Enhancement of norepinephrine-induced transient contraction in aortic smooth muscle of diabetic mice.

Changes in norepinephrine-induced transient contractions in Ca2+-deficient solution were investigated in the aortic smooth muscles of diabetic ALS (alloxan-induced diabetes susceptible) mice. The transient contractions in diabetic mice were significantly larger than those in normal mice. The longer incubation of the muscle preparations in Ca2+-deficient solution made the transient contractions smaller, probably due to the leakage and decrease in norepinephrine-releasable stored Ca2+. The rate of this reduction in contraction was slower in diabetic mice. These results suggest that the leakage of intracellular stored Ca2+ caused by extracellular Ca2+ deficiency is attenuated in diabetic mice, contributing to enhanced norepinephrine-induced transient contractions.</p

脱脂米糠抽出物のヒト由来食細胞に対する免疫賦活性の検討

The present report describes the effect of the three kinds of degreasing rice bran extracts (rice bran (Rb), rice bran extract E (RbE), and rice bran heat water extract (RbhS) on the immune response of human cells. Samples of Rb and RbE were processed by fermentation with Enterococcus sp. and Pichia sp. at 37°C for 3 to 5 days, subsequently, RbE samples were treated with hemicellulose enzyme. While RbhS samples were heattreated and used as the control. We studied the effect of inactive Staphylococcus aureus cells coated with the three kinds of rice bran extracts on phagocytosis and phagosome-lysosome fusion by human leukocytes in vitro. Rice bran extracts increased phagocytic activity and phagosome-lysosome fusion in neutrophils and monocytes in a doseresponse manner. The rate of phagocytosis of RbE-coated S. aureus by neutrophils was significantly higher, by approximately 3.6 times, than that of uncoated S. aureus. The rate of phagocytosis of RbE-coated S. aureus by monocytes was about 1.6 times higher than that of the control. The phagosome-lysosome fusion index (FI) for RbE-coated bacteria by neutrophils was higher, by approximately 6 times, than that of the control. The ratio of the FI to the phagocytic index (PI) in human neutrophils showed that 83% of phagocytosed cells were digested the fused phagolysosomes when the S. aureus coated with RbE at a concentration of 25 μg/ml. These results show that RbE has immune-enhancing activity. Although degreasing rice bran is traditionally discarded, if processed with Enterococcus sp., Pichia sp. and hemicellulose enzyme, it becomes consumable and can be used as a health food material to enhance immune activity in humans3種類の方法で調製した脱脂米糠抽出物について免疫賦活性(貧食活性と食胞融合性)の視点から,未利用資源としての活用可能性を検討した。　(1)ライフペップ(Rb)は乳酸菌・酵母処理(37℃ ・3～5日)。　(2)ライフペップE(RbE)は乳酸菌・酵母処理後　酵素(ヘミセルラーゼ)処理。　(3)熱水抽出物(RbhS)はオートクレープ処理。　何れの処理サンプルも,凍結乾燥後試験に供した。その結果,ヒト好中球に対する貧食活性は,3試料とも,濃度に依存して貧食作用が上昇した。RbEでは5μg/mlプラトーに達し,コントロールに比べ3.6倍(p<0.001)の活性を示した。また,単球に対する貧食活性では,RbEが1.6倍(p<0.001)活性を示した。RbEは,脱脂米糠の熱水抽出物(RbhS)とRbより高い活性を示した。　最も貧食活性の強かったRbEの食胞融合能は,コントロールに対して6倍の食胞融合性を示し,25μg/mlの濃度で貧食された菌のうち約8割が殺菌されることが確認できた。この結果,脱脂米糠を乳酸菌・酵母処理後酵素(ヘミセルラーゼ)処理すると,貧食活性を促進できることがわかり,免疫賦活作用を示す利用資源として役に立つことが明確となった

Simple method for large-scale production of macrophage activating factor GcMAF

Human group-specific component protein (Gc protein) is a multifunctional serum protein which has three common allelic variants, Gc1F, Gc1S and Gc2 in humans. Gc1 contains an O-linked trisaccharide [sialic acid-galactose-N-acetylgalactosamine (GalNAc)] on the threonine420 (Thr420) residue and can be converted to a potent macrophage activating factor (GcMAF) by selective removal of sialic acid and galactose, leaving GalNAc at Thr420. In contrast, Gc2 is not glycosylated. GcMAF is considered a promising candidate for immunotherapy and antiangiogenic therapy of cancers and has attracted great interest, but it remains difficult to compare findings among research groups because different procedures have been used to prepare GcMAF. Here, we present a simple, practical method to prepare high-quality GcMAF by overexpressing Gc-protein in a serum-free suspension culture of ExpiCHO-S cells, without the need for a de-glycosylation step. We believe this protocol is suitable for large-scale production of GcMAF for functional analysis and clinical testing

ジェネティック制御下にある発育鶏卵を用いた工学的in vivo 薬剤評価系の開発

The drug discovery research of clinical-use antioxidants, which may control various vascular disorders caused by the oxidative stress, is extremely important. We present the development of an in vivo evaluation system of antioxidants for their vascular protective activities using the chick embryonic chorioallantoic membrane (CAM). In the case of 12-days chick embryos, the topical administration of 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH) induced their vascular injuries against the CAM veins and venous capillaries but without substantial fatal damage. Artepillin C, a potent natural antioxidant, did not show the chick embryo’s venous injury, and pre-treatment with artepillin C would tend to protect the CAM veins injuries induced by the post-administration of AAPH. These results suggest that artepillin C might be able to protect the AAPH-induced vascular injury in in vivo. In conclusion, we showed the possibility of in vivo evaluation system of antioxidants for their vascular protective activities using the chick embryo

低酸素細胞特異的な蛍光イメージング剤の開発

We have designed and evaluated UTX-12 as a novel fluorescent pH probe for tumor hypoxia imaging. UTX-12 consists of a p-nitro benzyl moiety, which is a latent hypoxia-selective leaving group activated by nitro reduction, directly linked to SNARF. Although UTX-12 itself is colorless and non-fluorescent in aqueous solution, nitro reduction triggers the release of SNARF which has well-characterized long wavelength absorption and fluorescence that is sensitive to pH. The resultant SNARF, released intracellularly by enzymatic reduction of UTX-12, allows measurement of pH by pH-dependent dual emission shifts. UTX-12 showed clear differences in fluorescence behavior between hypoxic and aerobic conditions in liver microsomes and inside V79 cells. These data are confirmation that UTX-12 is biologically reduced inside tumor cells and the released SNARF should monitor intracellular pH of tumor cells selectively with reduced background signal

イソプレン ソクサ オ ユウスル ショクブツ セイブン ノ ex vivo ユウキ ゴウセイ ト ソノ セイリ カッセイ ノ ヒョウカ

Phytyl quinols, namely acyclic tocopherols, are key intermediates of tocopherol biosynthesis, but their biological activities remain unclear. We therefore investigated the structure-activity relationship of phytyl quinols to apply a chemical biosynthesis design for an antiatherosclerosis drug based on isoprenomics. We have achieved the biosynthesis-oriented synthesis of α- and β-phytyl quinol as an unnatural intermediate, other γ- and δ-phytyl quinol as a natural one. All four phytyl quinols showed almost the same moderate inhibitory activity against low-density lipoprotein oxidation instead of their different degree of C-methylation with character different from tocopherols. In vivo toxicities of phytyl quinols against chick embryo chorioallantoic membrane vasculature were hardly observed. We proposed phytyl quinols were possible antioxidants in plants and animals, like vitamin E

イソプレン ソクサ オ ユウスル ショクブツ セイブン ノ ex vivo ユウキ ゴウセイ ト ソノ セイリ カッセイ ノ ヒョウカ

Phytyl quinols, namely acyclic tocopherols, are key intermediates of tocopherol biosynthesis, but their biological activities remain unclear. We therefore investigated the structure-activity relationship of phytyl quinols to apply a chemical biosynthesis design for an antiatherosclerosis drug based on isoprenomics. We have achieved the biosynthesis-oriented synthesis of α- and β-phytyl quinol as an unnatural intermediate, other γ- and δ-phytyl quinol as a natural one. All four phytyl quinols showed almost the same moderate inhibitory activity against low-density lipoprotein oxidation instead of their different degree of C-methylation with character different from tocopherols. In vivo toxicities of phytyl quinols against chick embryo chorioallantoic membrane vasculature were hardly observed. We proposed phytyl quinols were possible antioxidants in plants and animals, like vitamin E