) showed that the quality of deciduous broad leaves decreased in November, and as a result the quality of hinoki became higher than that of the deciduous broad leaves. Browsing on hinoki cypress occurred when the availability of deciduous broad leaves decreased, and vice versa. These results suggest that the availability of preferred food plants influences browsing on hinoki cypress by sika deer.     

Antitumor agents 216. Synthesis and evaluation of paclitaxel-camptothecin conjugates as novel cytotoxic agents

Five conjugates (16-20) composed of a paclitaxel and a camptothecin derivative joined by an imine linkage were synthesized and evaluated as cytotoxic agents and as inhibitors of DNA topoisomerase I. All of the conjugates were potent inhibitors of tumor cell replication with improved activity relative to camptothecin. Significantly, compounds 16-18 were more active than paclitaxel and camptothecin against HCT-8 (colon adenocarcinoma) cell replication, and the spectrum of activity was different from a simple mixture of paclitaxel and camptothecin. All of the conjugates were significantly less potent than camptothecin as inhibitors of human topoisomerase I in vitro with 16, 18, and 19 showing only marginal activity at 50 microM. Based on activity against drug-resistant cell line replication, one could conclude that the conjugates are simply acting as 'weak taxanes', but the spectrum of activity, particularly against MCF-7 and HCT-8, strongly suggests that a novel mechanism of action has been achieved through conjugation.     

A new approach for syntheses of 2',3'-dideoxy-2',3'-dehydronucleosides using 2,2-difluoro-1,3-dimethylimidazolidine (DFI) as a dehydrating reagent

We found that 2,2-difluoro-1,3-dimethylimidazolidine (DFI) is useful for not only fluorination but also dehydrating reactions. This dehydrating ability of DFI was applied to the syntheses of dihydrofurans (2) that are possible starting materials for various anticancer or antiviral drugs.     

Thrombospondin-1 differentially regulates release of IL-6 and IL-10 by human monocytic cell line U937

We investigated possible regulatory effects of thrombospondin-1 (TSP-1), a multifunctional extracellular matrix protein, on cytokine release from macrophages. Immobilized TSP-1 enhanced IL-6 release from the human monocytic U937 cells stimulated with phorbol myristate acetate and LPS, whereas it inhibited IL-10 release. The 70-kDa fragment of TSP-1 containing the type 1 repeats showed the same regulatory effects. The enhanced IL-6 release by TSP-1 was inhibited by anti-CD36 antibody or antibody against the sequence of the binding site to CD36 in the type 1 repeats of TSP-1. Conversely, the decrease in IL-10 release by TSP-1 was strengthened by the blocking of the interaction between CD36 and TSP-1. Furthermore, the involvement of TGF-beta1 in the inhibition of IL-10 release by TSP-1 was indicated by the facts that (i) TSP-1 induced activation of TGF-beta1 produced by the U937 cells, (ii) exogenously added TGF-beta1 inhibited IL-10 release, and (iii) antibody against TGF-beta1 blocked the inhibition of IL-10 release by TSP-1. Together, the present findings suggest that TSP-1 enhances IL-6 release from macrophages by interaction with CD36, whereas IL-10 release is regulated by the balance between the enhancing effect of TSP-1 via CD36 and the suppressive effect by TSP-1-activated TGF-beta1.     

Two G12 family G proteins, G alpha12 and G alpha13, show different subcellular localization

The G alpha subunits of the G12 family of heterotrimeric G proteins, G alpha12 and G alpha13, are closely related in sequences and some effectors, but they often act through different pathways or bind to different proteins. We have examined subcellular distribution of these two G proteins and found that endogenous G alpha12 and G alpha13 localize in membrane and cytoplasmic fractions, respectively. Exogenously expressed G alpha12 and G alpha13 also localize in membrane and cytoplasmic fractions, respectively, in COS-7 cells. Stimulation of lysophosphatidic acid receptor coupled to G alpha13 markedly promotes the translocation of G alpha13 from cytoplasm to membrane. This different localization of G alpha12 and G alpha13 may explain some of the nonoverlapping actions of G alpha12 and G alpha13.     

Lysophospholipids transactivate HER2/neu (erbB-2) in human gastric cancer cells

The ligand-less receptor HER2/neu (erbB-2) has been proposed as a prognostic marker of gastric cancer that correlates with poor clinical outcome, indicating that HER2 signals play an important role in gastric cancer progression. This study demonstrated that two major natural lysophospholipids, lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P), induce rapid and transient phosphorylation of HER2 in two human gastric cancer cell lines, MKN28 and MKN74 cells. We also revealed that tyrosine phosphorylation of HER2 induced by both lysophospholipids was significantly attenuated by two inhibitors, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, AG1478, and a broad-spectrum matrix metalloproteinase inhibitor, GM6001. This suggests that the pathway of HER2 transactivation induced by these lysophospholipids is dependent on the proteolytically released EGFR ligands. Our results indicate that LPA and S1P act upstream of HER2 in gastric cancer cells, and thus may act as potent stimulators of gastric cancer.