Peritoneal dissemination of pancreatic cancer has a poor prognosis. We have reported that intraperitoneal radioimmunotherapy using a 64Cu-labeled antibody (64Cu-ipRIT) is a promising adjuvant therapy option to prevent this complication. To achieve personalized 64Cu-ipRIT, we developed a new in vitro tumor cell-binding assay (64Cu-TuBA) system with a panel containing nine candidate 64Cu-labeled antibodies targeting seven antigens (EGFR, HER2, HER3, TfR, EpCAM, LAT1, and CD98), which are reportedly overexpressed in patients with pancreatic cancer. We investigated the feasibility of 64Cu-TuBA to select the highest-binding antibody for individual cancer cell lines and predict the treatment response in vivo for 64Cu-ipRIT. 64Cu-TuBA was performed using six human pancreatic cancer cell lines. For three cell lines, an in vivo treatment study was performed with 64Cu-ipRIT using high-, middle-, or low-binding antibodies in each peritoneal dissemination mouse model. The high-binding antibodies significantly prolonged survival in each mouse model, while low-and middle-binding antibodies were ineffective. There was a correlation between in vitro cell binding and in vivo therapeutic efficacy. Our findings suggest that 64Cu-TuBA can be used for patient selection to enable personalized 64Cu-ipRIT. Tumor cells isolated from surgically resected tumor tissues would be suitable for analysis with the 64Cu-TuBA system in future clinical studies.

The demand for skin-whitening agents is high across the world, including Asian countries. An extensive screening using a tyrosinase inhibition assay was performed in order to discover novel plant materials. In our research program investigating a safe and effective agent, 50% ethanolic extracts prepared from discarded parts of Prunus persica were screened for in vitro tyrosinase inhibitory activity. Among the extracts tested, twig extract showed the most potent inhibitory activity: 38% inhibition at 500 µg/mL. The investigation of active compounds in twig extract found four flavanones that acted as moderate inhibitors, including (−)-prunin, persiconin, (+)-dihydrokaempferol, and (−)-naringenin. These compounds were only observed in the twig extract following preliminary quantification by HPLC, with the following concentrations: (−)-prunin, 1.8 mg/g sample; persiconin, 0.8 mg/g sample; (+)-dihydrokaempferol, 0.8 mg/g sample; (−)-naringenin, 1.7 mg/g sample. These results suggest that twig extracts can be more useful for skin-whitening compared with other parts of the plant. In addition, a new constituent of twig extract was identified, namely isoquercitrin, which suggests that twig extract can be a potent source of flavones and flavanones. Further studies on the identification of novel compounds from twig extract are now underway in our laboratory.

日ごろから鼻や目の不快感があり、症状緩和の目的で医薬品を常用していない成人男女80名を対象として、未熟ウンシュウミカン果実の乾燥粉末(CUP)を含有する食品(1日あたりhesperidin 215mg、narirutin 48mgを含有)を8週間連続摂取するランダム化二重盲検プラセボ対照並行群間比較試験を行い、被験食品の鼻や目の不快感に対する効果を検証した。摂取前、4週目、8週目のquality of life調査票、visual analogue scale調査票、医師所見、検査所見および鼻や目の不快感日誌を評価した。その結果、被験食品の8週間摂取によって鼻汁(鼻かみ回数)に起因する鼻の不快感が有意に軽減し、鼻汁中好酸球数の変化量は有意に大きかった。CUP含有食品は免疫応答を維持したまま鼻の不快感につながる炎症反応を抑制する可能性が示唆された。

HER1-and HER2-targeted drugs are effective in cancer therapy, especially against lung, breast and colon malignancies; however, resistance of cancer cells to HER1-and HER2-targeted therapies is becoming a serious problem. The avidity/affinity constant (KA) and growth inhibitory effect of anti-HER3 rat monoclonal antibodies (mAb, Ab1∼Ab6) in the presence of therapeutic mAb or low-molecular-weight inhibitors against HER family proteins were analyzed by flow cytometry-based Scatchard plots (Splot) and cell proliferation assay. The KA of Ab3 and Ab6, but not Ab1 or Ab4, split into dual (high and low) modes of KA, and Ab6 exhibited greater anti-proliferative effects against LS-174T colon cancer cells in the presence of Pertuzumab (anti-HER2 mAb). A high KA by Ab6 and Ab6-mediated increased growth inhibition were observed against NCI-H1838 lung or BT474 breast cancer cells, respectively, in the presence of Panitumumab (anti-HER1 mAb) or Perutuzumab. A high KA by Ab6 and Ab6-mediated increased anti-proliferative effects against NCI-H1838 or BT474 were also respectively observed in the presence of Erlotinib (HER1 inhibitor) or Lapatinib (HER1/HER2 inhibitor). In HER1-knockout (KO) NCI-H1838, the reactivity and KA of Ab4 increased compared with in parent NCI-H1838. In HER1-KO or HER3-KO SW1116 colon cancer cells, dual modes of KA with Pertuzumab were noted, and the combination Ab6 and Pertuzumab promoted growth inhibition of HER1-KO, but not of parent SW1116.

L-type amino acid transporter 1 (LAT1)/SLC7A5 is the first identified CD98 light chain disulfide linked to the CD98 heavy chain (CD98hc/SLC3A2). LAT1 transports large neutral amino acids, including leucine, which activates mTOR, and is highly expressed in human cancers. We investigated the oncogenicity of human LAT1 introduced to NIH/3T3 cells by retrovirus infection. NIH/3T3 cell lines stably expressing human native (164C) or mutant (164S) LAT1 (naLAT1/3T3 or muLAT1/3T3, respectively) were established. We confirmed that endogenous mouse CD98hc forms a disulfide bond with exogenous human LAT1 in naLAT1/3T3, but not in muLAT1/3T3. Endogenous mouse CD98hc mRNA increased in both naNIH/3T3 and muLAT1/3T3, and a similar amount of exogenous human LAT1 protein was detected in both cell lines. Furthermore, naLAT1/3T3 and muLAT1/3T3 cell lines were evaluated for cell growth-related phenotypes (phosphorylation of ERK, cell-cycle progression) and cell malignancy-related phenotypes (anchorage-independent cell growth, tumor formation in nude mice). naLAT1/3T3 had stronger growth- and malignancy- related phenotypes than NIH/3T3 and muLAT1/3T3, suggesting the oncogenicity of native LAT1 through its interaction with CD98hc. Anti-LAT1 monoclonal antibodies significantly inhibited in vitro cell proliferation and in vivo tumor growth of naLAT1/3T3 cells in nude mice, demonstrating LAT1 to be a promising anti-cancer target.

This study examines the effect of quantity sold (sales volume) on profitability of market participating smallholder farmers in northern Ghana. Market participation has been shown to be important for increasing incomes and improving production efficiency for farm households but still remains low in SSA. While agribusiness and development experts generally advocate for more intensive market participation, it is not clear if selling more results in more profits for smallholder farmers in remote markets that are prone to exorbitant transaction costs. The data used in this study is from the APS survey conducted in 2013 and 2014 in Northern Ghana which had a sample size of 527. The study is based on the theory of profit maximization, in which separability is inferred from observed market participation. OLS regression is used for empirical estimation after rejecting the hypothesis of endogeneity in the model. Mean gross margin/ kg across four groups of farmers ranked by quantity sold is also statistically examined. The results confirm the existence of economies of scale and also show that different crops have different effects on profitability. The results also show that although unambiguously positive, the relationship between quantity sold and profitability may not be linear.

During the course of Citrus unshiu fruit cultivation, large amounts of plant material including pruned leaves, thinned-out flowers and unripe fruit are obtained; these materials are generally considered unusable and discarded as plant wastes. We have previously attempted to establish an effective use of such plant wastes as beneficial natural resources and found that a methanolic extract of pruned Citrus leaves (CUL-ext) exhibited inhibitory activity against porcine pancreatic lipase. In this study, we sought to identify further the effective uses of C. unshiu plant wastes by determining the lipase inhibitory activity of methanolic extracts of thinning out flowers (CUF-ext) and unripe fruit (CUUF-ext). We accordingly found that the inhibitory activity of CUF-ext was superior to that of CUUF-ext and comparable to that previously observed using CUL-ext. Fractionation of CUF-ext and CUUF-ext, followed by chromatographic analyses, revealed that the pancreatic lipase inhibitory activities of these extracts could be attributed, at least in part, to the flavonoids hesperidin, nobiletin, narirutin and rutin. On the basis of present findings, we propose that, in addition to pruned Citrus leaves, the thinned-out flowers and unripe fruit of C. unshiu are natural resources which are suitable for preparing constituents with lipase inhibitory activity.

Dual-specificity phosphatase 6 (DUSP6) is a key negative feedback regulator of the member of the RAS-ERK MAPK signaling pathway that is associated with cellular proliferation and differentiation. Deterioration of DUSP6 expression could therefore result in deregulated growth activity. We have previously discovered ACA-28, a novel anticancer compound with a unique property to stimulate ERK phosphorylation and induce apoptosis in ERK-active melanoma cells. However, the mechanism of cancer cell-specific-apoptosis by ACA-28 remains obscure. Here, we investigated the involvement of DUSP6 in the mechanisms of the ACA-28-mediated apoptosis by using the NIH/3T3 cells overexpressing HER2/ErbB2 (A4-15 cells), as A4-15 exhibited higher ERK phosphorylation and are more susceptible to ACA-28 than NIH/3T3. We showed that A4-15 exhibited high DUSP6 protein levels, which require ERK activation. Notably, the silencing of the DUDSP6 gene by siRNA inhibited proliferation and induced apoptosis in A4-15, but not in NIH/3T3, indicating that A4-15 requires high DUSP6 expression for growth. Importantly, ACA-28 preferentially down-regulated the DUSP6 protein and proliferation in A4-15 via the proteasome, while it stimulated ERK phosphorylation. Collectively, the up-regulation of DUSP6 may exert a growth-promoting role in cancer cells overexpressing HER2. DUSP6 down-regulation in ERK-active cancer cells might have the potential as a novel cancer measure.

In this study, we aimed to identify the utility of pruned mango (Mangifera indica ‘Irwin’) leaves as a resource for ingredients with antioxidant activity. Firstly, we examined the antioxidant activity of extracts obtained from the pericarps, flesh, flowers, barks, seeds, young dark reddish brown leaves (YDL-ext), young yellow leaves (YYL-ext), and pruned old dark green leaves (OML-ext) obtained from ‘Irwin’ mango. Among them, methanolic extract of flower and OML-ext showed the most potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activity. The flesh extract showed weak DPPH radical scavenging activity, but did not show SOD-like activity. Secondly, we investigated the relationship between the maturation of leaves and their antioxidant activity by considering the contents of their two active polyphenolic components, 3-C-β-D-glucosyl-2,4,4’,6-tetrahydroxybenzophenone (1) and mangiferin (2), in addition to chlorophyll (3) and anthocyanins represented by cyanidin-3-O-glucoside (4). The DPPH radical scavenging activity of YDL-ext, YYL-ext and OML-ext were mainly attributable to 1, 2 and 3, whereas their SOD-like activity was partly attributable to 2. The DPPH radical scavenging and SOD-like activities of YDL-ext and YYL-ext were attributable to 1 and 2. These activities were also due to anthocyanins whose content is highest in YDL-ext. Considering the amounts of leaves obtained from pruning, old dark green leaves may be a reasonable natural resource for preparing cosmetics and/or supplemental ingredients with health-enhancing properties, antioxidant activity and inhibitory effect on AGEs formation and pancreatic lipase.

As a part of our ongoing research to find novel functions in mango leaves, we have reported that the methanolic extract of pruned old dark green mango leaf (Mangifera indica ‘Irwin’) exhibited inhibitory effects on the formation of advanced glycation end products (AGEs) in nonenzymatic glycation of albumin. The purpose of this study was to find other mango cultivars with more potent activity in this regard. We examined the inhibitory effect of seventeen mango (Mangifera indica) cultivar leaf extracts on AGEs formation. We also investigated the relationship between the inhibitory activity of the extracts and the contents of their active components, 3-C-β-D-glucosyl-2,4,4’,6-tetrahydroxybenzophenone (1), mangiferin (2) and chlorophyll (3). On the basis of the evaluation of the inhibitory activity of mango cultivar leaf extracts, the HPLC determination of the contents of 1 and 2, and the spectrophotometric determination of 3, it was found that almost all extract showed a significant activity, and the content of 2 and 3 detected in each was similar. In contrast, AGEs formation inhibition tended to be higher as the content of 1 in the leaf extracts increased. This is the first report of phytochemical analysis of compounds 1, 2 and 3 in various cultivars of mango leaf. From the phytochemical point of view, these results suggest that the pruned leaves of any cultivar of Mangifera indica except ‘Chiin Hwang No. 1’ and ‘Kyo Savoy’ may be useful for the preparation of natural ingredients with inhibitory activity of AGEs formation.

In the cultivation of Citrus fruit, pruning in late winter after harvest is important to obtain excellent flower buds in the next spring and a rich harvest of superior fruit in the following winter; a large number of pruned leaves are usually discarded. The purpose of this study was to determine the utility value of pruned leaves of Citrus unshiu. A methanol extract of pruned C. unshiu leaves showed a porcine pancreatic lipase inhibitory activity. Fractionation of C. unshiu leaf extract (CUL-ext) followed by bibliographic and chromatographic analyses revealed that a part of the pancreatic lipase inhibitory activity of CUL-ext was attributable to nobiletin (1), rutin (2), and hesperidin (3). Thus, pruned C. unshiu leaves may be a reasonable natural resource for the preparation of ingredients with lipase inhibitory activity.

大腸菌、サルモネラの定性試験における製品存在下での試験法の適合性確認(生薬の抗菌性による発育阻害の検証と來雑菌の影響の検証)、胆汁酸抵抗性グラム陰性菌試験における新旧試験法(定量試験)での比較結果を報告した。供試した18生薬すべてにおいて選択培地上に添加した大腸菌の集落を認め、生薬の抗菌性による添加菌の発育阻害は認められなかった。來雑菌の影響の検証では、14検体について各2回実施した試験において、3種の選択培地のいずれを使用しても添加した大腸菌が検出できる陽性の結果を得た。また、胆汁酸抵抗性グラム陰性菌試験の差異の検証に関しては、JP16<5.02>法とJP17<5.02>法を比較した結果、14検体が同等、7検体でJP17<3.02>法の検出感度が高いという結果となった。JP16<5.02>法の検出感度が高かったのは1検体のみであり、JP17<5.02>法の検出感度が高いことが示唆された。

日本漢方生薬製剤協会(日漢協)では、漢方生薬製剤の安全性を保証するため、1996年に『漢方製剤、生薬製剤及び生薬についての残留農薬に関する自主基準』(以下;自主基準)を設定し、2度の改変を経て、現在では有機塩素系・有機リン系・ピレスロイド系を対象に実施している。自主基準の運用状況を調べるため2006年と2010年に日漢協の会員会社に対してアンケートを行った結果、概ね良好な管理がなされていることが明らかになった。今回、同様のアンケートを日漢協会員会社71社に行った。質問内容は、自主基準の対象製剤を販売しているか否かを尋ね、販売していると答えた会社に対して、自主基準対象製剤における残留農薬の試験頻度」「自主基準の基準値を超える残留農薬検出の有無」などについて尋ねた。調査期間は2012年4月〜2013年3月とし、アンケート票は電子メールで配布した。57社から回答が得られ、自主基準の対象製剤を販売しているのは47社(82%)であった。残留農薬の試験頻度は、「全ロット管理している」と答えた会社が最も多く25社(53%)、次いで「一部のロットについて管理している又はスキップテストを実施」10社(21%)であった。基準値を超える残留農薬が検出された事例はなく、自主基準の運用効果が現在も継続していることが示唆された。

We have developed a simple method using a lateral flow kit for the screening of aflatoxins (AFLs) in Kampo extracts. We selected 7 kinds of Kampo extract, differing in terms of color, pH level, and viscosity, listed in the Japanese Pharmacopoeia. Since most of the Kampo extract samples were colored, two decolorization pretreatment methods, namely multifunctional column and immunoaffinity column, were used to examine AFL-spiked extracts (each 10 ppb total AFLs). Using the multifunctional column method and the lateral flow kit, the results revealed that visual judgment of two AFL spiked Kampo extracts (Orengedokuto and Daiokanzoto extracts) was not possible. However, by using the immunoaffinity column method together with the lateral flow kit, good visual judgment of all of the spiked Kampo extracts was possible. Thereafter, HPLC analyses were performed for the cleaned-up samples by the immunoaffinity column method. The results showed that most of the recoveries were in the range of 70-110%, except for the two aforementioned Kampo extracts. Since our method, which combines the application of a lateral flow kit and the immunoaffinity column pretreatment method, does not involve the use of advanced equipment like an HPLC instrument, it may be useful for the screening of AFLs in Kampo extracts.

The Japan Kampo Medicines Manufacturers Association set up self-imposed limits of residual pesticides for Kampo products (organochlorine, organophosphorus and pyrethroid) and conventional crude drug products(organochlorine) in June 2005, and additional limits on organophosphorus and pyrethroid insecticides in conventional crude drug products were set in June 2006. As reported in previous papers, we have been continuously surveying the actual implementation of these self-regulations of residual pesticides by member companies since 2005, and this report deals with our survey result from August 2009. From April 2008 to March 2009, 82.5% of the member companies implemented pesticide tests on the basis of self-regulation, and at least 13,844 samples were tested (Table 10 and 11). As a result, the implementation rate has improved compared to the previous survey. On the other hand, several pesticides and herbal medicines outside of self-regulations were detected, as shown in Table 13 and 14; however, by comparison of the number of detections and various criteria, no results have indicated an increase in risk level.

This investigation was carried out to clarify various chemical effects of Gypsum Fibrosum (石膏) on a representative Kampo medicine, makyokansekito (麻杏甘石湯) , prescribing Gypsum. Based on the combination experiments, Gypsum not only increased the content of inorganic ions, it also stabilized pH and lowered turbidity. Furthermore, it influenced the contents of organic constituents, l-ephedrine, d-pseudoephedrine, amygdalin, liquiritin and glycyrrhizin. From the result of the enzymatic experiments, it was found that Gypsum prevents the decrease in the contents of amygdalin and liquiritin due to its suppression of the hydrolytic potency of emulsin.

The purpose of this study is to clarify the effect of the traditional Japanese medicine Sairei-to (TJ-114) on edema in mice with anti-glomerular basement membrane (GBM) nephritis. Sairei-to (0.38-1.5 g/kg/day) and furosemide (50mg/kg/day) were orally administered consecutively for 4 days from day 6 after an anti-GBM serum injection. Plasma volume (PV) was determined with Evans blue dye and extracellular fluid volume (ECFV) was measured using a single-injection potassium bromide technique. In the nephritic control mice, proteinuria and hyponatreuria were induced and PV, ECFV and calculated interstitial fluid volume (ISFV) were increased. Sairei-to significantly decreased ECFV and ISFV in a dose-dependent manner, but did not change PV or urinary protein excretion. A similar result was observed in the furosemide-treated mice. These results suggest that the treatment with Sairei-to was useful against nephrotic edema. © 2004, Medical and Pharmaceutical Society for WAKAN-YAKU. All rights reserved.

Hoelen, sclederma of Poria cocos Wolf, has long been used as a sedative and diuretic in traditional medicine. Formerly, we demonstrated that Hoelen in vitro protects red blood cells from AAPH-induced hemolysis. In this study, tests were carried out to identify the main ingredient of Hoelen that has the scavenging effect on free-radicals. Triterpene carboxylic acids isolated from the methanol extract of Hoelen, i.e. pachymic acid, polyporenic acid, 3-epidehydrotumulosic acid, 3beta-hydroxylanosta-7,9(11), 24-trien-21-oic acid and 3-o-acetyl-16 alpha -hydroxytrametenolic acid, were found to have inhibitory activities against AAPH-induced lysis of red blood cells. Copyright (C) 2003 John Wiley Sons, Ltd.

Monoamine oxidase B (MAO-B) inhibitors were isolated from the bark of a Brazilian plant, Himatanthus sucuuba (SPR.). Assignments of the H-1- and C-13-NMR data using two dimensional (2D)-NMR techniques showed the active components to be known lichen depsides, confluentic acid (1) and 2'-O-methylperlatolic acid (2). The depside (1) showed selective inhibition of MAO-B with IC50 value of 0.22 mu M.

Three cytotoxic 6-substituted 5, 6-dihydro-2H-pyran-2-ones (1 - 3) were isolated from a Brazilian medicinal plant, Chorisia crispiflora. 1 was identified as argentilactone, and the structures of two new compounds (2 and 3) were determined on the basis of the physicochemical properties.

Three new benzocycloheptenone derivatives, favelol (1), isofavelol (2), and favelone (3), were isolated from the bark of Cnidoscolus phyllacanthus. Their structures were elucidated on the basis of the spectroscopic analysis. Application of the CD exciton chirality method to 4-methoxybenzoates of 1 and 2 led to determination of the R absolute configuration at C-9, which was confirmed by X-ray crystallographic analysis of the 4-bromobenzoyl derivative of 2. © 1994.

A new isoprenoid derivative, faveloxide, was isolated from the bark of Cnidoscolus phyllacanthus. The relative stereostructure (1) was determined on the basis of a spectroscopic analysis and a chemical transformation.

A novel tetracyclic cyclobutene derivative, neofavelanone, was isolated from the bark of Cnidoscolus phyllacanthus. The structure (1) was determined on the basis of the spectroscopic analysis.

A novel tetracyclic cyclopropane derivative, favelanone (1), having cytotoxic activities, was isolated from the bark of Cnidoscolus phyllacanthus and the structure was elucidated on the basis of its spectroscopic data.

Novel cytotoxic benzocycloheptene derivatives, faveline methyl ether (1), faveline (2), and deoxofaveline (3), have been isolated from the bark of Cnidoscolus phyllacanthus. Those structures were determined on the basis of spectroscopic analysis.