Subchronic safety evaluation of hot-water extract from thinned immature mangos (Mangifera indica ‘Irwin’): 90-days oral toxicity study in rats

application/pdfThinned immature fruit of the mango tree （Mangifera indica ‘Irwin’） are handled as waste. In this study, we conducted a 90-days toxicity study in male and female Sprague Dawley rats to evaluate the safety of a hot-water extract of thinned immature mango fruits （TIMEx） administered by oral gavage at doses of 500, 1000 and 2500 mg/kg body weight/day. Treatment did not result in death or changes in the behavior or external appearance of the animals. No alterations were observed in hematological or serum chemical parameters, urinalysis, food consumption, body weight gain or organ weights at the end of the treatment period, with the exception of higher mean corpuscular volume in male rats that received high doses and lower serum creatine phosphokinase levels in female rats that received medium doses. Under the conditions of this study and based on the toxicological endpoints evaluated, the no-observed-adverse-effect level （NOAEL） for TIMEx was 2500 mg/kg/day. The findings indicate that TIMEx is safe for consumption and should be investigated as a candidate food

Social confrontation stress decreases hepatic fibroblast growth factor-21 expression in aged mice

application/pdfWe previously showed that social stress exposure in mature adult mice increased blood corticosterone concentrations at 2 days, disrupted hepatic lipid metabolism-related pathway at 30 days, and increased the risk of overweight with hepatic hypertrophy at 90 days. To further investigate the effects of aging on the physiological responses to social stress, we conducted a study using male BALB/c mice at the ages of 2 months （mature age）, 14 months （middle age） and 26 months （old age）, and exposed them to confrontation stress for 2 or 7 days. Blood corticosterone concentrations were increased at 2 days of stress, and then returned to baseline concentrations. This change was observed only at 2 months of age. We further examined the effect of aging on hepatic gene expression of fibroblast growth factor-21 （Fgf21） and found that its expression was significantly decreased after 7 days of stress at 14 months of age and after 2 days of stress at 26 months of age, indicating these decreasing effects became more pronounced with age. In conclusion, our study suggests that hepatic Fgf21 expression decrease under exposure to confrontation stress at middle or more age, indicating that stress response on Fgf21 related pathway might be more pronounced with age when exposed to stress

２段連続バイオリアクターを用いたタマネギ食酢製造の最適操作

application/pdfOnion vinegar was produced using a 2-stage continuously stirred tank reactor. Regarding the alcohol fermentation and the acetic acid fermentation examined in this study, the immobilized cells on porous ceramics offered stable production of alcohol and acetic acid for long periods of 300 and 700 days, respectively. Compared with the steady-state operation method, the temperature-change forced-cyclic operation method increased ethanol yield of alcohol fermentation by a maximum of 15％. Acetic acid yield in the vinegar showed no difference between the steady-state operation and temperature-change forced-cyclic operation. The onion vinegar produced in this study contained 3.5-11.5 times the amount of organic acids and 1.6-6.9 times the amount of amino acids as commercially sold vinegars.departmental bulletin pape

Fundamental Studies on the Rheological Properties of Fresh Concrete : The Propagation Characteristics of the Elastic Wave in Mortar

departmental bulletin pape

Study of CP-Violating Asymmetries in B0→π+π- Decays

journal articl

昆虫の変態・休眠の分子機構

平成8年度から平成11年度までの4年間に実施された特定領域研究(A)(1)「昆虫の変態・休眠の分子機構(略称:変態・休眠機構)」は,昆虫の有する旺盛な生存,生活戦略を変態と休眠という高次の生命現象に焦点を合わせ,この分子機構を多面的に解明することを目的として,延べ62名の研究者の参加を得て進められ,多くの研究成果を挙げてきた.「変態・休眠を支配するホルモン分子の動態と環境応答」に関する研究では,脳神経ペプチドホルモンの遺伝子の同定,合成分泌とその調節機構,ホルモンの血中動態及びホルモン受容体の解析を進め,新たな知見を得た.また「変態における自己・非自己の認識転換と形作りの分子機構」に関する研究では,幼虫組織の識別と排除機構,成虫組織の分化発生機構,ミツバチの社会性分化をもたらす遺伝子の同定とその発現機構を解明した.これらの成果の一端は平成12年12月神戸市にいて開催された平成12年度文部省主催の「大学と科学」公開シンポジウム「昆虫から学ぶ生きる知恵」において,研究者のみならず一般市民約500名に対し,公表された.一方,総括班員を中心とした研究成果の最終まとめに関する検討をもとにして,学術審議会科学研究費分科会審査第一部会生物系小委員会による特定領域研究研究終了ヒアリングを受け,その結果,Aの評価を得た.また,本研究の成果は,研究目的や研究経過に加え,本研究参加者の研究結果と発表論文リストを編集した研究成果報告書として発刊し,関連機関や研究者等に配布した.科学研究費補助金 研究種目:特定領域研究(A)(1) 課題番号:08276103 研究代表者:山下 興亜 研究期間:1996-2000年度research repor

Increased bone mass due to the GSK-3β insufficiency by radiological and histological comparisons of <i>Gsk-3β^+/+</i> and <i>Gsk-3β^+/–</i> littermates at 12 weeks of age.

<p>(A) Plain X-ray of the whole body. (B) Plain X-ray of the entire femur. (C) 3D-CT image of the distal femur. (D) von Kossa staining of the proximal tibia (bar, 200 µm) and toluidine blue staining of the growth plate indicated by the inset box above (bar, 20 µm). (E) Histomorphometric analyses of bone volume and bone formation parameters in the proximal tibia. BV/TV, trabecular bone volume per tissue volume; C.Th, cortical thickness; Ob.S/BS, osteoblast surface per trabecular bone surface; Ob.S/B.Pm, osteoblast surface per trabecular bone perimeter; MAR, mineral apposition rate; BFR/BS, bone formation rate per trabescular bone surface. Lower right panel shows fluorescent micrographs of calcein-labeled mineralization fronts of the trabecular bones (bar, 10 µm). (F) Histomorphometric analyses of bone resorption parameters in the proximal tibia. N.Oc/B.Pm, number of osteoclasts per 100 mm of bone perimeter; Oc.S/BS, osteoclast surface per bone surface; ES/BS, eroded surface per bone surface. For (E) and (F), data are mean (bars)±SEM (error bars) of 10 mice per genotype. *<i>P<</i>0.05, **<i>P<</i>0.01 vs. <i>Gsk-3</i>β^+/+. (G) Formation of TRAP-positive multinucleated osteoclasts by the co-culture of calvarial primary osteoblasts (POB) and bone marrow macrophages (BMMφ) derived from either <i>Gsk-3</i>β^+/+ or <i>Gsk-3</i>β^+/– mice. Representative pictures (left; bar, 200 µm) and the number of osteoclasts expressed as mean (bars)±SEM (error bars) of 8 wells per group. *<i>P<</i>0.05 vs. <i>Gsk-3</i>β^+/+ X <i>Gsk-3</i>β^+/+.</p

Inactivation through phosphorylation of Runx2 by GSK-3β.

<p>(A) Subcellular nuclear (N) and cytoplasmic (C) localizations of Runx2 by immunoblot analysis (top) and Runx2 mRNA level determined by real-time RT-PCR (bottom) in <i>Gsk-3</i>β^+/+ and <i>Gsk-3</i>β^+/– calvarial osteoblasts overexpressing CA-GSK-3β or treated with LiCl, and cultured for 3 days. The mRNA levels are mean (bars)±SEM (error bars) of the relative amount compared to the control culture of 6 wells per group. (B) EMSA for specific binding (arrowheads) of a labeled OSE2 oligonucleotide probe with the nuclear extracts (N.E.) from <i>Gsk-3</i>β^+/+ or <i>Gsk-3</i>β^+/– osteoblasts overexpressing Runx2. Cold competition (Comp.) was performed with 50-fold excess of unlabeled wild-type OSE2 probe (wt) and the mutated probe lacking the Runx2 binding sequence (mut). For controls, incubations without the probe (the 1st lane) and using nuclear extracts from osteoblasts without Runx2 transfection (the last lane) were performed. (C, D) Co-immunoprecipitation (co-IP) analysis of GSK-3b and Runx2. (C) Whole cell lysate (CL) and co-IP precipitant by anti-FLAG antibody-immobilized beads were immunoblotted with either anti-HA tag or anti FLAG tag antibodies. Filled arrowhead indicates non-specific band, and blank arrowhead indicates specific band. (D) Whole cell lysate (CL) and co-IP precipitant by anti-HA tag antibody or IgG (as a negative control) were immunoblotted with either anti-FLAG tag or anti-HA tag antibodies. (E) Luciferase reporter analysis of the effects of Runx2 mutations at the five consensus sites for the phosphorylation by GSK-3β on the Runx2 transcriptional activity. Mutations were created by three to four amino acid replacements as follows; S92A-S96A-S100A [M(96)3], S369A-S373A-S377A [M(373)3], S389A-T393A-S397A [M(393)3], T394A-S398A-T402A [M(398)3], and T476A-T480A-S484A-S488A [M(480)4]. HuH-7 cells were transfected with 1,050 OC-Luc alone or in combination with the plasmids expressing wild-type Runx2 (WT) or the mutants above, then cultured for 2 days. Data are mean (bars)±SEM (error bars) of the relative activity compared to control of 6 wells per group. *<i>P<</i>0.01 vs. WT-Runx2. (F) <i>In vitro</i> kinase assay. WT-Runx2 and M(373)3-Runx2 proteins were extracted by immunoprecipitation of the overexpresssing HeLa cells, and were incubated with recombinant GSK-3β. Reaction products were analyzed by immunoblotting using an antibody to phosphoserine. (G) EMSA for specific binding (arrowheads) of a labeled OSE2 probe with the nuclear extracts (N.E.) from HeLa cells transfected with wild-type Runx2 (WT) and M(373)3 Runx2 (M). Cold competition (Comp.) was performed as above. (H) Luciferase reporter analysis of the effects of GSK-3β signaling on the Runx2 transcriptional activity induced by WT-Runx2 and M(373)3-Runx2. HuH-7 cells were transfected with 1,050 OC-Luc alone or in combination with the plasmid expressing WT-Runx2 or M(373)3-Runx2 in the presence or absence of CA-GSK-3β overexpression or LiCl, then cultured for 2 days. Data are mean (bars)±SEM (error bars) of the relative activity compared to control of 6 wells per group. *<i>P<</i>0.01, significant effect of CA-GSK-3β overexpression or LiCl.</p

Suppression of Runx2 transcriptional activity by GSK-3β.

<p>(A) Expressions of GSK-3β and Runx2 determined by immunoblot analysis in mouse calvaria, tibia, and cultured calvarial primary osteoblasts (POB). (B) von Kossa staining (left) and osteocalcin mRNA level determined by real-time RT-PCR analysis (right) of <i>Gsk-3</i>β^+/+ and <i>Gsk-3</i>β^+/– osteoblasts transfected with the adenovirus expressing GFP or Runx2, and cultured for 2 weeks. The mRNA levels are mean (bars)±SEM (error bars) of the relative amount of mRNA compared to that of the <i>Gsk-3</i>β^+/+ with GFP culture of 6 wells per group. *<i>P<</i>0.01, significant stimulation by the Runx2 overexpression. (C) Luciferase reporter analysis of the effects of GSK-3β overexpression on the Runx2 transcriptional activity. HuH-7 cells were transfected with 1,050 OC-Luc alone or in combination with the plasmid expressing Runx2, and co-transfected with 0.1 or 0.2 µg plasmid expressing wild-type GSK-3β, CA-GSK-3β, or KI-GSK-3β, and cultured for 2 weeks. (D) Luciferase reporter analysis of the effects of GSK-3β inhibitors on the Runx2 transcriptional activity. HuH-7 cells were transfected with 1,050 OC-Luc alone or with the plasmid expressing Runx2, and cultured in the presence or absence of two doses of lithium chloride (LiCl) or SB21673 for 2 days. For (C) and (D), data are mean (bars)±SEM (error bars) of the relative activity compared to control culture of 6 wells per group. *<i>P<</i>0.01 vs. Runx2 alone.</p