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Reportedly, the intake of koji amazake, a beverage made from steamed rice fermented by Aspergillus oryzae, improves defecation frequency. However, its functional ingredients and mechanism of action remain unclear. To compare the effects of koji amazake and a placebo beverage on defecation frequency and to identify the functional ingredients and mechanism of action, a randomized, placebo-controlled, double-blind parallel-group comparative trial was performed on two groups. The koji amazake had 302 ± 15.5 mg/118 g of A. oryzae cells, which was not in the placebo. Compared with the placebo group, the koji amazake group showed a significant increase in weekly defecation frequency at 2 weeks (5.09 days vs. 4.14 days), 3 weeks (5.41 days vs. 4.18 days), and 4 weeks (5.09 days vs. 3.95 days), along with an increase in the weekly fecal weight at 4 weeks (724 g vs. 501 g). The intake of koji amazake did not induce significant intergroup differences in the fecal SCFA concentration, whereas it significantly decreased the relative abundance of Blautia and significantly increased that of Bacteroides at 3 weeks. Therefore, koji amazake intake improved defecation frequency, and A. oryzae cells played potentially important roles as functional ingredients.

Rice with double mutation of starch synthase IIIa and branching enzyme IIb (ss3a/be2b) has much higher amounts of apparent amylose and resistant starch (RS) than usual varieties. In this study, we conducted two randomized, single-blind, crossover trials to investigate the effect of single ingestion of two processed foods composed of ss3a/be2b mutant rice on postprandial blood glucose and insulin response in healthy adults, compared to those of usual cultivar. In trial 1, of ingestion of rice crackers, the incremental area under the curves of glucose (IAUCglc) and insulin (IAUCins) in RS group was significantly lower than in the control group. In trial 2, of the ingestion of cooked rice, IAUCglc in the RS group was significantly lower than in the control group. These results showed that the ss3a/be2b-mutant rice cracker and cooked rice having high RS can attenuate postprandial blood glucose and insulin response.

Introduction Oxygen has positive benefits and potentially damaging properties in biological systems. It is a highly reactive molecule, which participates in a high-energy electron transfer and through oxidative phosphorylation thereby supports generation of adenosine-5-triphosphate (ATP)

The cell cycle checkpoint system play a pivotal role in the cellular DNA damage response, and the discovery of checkpoint inhibitors is expected to sensitize current cancer therapies. Checkpoint signaling cascades are critically modulated by ATM (ataxia telangiectasia-mutated) and its related molecules. Generally, ATM primarily responds to ionizing irradiation-induced DNA double-strand breaks. Heavy ions from an accelerated carbon ion beam have been used to cure cancer because they are more effective than ionizing irradiation such as X-ray and γ-radiation in terms of biological damage. In a previous study, we demonstrated that a persimmon leaf flavonol (PLF) promoted the cytotoxic effect of chemotherapeutic agents on cancer cells through inhibition of checkpoint activities, especially in the ATM dependent pathway. The present study investigated whether PLF inhibits checkpoint activity during the DNA damage response induced by heavy ion irradiation. Treatment with PLF significantly increased the cytotoxicity of heavy ion irradiation in A549 adenocarcinoma cells. The phosphorylation of checkpoint proteins such as p53, SMC1, and Chk1 was increased by heavy ions. PLF reduced the phosphorylation of checkpoint proteins. Pre-treatment with PLF significantly prevented the decrease of mitotic cells in heavy ion-exposed cells. We further evaluated tumor volume in SCID mice inoculated with human lung adenocarcinoma A549 cells. The combination treatment of PLF and heavy ion resulted in a decrease of tumor volume compared with controls, although PLF itself did not exhibit any effect. These results indicate that PLF inhibits tumor growth through modulation of the DNA damage response. PLF may be useful for clinical application in combination with heavy ion radiotherapy.

The antihypertensive and vasorelaxant effects of water-soluble proanthocyanidins, extracted in persimmon leaf tea, were investigated in spontaneously hypertensive rats, rat aortas, and human umbilical vein endothelial cells. Oral administration of proanthocyanidins significantly decreased the systolic blood pressure of the rats after 4 h, as compared with distilled water controls. A vasorelaxant effect on rat aortas was induced by proanthocyanidins, and it was abolished by removal of the endothelium and inhibition of endothelial nitric oxide synthase and soluble guanylyl cyclase activity. The phosphorylation levels of endothelial nitric oxide synthase (Ser-1177) and the upstream kinase Akt (Ser-473) in umbilical cells also increased in a time-dependent manner after the addition of a proanthocyanidin-rich fraction. These results suggest that the antihypertensive effect of proanthocyanidins in persimmon leaf tea is due to vasorelaxation via an endothelium-dependent nitric oxide/cGMP pathway, and that proanthocyanidins might be useful in dietary lowering of blood pressure.

Persimmon leaf (PL) has been commonly recognized for its wide variety of health benefits. A previous study has reported that persimmon leaf extract (PLE) contained flavonols with the 2″-galloly moiety (PLEg). Galloylated homologues generically show stronger activity in their biological function, so enhanced functions can be expected for PLEg. We investigated in this present study the effect of PLEg on the cellular DNA damage checkpoint signaling to sensitize cancer chemotherapy. Treatment with PLE and PLEg significantly increased the cytotoxicity of doxorubicin (DOX) in A549 adenocarcinoma cells. PLE and PLEg reduced the phosphorylation of checkpoint proteins such as structural maintenance of chromosomes 1 (SMC1), checkpoint kinase 1 (Chk1), and p53 in DOX-treated cells. Moreover, PLE decreased the phosphorylation of ATM (ataxia telangiectasia mutated) in a dose-dependent manner. PLE, and especially PLEg, abrogated the G2/M checkpoint during DOX-induced DNA damage. These results suggest that PLEg specifically inhibited ATM-dependent checkpoint activation by DOX, and that PLEg might be a useful sensitizer in cancer chemotherapy.

The amounts and compositions of polyphenol in persimmon leaves and persimmon leaf tea were investigated. The predominant polyphenols in fresh leaves were water-soluble, and the contents reached a maximum (2.40% w/w) in June, and then gradually decreased. Separation of them followed by thiolytic degradation revealed that the major components were unique proanthocyanidin oligomers consisting of four heterogeneous extension units, including epigallocatechin-3-O-gallate. Persimmon leaf tea also contained similar proanthocyanidins with similar compositional units. Oral administration of starch with polyphenol concentrate of persimmon leaf tea resulted in a significant and dose-dependent decrease in the blood glucose level in Wistar rats. This effect is considered to be due to inhibition of pancreas alpha-amylase. These results indicate that persimmon leaf tea containing peculiar proanthocyanidins has a significant role in suppressing blood glucose elevation after starch intake, and that the best harvest time is June.

The purpose of this study was to examine the effects of gamma-aminobutyric acid (GABA) in fermented drinking water prepared from sodium glutamate, vinegar, and dried bonito (FDWG) compared with placebo [vinegar and dried bonito without GABA (FDW)] and its safety in normotensive and mildly or moderately hypertensive volunteers. A double-blind, placebo-controlled, randomized study was conducted involving volunteers with normal (group-N) and mildly or moderately high (group-H) blood pressure (BP). After a pretreatment period of 2 weeks (weeks -2), the subjects received FDWG or FDW for 12 weeks followed by 4 weeks of no intake (weeks 16). In group-H, both FDWG and FDW significantly decreased systolic (SBP, -7.6 +/- 4.0 and -5.5 +/- 1.5 mmHg, p0.05, respectively) and diastolic (DBP, -10.6 +/- 4.0 and -7.6 +/- 1.7 mmHg, p0.01, respectively) BP compared to the baseline (0-week) value at 12 weeks, respectively. There were no abnormal changes in hematological or blood chemistry variables, urinalysis, heart rate, or body weight in the study groups. These findings indicated that vinegar and dried bonito with or without GABA might have an effect on BP in mildly or moderately hypertensive patients.

The purpose of this study was to investigate body composition focusing on motor functional capacity and to correlate body composition with functional outcomes with subacute stroke patients. The subjects of this study were 34 inpatients with hemiparetic stroke who could walk. Motor functional capacity of patients was assessed on the basis of the Brunnstrom recovery stage (BRS), and functional status was measured using the Functional Independence Measure (FIM) instrument scores and gait velocity. Body composition was assessed by bioelectrical impedance analysis. Significant correlations were observed between FIM scores and the BRS of the lower extremity or lean body mass. The correlations for body water in affected and unaffected legs with the FIM subscores were statistically significant (p

This paper reviews the evidence for novel physiological functions of oligosaccharides. A history of the research and development and the types of health benefits are discussed. A key property of the physiological functions lies in the indigestibility of oligosaccharides, which gives rise to fermentation in the large intestine, followed by an increase of Bifidobacteria and short-chain fatty acid (SCFA) production. This property leads to several kinds of physiological functions, classified into three types of health benefits. The first is the improvement of gastrointestinal conditions, including a normal stool frequency, less constipation, and healthy intestinal microflora. The second is the promotion of mineral absorption, including an increase of bone density and relief of anemia. The third is an immunomodulation effect, such as allergy and cancer prevention.

The purpose of this study was to investigate the relationship between daily physical activity and physical fitness in twenty ambulatory post-stroke patients. The pedometric measurement was made using a caloriecounter in daily life at home and assessed to quantify the time constant (τ on) of oxygen uptake (VO2) and oxygen deficit with gas analysis using a bicycle ergometer. There were negative correlations between the amount of daily walking steps and τ on, as well as the oxygen deficit at the onset of exercise (τ on; r=-0.52, p

Hemiparetic stroke patients have both decreased oxygen consumption and low endurance to exercise. This study examined their physical fitness to quantify the initial deficit and change in oxygen uptake kinetics (VO2 kinetics) during inpatients rehabilitation following stroke. Twenty-nine ambulatory hemiparetic stroke patients were treated in an 8-week program of low intensity aerobic exercise. They undertook a bicycle ergometer test with gas analysis before and after the exercise program to obtain maximum oxygen uptake (VO2 peak), maximum work load (max WR), and VO2 kinetics data. The time constant (τon) and O2 deficit on constant load after exercise training were significantly lower than those in initial measurement by 17% and 10%, respectively. On the incremental load, VO2 peak and max WR were significantly increased by 16% and 17% respectively and ΔVO 2/ΔWR was decreased by 11%. These results show that low-intensity endurance training produces energy efficiency of oxygen uptake kinetics and improves the physical fitness of ambulatory hemiparetic stroke patients.

The effects of acute or chronic intake of boysenberry juice or artificial vinegar on blood pressure (BP) and endothelial function were investigated in spontaneous hypertensive rats (SHR). A single administration of boysenberry juice (BJ, equivalent to 0.5 mL/kg body weight) or artificial boysenberry juice vinegar (BJV, equivalent to 0.5 mL BJ and 0.10 g acetic acid/kg body weight) decreased both systolic blood pressure (SBP) and diastolic blood pressure (DBP) significantly. Reductions in SBP of the control group compared with the BJ and BJV groups reached maxima of -16.8±4.3 and -28.4±7.3 mmHg 8 h after administration, respectively. Chronic SBP- and DBP-lowering effects were also observed upon daily feedings of both BJ and BJV at 4 wk. No significant differences were found in SBP or DBP between respective acute and chronic intake of BJ and BJV, except for the decrease in DBP after 4 wk of BJV intake. This suggests that the polyphenol constituents in BJ and BJV likely play a major role in lowering SBP and DBP under these conditions and that acetic acid added to BJ exerts a DBP-lowering effect after 4 wk of BJV intake. The polyphenolic constituents of these beverages might elevate plasma NO concentration via aortic endothelial nitric oxide synthase activation, but the effects of chronic intake on blood pressure might also be at least partly mediated by the renin-angiotensin system. These results may help explain the beneficial effects of boysenberry intake on cardiovascular health, such as reduced blood pressure and improved endothelial function.

Endoinulinase was partially purified from the culture filtrate of a filamentous fungus Aspergillus niger mutant 817. The enzyme preparation was immobilized covalently onto a porous cellulose derivative, Amino-Cellulofine. A 5% (w/v) solution (pH 5.0) of pure dahlia inulin was fed continuously into a packed-bed column reactor containing the immobilized enzyme. The operating conditions were studied to produce a mixture of oligomers with a degree of polymerization of 3 to 5 by partial hydrolysis of inulin. Inulotriose (F3) and -tetraose (F4) were purified from the hydrolysis products by preparative high-performance liquid chromatography. In vitro studies indicated that both the F3 and F4 were preferentially utilized by Bifidobacterium spp., but not by Escherichia coli or Clostridium perfringens.

The fopA gene encoding a fructooligosaccharide-producing beta-fructofuranosidase was isolated from Aspergillus niger ATCC 20611. The primary structure deduced from the nucleotide sequence showed considerable similarity to those of two other beta-fructofuranosidases from A. niger, but the fopA gene product had several amino acid insertions and an extra C-terminal polypeptide consisting of 38 amino acids that could not be found in the two others. We could successfully express the fopA gene in S. cerevisiae and the fopA gene product obtained from the culture supernatant of the S. cerevisiae transformant had similar characteristics to the beta-fructofuranosidase purified from A. niger ATCC 20611. However, we could not detect any beta-fructofuranosidase activity in either the culture supernatant or cell lysate when the C-terminal truncated fopA gene product by 38 amino acids was used to transform S. cerevisiae. In western analysis of those samples, there was no protein product that is cross-reacted with anti-beta-fructofuranosidase antibody. These results suggested that the C-terminal region of the fopA gene product consisting of 38 amino acids was essential for the enzyme production.

Previously, we confirmed that dietary fructooligosaccharides (FOS) increase calcium absorption in rats. In this study, we examined the influence of FOS feeding on the concentration of calbindin-D9k of several intestinal segments in rats. Rats in the control group were fed a diet without FOS. Rats in the other two groups were fed the diet containing FOS at either 50 or 100 g/kg for 10 d and subjected to a calcium absorption study. On the final day of feeding, the rats were killed and the entire intestine was removed. The intestinal mucosa was collected from four segments, i.e., the proximal and distal segments of the small intestine, the cecum and the colorectum, respectively. The apparent absorption of calcium increased dose dependently (r = 0.9256, P < 0.0001). Significant positive correlations between apparent calcium absorption and the relative amounts of calbindin in both large intestinal segments were observed (cecum, r = 0.8956, P = 0.0011; colorectum, r = 0.8828, P = 0.0016). Also, significant negative correlations between apparent calcium absorption and the relative amounts of calbindin-D9k in both small intestinal segments were observed (proximal, r = -0.7149, P = 0. 0304; distal, r = -0.8740, P = 0.0021). In conclusion, FOS feeding increases levels of calbindin-D9k in the large intestine, but decreases those in the small intestine. Moreover, these results suggest that part of the stimulatory effect of fructooligosaccharides relates to the transcellular route of calcium absorption in the large intestine of rats.

Gastrectomized rats develop anemia and osteopenia, and ingestion of fructooligosaccharides leads to an increase in iron absorption and promotes recovery from anemia in iron-deficient rats. Laparotomized (sham-operated control) rats and totally gastrectomized (Billoth II) rats, in groups of 14 each, were fed a control diet without fructooligosaccharides or a diet containing fructooligosaccharides (75 g/kg of diet) for 6 wk. All rats received an intramuscular injection of vitamin B-12 every 2 wk. Tail blood was collected every week for determination of hematocrit and hemoglobin concentration. At the end of the experiment, the rats were killed and the femur and tibia were collected for measurement of bone mineral density (BMD). The hematocrit, hemoglobin concentration, hemoglobin regeneration efficiency, and BMD of both femurs and tibias were significantly lower in gastrectomized rats fed the control diet than in the other three groups. Dietary fructooligosaccharides prevented anemia and osteopenia in totally gastrectomized rats.

カゼインホスホペプチド(CPPs:α-CPP及びβ-CPPの混合物)が,ウシ精子のウシ体外成熟卵子への侵入(体外受精)およびその後の胚発生に及ぼす影響について検討した.受精試験には,通常の体外受精方法では受精率が低いウシAおよびBならびに受精率が高いウシCの凍結融解精子を用い,体外成熟卵子への精子侵入率(受精率)を調べた.体外受精用培養液には,カフェイン(5mM),ヘパリン(10単位/ml)を添加し,実験区にはさらにCPPs(1mg/ml)を添加した.その結果,体外受精培養液にCPPsを添加することによって,ウシAおよびBにおいて有意に高い受精率が得られた:ウシA(添加79%,無添加38%),ウシB(添加45%,無添加13%)(P

A plasmid pLC-bar containing the bialaphos resistance gene derived from Streptomyces hygroscopicus between the Lentinus edodes ras gene promoter and priA gene terminator was constructed. When protoplasts of Pleurotus ostreatus were mixed with the plasmid DNA in the presence of polyethylene glycol and CaCl2, bialaphos-resistant colonies were obtained. This indicated that transformation was successful. Southern blot analysis of total DNAs from transformants showed that the introduced plasmid DNA was integrated into the host chromosome and partly rearranged. A plasmid, pLC-GUS, containing the Escherichia coli beta-glucuronidase (GUS) gene under the control of the L. edodes ras gene promoter and priA gene terminator was constructed and introduced into protoplasts of P. ostreatus with pLC-bar by co-transformation. Two of 5 transformants obtained as bialaphos-resistant colonies showed two to twenty times higher specific activity of GUS than the recipient. Southern blot analysis of total DNAs from transformants indicated the presence of the GUS gene only in the two transformants. These results indicated that co-transformation of P. ostreatus was successful, and that the GUS gene was expressed in P. ostreatus. This transformation system will enable us to breed commercial strains of P. ostreatus at the molecular level.

The antihypertensive effect of a single oral administration of a boysenberry seed polyphenol extract to spontaneously hypertensive rats was evaluated at different doses (100 and 200 mg/kg), and a significant decrease in systolic blood pressure (SBP) was observed up to 6 h post administration. The extract was separated into proanthocyanidin-rich and ellagitannin fractions by solvent partition. A significant decrease in SBP was observed only after administering the proanthocyanidin-rich fraction, and this decrease was abolished by an N(G)-nitro-L-arginine methyl ester (L-NAME) injection. An analysis of the orally absorbable components showed that intact dimeric and trimeric procyanidins and propelargonidins were detectable in the plasma with a maximal concentration 2 h post administration. The vasorelaxant activity of the extract was also confirmed by in vitro assay using rat aorta rings. These results suggest that proanthocyanidins (PAs) in boysenberry seeds may have played an important role in the observed antihypertensive effect.

Using microplates as pressure and cultivation vessels, a high-throughput method was developed for analyzing the high-pressure inactivation kinetics of microorganisms. The loss of viability from a high-pressure treatment, measured based on the growth delay during microplate cultivation, showed reproducibility with the conventional agar plate method and was applicable for the kinetics analysis.

豚精子の凍結・融解後 (以下, 凍融後と略記) の生存および正常頭帽の維持に対するフラクトオリゴ糖の効果について比較検討し, 次の結果を得た。メイオリゴP, GF2, GF3及びGF4のうち, GF3およびGF4は凍融後37℃, 15～270分における豚精子の生存並びに正常頭帽の維推に優れた効果を示し, その効果は対照の乳糖を有意に上まわった。

Exercise testing was used to examine 38 type 2 diabetes mellitus (DM) patients without cardiovascular complications, aged 47-74 years (body mass index (BMI), 23.8 ± 1.6 kg/m 2 ), and 18 healthy volunteers, aged 45-68 years (BMI, 22.5 ± 1.5 kg/m 2 ). A graded cycling exercise test was performed, monitoring gas exchange to evaluate physical fitness and exercise endurance by oxygen uptake (V • O2) kinetics. The following results were obtained. (1) Oxygen deficit (O2deficit) and time constant (τ on) at the onset of exercise were significantly higher in subjects with type 2 DM compared with controls (p

今回,我々は呼気ガス分析による運動負荷試験を試行してCVA片麻痺患者の回復期リハビリテーション期における運動耐容能と運動エネルギー効率の改善の有無について検討を行った。対象は,当センターに入院した歩行可能なCVA片麻痺患者29名(男性23名,女性6名,年齢55.7歳)とした。運動麻痺の程度による比較検討するために,下肢のBrunnstrom StageによりIII∼IVの重度障害群とV∼VIの軽度障害群の2群に分類した。入院時と退院時のVO2 peakを比較すると、重度障害群では,有意に改善したのに対して,軽度障害群では改善傾向を示した。また,max WRでは,重度及び軽度障害群で有意に改善した。重度障害群と比較して軽度障害群では,患側下肢を含めた運動遂行能力が高く,早期歩行自立の獲得ができ,末梢運動骨格筋機能の向上による運動エネルギー効率の改善につながったものと考えられる。

ラット小腸粘膜から調製した生体酵素(小腸粘膜ホモジネート,微絨毛膜)とSigma社市販小腸アセトン粉末酵素(ラット)の糖質水解活性を比較検討し,更に各種動物の市販小腸アセトン粉末酵素を用いて糖質の水解活性を比較した. 1)水解活性は微絨毛膜,小腸粘膜ホモジネート,市販アセトン粉末酵素の順に低下した. 2)マルトースの水解活性を100としたとき,微絨毛膜と小腸粘膜ホモジネートの糖質に対する相対活性はほぼ一致した.市販小腸粉末酵素では,イソマルターゼ活性,トレハラーゼ活性が生体酵素に比べやや低い値ではあったが,糖質の水解活性比較の簡便な評価系として有効と考えられた. 3)水解活性はグルコースがグルコースとα結合した糖質,グルコースがフラクトースとα結合した糖質,ガラクトースがグルコースとβ結合した糖質,グルコースがグルコースとβ結合した糖質の順に低くなり,水解活性が糖質の結合様式に大きく依存していることが示唆された.また反すう類(ウシ,ヒツジ)および鳥類(ハト)の酵素において水解特性に差異がみられた。

ATM (ataxia telangiectasia mutated) and ATR (AT and Rad3 related) protein kinases play a crucial role in cellular DNA damage responses. The inhibition of these kinases and/or related signaling molecules lead to an abolishment of one such signal pathways termed as checkpoints. It is expected that the discovery of checkpoint modulator such as natural products will be an effective assistance of anti-cancer therapy with lower side effect. In the present study, we demonstrated the inhibitory effect of Squalene (SQ), flavonoids derived from Persimmon leaves (PL) and Schisandrin-B (SchB) on ATM/ATR-dependent checkpoint signaling pathway. The cell viability was decreased by SQ, PL or SchB treated A549 adenocarcinoma after gamma-irradiation (IR) or UV exposure. In each SQ, PL and SchB treated cells, G2/M checkpoint activity was abolished in DNA damaged cells. In vitro kinase assay revealed that PL and SchB inhibits ATM and ATR, respectively. SQ, on the other hand, showed no effect on both kinases. Interestingly, SQ increased Wip1 protein expression in unperturbed cells, and suppressed ATM activation in IR treated cells. Consistent with potential inhibition of ATM by SQ or PL, IR-induced phosphorylations of ATM effectors such as p53 (S15) and Chk1 (S317) were remarkably inhibited by the treatment of SQ or PL. SchB inhibited the specific phosphorylation of these effectors by ATR. Moreover, Xenograft and allograft studies with natural products were demonstrated that the tumor growth of both cancer cells in vivo were obviously inhibited by SQ, PL or SchB treatment. These results suggest that some natural products such as SQ, PL and SchB might be applicable to the sensitization of anti-cancer therapies such as radio- /chemotherapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2971. doi:10.1158/1538-7445.AM2011-2971

Persimmon leaves have been commonly used as traditional medicines in Asia. They have variety kinds of health benefits in such prevention of cancer, hypertension, and inflammation. However, a few of scientific evidences are reported to date. Recent study reported that the cell cycle checkpoint system is playing a pivotal role in DNA damage response, and it is expected that the discovery of checkpoint inhibitor will be an effective assistance of anti-cancer therapy. Checkpoint signaling cascades are critically modulated by ATM (Ataxia telangiectasia-mutated) and ATR (ATM-Rad3-related) protein kinases. Generally, ATM primarily responds to ionizing radiation-induced DNA double-strand breaks, whereas ATR is activated by stalled replication resulting from UV and some genotoxic drugs. In the previous study, we demonstrated that a flavonoid-enriched persimmon leaf extract (FEPLE) promoted cytotoxic effect of cancer cells by chemotherapeutic agents inhibiting the checkpoint activity, especially in ATM dependent pathway. Here, we investigated whether FEPLE inhibits checkpoint activity during DNA damage response induced by radiation such as gamma irrradiation (γIR) and heavy ion (HI) treatment. Dried persimmon leaves were prepared by a decoction for 30 min. A soluble extract was subsequently partitioned into ethyl acetate layer which is flavonoid-enriched persimmon leaves extract (FEPLE). FEPLE included eight flavonol components, non-galloylated flavonoids (kaempferol 3-O-glucoside, kaempferol 3-O-galactoside, quercetin 3-O-glucoside, quercetin 3-O-galactoside) and galloylated flavonoids (their 2″-gallated flavonol glucoside). γIR (2.5 to 20 Gy) and HI (2.5 to 20 Gy) were performed in National Institute of Radiological Sciences (NIRS) and Heavy Ion Medical Accelerator in Chiba (HIMAC), respectively. Human lung adenocarcinoma A549 cells were incubated with FEPLE (0, 1, 10, 30 μg/mL) at one hour prior to γIR or HI treatment. Intracellular levels of phosphorylation in each checkpoint molecule such as p53, SMC1 and Chk1 were determined by western blot using phospho-specific antibodies of Ser15, Ser966 and Ser345, respectively. Phosphorylation on Ser10 of histone H3 was measured by flow-cytometry as a marker of cells in mitosis. The phosphorylation of p53, SMC1 and Chk1 were increased by the treatment of γIR and HI. FEPLE reduced the phosphorylation of checkpoint proteins in both γIR and HI. Furthermore, ATM activity, estimated by the phosphorylation of Ser1981, was inhibited by the treatment of FEPLE. The pre-treatment of FEPLE significantly prevented the decrease of mitotic cells in HI exposed cells. These results indicated that DNA damage checkpoint system in γIR or HI exposed cells were abrogated by FEPLE treatment through the inhibition of ATM-dependent signaling pathway. FEPLE might be useful for clinical application by combining γIR or HI cancer therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3875.

The fructosyl transfer to sucrose was investigated, and Aspergillus niger ATCC 20611 was selected as the most suitable strain for fructooligosaccharide production. This strain showed very high enzyme productivity, and its transfructosylating activity was very strong compared to its hydrolyzing activity. Fructooligosaccharides could be more effectively prepared with a higher concentration of sucrose if use could be made of an enzyme having higher transfructosylating ability. Treatment of 50% (w/v) sucrose with the A. niger enzyme afforded a mixture of fructooligosaccharides with inulin-type structures of 1F(1-β-fructofuranosyl)n-sucrose (n = 1 to 3). The individual saccharides could be separated by the combination of a carbon column chromatography and preparative HPLC.

A fructooligosaccharide-producing β-fruetofuranosidase was purified from cells of Aspergillus niger ATCC 20611. The molecular weight was 340,000 by gel filtration. The optimum pH of the enzyme was 5.0 ~ 6.0 and the optimum temperature was 50~60°C. The enzyme was rendered inactive by 1 mM Hg2+ and the Km value for sucrose was 0.29 m. The enzyme catalyzed an almost exclusively fructosyl transfer reaction in a 50% sucrose solution to produce a mixture of fructooligosaccharides and glucose, but both fructosyl transfer and hydrolytic action were observed in a 0.5 % solution. The β-fructofuranosidase showed a high regiospecificity to transfer the fructosyl moiety for the 1-OH group of terminal fructofuranosides.

(S)-(+)-Phosphinothricin was prepared in good optical yield by the Michael addition of chiral glycine Schiff base derived from (S)-2-hydroxy-3-pinanone to vinyl phosphorus compound. (R)-(−)-Phosphinothricin, an enantiomeric isomer, can also be prepared from the same chiral glycine Schiff base by choosing suitable reaction temperature.

The preparation of pent-2-en-4-ynylamine and pent-2-en-4-ynylthiol and their derivatives from 5-bromopent-3-en-1-yne was studied. Moreover, in connection with the preparation of the bromide, the rearrangement-bromination of vinylethynylcarbinol was found to give three isomeric bromides, 5-bromopent-trans-3-en-1-yne, 5-bromopent-cis-3-en-1-yne, and 3-bromopent-1-en-4-yne. Pent-2-en-4-ynylamine was obtained from 5-bromopent-3-en-1-yne by two methods, by the hydrolysis of the hexaminium salt and by the cleavage of N-(pent-2-en-4-ynyl)phthalimide. The reaction of 5-bromopent-3-en-1-yne with potassium hydrosulfide and the decomposition of ethyl pent-2-en-4-ynylxanthate by alcoholic ammonia gave pent-2-en-4-ynylthiol.