Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
kuniyoshi shimizu Last modified date:2019.09.18

Associate Professor / Systematic Forest and Forest Products Science / Department of Agro-environmental Sciences / Faculty of Agriculture


Papers
1. Jun Hata, Tomoyuki Ohara, Yoshinori Katakura, Kuniyoshi Shimizu, Shuntaro Yamashita, Daigo Yoshida, Takanori Honda, Yoichiro Hirakawa, Mao Shibata, Satoko Sakata, Takanari Kitazono, Satoru Kuhara, and Toshiharu Ninomiya, Association Between Serum β-Alanine and Risk of Dementia
The Hisayama Study, American Journal of Epidemiology, 10.1093/aje/kwz116, 188, 9, 1637-1645, 2019.05, We examined the association between serum concentrations of β-alanine, a metabolite of carnosine and anser-
ine, and the risk of dementia in a general population of elderly Japanese persons. In 2007, 1,475 residents of Hi-
sayama, Japan, aged 60–79 years and without dementia were divided into 4 groups according to quartiles of

serum β-alanine concentrations (quartile 1, lowest; quartile 4, highest) and followed for a median of 5.3 years. Dur-
ing follow-up, 117 subjects developed all-cause dementia (Alzheimer in 77 cases and vascular dementia in 31).

The risk of all-cause dementia decreased with increasing serum β-alanine levels after adjustment for potential con-
founding factors (quartile 2, hazard ratio (HR) = 0.73 (95% confidence interval (CI): 0.45, 1.18); quartile 3, HR =

0.50 (95% CI: 0.28, 0.89); quartile 4, HR = 0.50 (95% CI: 0.27, 0.92); P = 0.01 for trend). A similar inverse associa-
tion was observed for Alzheimer disease (quartile 2, HR = 0.78 (95% CI: 0.44, 1.38); quartile 3, HR = 0.53 (95% CI:

0.26, 1.06); quartile 4, HR = 0.53 (95% CI: 0.25, 1.10); P = 0.04 for trend) but not for vascular dementia. We found

that higher serum β-alanine levels were significantly associated with lower risks of all-cause dementia and Alzhei-
mer disease. Because serum β-alanine levels reflect intakes of carnosine/anserine, higher intakes of carnosine/an-
serine might be beneficial for the prevention of dementia.

β-alanine; Alzheimer disease; cohort study; dementia; imidazole dipeptides.
2. Dina I.A. Othman,Khalid B. Selim, Magda A.-A. El-Saye, Atif S. Tantawy, Yhiya Amen Kuniyoshi Shimizu, Tatsuo Okauchi, Mitsuru Kitamura, Design, Synthesis and Anticancer Evaluation of New Substituted Thiophene-
Quinoline Derivatives, Bioorganic & Medicinal Chemistry, https://doi.org/10.1016/j.bmc.2019.07.042, 27, 19, 2019.10, A series of new isoxazolyl, triazolyl and phenyl based 3-thiophen-2-yl-quinoline derivatives were synthesized
adopting click chemistry approach. In addition, the synthesis of new useful synthon, (2-chloroquinolin-3-yl)
(thiophen-2-yl) methanol, is reported. The obtained compounds were characterized by spectral data analysis and
evaluated for their anticancer activity. All the derivatives were subjected to in vitro MTT cytotoxicity screening
assay against a panel of four different human cancer cell lines, liver (HepG-2), colon (HCT-116), human cervical
cancer (HeLa) and breast (MCF-7). Out of a library of 17 compounds, two compounds have been identified as
potent and selective cytotoxic agents against HeLa and MCF-7 cell lines. SAR studies for such hybridized analogues
were investigated and phenyl derivatives were proved to be more potent than isoxazole and triazole
derivatives. Furthermore, the promising compounds were selected for in vitro inhibition of EGFR-TK and Topo II
enzymes. Also, they were subjected to cell cycle arrest analysis and apoptosis assay on MCF-7 cells. Our recent
finding highlights these thiophene-quinoline analogues as a promising class of compounds for further studies
concerning new anticancer therapies..
3. Asmaa KamalEl-Deen,KuniyoshiShimizu, Deep eutectic solvent as a novel disperser in dispersive liquid-liquid microextraction based on solidification of floating organic droplet (DLLME-SFOD) for preconcentration of steroids in water samples: Assessment of the method deleterious impact on the environment using Analytical Eco-Scale and Green Analytical Procedure Index, Microchemical Journal, https://doi.org/10.1016/j.microc.2019.103988, Volume 149, 103988, 2019.11, Herein, we seek clues to investigate the effectiveness of deep eutectic solvent (DES) as a novel dispersive solvent in DLLME-SFOD for the enrichment of nine steroids in water for the first time. The novel DLLME-SFOD method is premised on using 2-dodecanol as extractant and DES (tetrabutylammonium bromide: acetic acid) as a dispersive solvent. Using DES as disperser was compared with other conventional dispersers like (ethanol, methanol, and acetone) attaining a higher recovery than the conventional dispersers. Both HPLC-DAD separation and DLLME-SFOD procedure were optimized by one-factor-at-a-time (OFAT) succeeded by experimental design applying response surface methodology (RSM) to check the optimal conditions of both HPLC separation and extraction. Under the optimal conditions, the proposed method revealed good linearity (correlation coefficients ≥0.998), low detection limits (in the range of 1.0–9.7 ng/mL), and high inter- and intra-day precision (%RSD <5) with high preconcentration factors in the ranges of (44–112) for the nine concerned steroids. Moreover, the method was successfully applied to determine the concerned steroids in different real aqueous samples with expanded measurement uncertainty of the results <17% at 95% confidence level. Finally, the developed procedure impact on the environment was assessed by applying the Analytical Eco-Scale and Green Analytical Procedure Index (GAPI) tools, proving the greenness of the method. The novel disperser DESs have the merits of simple preparation, low cost, enhancing extraction efficiency and being eco-friendly. These DESs can be used to replace conventional organic disperser solvents..
4. Tamrakar, S., Tran, H.B., Nishida, M., Kaifuchi, S., Suhara, H., Doi, K., Fukami, K., Parajuli, G.P., Shimizu, K., Antioxidative activities of 62 wild mushrooms from Nepal and the phenolic profile of some selected species, Journal of Natural Medicines, 10.1007/s11418-016-1013-1. Epub 2016 Jun 4., 70, 4, 769-779, 2019.10, Mushrooms have garnered immense popularity for their nutritional as well as medicinal values. The therapeutic potential of mushrooms in Nepal, a country well known for its biodiversity and natural medicinal resources, remains largely unstudied. Therefore, this study attempts to unveil the antioxidative properties of Nepalese wild mushrooms. Sixty-two wild mushroom samples were collected from several forests in different parts of Nepal. Ethanol and water extracts of the dried samples were tested for their antioxidative activities using total phenolic content (TPC), oxygen radical absorbance capacity (ORAC) assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and reducing power (RP) assays. Ethanol extracts of samples belonging to the order Hymenochaetales showed significantly high activity in all the assays. Inonotus clemensiae had an exceptionally high TPC of 643.2 mg gallic acid equivalent (GAE)/g extract and also exhibited the lowest EC50 values in DPPH (0.081 mg/mL), ABTS (0.409 mg/mL), and EC0.5 value in reducing power (RP; 0.031 mg/mL) assays. High-performance liquid chromatography (HPLC) analysis of the top ten samples with the highest TPC was done to identify the phenolic compounds in the extracts, followed by liquid chromatography-mass spectrometry (LC-MS) analysis for some unknown compounds. These findings highlight the very strong antioxidative activity of Nepalese mushrooms, and paves the way for further research to explore their economic potential..
5. Ahmed Ashour,Yhiya Amen,Ahmed E.Allam,Takashi Kudo,Maki Nagata, Koichiro Ohnuki,Kuniyoshi Shimizu, New isoindolinones from the fruiting bodies of the fungus Hericium erinaceus, Phytochemistry Letters, 10.1016/j.phytol.2019.04.017, 32, 10-14, 2019.08.
6. Mayumi Kaneyasu, Maki Nagata, Hiromi Ikeda, Koichiro Ohnuki , Kuniyoshi Shimizu, Anti-allergic activity of lotus root (Nelumbo nucifera) powder in TDI-sensitized nasal allergy model mice, Food and Agricultural Immunology , 30, 1, 968-978, 2019.08, Lotus root (Nelumbo nucifera) is a popular food in Japan and southeast Asia. It is reported that lotus root includes dietary fiber, vitamin C, and abundant polyphenolic compounds. In Chinese medicine, lotus powder is also utilized in anti-tussive or anti- inflammatory drugs. In this study, we focused on the effects of lotus root powder on allergy symptoms in mice. First, we assessed nasal allergy symptoms in 2,4-diisocyanic acid toluene (TDI)- sensitized nasal allergy model mice. We found that lotus root powder alleviated nasal allergy symptoms in the mice. In addition, we estimated serum inflammatory parameters such as serum total IgE, histamine, and leukotriene B4. Lotus root powder intake significantly decreased serum total IgE and leukotriene B4 compared with standard food intake. We suggest that lotus root powder intake may alleviate allergic symptoms..
7. Rogers Mwakalukwa,Ahmed Ashour,Yhiya Amenac,Yasuharu Niwa,Sonam Tamrakara,Tomofumi Miyamoto,Kuniyoshi Shimizu, Anti-allergic activity of polyphenolic compounds isolated from olive mill wastes, Journal of Functional Foods, 10.1016/j.jff.2019.04.058, 58, 207-217, 2019.07.
8. Yusuke Komiya Toshiya Nakamura Momoko Ishii Kuniyoshi Shimizu Eri Hiraki Fuminori Kawabata Mako Nakamura Ryuichi Tatsumi Yoshihide Ikeuchi Wataru Mizunoya, Increase in muscle endurance in mice by dietary Yamabushitake mushroom (Hericium erinaceus) possibly via activation of PPARδ, Animal Science Journal, 10.1111/asj.13199, 90, 6, 781-789, 2019.06, Skeletal muscle fiber is largely classified into two types: type 1 (slow‐twitch) and type 2 (fast‐twitch) fibers. Meat quality and composition of fiber types are thought to be closely related. Previous research showed that overexpression of constitutively active peroxisome proliferator‐activated receptor (PPAR)δ, a nuclear receptor present in skeletal muscle, increased type 1 fibers in mice. In this study, we found that hexane extracts of Yamabushitake mushroom (Hericium erinaceus) showed PPARδ agonistic activity in vitro. Eight‐week‐old C57BL/6J mice were fed a diet supplemented with 5% (w/w) freeze‐dried Yamabushitake mushroom for 24 hr. After the treatment period, the extensor digitorum longus (EDL) muscles were excised. The Yamabushitake‐supplemented diet up‐regulated the PPARδ target genes Pdk4 and Ucp3 in mouse skeletal muscles in vivo. Furthermore, feeding the Yamabushitake‐supplemented diet to mice for 8 weeks resulted in a significant increase in muscle endurance. These results indicate that Yamabushitake mushroom contains PPARδ agonistic ligands and that dietary intake of Yamabushitake mushroom could activate PPARδ in skeletal muscle of mice. Unexpectedly, we observed no significant alterations in composition of muscle fiber types between the mice fed control and Yamabushitake‐supplemented diets..
9. Ahmed Elbermawi, Ahmed F. Halim, El-Sayed S. Mansour, Kadria F. Ahmad, Ahmed Ashour, Yhiya Amen , Kuniyoshi Shimizu, A new glucoside with a potent α-glucosidase inhibitory activity from Lycium schweinfurthii, Natural Product Research, https://doi.org/10.1080/14786419.2019.1616730, 2019.06.
10. Kilonzi Francisca, Xuting Shen, Toshihori Nakagawa, Thomas Gianfagna, Kuniyoshi Shimizu, Seiko Goto, A Pilot Study on the Effects of Chrysanthemum Scent on Memory and Mood, JOURNAL OF THERAPEUTIC HORTICULTURE, XXIX, 1, 11-23, 2019.06, This pilot study aims to analyze effects on the
short-term memory and mood of the scent of
bornyl acetate and eucalyptol, two volatile
which are commonly used in many essential
oils. The 44 participants in this study were from
Nagasaki University’s School of Fisheries and
Environmental Sciences (19 male, 25 female),
and ranged in age from 18-20 with a mean
age of 19. Effects on short-term memory by a
memory test and changes in mood by the Profile
of Mood States (POMS) questionnaire were
examined. Results showed that the average
score of females and the combination of males
and females was significantly better in the
room with eucalyptol scent compared to the
unscented room. The scent of eucalyptol was
more effective than bornyl acetate in improving
short-term memory, whereas bornyl acetate
was effective in reducing vigor. This effect was
particularly apparent among female subjects..
11. Nakagawa,T., Ashour, A., Amen, Y., Koba, Y., Ohnuki,K., Shimizu, K., α-Glucosidase inhibitory activity of resin from sakhalin fir tree (abies sachalinensis) and its bioactive compounds, Natural Product Communications, 10.1177/1934578X19858460, 14, (6), 2019.06, This study investigated in vitro the α-glucosidase inhibitory activity of resin from the Sakhalin fir tree (Abies sachalinensis). The resin showed extremely high activity (IC50 of 17.3 µg/mL). We isolated 8 compounds from the resin and identified them. All of the compounds isolated from A. sachalinensis resin are reported for the first time in the present study. In an α-glucosidase inhibitory assay, 6 compounds—(holophyllane C (1), (−) -(24 E)-23-oxo-3,4- seco-9βH-lanosta-4(28),6,8(14),24-tetraen-3,26-dioic acid (2), abiesonic acid (3), (−)-(24 E)-23-oxo-3,4- seco-9β H-lanosta-4(28),7,24-triene-3,26-dioic acid (4), 3,4-seco-4(28),6,8(14),22Z,24-mariesapentaen-26,23-olide-3-oic acid (5), and (−)-abiesonic acid 3-methyl ester (6))—showed high activity (their IC50 values were 25.1, 20.9, 82.4, 20.5, 27.3, and 20.5 µg/mL, respectively). These compounds should contribute to the resin’s α-glucosidase inhibitory activity. From an industrial point of view, these findings provide new data to support that the resin of A. sachalinensis is a rich source of α-glucosidase inhibitors, which are highly valuable for their potential to be developed as functional health foods..
12. Satria, D., Tamrakar, S., Suhara, H., Kaneko, S.,Shimizu, K., Mass spectrometry-based untargeted metabolomics and α-glucosidase inhibitory activity of lingzhi (ganoderma lingzhi) during the developmental stages, Molecules, 10.3390/molecules24112044., 24, (11), 2019.05, Lingzhi is a Ganoderma mushroom species which has a wide range of bioactivities. Analysis of the changes in metabolites during the developmental stages of lingzhi is important to understand the underlying mechanism of its biosynthesis, as well as its bioactivity. It may also provide valuable information for the cultivation efficiency of lingzhi. In this study, mass spectrometry based untargeted metabolomics was carried out to analyze the alteration of metabolites during developmental stages of lingzhi. Eight developmental stages were categorized on the basis of morphological changes; starting from mycelium stage to post-mature stage. GC/MS and LC/MS analyses along with multivariate analysis of lingzhi developmental stages were performed. Amino acids, organic acids, sugars, polyols, fatty acids, fatty alcohols, and some small polar metabolites were extracted as marker metabolites from GC/MS analysis, while, lanostane-type triterpenoids were observed in LC/MS analysis of lingzhi. The marker metabolites from untargeted analysis of lingzhi developmental stages were correlated with the α-glucosidase inhibitory activity. Two metabolites, compounds 34 and 35, were identified as potential contributors of the α-glucosidase inhibitory activity. The current result shows that some metabolites are involved in the developmental process and α-glucosidase inhibitory activity of lingzhi..
13. @Nakagawa, T., Ashour, A., Amen, Y., Ohnuki, K., Fujimoto, N., @Shimizu, K., Antioxidant and anti-lipase compounds isolated from heartwood of Yakushima native cedar (Cryptomeria japonica), Journal of Wood Chemistry and Technology, https://doi.org/10.1080/02773813.2019.1578806, 2019.04, This article describes the biological activities of heartwood extracts from Yakushima native cedar (Cryptomeria japonica) and identifies their main active compounds. Black heartwood and red heartwood were each divided into two parts from the center (the pith) to outer heartwood (the cambium). The methanol extracts of those parts were screened for antioxidant and anti-lipase activities. The extract from the pith of the red heartwood showed the highest ORAC activity among the samples. Regarding anti-lipase activity, all the extracts showed relatively high activity (the IC50 value were 9.71–37.24 μg mL−1). In addition, four compounds were isolated and identified from the pith of red heartwood by NMR and MS analyses. Agatharesinol (3) and sequirin C (4) were thought to be the main active compounds responsible for the antioxidant and anti-lipase activities..
14. Hui Tan,Chong Zhao,Qinchang Zhu,Yoshinori Katakura,Hiroyuki Tanaka,,Koichiro Ohnuki,Kuniyoshi Shimizu, Ursolic Acid Isolated from the Leaves of Loquat (Eriobotrya japonica) Inhibited Osteoclast Differentiation through Targeting Exportin 5, Journal of Agricaltural and Food Chemistry, 10.1021/acs.jafc.8b06954, 67, 12, 3333-3340, 2019.03, One of the conventional strategies for treating osteoporosis is to eliminate the multinucleated osteoclasts that are responsible for bone resorption. Our previous study revealed that ursolic acid, isolated from leaves of loquat that is used as tasty tea in Japan, suppressed osteoclastogenesis. We confirmed that ursolic acid exhibited osteoclast differentiation inhibitory activity with an 50% inhibitory concentration (IC50) value of 5.4 ± 0.96 μM. To disclose its mechanism of action, this study first uses polymer-coated magnetic nanobeads to identify potential target proteins. As a result, we identified a nuclear exporter protein named exportin 5 (XPO5). Further studies demonstrated that knockdown of XPO5 significantly blocks osteoclast differentiation (P < 0.01). Expression profiling of mature microRNAs in the cells revealed that downregulation of XPO5 by small interfering RNA or by ursolic acid could downregulate the expression of mature microRNA let-7g-5p during osteoclast differentiation (P < 0.01). Collectively, our findings suggest that ursolic acid inhibits osteoclast differentiation through targeting XPO5, which provides further evidence for the healthy function of the tea. This study also provides new insights into the role of XPO5 and its mediated microRNAs in treatment for bone resorption diseases..
15. A.E. Allam, A.M. Nafady, T. Nakagawa, K. Shimizu, Effect of polyphenols fromVicia faba on lipase activity and melanogenesis, Natral Prodct Research, 32, 16, 1920-1925, 2019.03, Two new flavonoid glycosides, kaempferol 3-O-α-L-rhamnopyranosyl (1→6) (3′′-acetyl)-β-D-galactopyranoside 1 and kaempferol 3-O-α-L-arabinopyranosyl-5-O-α-L-rhamnopyranoside 2, along with six known ones 3–8 were isolated from the flowers of Vicia faba L. (Fabaceae). Methanol extract and the isolated compounds were tested against lipase and melanogenesis inhibition activities and resulted in that compound 2 showed 53 and 77% lipase inhibition activity in concentrations of 400 and 800 μg/mL, respectively. For melanogenesis, compounds 2, 3 and 4 exhibited potent melanogenesis inhibition activity where the melanin content in melanoma cells was decreased to be about 57.5, 56 and 61%, respectively, with no obvious melanocytotoxicity. The rest of compounds showed weak to moderate activity. The results of melanogenesis inhibition activity of this study suggested the potential use of Vicia faba flowers as a skin-whitening agent and reveal the flowers to be a rich source of important phytochemicals with antilipase and melanogenesis inhibitory activity..
16. @Sonam Tamrakar, @Katsuya Fukami, Gopal Prasad Parajuli, @Kuniyoshi Shimizu, Antiallergic Activity of the Wild Mushrooms of Nepal and the Pure Compound Hispidin, Journal of Medicinal Food, Volume 22, Issue 2, February 2019, Pages 225-227, 2019.02, In the present study, ethanol extracts of 90 wild mushroom samples from Nepal, and the pure compound hispidin, were screened for their ability to inhibit β-hexosaminidase release (BHR) from rat basophilic leukemia-2H3 cells. Simultaneously, the toxicity of the extracts toward the cells was also determined, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Samples belonging to the groups Hymenochaetales and Polyporales showed promising anti-allergic activity, with Phellinus adamantinus and Ganoderma lingzhi 3 allowing a mere 19.4% and 16.7% BHR, respectively, without any cell cytotoxicity. Moreover, the 50% inhibitory concentration (IC50) values for Inonotus clemensiae and P. adamantinus were determined to be 51.24 and 50.65 μg/mL, respectively; whereas hispidin, the major bioactive compound in I. clemensiae showed an IC50 value of 82.47 μg/mL. These findings are crucial in underscoring the medicinal value of the wild mushrooms of Nepal, as a source of strong antiallergic agents..
17. Fawzia Ibrahim, Mohie K. Sharaf El-Din, @Asmaa Kamal El-Deen,Kuniyoshi Shimizu, A New HPLC-DAD Method for the Concurrent
Determination of Hydroquinone, Hydrocortisone
Acetate and Tretinoin in Different
Pharmaceuticals for Melasma Treatment, Journal of Chromatographic Science, 2019.01, A new simple and robust HPLC-DAD method was developed for the concurrent analysis of hydroquinone (HQ), hydrocortisone acetate (HCA) and tretinoin (TRN) triple combination for the first time using an Inertsil ODS 3-C18 column (150 mm × 4.6 mm, 5 μm particle size) column with 0.05 M phosphate buffer (pH 5.0) and acetonitrile at a ratio of (10:90, v/v) as a mobile phase, eluted by an isocratic elution mode at a flow rate of 1.0 mL/min and detected at 265 nm. Mefenamic acid was used as an internal standard (I.S.). The method produced linear responses in the concentration range of 10-200, 5-100 and 1-40 μg/mL, with detection limits of 2.01, 1.13 and 0.28 × 10-3 and quantitation limits of 6.11, 3.41 and 0.87 × 10-3 μg/mL for HQ, HCA and TRN, respectively, and a correlation coefficient higher than 0.9998. All validation requirements were satisfied by proving its linearity, precision, accuracy, robustness and specificity. The method was extended for application in triple combination cream, HQ/TRN co-formulated cream and HQ and TRN single ingredient cream with a recovery >97%..
18. Takae, R., Goto, J., Ohnuki, K., Nishide, A., Ohnuki, K., Sato, D., Shimizu, K., Effects of panax ginseng-containing tea on cognitive performance in adolescence-a randomized, double-blinded, placebo-controlled parallel comparison trial-, Japanese Pharmacology and Therapeutics, 47, 3, 485-491, 2019.01, Objectives The purpose of this present study was to investigated effect of ginseng-containing tea intake on cognitive performance. (UMIN000034367) Methods Subjects was 40 volunteers, aged between 13 and 18 years, were randomly allocated to either the placebo group, in which subjects consumed placebo tea, or the ginseng group, in which subjects consumed the ginseng-containing tea. The cognitive performances measured by Benton Visual Retention Test (BVRT), Uchida-Kraepelin psychodiagnostic test (UKT) and TK system Tanaka AB style intelligence test that before and after a 4 week. Results There was a significant interaction over 4 weeks in BVRT score and between the ginseng group and the placebo group. Conclusion These results suggest that Panax ginseng containing tea was beneficial in the visual cognitive performance improvement among adolescence.
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19. Abdelkarem, F.M., Desoky, E.-E.K., Nafady, A.M., Allam, A.E., Mahdy, A., Ashour, A., Mohamed, G.A., Miyamoto, T., Shimizu, K., Two new polyhydroxylated steroids from Egyptian soft coral Heteroxenia fuscescens (Fam.; Xeniidae), Natural Product Research, 10.1080/14786419.2019.1624958. , 2019.01, Two new polyhydroxylated steroids, 3β-acetoxy-gorgost-5α,6β,11α-triol (3) and (23 R) methylergosta-20-ene-3β,5α,6β,17α-tetrol (4), together with three known gorgosteroid compounds, gorgost-3β, 5α,6β,11α- tetrol (1), 11α-acetoxy-gorgost- 3β,5α, 6β- triol (2), and gorgost-5 (E) ene-3-β-ol (5), as well as batyl alcohol (6), were isolated from the Egyptian soft coral Heteroxenia fuscescens. The structures of these compounds were elucidated based on NMR spectroscopic analyses, HR-FAB-MS, and comparisons with published data. The cytotoxic activities of the fractions and compounds were evaluated against MCF-7 cancer cell lines using MTT colorimetric assay. Compounds 2 and 4 showed moderate cytotoxic activity, with IC50 values equal to 33.2 and 25.1 µM, respectively, in comparison with the IC50 of 5-fluorouracil 18.7 µM..
20. Ahmed Ashour, Yhiya Amen, Toshinori Nakagawa, Yasuharu Niwa, Amira Mira, Koichiro Ohnuki, Shinki Murakami, Mitsuko Imao Kuniyoshi Shimizu, A new aliphatic ester of hydroxysalicylic acid from fermented Carica papaya L. preparation with a potential hair growth stimulating activity, Natural Product Research, 2018.12, An aliphatic ester of hydroxysalicylic acid (6), reported for the first time from a natural source in addition to five known compounds were isolated from the fermented Carica papaya L. preparation, a commercialized functional food. The known compounds were identified as 5-hydroxymethylfurfuraldehyde (1), trans-caffeic acid (2), butyl 4-hydroxybenzoate (butylparaben) (3), lycopene (4), benzyl isothiocyanate (5). Compounds 1 and 3 were reported for the first time from Papaya fruits through this study. The new compound showed a moderate antioxidant activity and a potent hair growth stimulating activity in vitro..
21. Enos Tangke Arung, Rudianto Amirta ,Qinchang Zhu, Yhiya Amen,Kuniyoshi Shimizu, Effect of wood, bark and leaf extracts of Macaranga trees on cytotoxic activity in some cancer and normal cell lines, Journal of the Indian Academy of Wood Science, 10.1007/s13196-018-0215-4, 15, 2, 115-119, 2018.11, The genus of Macaranga (Euphorbiacaceae) has
250 species of which 160 species are endemic in Kalimantan
and New Guinea. They grow as pioneer trees and
are used as traditional medicines in the Asian regions. This
experiment concerns cytotoxicity in both cancer and normal
cells of their methanol extracts from wood, bark and
leaf parts. Some of them have not yet reported its cytotoxic
activity in those cell lines. The 21 methanol extracts were
prepared from seven Macaranga tree species (Macaranga
bancana, Macaranga gigantea, Macaranga hullettii,
Macaranga pruinosa, Macaranga tanarius, Macaranga
trichocarpa and Macaranga triloba). The MTT assay was
used to evaluate cytotoxic activity of extracts in cancer cell
lines [human breast cancer (MCF-7), mouse melanoma
(B16 melanoma), human colon cancer (HCT116), human
cervical adenocarcinoma (HeLa)] and normal cell lines
[human normal fibroblast (TIG-1) and normal human
dermal fibroblast (NHDF)]. The leaf extracts of M. pruinosa,
M. tanarius, M. trichocarpa showed more cytotoxicity
than wood and bark in all cancer cell lines. In
addition, the 5-fluorouracil was used as a positive control.
These findings indicated the extracts from leaves of
M. pruinosa, M. tanarius and M. trichocarpa had more
potential than wood and bark parts to be developed as anticancer
agents..
22. H.K. Assaf, A.M. Nafady, A. E. Allam, A.N.E. Hamed, M.S. Kamel,K. Shimizu, Forsskamide, A New Ceramide from Aerial Parts of Forsskaole, Natural Product Research, 10.1080/14786419.2017.1419234, 32, 20, 2452-2456, 2018.10, Although the various folk medicine uses and the biological activity of Forsskaolea tenacissima L., few chemical constituents of this plant have been reported, this provoked us to make our study. Forsskamide, a new ceramide was isolated from aerial parts of F. tenacissima L. (Urticaceae). The chemical structure was established by different spectroscopic methods (1H, 13C-NMR, HMBC, HSQC, ROESY, FAB-MS and HR-FAB-MS). Forsskamide showed a moderate cytotoxic activity by (MTT) method against human colorectal carcinoma cell line (HCT-116) with IC50 33.25 μM in comparison with 5-fluorouracil IC50 26.42 μM. While, it did not show any activity against human hepatocarcinaoma cell line (HepG-2)..
23. El-Tantawy, N.L., Soliman, A.F., Abdel-Magied, A., Ghorab, D., Khalil, A.T., Naeem, Z.M.,Shimizu, K., El-Sharkawy, S.H. , Could Araucaria heterophylla resin extract be used as a new treatment for toxoplasmosis?, Experimental Parasitology, 10.1016/j.exppara.2018.10.003. Epub 2018 Oct 16., 195, 44-53, 2018.10, Toxoplasmosis is a worldwide parasitic disease responsible for serious health problems to human. The currently available drugs used for toxoplasmosis treatment showed a limited efficacy and cause serious host toxicity. The in vitro screening for toxoplasmicidal activity of Araucaria heterophylla resin (AHR) extract and its major component 13-epi-cupressic acid (CUP) showed that both AHR (EC50 = 3.90) and CUP (EC50 = 3.69) have high toxoplasmicidal activity in comparison with standard cotrimoxazole (EC50 = 4.28). The antiprotozoal effects of AHR and CUP were investigated against acute and chronic toxoplasmosis using mice models. Two groups of Swiss albino mice were infected by RH Toxoplasma strain intraperitoneally and by Me49 strain orally. Both groups were treated with AHR and CUP in different doses. Their effects were evaluated by survival rate, peritoneal, spleen and liver parasite burdens, brain cyst burden, NO serum level and histopathological lesions. The ultrastructural changes of tachyzoites of acutely infected mice were studied using scanning electron microscopy (SEM). There is an evidence of toxoplasmicidal activity of AHR and CUP in acute and chronic experimental toxoplasmosis. In the acute model, mice treated with AHR and CUP showed prolonged survival rates, a significant decrease in the parasite density in peritoneal lavage and pathological insult in both liver and spleen compared with that of untreated ones. SEM results denote evident morphological alterations of treated tachyzoites. In chronic experimental toxoplasmosis, AHR and CUP treated groups could significantly reduce brain cyst burden by 96.05% and 98.02% respectively. This study indicates that AHR and CUP showed potent toxoplasmicidal activities experimentally and could be used as a potential natural nontoxic agent for treatment of toxoplasmosis..
24. Yuki Taga, Osamu Hayashida, Ahmed Ashour, Yhiya Amen, Masashi Kusubata, Kiyoko Ogawa-Goto, Kuniyoshi Shimizu, Shunji Hattori, Characterization of Angiotensin-Converting Enzyme Inhibitory Activity of X-Hyp-Gly-Type Tripeptides
Importance of Collagen-Specific Prolyl Hydroxylation, Journal of Agricultural and Food Chemistry, 10.1021/acs.jafc.8b03648, 66, 33, 8737-8743, 2018.08, Hydroxyproline (Hyp) is a collagen-specific amino acid formed by post-translational hydroxylation of Pro residues. Various Hyp-containing oligopeptides are transported into the blood at high concentrations after oral ingestion of collagen hydrolysate. Here we investigated the angiotensin-converting enzyme (ACE) inhibitory activity of X-Hyp-Gly-type tripeptides. In an in vitro assay, ginger-degraded collagen hydrolysate enriched with X-Hyp-Gly-type tripeptides dose-dependently inhibited ACE and various synthetic X-Hyp-Gly-type tripeptides showed ACE-inhibitory activity. In particular, strong inhibition was observed for Leu-Hyp-Gly, Ile-Hyp-Gly, and Val-Hyp-Gly with IC50 values of 5.5, 9.4, and 12.8 μM, respectively. Surprisingly, substitution of Hyp with Pro dramatically decreased inhibitory activity of X-Hyp-Gly, indicating that Hyp is important for ACE inhibition. This finding was supported by molecular docking experiments using Leu-Hyp-Gly/Leu-Pro-Gly. We further demonstrated that prolyl hydroxylation significantly enhanced resistance to enzymatic degradation by incubation with mouse plasma. The strong ACE-inhibitory activity and high stability of X-Hyp-Gly-type tripeptides highlight their potential for hypertension control..
25. F. Ibrahim, A.K. EI-Deen,K. Shimizu, Comparative study of two different chromatographic approaches for quantification of hydrocortisone acetate and pramoxine hydrochloride in presence of their impurities, Journal of Food and Drug Analysis, https://doi.org/10.1016/j.jfda.2017.12.008, 26, 3, 1160-1170, 2018.07, In the present study, we compare the performance of two reversed-phase liquid chromatographic approaches using different eluents either conventional hydro-organic eluent or micellar one for simultaneous estimation of hydrocortisone acetate and pramoxine hydrochloride in presence of their degradants and process-related impurities; hydrocortisone and 4-butoxyphenol, respectively. For conventional reversed-phase liquid chromatography (RPLC), separation of the studied compounds was completed on an Inertsil ODS 3-C18 column (150 mm × 4.6 mm, 5 μm particle size) with a mobile phase consists of 50 mM phosphate buffer (pH 5.0): acetonitrile (50: 50, v/v). For micellar liquid chromatography (MLC), an Eclipse XDB-C8 column (150 mm × 4.6 mm, 5 μm particle size) was chosen for the separation with a green mobile phase consists of 0.15 M sodium dodecyl sulfate, 0.3% triethylamine and 10% n-butanol in 20 mM orthophosphoric acid (pH 5.0). Both methods were extended to analyze hydrocortisone acetate and pramoxine hydrochloride in their co-formulated cream. RPLC was superior to MLC with regard to sensitivity for the estimation of impurities. While, MLC represents an eco-friendly, less hazardous and biodegradable approach. Furthermore, the direct injection of the cream to the system without the need to laborious samples pretreatment, excessive amount of analysis time and/or use of large amount of toxic organic solvents is one of the outstanding advantages of MLC..
26. Zhao Chong, Haruka Matsuo, Mai Kuroda, Shuntaro Yamashita, Gopal Prasad Parajuli, Hira Kaji Manandhar, Kuniyoshi Shimizu, Yoshinori Katakura, Mushroom extract inhibits ultraviolet B-induced cellular senescence in human keratinocytes, Cytotechnology, 10.1007/s10616-018-0229-1, 1-8, 2018.06, Mushrooms possess various bioactivities and are used as nutritional supplements and medicinal products. Twenty-nine bioactive components have been extracted recently from mushrooms grown in Nepal. In this study, we evaluated the ability of these mushroom extracts to augment SIRT1, a mammalian SIR2 homologue localized in cytosol and nuclei. We established a system for screening food ingredients that augment the SIRT1 promoter in HaCaT cells, and identified a SIRT1-augmenting mushroom extract (number 28, Trametes versicolor). UVB irradiation induced cellular senescence in HaCaT cells, as evidenced by increased activity and expression of cellular senescence markers including senescence-associated β-galactosidase, p21, p16, phosphorylated p38, and γH2AX. Results clearly showed that the mushroom extract (No. 28) suppressed the ultraviolet B irradiation-induced cellular senescence in HaCaT cells possibly through augmenting SIRT1 expression..
27. T. Nakagawa, Q. Zhu, S. Tamrakar, Y. Amen, Y. Mori, H. Suhara, S. Kaneko, H. Kawashima, K. Okuzono, Y. Inoue, K. Ohnuki, K. Shimizu , Changes in triterpenoids and polysaccharides content in Ganoderma lingzhi at different growth stages, Journal of Natural Medicines, 72, 3, 734-744, 2018.06, Ganoderma lingzhi is a traditional medicinal mushroom, and its extract contains many bioactive compounds. Triterpenoids and polysaccharides are the primary bioactive components that contribute to its medicinal properties. In this study, we quantified 18 triterpenoids, total triterpenoid content and total polysaccharide content in the ethanol and water extracts of G. lingzhi at different growth stages. Triterpenoids were quantified by liquid chromatograph–tandem mass spectrometry in the multiple-reaction-monitoring mode. Total triterpenoid and total polysaccharide content were determined by colorimetric analysis. The results indicated that the fruit bodies at an early growth stage had a higher content of ganoderic acid A, C2, I and LM2, as well as of ganoderenic acid C and D, than those at a later growth stage. In contrast, ganoderic acid K, TN and T–Q contents were higher in mature fruit bodies (maturation stage). The highest total triterpenoid and total polysaccharide contents were found in fruit bodies before maturity (stipe elongation stage or early stage of pileus formation). Our results provide information which will contribute to the establishment of an efficient cultivation system for G. lingzhi with a higher content of triterpenoids..
28. Yang Hu, Jie Ren, Lei Wang, Xin Zhao, Mian Zhang, Kuniyoshi Shimizu, Chaofeng Zhang, Protective effects of total alkaloids from Dendrobium crepidatum against LPS-induced acute lung injury in mice and its chemical components, Phytochemistry, 10.1016/j.phytochem.2018.02.006, 149, 12-23, 2018.05, Dendrobium crepidatum was one of the sources of Herba Dendrobii, a famous and precious traditional Chinese medicine. Indolizine-type alkaloids are the main characteristic ingredients of D. crepidatum, which possesses a variety of changeable skeletons. In the present study, we found that the total alkaloids of D. crepidatum (TAD) can inhibit the production of nitric oxide (NO) in lipopolysaccharide (LPS)-activated macrophages and showed protective effects against LPS-induced acute lung injury (ALI) in mice through downregulating the TLR4-mediated MyD88/MAPK signaling pathway. Further phytochemical study showed that six previously undescribed indolizine-type compounds, including a racemic mixture (dendrocrepidine A-E) were isolated from TAD. Meanwhile, dendrocrepidine F was separated into a pair of enantiomers by a chiral chromatography, and their absolute configurations were assigned by single-crystal X-ray diffraction analysis. The isomer (−)-dendrocrepidine F showed higher anti-inflammatory effects by inhibiting NO production in LPS-treated macrophages with an IC50 value of 13.3 μM. Taken together, indolizine-type alkaloids are the active components of D. crepidatum through downregulating the TLR4-mediated pathway, indicating some kind of therapy of TAD for ALI treatment..
29. A.F. Soliman, Z.M. Naeem, A.T. Khalil, K. Shimizu, S. H. El-Sharkawy, Microbial transformation of the labdane diterpene 13-epi-cupressic acid, World J. Pharm. Sci, 6, 5, 61-69, 2018.05, The labdane diterpene, 13-epi-cupressic acid 1, was isolated from the n-hexane-chloroform extract of Araucaria heterophylla resin. Of the 20 fungi screened for possible transformation, only four showed to produce metabolites including 13, 14, 15-trihydroxy-8(17)-labden-19-oic acid 2, 7α,13-dihydroxy-8(17),14-labdadien-19-oic acid 3,13-epi-cupressic acid β-glucosyl ester 4 and iso-communic acid 5 by Cunninghamilla echiulata NRRL 1382 , Aspergillus restrictus NRRL 2869, Coriolus hirsutus ATCC MYA-828 and Cordyceps sinclairii ATCC 24400, respectively. The identity of the isolated compounds was confirmed using 1D and 2D NMR and MS spectroscopies. Compound 4 is a new metabolite while 5 is first time being produced by biotransformation technique. Compound 3 was proved to be a potent anti-inflammatory agent through selective inhibition of COX-2 enzyme assay..
30. Mohie Khaled Sharaf El-Din, Fawzia Ibrahim, Asmaa Kamal El-Deen, Kuniyoshi Shimizu, Stability-indicating spectrofluorimetric method with enhanced sensitivity for determination of vancomycin hydrochloride in pharmaceuticals and spiked human plasma
Application to degradation kinetics, Journal of Food and Drug Analysis, 10.1016/j.jfda.2017.06.005, 26, 2, 834-841, 2018.04, Based on investigating the relative fluorescence intensity of vancomycin hydrochloride (VCM) in methanol, a simple, highly sensitive, time-saving and specific spectrofluorimetric method was developed and validated. VCM fluorescence was measured at 335 nm when excited at 268 nm. Excellent linearity is obeyed in the concentration range 1–100 ng/mL with a detection limit of 5.94 pg/mL, a quantitation limit of 18.03 pg/mL and a very good correlation coefficient (r = 0.9999). Our method was applied to analyze VCM in pharmaceuticals as well as spiked human plasma. Moreover, VCM stability was studied when exposed to various degradation conditions such as oxidative, alkaline as well as acidic stress. Acidic and alkaline degradation kinetics of VCM was studied for the first time. The degradation follows pseudo-first-order kinetics. The apparent rate constants and half-life times were calculated. The Arrhenius equation was assessed and the activation energies of the degradation were also calculated. The developed method can be easily applied in quality control laboratories due to its sensitivity, specificity, simplicity and low cost..
31. Toshinori Nakagawa, Qinchang Zhu, Sonam Tamrakar, Yhiya Amen, Yasuhiro Mori, Hiroto Suhara, Shuhei Kaneko, Hiroko Kawashima, Kotaro Okuzono, Yoshiyuki Inoue, Koichiro Ohnuki, Kuniyoshi Shimizu, Changes in content of triterpenoids and polysaccharides in Ganoderma lingzhi at different growth stages, Journal of Natural Medicines, 10.1007/s11418-018-1213-y, 1-11, 2018.04, Ganoderma lingzhi is a traditional medicinal mushroom, and its extract contains many bioactive compounds. Triterpenoids and polysaccharides are the primary bioactive components that contribute to its medicinal properties. In this study, we quantified 18 triterpenoids, total triterpenoid content and total polysaccharide content in the ethanol and water extracts of G. lingzhi at different growth stages. Triterpenoids were quantified by liquid chromatograph–tandem mass spectrometry in the multiple-reaction-monitoring mode. Total triterpenoid and total polysaccharide content were determined by colorimetric analysis. The results indicated that the fruit bodies at an early growth stage had a higher content of ganoderic acid A, C2, I and LM2, as well as of ganoderenic acid C and D, than those at a later growth stage. In contrast, ganoderic acid K, TN and T–Q contents were higher in mature fruit bodies (maturation stage). The highest total triterpenoid and total polysaccharide contents were found in fruit bodies before maturity (stipe elongation stage or early stage of pileus formation). Our results provide information which will contribute to the establishment of an efficient cultivation system for G. lingzhi with a higher content of triterpenoids..
32. Fawzia Ibrahim, Asmaa Kamal El-Deen, Kuniyoshi Shimizu, Application of quinone-based fluorophore and native fluorescence for the spectrofluorimetric determination of agomelatine in dosage form
Identification of acidic and alkaline- induced degradation products by LC–MS/TOF, Luminescence, 10.1002/bio.3405, 33, 1, 225-231, 2018.02, Three spectrofluorimetric methods were developed for agomelatine (AGM) determination in commercial tablets. Method A is based on measuring the native fluorescence of AGM aqueous solution at 230/360 nm. Methods B and C are based on the formation of a charge transfer complex between AGM and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and 7,7,8,8-tetracyanoquinodimethane (TCNQ) with measurement of the formed fluorophore at 365/475 nm and 250/304 nm, respectively. The relative fluorescence intensity (RFI) of AGM–DDQ complex was greatly enhanced in the presence of methyl-β-cyclodextrin (CD). The methods were linear over the concentration ranges of 0.015–0.5, 0.5–8.0, 0.09–6.0 and 0.05–0.2 μg/ml for AGM-native fluorescence, AGM–DDQ, AGM–DDQ–CD and AGM–TCNQ complexes, respectively with excellent correlation coefficients (r = 0.9999). The methods were validated as per the International Conference on Harmonization (ICH) guidelines and all validation requirements were satisfied. The developed methods were extended to the analysis of AGM in commercial tablets. Furthermore, the stability of AGM was studied under different stress conditions (alkaline, acidic, oxidative and photolytic). The potential alkaline and acidic degradation products were identified by LC–MS/TOF..
33. Dedi Satria, Yhiya Amen, Yasuharu Niwa, Ahmed Ashour, Ahmed E. Allam, Kuniyoshi Shimizu, Lucidumol D, a new lanostane-type triterpene from fruiting bodies of Reishi (Ganoderma lingzhi), Natural Product Research, 10.1080/14786419.2018.1440229, 1-6, 2018.02, A new lanostane-type triterpenoid, lucidumol D (1) was isolated from the fruiting bodies of Ganoderma lingzhi. Its structure was elucidated on the basis of extensive 1D- and 2D-NMR studies as well as mass spectrometry. The cytotoxicity of lucidumol D against proliferation of several cancer cells were assayed by using MTT method and the obtained result suggested selective anti-proliferative and cytotoxic effects against MCF-7, HepG2, HeLa, Caco-2, and HCT-116. In comparison to lucidumol C (2) isolated previously by our group, the structure-activity relationship indicated that carbonyl function at C-11 is necessary to enhance the cytotoxicity..
34. Fengyan Cao, Yunfang Zhang, Weiguang Li, Kuniyoshi Shimizu, Haifeng Xie, Chaofeng Zhang, Mogroside IVE attenuates experimental liver fibrosis in mice and inhibits HSC activation through downregulating TLR4-mediated pathways, International Immunopharmacology, 10.1016/j.intimp.2017.12.023, 55, 183-192, 2018.02, Liver fibrosis has been emphasized as a serious threat to human health. There is currently no effective clinical drug treatment. Although mogrosides (MGs) have extensive pharmacological effects with minimal toxicity, their effects on liver function, inflammation, matrix metalloproteinases and hepatic stellate cell (HSC) activation remain to be researched. In the current study, we investigated whether mogroside IVE (MGIVE), a main compound isolated from MGs, provided protection against liver fibrosis in mice. MGIVE (25 mg/kg) significantly reduced carbon tetrachloride (CCl4)-induced inflammatory infiltration, pro-inflammatory cytokine release, and myeloperioxide (MPO) activity, as well as improved liver function in CCl4-treated mice. Additionally, MGIVE also significantly impaired CCl4-induced increases in liver fibrotic marker expression, such as collagen type I and hypoxia inducible factor-1α (HIF-1α). Further investigation indicated that the possible molecular target of MGIVE is the toll-like receptor 4 (TLR4)-mediated pathway, and MGIVE treatment significantly prevented CCl4-induced transforming growth factor-β1 (TGF-β1) overexpression and the phosphorylation of mitogen activated protein kinase (MAPK) in vivo. In vitro tests of HSCs or RAW 264.7 cells challenged with TGF-β1 or lipopolysaccharide (LPS) demonstrated that TLR4 expression partly mediated the anti-fibrotic effects of MGIVE. In conclusion, supplementation with MGIVE may attenuate liver fibrosis through inhibiting the TLR4 signaling pathway, including MyD88 and MAPKs, as well as HIF-1α. MGIVE may act as a therapeutic potential drug for the treatment of liver fibrosis via the TLR4/HIF-1α cohort signaling pathway..
35. Tukiran,Andika Pramudya Wardana,Nurul Hidajati,Kuniyoshi Shimizu, Chemical components and antioxidant activities of methanol extract of syzygium polycephalum miq. stem bark (Myrtaceae), Indonesian Journal of Chemisty, 10.22146/ijc.25467, 18, 1, 26-34, 2018.02.
36. Yhiya M. Amen, Satoru Kaifuchi, Kuniyoshi Shimizu., Methoxyflavones from New Lingzhi Medicinal Mushroom, Ganoderma lingzhi (Agaricomycetes), International Journal of Medicinal Mushrooms, 10.1002/bio.3405. Epub 2017 Oct 2., 33, 1, 225-231, 2018.02, Ganoderma lingzhi is one of the most famous medicinal fungi in the world. It has been used in folk medicine, especially in East Asian countries. It is also a white-rot fungus with strong wood degradation ability, especially against lignin. Different classes of bioactive natural products have been reported in Ganoderma, including triterpenes, polysaccharides, sterols, and peptides. The triterpenes and polysaccharides are the primary bioactive compounds of Ganoderma. We report for the first time the presence of 3 methoxyflavones as minor constituents in G. linghzi. The 3 compounds were identified based on different spectroscopic techniques, including 1- and 2-dimensional nuclear magnetic resonance (1H-1H correlation spectroscopy, heteronuclear single quantum coherence, and heteronuclear multiple bond correlation) and mass spectrometry (high-resolution electrospray ionization mass spectrometry). Our report provides an approach to a possible biosynthetic pathway for biosynthetic genes in the mushrooms. Another great possibility is that these compounds may exist or be formed through degradation of the components in the woody substrate, such as lignin, and then subsequently translocate to the fruiting bodies..
37. Tukiran, Andika Pramudya Wardana, Nurul Hidayati, Kuniyoshi Shimizu, An ellagic acid derivative and its antioxidant activity of chloroform extract of stem bark of Syzygium polycephalum Miq. (Myrtaceae), Indonesian Journal of Chemistry, 10.22146/ijc.25467, 18, 1, 26-34, 2018.01, The investigation of the Syzygium polycephalum Miq. (Myrtaceae) aimed to assess the phytochemical contents and antioxidant activity of the chloroform fraction. In this study, the fraction was obtained from methanol extract of S. polycephalum stem bark partitionated by chloroform. An ellagic acid derivative was successively isolated from the chloroform fraction. The molecular structure of isolated compound was elucidated and established as 3,4,3’-tri-O-methylellagic acid through extensive spectroscopic studies including UV-Vis, FTIR, NMR and LC-MS analyses and by comparison with literature data. The finding of the isolated compound is the first time from the plant, although the isolated compound previously have been found in the other Syzygium species such as S. cumini together with ellagic acid and 3,3’-di-O-methylellagic acid. The chloroform fraction, isolated compound, and vitamin C showed antioxidant activity against 2,2’-diphenyl-1-picrylhydrazyl (DPPH) with IC50 value of 163.6, 72.1, and 11.5 μg/mL, respectively..
38. Yanisa Mittraphab, Nattaya Ngamrojanavanich, Kuniyoshi Shimizu, Kiminori Matsubara, Khanitha Pudhom, Anti-Angiogenic Activity of Rotenoids from the Stems of Derris trifoliata, Planta Medica, 10.1055/s-0044-100797, 2018.01, The plants in the genus Derris have proven to be a rich source of rotenoids, of which cytotoxic effect against cancer cells seem to be pronounced. However, their effect on angiogenesis playing a crucial role in both cancer growth and metastasis has been seldom investigated. This study aimed at investigating the effect of the eight rotenoids ( 1 – 8 ) isolated from Derris trifoliata stems on three cancer cells and angiogenesis. Among them, 12a-hydroxyrotenone ( 2 ) exhibited potent inhibition on both cell growth and migration of HCT116 colon cancer cells. Further, anti-angiogenic assay in an ex vivo model was carried out to determine the effect of the isolated rotenoids on angiogenesis. Results revealed that 12a-hydroxyrotenone ( 2 ) displayed the most potent suppression of microvessel sprouting. The in vitro assay on human umbilical vein endothelial cells was performed to determine whether compound 2 elicits anti-angiogenic effect and its effect was found to occur via suppression of endothelial cells proliferation and tube formation, but not endothelial cells migration. This study provides the first evidence that compound 2 could potently inhibit HCT116 cancer migration and anti-angiogenic activity, demonstrating that 2 might be a potential agent or a lead compound for cancer therapy..
39. Fawzia Ibrahim, Asmaa Kamal El-Deen, Kuniyoshi Shimizu, Comparative study of two different chromatographic approaches for quantitation of hydrocortisone acetate and pramoxine hydrochloride in presence of their impurities, Journal of Food and Drug Analysis, 10.1016/j.jfda.2017.12.008, 2018.01, In the present study, we compare the performance of two reversed-phase liquid chromatographic approaches using different eluents either conventional hydro-organic eluent or micellar one for simultaneous estimation of hydrocortisone acetate and pramoxine hydrochloride in presence of their degradants and process-related impurities; hydrocortisone and 4-butoxyphenol, respectively. For conventional reversed-phase liquid chromatography (RPLC), separation of the studied compounds was completed on an Inertsil ODS 3-C18 column (150 mm × 4.6 mm, 5 μm particle size) with a mobile phase consists of 50 mM phosphate buffer (pH 5.0): acetonitrile (50: 50, v/v). For micellar liquid chromatography (MLC), an Eclipse XDB-C8 column (150 mm × 4.6 mm, 5 μm particle size) was chosen for the separation with a green mobile phase consists of 0.15 M sodium dodecyl sulfate, 0.3% triethylamine and 10% n-butanol in 20 mM orthophosphoric acid (pH 5.0). Both methods were extended to analyze hydrocortisone acetate and pramoxine hydrochloride in their co-formulated cream. RPLC was superior to MLC with regard to sensitivity for the estimation of impurities. While, MLC represents an eco-friendly, less hazardous and biodegradable approach. Furthermore, the direct injection of the cream to the system without the need to laborious samples pretreatment, excessive amount of analysis time and/or use of large amount of toxic organic solvents is one of the outstanding advantages of MLC..
40. Toshinori Nakagawa, Ahmed E. Allam, Koichiro Ohnuki, Kuniyoshi Shimizu, Biological activities of extracts from different parts of two cultivars of Prunus persica 'akatsuki' and 'fastigiata', Natural Product Communications, 13, 10, 1293-1296, 2018.01, We investigated the antioxidant, anti-lipase and anti-dementia activities of peach (Prunus persica (L.) Batsch) fruit and its by-products. The ethanol extracts of branch showed relatively high activity in all biological activities. Then, the extract was fractionated, and eight compounds were isolated from the ethyl acetate fraction. Results showed 4,2',4'-trihydroxy-6'-methoxychalcone 4'-O-β-D-glucopyranoside (5) and quercetin 3-O-β-D-glucopyranoside (7) as newly identified compounds in P. persica. From the biological investigation, it was considered that quercetin 3-O-β-D-glucopyranoside (7) was the main active compound of antioxidant activity. The main active compound of anti-lipase activity in these was oleanolic acid (1). In addition, (+)-4'-O-methylcatechin (4), 4,2',4'- trihydroxy-6'-methoxychalcone 4'-O-β-D-glucopyranoside (5) and ferulic acid (6) were the main active compounds of anti-dementia activity with acetylcholinesterase inhibitory assay. The results obtained suggested that these active compounds from peach branches of P. persica could be exploited as natural antioxidants, anti-lipase and anti-dementia materials in the future..
41. Akemi Nishide, Takashi Fujita, Yoshiteru Nagaregawa, Yukihiro Shoyama, Keiko Ohnuki, Kuniyoshi Shimizu, Tuneyuki Yamamoto, Tomonari Watanabe, Koichiro Ohnuki, Sleep enhancement by saffron extract in randomized control trial, Japanese Pharmacology and Therapeutics, 46, 8, 1407-1415, 2018.01, Background A clinical trial was conducted using 21 healthy adults randomly assigned to a saffron extract group or placebo group in order to confirm the increase of sleep quality by taking saffron extract. Objective To determine the effect of saffron extract on the sleep quality of 21 healthy adults. Trial design A clinical parallel, double-blind, randomized control trial (RCT) was conducted. Subgroups were defined by good sleeper or poor sleeper using a PSQI test for analysis. Method The participants, who gave self-report information, were under a randomized controlled trial (RCT) study designed for all participants receiving the saffron extract (crocin: 0.6mg/day) or placebo. Result There was a significant reduction in score for the Pittsburgh sleep quality index (PSQI) only in the group treated with the saffron extract. In addition, a significant saffron extract effect on daytime dysfunction over 4 weeks was apparent between the extract group and the placebo group in the subjects with poor sleeper at baseline..
42. Tukiran,A. P. Wardana, N. Hidayati,K. Shimizu, An Ellagic Acid Derivative and Its Antioxidant Activity of Chloroform Extract of Stem Bark of Syzygium polycephalumMiq. (Myrtaceae)
, Indones. J. Chem, 18, 1, 26-34, 2018.01, The investigation of the Syzygium polycephalum Miq. (Myrtaceae) aimed to assess the phytochemical contents and antioxidant activity of the chloroform fraction. In this study, the fraction was obtained from methanol extract of S. polycephalum stem bark partitionated by chloroform. An ellagic acid derivative was successively isolated from the chloroform fraction. The molecular structure of isolated compound was elucidated and established as 3,4,3’-tri-O-methylellagic acid through extensive spectroscopic studies including UV-Vis, FTIR, NMR and LC-MS analyses and by comparison with literature data. The finding of the isolated compound is the first time from the plant, although the isolated compound previously have been found in the other Syzygium species such as S. cumini together with ellagic acid and 3,3’-di-O-methylellagic acid. The chloroform fraction, isolated compound, and vitamin C showed antioxidant activity against 2,2’-diphenyl-1-picrylhydrazyl (DPPH) with IC50 value of 163.6, 72.1, and 11.5 μg/mL, respectively..
43. Hamdy K. Assaf, Alaa M. Nafady, Ahmed E. Allam, Ashraf N.E. Hamed, Mohamed S. Kamel, Kuniyoshi Shimizu, Forsskamide, a new ceramide from aerial parts of Forsskaolea tenacissima Linn., Natural Product Research, 10.1080/14786419.2017.1419234, 1-5, 2017.12, Although the various folk medicine uses and the biological activity of Forsskaolea tenacissima L., few chemical constituents of this plant have been reported, this provoked us to make our study. Forsskamide, a new ceramide was isolated from aerial parts of F. tenacissima L. (Urticaceae). The chemical structure was established by different spectroscopic methods (1H, 13C-NMR, HMBC, HSQC, ROESY, FAB-MS and HR-FAB-MS). Forsskamide showed a moderate cytotoxic activity by (MTT) method against human colorectal carcinoma cell line (HCT-116) with IC50 33.25 μM in comparison with 5-fluorouracil IC50 26.42 μM. While, it did not show any activity against human hepatocarcinaoma cell line (HepG-2)..
44. Yhiya Amen, Enos Tangke Arung, Mohamed S. Afifi, Ahmed F. Halim, Ahmed Ashour, Ryoji Fujimoto, Takahiro Goto, Kuniyoshi Shimizu, Melanogenesis inhibitors from Coix lacryma-jobi seeds in B16-F10 melanoma cells, Natural Product Research, 10.1080/14786419.2017.1292270, 31, 23, 2712-2718, 2017.12, An adenine derivative, 9-β-D-glucopyranosyl adenine, reported for the first time from a natural source, in addition to nine known compounds were isolated from the seeds of Coix lacryma-jobi. Their structures were elucidated based on extensive spectroscopic and chemical studies. The isolated compounds and the ethanol extract have been assayed for melanin inhibition using B16-F10 melanoma cell line. The results of our study suggested the potential use of Coix lacryma-jobi seeds as a skin whitening agent and reveal the seeds to be a rich source of important phytochemicals with melanogenesis inhibitory activity. Among the isolated compounds, coixol (2) and 2-O-β-glucopyranosyl-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (8) exhibited potent melanogenesis inhibitory activity with no obvious melanocytotoxicity. The rest of the compounds showed weak to moderate activity..
45. Ibrahim, F., El-Deen, A.K.,Shimizu, K., Application of quinone-based fluorophore and native fluorescence for the spectrofluorimetric determination of agomelatine in dosage form: Identification of acidic and alkaline- induced degradation products by LC–MS/TOF, Luminescence, 33, 1, 223-231, 2017.10, Three spectrofluorimetric methods were developed for agomelatine (AGM) determination in commercial tablets. Method A is based on measuring the native fluorescence of AGM aqueous solution at 230/360 nm. Methods B and C are based on the formation of a charge transfer complex between AGM and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and 7,7,8,8-tetracyanoquinodimethane (TCNQ) with measurement of the formed fluorophore at 365/475 nm and 250/304 nm, respectively. The relative fluorescence intensity (RFI) of AGM-DDQ complex was greatly enhanced in the presence of methyl-β-cyclodextrin (CD). The methods were linear over the concentration ranges of 0.015-0.5, 0.5-8.0, 0.09-6.0 and 0.05-0.2 μg/ml for AGM-native fluorescence, AGM-DDQ, AGM-DDQ-CD and AGM-TCNQ complexes, respectively with excellent correlation coefficients (r = 0.9999). The methods were validated as per the International Conference on Harmonization (ICH) guidelines and all validation requirements were satisfied. The developed methods were extended to the analysis of AGM in commercial tablets. Furthermore, the stability of AGM was studied under different stress conditions (alkaline, acidic, oxidative and photolytic). The potential alkaline and acidic degradation products were identified by LC-MS/TOF..
46. Yang Hu, Lijun Tao, Hui Tan, Mian Zhang, Kuniyoshi Shimizu, Fei Zhang, Chaofeng Zhang, An Active Drimane-Type Lactone from Polygonum jucundum Attenuates Lipopolysaccharide-Induced Acute Lung Injury in Mice Through TLR4-MAPKs Signaling Pathway, Inflammation, 10.1007/s10753-017-0563-z, 40, 4, 1204-1213, 2017.08, The herbs of Polygonum jucundum Lindex. (Polygonaceae) is a traditional Chinese medicine for inflammatory diseases. 2α-Hydroxyl-3β-angeloylcinnamolide (HAC), a drimane-type sesquiterpenoid, was the major active compound of the ethanol extract of P. jucundum which inhibited the production of inflammatory mediators. However, the biological mechanism of HAC for anti-inflammatory activity has not been reported. In the current study, we investigated whether HAC could suppress the production of inflammatory mediators in lipopolysaccharide (LPS)-induced acute lung injury in mice (ALI) through downregulation of Toll-like receptor 4 (TLR4) and activations of mitogen-activated protein kinases (MAPKs) and inducible protein nitric oxide synthase (iNOS). Moreover, our data indicated that HAC inhibits the overexpression of iNOS and TLR4 in LPS-treated RAW264.7, and also inhibits MAPK signal. These findings suggest that HAC shows anti-inflammatory effects in ALI mice through suppressing TLR4-mediated MAPK pathway in activated macrophages. In addition, six derivatives of HAC obtained by structure modification were investigated for their inhibitory effects on the production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α), suggesting that the acetylation could increase the inhibition of HAC on TNF-α release in LPS-treated RAW264.7 cells. In summary, all these results showed that HAC may be a potential anti-inflammatory lead compound for the treatment of acute lung injury..
47. Sonam Tamrakar, Marina Nishida, Yhiya Amen, Hai Bang Tran, Hiroto Suhara, Katsuya Fukami, Gopal Prasad Parajuli, Kuniyoshi Shimizu, Antibacterial activity of Nepalese wild mushrooms against Staphylococcus aureus and Propionibacterium acnes, Journal of Wood Science, 10.1007/s10086-017-1636-1, 63, 4, 379-387, 2017.08, The present study aims to reveal the antibacterial potential of the wild mushrooms of Nepal. Despite the recognition of the medicinal potential of the natural resources in this country, a systematic study on the bioactivities of the wild mushrooms is still lacking. Therefore, in an attempt to fill this gap, ethanol extracts of 90 Nepalese wild mushroom samples were tested for antibacterial activity against Staphylococcus aureus and Propionibacterium acnes. Staphylococcus aureus was comparatively more susceptible with Inonotus clemensiae exhibiting the least minimum bactericidal concentration (MBC) of 100 μg/mL. The major compound in I. clemensiae was identified to be hispidin using high resolution liquid chromatography–electron spray ionization mass spectrometry (LC–ESI-MS) and nuclear magnetic resonance (NMR) analyses. Furthermore, the minimum inhibitory concentration (MIC) and MBC values of hispidin were determined to be 25 and 100 µg/mL for S. aureus and P. acnes, respectively. These findings show that the Nepalese wild mushrooms have the potential to be a novel addition to the functional ingredients industry due to their strong antibacterial potential..
48. Ahmed E. Allam, Alaa M. Nafady, Toshinori Nakagawa, Naomichi Takemoto, Kuniyoshi Shimizu, Effect of polyphenols from Vicia faba L on lipase activity and melanogenesis, Natural Product Research, 10.1080/14786419.2017.1359169, 1-6, 2017.08, Two new flavonoid glycosides, kaempferol 3-O-α-L-rhamnopyranosyl (1→6) (3′′-acetyl)-β-D-galactopyranoside 1 and kaempferol 3-O-α-L-arabinopyranosyl-5-O-α-L-rhamnopyranoside 2, along with six known ones 3–8 were isolated from the flowers of Vicia faba L. (Fabaceae). Methanol extract and the isolated compounds were tested against lipase and melanogenesis inhibition activities and resulted in that compound 2 showed 53 and 77% lipase inhibition activity in concentrations of 400 and 800 μg/mL, respectively. For melanogenesis, compounds 2, 3 and 4 exhibited potent melanogenesis inhibition activity where the melanin content in melanoma cells was decreased to be about 57.5, 56 and 61%, respectively, with no obvious melanocytotoxicity. The rest of compounds showed weak to moderate activity. The results of melanogenesis inhibition activity of this study suggested the potential use of Vicia faba flowers as a skin-whitening agent and reveal the flowers to be a rich source of important phytochemicals with antilipase and melanogenesis inhibitory activity..
49. Fawzia Ibrahim, Asmaa Kamal El-Deen, Samah Abo El Abass, Kuniyoshi Shimizu, An ecofriendly green liquid chromatographic method for simultaneous determination of nicotinamide and clindamycin phosphate in pharmaceutical gel for acne treatment, Journal of Food and Drug Analysis, 10.1016/j.jfda.2016.09.009, 25, 3, 741-747, 2017.07, A new green micellar liquid chromatographic method was developed and validated for the quantitative estimation of nicotinamide (NICO) and clindamycin phosphate (CLD) in bulk and pharmaceutical gel formulation. The analytes are well resolved in less than 6.0 minutes using micellar mobile phase consisting of 0.10M sodium dodecyl sulfate (SDS), 0.3% triethylamine, and 10% 2-propanol in 0.02M orthophosphoric acid at pH 3.0, running through an Eclipse XDB-C8 column (150 mm × 4.6 mm, 5 μm particle size) with flow rate 1.0 mL/min. The effluent was monitored with diode array detection at 210 nm. The retention times of NICO and CLD were 3.8 minutes and 5.6 minutes, respectively. The method was validated according to the International Conference on Harmonisation (ICH) guidelines in terms of linearity, limit of detection, limit of quantification, accuracy, precision, robustness, and specificity to prove its reliability. Linear correlation was achieved by plotting the peak area of each drug against its concentration. It was found to be rectilinear in the ranges of 1.0–40.0 μg/mL and 0.5–15.0 μg/mL with limits of detection of 0.06 μg/mL and 0.03 μg/mL and limits of quantification of 0.19 μg/mL and 0.09 μg/mL for NICO and CLD, respectively. The method was successfully implemented for the simultaneous determination of the analytes in their bulk powder and combined gel formulation with high % recoveries. The ease of sample treatment facilitates and greatly expedites the treatment with reduced cost and improved accuracy of the procedure..
50. Qinchang Zhu, Yhiya M. Amen, Koichiro Ohnuki, Kuniyoshi Shimizu, Anti-influenza effects of Ganoderma lingzhi
An animal study, Journal of Functional Foods, 10.1016/j.jff.2017.04.040, 34, 224-228, 2017.07, The consumption of mushroom Ganoderma lingzhi (G. lingzhi) is believed to help people fight influenza. However, few scientific studies, particularly in animals, has addressed that. Here, a hot-water extract of G. lingzhi was orally or intranasally administered to mice infected with influenza A virus, following by a 21-day post-infection observation. An in vitro neuraminidase (NA) assay, with four subtypes of NA, was used to assess inhibition. The total content of triterpenoids and carbohydrates in the extract were determined with colorimetric assays. Results showed that intranasal administration of the extract reduced the severe weight loss in infected mice by 55.1%. Oral administration of the extract did not significantly interfere the virus infection. In addition, the extract strongly inhibited NA from influenza virus H1N1 and H5N1. These findings suggest that short-term oral consumption of hot-water extract of G. lingzhi has limited anti-influenza function, which also inform further research for its activity observed.
霊芝には、古来よりガンを初め、様々な疾病に関する効能が伝承されている。その効能を裏付けようと多くの基礎研究が実施されており、霊芝の薬理活性に関する研究は、近年、劇的に増大している。その薬理活性は、含有されるβ-グルカンに代表される多糖類と、特徴的な構造を有するラノスタン型トリテルペノイド類に由来すると言われている。抗インフルエンザ薬のターゲットして、ノイラミニダーゼ阻害剤が知られている。すでに、我々は、霊芝に含有されるラノスタン型トリテルペノイドのいくつかが、強力にノイラミニダーゼ活性を阻害することを見出している(SciRep2016).
そこで、その効果をin vivoで検証するために、インフルエンザ罹患動物を用いて、その効果を検討したところ、霊芝には、インフルエンザ予防効果を有することが明らかとなった。.
51. Eri Hiraki, Shoko Furuta, Rika Kuwahara, Naomichi Takemoto, Toshiro Nagata, Taiki Akasaka, Bungo Shirouchi, Masao Sato, Koichiro Ohnuki, Kuniyoshi Shimizu, Anti-obesity activity of Yamabushitake (Hericium erinaceus) powder in ovariectomized mice, and its potentially active compounds, Journal of Natural Medicines, 10.1007/s11418-017-1075-8, 71, 3, 482-491, 2017.07, Hericium erinaceus (H. erinaceus) improves the symptoms of menopause. In this study, using ovariectomized mice as a model of menopause, we investigated the anti-obesity effect of this mushroom in menopause. Mice fed diets containing H. erinaceus powder showed significant decreases in the amounts of fat tissue, plasma levels of total cholesterol, and leptin. To determine the mechanism, groups of mice were respectively fed a diet containing H. erinaceus powder, a diet containing ethanol extract of H. erinaceus, and a diet containing a residue of the extract. As a result, H. erinaceus powder was found to increase fecal lipid levels in excreted matter. Further in vitro investigation showed that ethanol extract inhibited the activity of lipase, and four lipase-inhibitory compounds were isolated from the extract: hericenone C, hericenone D, hericenone F, and hericenone G. In short, we suggest that H. erinaceus has an anti-obesity effect during menopause because it decreases the ability to absorb lipids..
52. Adi Setyo Purnomo, Refdinal Nawfa, Fahimah Martak, Kuniyoshi Shimizu, Ichiro Kamei, Erratum to
Biodegradation of Aldrin and Dieldrin by the White-Rot Fungus Pleurotus ostreatus (Current Microbiology, (2017), 74, 3, (320-324), 10.1007/s00284-016-1184-8), Current Microbiology, 10.1007/s00284-017-1222-1, 74, 7, 2017.07, The original version of this article unfortunately contained a mistake. In Fig. 3 the structure of 9-hydroxyaldrin was incorrect. The correct Fig. 3 is given below. (Figure presented.)..
53. Yhiya Amen, Qinchang Zhu, Hai Bang Tran, Mohamed S. Afifi, Ahmed F. Halim, Ahmed Ashour, Ryoji Fujimoto, Takahiro Goto, Kuniyoshi Shimizu, Rho-kinase inhibitors from adlay seeds, Natural Product Research, 10.1080/14786419.2017.1354183, 1-5, 2017.07, Rho-kinase enzymes are one of the most important targets recently identified in our bodies. Several lines of evidence indicate that these enzymes are involved in many diseases and cellular disorders. ROCK inhibitors may have clinical applications for cancer, hypertension, glaucoma, etc. Our study aims to identify the possible involvement of Rho-kinase inhibition to the multiple biological activities of adlay seeds and provide a rationale for their folkloric medicines. Hence, we evaluated Rho-kinase I and II inhibitory activity of the ethanol extract and 28 compounds derived from the seeds. A molecular docking assay was designed to estimate the binding affinity of the tested compounds with the target enzymes. The results of our study suggest a possible involvement of Rho-kinase inhibition to the multiple biological activities of the seeds. Furthermore, the results obtained with the tested compounds revealed some interesting skeletons as a scaffold for design and development of natural Rho-kinase inhibitors..
54. Hui Tan, Tamrakar Sonam, Kuniyoshi Shimizu, The potential of triterpenoids from loquat leaves (Eriobotrya japonica) for prevention and treatment of skin disorder, International Journal of Molecular Sciences, 10.3390/ijms18051030, 18, 5, 2017.05, The leaves of loquat (Eriobotrya japonica) possess high medicinal value and have been used as traditional medicines. However, there are no evidence-based studies on the skin-care effects of E. japonica leaves. To explore new biological activities of E. japonica leaves against skin disorder and to gain a better understanding of the chemical components associated with bioactivities, we evaluated 18 triterpenoids from E. japonica leaves on anti-melanogenesis, anti-acne, anti-allergy and anti-aging activities. Our results revealed that eight compounds showed anti-melanogenesis activity, of which ursolic acid (1) and maslinic acid (7) were the most potent with the similar selective index to that of arbutin. Structure–activity relationship and possible mechanism of active compounds were proposed. Twelve compounds exhibited anti-acne effect; ursolic acid (1), maslinic acid (7), corosolic acid (8) and euscaphic acid (12) showed highest activities against P. acnes. Four compounds displayed anti-allergy and anti-inflammatory activity; 3-epicorosolic acid (9) and euscaphic acid (12) showed marked activity against β-hexosaminidase release. Finally, ursolic acid (1), pomolic acid (10), colosolic acid (8) and its methylated derivative (6) exhibited the highest anti-aging activity by stimulating collagen and hyaluronic acid (HA) production. Our findings provide valuable evidence that E. japonica leaves have potential applications as ingredients of function foods or cosmetics for health benefits and a number of triterpenoids may play an important role in these bioactivities..
55. Yhiya Amen, Qinchang Zhu, Hai Bang Tran, Mohamed S. Afifi, Ahmed F. Halim, Ahmed Ashour, Kuniyoshi Shimizu, Partial contribution of Rho-kinase inhibition to the bioactivity of Ganoderma lingzhi and its isolated compounds
insights on discovery of natural Rho-kinase inhibitors, Journal of Natural Medicines, 10.1007/s11418-016-1069-y, 71, 2, 380-388, 2017.04, Recent studies identified Rho-kinase enzymes (ROCK-I and ROCK-II) as important targets that are involved in a variety of diseases. Synthetic Rho-kinase inhibitors have emerged as potential therapeutic agents to treat disorders such as hypertension, stroke, cancer, diabetes, glaucoma, etc. Our study is the first to screen the total ethanol extract of the medicinal mushroom Ganoderma lingzhi with thirty-five compounds for Rho-kinase inhibitory activity. Moreover, a molecular binding experiment was designed to investigate the binding affinity of the compounds at the active sites of Rho-kinase enzymes. The structure–activity relationship analysis was investigated. Our results suggest that the traditional uses of G. lingzhi might be in part due to the ROCK-I and ROCK-II inhibitory potential of this mushroom. Structure–activity relationship studies revealed some interesting features of the lanostane triterpenes that potentiate their Rho-kinase inhibition. These findings would be helpful for further studies on the design of Rho-kinase inhibitors from natural sources and open the door for contributions from other researchers for optimizing the development of natural Rho-kinase inhibitors..
56. Toshitaka Kohno, Tran Hai-Bang, Qinchang Zhu, Yhiya Amen, Seiichi Sakamoto, Hiroyuki Tanaka, Satoshi Morimoto, Kuniyoshi Shimizu, Tubulin polymerization-stimulating activity of Ganoderma triterpenoids, Journal of Natural Medicines, 10.1007/s11418-017-1072-y, 71, 2, 457-462, 2017.04, Tubulin polymerization is an important target for anticancer therapies. Even though the potential of Ganoderma triterpenoids against various cancer targets had been well documented, studies on their tubulin polymerization-stimulating activity are scarce. This study was conducted to evaluate the effect of Ganoderma triterpenoids on tubulin polymerization. A total of twenty-four compounds were investigated using an in vitro tubulin polymerization assay. Results showed that most of the studied triterpenoids exhibited microtuble-stabilizing activity to different degrees. Among the investigated compounds, ganoderic acid T-Q, ganoderiol F, ganoderic acid S, ganodermanontriol and ganoderic acid TR were found to have the highest activities. A structure–activity relationship (SAR) analysis was performed. Extensive investigation of the SAR suggests the favorable structural features for the tubulin polymerization-stimulating activity of lanostane triterpenes. These findings would be helpful for further studies on the potential mechanisms of the anticancer activity of Ganoderma triterpenoids and give some indications on the design of tubulin-targeting anticancer agents..
57. Asuka Kishikawa, Yhiya Amen, Kuniyoshi Shimizu, Anti-allergic triterpenes isolated from olive milled waste, Cytotechnology, 10.1007/s10616-016-0058-z, 69, 2, 307-315, 2017.04, The degranulation of basophils stimulated by antigen is one of the allergic mechanisms of immediate hypersensitivity reactions. In a previous study, the β-hexosaminidase release of rat basophilic leukemia (RBL-2H3) cells stimulated by IgE cross-linking was specifically inhibited by ethanol extract of olive milled waste (OMW). OMW is a potential source of triterpenes, which show several biological activities. However, little is known about the effect of triterpenes in OMW on β-hexosaminidase release. Here we investigated anti-allergic triterpenes from the ethanol extract of OMW, and we discuss the structure–activity relationship. We subjected ethanol extract of OMW to activity-guided fractionation; this led to the isolation of five known triterpenes, i.e., maslinic acid (1), oleanolic acid (2), punicanolic acid (3), 2-O-acetylmaslinic acid (4), and epimaslinic acid (5) as well as one new triterpene, 11-oxo-maslinic acid (6). An anti-allergic assay revealed that 2-O-acetylmaslinic acid (4) and 11-oxo-maslinic acid (6) showed anti-allergic activity. Considering the structure–activity relationship of these triterpenes, it appears that the existence of a ketone group at C-11 or an acetyl group at C-2 would influence the anti-allergic activity..
58. Adi Setyo Purnomo, Refdinal Nawfa, Fahimah Martak, Kuniyoshi Shimizu, Ichiro Kamei, Biodegradation of Aldrin and Dieldrin by the White-Rot Fungus Pleurotus ostreatus, Current Microbiology, 10.1007/s00284-016-1184-8, 74, 3, 320-324, 2017.03, Aldrin and its metabolite dieldrin are persistent organic pollutants that contaminate soil in many parts of the world. Given the potential hazards associated with these pollutants, an efficient degradation method is required. In this study, we investigated the ability of Pleurotus ostreatus to transform aldrin as well as dieldrin in pure liquid cultures. This fungus completely eliminated aldrin in potato dextrose broth (PDB) medium during a 14-day incubation period. Dieldrin was detected as the main metabolite, and 9-hydroxylaldrin and 9-hydroxyldieldrin were less abundant metabolites. The proposed route of aldrin biotransformation is initial metabolism by epoxidation, followed by hydroxylation. The fungus was also capable of degrading dieldrin, a recalcitrant metabolite of aldrin. Approximately 3, 9, and 18% of dieldrin were eliminated by P. ostreatus in low-nitrogen, high-nitrogen, and PDB media, respectively, during a 14-day incubation period. 9-Dihydroxydieldrin was detected as a metabolite in the PDB culture, suggesting that the hydroxylation reaction occurred in the epoxide ring. These results indicate that P. ostreatus has potential applications in the transformation of aldrin as well as dieldrin..
59. Yhiya Amen, Enos Tangke Arung, Mohamed S. Afifi, Ahmed F. Halim, Ahmed,Ashour, Ryoji Fujimoto, Takahiro Goto, Kuniyoshi Shimizu, Melanogenesis inhibitors from Coix lacryma-jobi seeds in B16-F10 melanoma cells, Natural Product Research, 31, 23, 2712-2718, 2017.02, An adenine derivative, 9-β-D-glucopyranosyl adenine, reported for the first time from a natural source, in addition to nine known compounds were isolated from the seeds of Coix lacryma-jobi. Their structures were elucidated based on extensive spectroscopic and chemical studies. The isolated compounds and the ethanol extract have been assayed for melanin inhibition using B16-F10 melanoma cell line. The results of our study suggested the potential use of Coix lacryma-jobi seeds as a skin whitening agent and reveal the seeds to be a rich source of important phytochemicals with melanogenesis inhibitory activity. Among the isolated compounds, coixol (2) and 2-O-β-glucopyranosyl-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (8) exhibited potent melanogenesis inhibitory activity with no obvious melanocytotoxicity. The rest of the compounds showed weak to moderate activity..
60. Yhiya Amen, Qinchang Zhu, Hai-Bang Tran, Mohamed S. Afifi, Ahmed F. Halim,, Ahmed Ashour, Kuniyoshi Shimizu, Partial contribution of Rho-kinase inhibition to the bioactivity of Ganoderma lingzhi and its isolated compounds; Insights on discovery of natural Rho-kinase inhibitors, Journal of Natural Medicines, in press, 2017.01.
61. Amira Mira, Wael Alkhiary, Kuniyoshi Shimizu, Antiplatelet and Anticoagulant Activities of Angelica shikokiana Extract and Its Isolated Compounds, Clinical and Applied Thrombosis/Hemostasis, 10.1177/1076029615595879, 23, 1, 91-99, 2017.01, Angelica shikokiana is a Japanese medicinal plant that is used traditionally in several ailments of cardiovascular diseases. However, there is no report regarding its anticoagulant or antiplatelet activities. So this study was designed to screen for such activities (anticoagulant by prothrombin time [PT], activated partial thromboplastin time, and thrombin time assays and antiplatelet activities against adenosine 5′-diphosphate [ADP] and arachidonic acid-induced platelet aggregations) for the methanol extract of the aerial part (Angelica methanol extract [AME]), its isolated coumarins, flavonoids, and flavonoid metabolites. The AME had potent anticoagulant and antiplatelet activities, and the flavonoid compounds were evidenced to be responsible for such activities. Among coumarins compounds, hyuganin C showed significant prolongation of only PT, while other coumarins were inactive. Similarly, hyuganin C and bergapten were the only active coumarins against ADP-induced platelet aggregation. Compared to the parent compounds, colonic metabolites of the flavonoids had similar anticoagulant and antiplatelet activities, while glucuronides showed sharp decreases in all studied activities. This is the first report showing that the medicinal plant A shikokiana has potent antiplatelet and anticoagulant activities..
62. A. Mira, N. Elsherbiny, W. Alkhiary, A. Shebl, H. Tran, Kuniyoshi Shimizu, Hepatoprotective activities of the methanol extract of Angelica shikokiana and isoepoxypteryxin against hepatocellular carcinoma, Indian Journal of Pharmaceutical Sciences, 10.4172/pharmaceutical-sciences.1000265, 79, 4, 576-584, 2017.01, Angelica shikokiana (Apiaceae) is a Japanese traditional herb and widely marketed as a dietary food supplement as a health tea preparation consumed as a daily beverage for its health benefits. Our previous research showed that the methanol extract of the aerial parts of A. shikokiana and its major coumarin, isoepoxypteryxin had a significant cytotoxicity against HepG2 cell line. In this study, we investigated the hepatoprotective activity more deeply by the in vitro inhibition of HepG2 cells invasion assay and in vivo using thioacetamide-induced hepatocellular carcinoma model in rats. The ex vivo levels of nitric oxide production, inducible nitric oxide synthase, vascular endothelial cell growth factor-C and caspase-3 were also estimated to investigate the possible mechanisms of hepatoprotection. The methanol extract of A. shikokiana and isoepoxypteryxin concentration-dependently depressed the invasion of HepG2 cells induced by 12-O-tetradecanoylphorbol 13-acetate. In vivo, they significantly reduced the levels of alanine transaminase, aspartate transaminase, gamma-glutamyl transpeptidase, total bilirubin and nitric oxide production when compared with hepatocellular carcinoma group. Besides, the thioacetamide-injected groups treated with methanol extract of A. shikokiana and isoepoxypteryxin exhibited a significant increase in the level of caspase-3 protein and a significant decrease of inducible nitric oxide synthase and vascular endothelial cell growth factor C levels when compared with hepatocellular carcinoma group. To the best of our knowledge, this is the first report of the hepatoprotective effect of A. shikokiana and isoepoxypteryxin against hepatocellular carcinoma..
63. Fawzia Ibrahim, Mohie K.Sharaf El-Din, Asmaa Kamal El-Deen, Kuniyoshi Shimizu, Micellar HPLC method for simultaneous determination of ethamsylate and mefenamic acid in presence of their main impurities and degradation products, Journal of Chromatographic Science, 10.1093/chromsci/bmw143, 55, 1, 23-29, 2017.01, An eco-friendly sensitive, rapid and less hazardous micellar liquid chromatographic method was developed and validated for the simultaneous analysis of ethamsylate (ETM) and mefenamic acid (MFA) in the presence of hydroquinone (HQ) and 2,3-dimethylaniline (DMA) the main impurities of ETM and MFA, respectively. Good chromatographic separation was attained using Eclipse XDBC8 column (150mm × 4.6 mm, 5 μm particle size) adopting UV detection at 300 nm with micellar mobile phase consisting of 0.12M sodium dodecyl sulfate, 0.3% triethylamine and 15% 2- propanol in 0.02M orthophosphoric acid (pH 7.0) at 1.0 mL/min. The analytes were well resolved in <6.0 min, ETM (tR = 1.55 min), HQ (tR = 1.95 min), MFA (tR = 4.55 min) and DMA (tR = 5.80 min). Different validation parameters were examined as recommended by international conference on harmonization (ICH) guidelines. The method was linear over the concentration ranges of 0.5-18.0, 0.5-20.0, 0.01-0.5 and 0.02-0.2 μg/mL with limits of detection of 0.118, 0.159, 0.005 and 0.005 μg/ mL and limits of quantification of 0.358, 0.482, 0.014 and 0.015 μg/mL for ETM, MFA, HQ and DMA, respectively. The suggested method was successfully applied for the determination of the two drugs in their bulk powder, laboratory-prepared mixtures, single-ingredient and co-formulated tablets. The obtained results were in accordance with those of the comparison method. The method can also detect trace amounts of HQ and DMA as the main impurities of ETM and MFA, respectively, within the BP limit (0.1%) for both impurities. Furthermore, it is a stability-indicating one for the determination of ETM in its pure form, single-component tablet and co-formulated tablets with other drugs..
64. Mutia Devi Hidayati, Taslim Ersam, Kuniyoshi Shimizu, Sri Fatmawati, Antioxidant activity of Syzygium polynthum extracts, Indonesian Journal of Chemistry, 10.22146/ijc.23545, 17, 1, 49-53, 2017.01, Antioxidant activities of Syzygium polyanthum leaves extracts (methanol, ethyl acetate, dichloromethane and n-hexane) were evaluated by using DPPH (2,2-diphenyl-2-picrylhydrazyl) and ABTS (2,2’-azinobis (3-ethylbenzothiazole-6-sulfonic acid) methods. The methanol extract showed the highest antioxidant activity of DPPH assay among extract with IC50 value of 44.35 μg/mL. In addition, methanol extract also showed the highest antioxidant activity of ABTS assay among extracts with IC50 value of 17.69 μg/mL. This study indicated that the methanol extract of S. polyanthum leaves is potential as antioxidant..
65. Gorawit Yusakul, Poomraphie Nuntawong, Seiichi Sakamoto, Pahweenvaj Ratnatilaka Na Bhuket, Toshitaka Kohno, Nao Kikkawa, Pornchai Rojsitthisak, Kuniyoshi Shimizu, Hiroyuki Tanaka, Satoshi Morimoto, Bacterial expression of a single-chain variable fragment (scFv) antibody against ganoderic acid a
A cost-effective approach for quantitative analysis using the scfv-based enzyme-linked immunosorbent assay, Biological and Pharmaceutical Bulletin, 40, 10, 1767-1774, 2017, Due to the highly specific binding between an antibody and its target, superior analytical performances was obtained by immunoassays for phytochemical analysis over conventional chromatographic techniques. Here, we describe a simple method for producing a functional single-chain variable fragment (scFv) antibody against ganoderic acid A (GAA), a pharmacologically active metabolite from Ganoderma lingzhi. The Escherichia coli BL21(DE3) strain produced a large amount of anti-GAA scFv. However, in vitro refolding steps, which partially recovered the reactivity of the scFv, were required. Interestingly, the functional scFv was expressed as a soluble and active form in the cytoplasm of an engineered E. coli SHuffle® strain. Purified anti-GAA scFv, which yielded 2.56mg from 1L of culture medium, was obtained from simple and inexpensive procedures for expression and purification. The anti-GAA scFv-based indirect competitive enzyme-linked immunosorbent assay (icELISA) exhibited high sensitivity (linearity: 0.078–1.25µg/mL) with precision (CV: ≤6.20%) and reliability (recovery: 100.1–101.8%) for GAA determination. In summary, the approach described here is an inexpensive, simple, and efficient expression system that extends the application of anti-GAA scFv-based immunoassays. In addition, when in vitro refolding steps can be skipped, the cost and complexity of scFv antibody production can be minimized..
66. Mai H. El-Naggar, Amira Mira, Fatma M. Abdel Bar, Kuniyoshi Shimizu, Mohamed M. Amer, Farid A. Badria, Synthesis, Docking, Cytotoxicity, and LTA4H Inhibitory Activity of New Gingerol Derivatives as Potential Colorectal Cancer Therapy, Bioorganic & Medicinal Chemistry, in press, 2016.12.
67. Tukiran, F.Mahmudah, N. Hidayati, K.Shimizu, A phenolic acid and its antioxidant activity from stem bark of chloroform fraction of Syzygium littorale (Blume) amshoff (Myrtaceae), Molekul, 2016.11.
68. Amira Mira,Wael Alkhiary,Qinchang Zhu, Toshinori Nakagawa,Hai-Bang Tran, Yhiya M. Amen,Kuniyoshi Shimizu, Improved Biological Activities of Isoepoxypteryxin by Biotransformation, Chemistry and Biodiversity, 1307-1315, 2016.10.
69. Mira, A., Shimizu, K. , An update on antitumor activity of Angelica species, Medicinal Plants - Recent Advances in Research and Development, 363-371, 2016.10, Genus Angelica (Family: Apiaceae) comprises more than 60 species of medicinal plants. Many of these species have long been used in ancient traditional medicine systems, especially in the Eastern countries. For many years, several species of Angelica, e.g., A. archangelica, A. dahurica, A. keiskei, A. gigas, A. pubescens, A. sinensis, A. shikokiana have been used traditionally as anti-inflammatory, diuretic, expectorant, and diaphoretic. Also, these have been used as remedies for hepatitis, arthritis, indigestion, coughs, chronic bronchitis, pleurisy, fever, rheumatism, bacterial and fungal infections, and diseases of the urinary organs. Several classes of compounds such as coumarins, acetylenic compounds, chalcones, sesquiterpenes, and polysaccharides have been reported from these plants. This chapter outlines the results of various scientific studies on Angelica species that were reported to have anticancer and antitumor activities..
70. F.Ibrahim, M.K.S.El-Din, A.K.El-Deen, Shimizu K, Micellar HPLC method for simultaneous determination of ethamsylate and mefenamic acid in presence of their main impurities and degradation products, journal of Choromatographic Science, in press, 2016.09.
71. F.Ibrahim, A.K.El-Deen, S.A.El Abass, Shimizu K, An Eco-friendlly Green Liquid Chromatographic Method for Simultaneous Determination of Nicotinamide and Clindamycin phosphate in Pharmaceutical Gel for Acne Treatment, Jounal of Food and Drug Analysis, in press, 2016.09.
72. S.Sakamoto, N.Kikkawa, T.Kohno, Shimizu K, H.Tanaka, S.Morimoto, Immunochromatographic strip assay for detection of bioactive Ganoderma triterpenoid, ganoderic acid A in Ganoderma Lingzhi, Fitoterapia, 2016.08.
73. Shimizu, K., Amen, Y.M., Kaifuchi, S., Methoxyflavones from New Lingzhi medicinal mushroom, Ganoderma lingzhi (agaricomycetes), International Journal of Medicinal Mushrooms, 18, 8, 713-719, 2016.08, Ganoderma lingzhi is one of the most famous medicinal fungi in the world. It has been used in folk medicine, especially in East Asian countries. It is also a white-rot fungus with strong wood degradation ability, especially against lignin. Different classes of bioactive natural products have been reported in Ganoderma, including triterpenes, polysaccharides, sterols, and peptides. The triterpenes and polysaccharides are the primary bioactive compounds of Ganoderma. We report for the first time the presence of 3 methoxyflavones as minor constituents in G. linghzi. The 3 compounds were identified based on different spectroscopic techniques, including 1- and 2-dimensional nuclear magnetic resonance (1H-1H correlation spectroscopy, heteronuclear single quantum coherence, and heteronuclear multiple bond correlation) and mass spectrometry (high-resolution electrospray ionization mass spectrometry). Our report provides an approach to a possible biosynthetic pathway for biosynthetic genes in the mushrooms. Another great possibility is that these compounds may exist or be formed through degradation of the components in the woody substrate, such as lignin, and then subsequently translocate to the fruiting bodies..
74. W.D. Ditriana, T. Ersam, K. Shimizu, S. Fatmawati, Antioxidant activity of Moringa oleifera extracts, Indones. J. Chem, https://doi.org/10.22146/ijc.21145 , 16, 3, 297-301, 2016.08, Moringa oleifera have been evaluated for its antioxidant activity. M. oleifera leaves were extracted with methanol, ethyl acetate, dichloromethane and n-hexane. The antioxidant activity of extracts were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity assay and an improved 2,2’-azino-bis-[3-ethylbenzothiazoline sulphonate] (ABTS) radical cation decolorization assay in vitro. Trolox was used as standard with IC50 5.89 μg/mL in DPPH assay and 3.06 μg/mL in ABTS assay. The methanol extract showed the highest free radical scavenging activity with IC50 value of 49.30 μg/mL in DPPH assay and 11.73 μg/mL in ABTS assay. This study provided that M. oleifera leaves possess antioxidant..
75. Y.M.Amen, Q.Zhu, M.S.Afifi, A.Ashour, Shimizu K, New cytotoxic lanostanoid triterpenes from Ganoderma lingzhi, Phytochemistry Letterss, 2016.07.
76. Yhiya M. Amen, Amira Mira, Hai-Bang Tran, Mohamed S. Afifi, Ahmed F. Halim, Ahmed Ashour,Kuniyoshi Shimizu, Lucidumol C, a new cytotoxic lanostanoid triterpene from Ganoderma lingzhi against human cancer cells, Journal of Natural Medicines, 70, 3, 661-666, 2016.07, A new oxygenated lanostane-type triterpene, named lucidumol C, together with six known compounds, was isolated from the chloroform extract of the fruiting bodies of Ganoderma lingzhi. Structures were established based on extensive spectroscopic and chemical studies. Potential cytotoxic activities of the isolated compounds were evaluated against human colorectal carcinoma (HCT-116, Caco-2), human liver carcinoma (HepG2), and human cervical carcinoma (HeLa) cell lines using WST-1 reagent. Selectivity was evaluated using normal human fibroblast cells (TIG-1 and HF19). Among the compounds, lucidumol C showed potent selective cytotoxicity against HCT-116 cells with an IC50 value of 7.86 ± 4.56 µM and selectivity index (SI) >10 with remarkable cytotoxic activities against Caco-2, HepG2 and HeLa cell lines..
77. S. Tamrakar, H. B. Tran, M. Nishida, S. Kaifuchi, H. Suhara, K. Doi, K. Fukami, G. P. Parajuli, Shimizu K, Anti-oxidative Activities of 62 Wild Mushrooms from Nepal and the Phenolic Profile of Some Selected Species, Journal of Natural Medicines, 1-11, 2016.06.
78. M.Amira, Shimizu K, In vitro Cytotoxic Activities and Molecular Mechanisms of Angelica shikokiana Extract and Its isolated compounds, Pharmacognosy Magazine, 11, 4, 564-569, 2016.06.
79. A.Sallam, A.Mira, A.Ashour, Shimizu K, Acetyl choline esterase inhibitors and melanin synthesis inhibitors from Salvia officinalis, Phytomedicine in Press, 2016.06.
80. T. Kohno, T. Hai-Bang, Q. Zhu, S. Sakamoto, H. Tanaka, S. Morimoto, Shimizu K, Tubulin polymerization stimulating-activity of Ganoderma triterpenoids(in press), in press, 2016.05.
81. Yhiya M. Amen, Amira Mira, Hai-Bang Tran, Mohamed S. Afifi, Ahmed F. Halim, Ahmed Ashour, Shimizu K, Lucidumol C, a new lanostanoid triterpene from Ganoderma lingzhi with potential activity against colon cancer (in press), Journal of Natural Medicines, 2016.05.
82. Yhiya M. Amen, Satoru Kaifuchi, Shimizu K, Methoxyflavones from Ganoderma lingzhi (in press)
, International Journal of Medicinal Mushrooms, 2016.05.
83. Q. Zhu, K. Kakino, C. Nogami, K. Ohnuki, Shimizu K, An LC-MS/MS-SRM method for simultaneous quantification of four representative organosulfur compounds in garlic products, Food Analytical Methods , 2016.05.
84. S.Furuta, R.Kuwahara, E.Hiraki, K.Ohnuki, S.Yasuo, K.Shimizu, Hericium erinaceus extracts alter behavioral rhythm in mice, Biomedical Research, 37, 4, 2016.05, Hericium erinaceus (HE), an edible mushroom, has been used as a herbal medicine in several Asian countries since ancient times. HE has potential as a medicine for the treatment and prevention of dementia, a disorder closely linked with circadian rhythm. This study investigated the effects of the intake of HE extracts on behavioral rhythm, photosensitivity of the circadian clock, and clock gene mRNA expression in the suprachiasmatic nucleus (SCN), a central clock, in mice. Although the HE ethanol extract only affected the offset time of activity, the HE water extract advanced the sleep-wake cycle without affecting the free-running period, photosensitivity, or the clock gene mRNA expression in SCN. In addition, both extracts decreased wakefulness around end of active phase. The findings of the present study suggest that HE may serve as a functional food in the prevention and treatment of Alzheimer's disease and delayed sleep phase syndrome..
85. W.D. Ditriana, T. Ersam, K.Shimizu, S. Fatmawati, Antioxidant activity of Moringa oleifera extracts, Indones. J. Chem. , 2016.05.
86. Ahmed Zaki,Ahmed Ashour,Amira Mira,Asuka Kishikawa,Toshinori Nakagawa, Qinchang Zhu,Kuniyoshi Shimizu1*, Biological Activities of Oleanolic Acid Derivatives from Calendula officinalis Seeds, Phytotherapy Research, 30, 5, 835-841, 2016.05.
87. Horiba Hiroki,Nakagawa Toshinori,Zhu Qinchang,Ashour Ahmed,Watanabe Atsushi,Shimizu Kuniyoshi, Biological activities of extracts from different parts of cryptomeria japonica, Natural Product Communications, 11, 9, 1337-1342, 2016.04.
88. R. Abdelhameed, M. S. Elgawish, A. Mira, A. K. Ibrahim, S. A. Ahmed, Shimizu K, K. Yamada, Anti-choline esterase activity of ceramides from the Red Sea Marine Sponge Mycale euplectellioides, RSC Advances , 10.1039/C5RA26424C , 6, 20422-20430, 2016.02.
89. F. Ibrahim, M. K. S. El-Din, R. N. El-Shaheny, A. K. El-Deen, Shimizu K, Simultaneous determination of four vasoactive phytochemicals in different pharmaceutical preparations by a simple HPLC-DAD method, Analytical Methods , 10.1039/C5AY03003J , 8, 1858-1866, 2016.02.
90. T. Nakagawa, Q. Zhu, H. Ishikawa, K. Ohnuki, K. Kakino, N. Horiuchi, H. Shinotsuka, T. Naito, T. Matsumoto, N. Minamisawa, M. Yatagai, Shimizu, K, Multiple uses of essential oil and by-products from various parts of the Yakushima native cedar (Cryptomeria japonica), Journal of Wood Chemistry and Technology, DOI:10.1080/02773813.2015.1057648, 36, 1, 42-55, 2016.01, This article presents multiple potential uses of the Yakushima native cedar (Cryptomeria japonica), known in Japan as the Yaku-sugi tree. The Yaku-sugi was divided into four parts: leaves, branches with leaves, branches, and stems. We obtained the the ess.
91. Sakamoto, S, Kohno, T, Shimizu K, Tanaka, H, Morimoto, S., Detection of ganoderic acid A in Ganoderma lingzhi by an indirect competitive enzyme-linked immunosorbent assay, Planta Med, 10.1055/s-0042-104202, 2015.11.
92. Chisato Inoue, Tomomi Kozaki, Yukiko Morita, Bungo Shirouchi, Katsuya Fukami, Kuniyoshi Shimizu, Masao Sato, Yoshinori Katakura, Kibizu concentrated liquid suppresses the accumulation of lipid droplets in 3T3-L1 cells, Cytotechnology, 10.1007/s10616-015-9849-x, 67, 4, 721-725, 2015.08, Adipocyte size is closely related to the occurrence of diabetes, metabolic syndrome, and insulin resistance. Thus, researchers are searching for active substances that function to reduce adipocyte size. In the present study, we focused on sugar cane vinegar, Kibizu, and evaluated the function of Kibizu to reduce adipocyte size by using an in vitro model system, because people in Amami Oshima famous for longevity regularly consume Kibizu. Results showed that Kibizu treatment significantly reduced the size and number of lipid droplets in 3T3-L1 cells, relative to treatment with Kurozu, another traditional vinegar. Results of an extraction experiment suggest that the active components in Kibizu are lipophilic and hydrophobic. In addition, an in vivo experiment on rats treated with Kibizu showed that the active components were contained in large vein blood. Results of an additional in vivo experiment suggest that metabolites generated by Kibizu-treated rats are primarily contained or modified specifically in the large vein blood..
93. A. Mira, W, Alkhiary, Shimizu, K, Antiplatelet and Anticoagulant Activities of Angelica shikokiana Extract and its Isolated Compounds, Wood Research Journal, doi: 10.1177/1076029615595879, 6, 1, 2015.07, Angelica shikokiana is a Japanese medicinal plant that is used traditionally in several ailments of cardiovascular diseases. However, there is no report regarding its anticoagulant or antiplatelet activities. So this study was designed to screen for such.
94. Shimizu, K, T. H. Bang, Structure-Activity Relationship and Inhibition Pattern of Reishi-derived (Ganoderma lingzhi) Triterpenoids against Angiotensin-Converting Enzyme, Phytochemistry Letters, doi:10.1016/j.phytol.2015.04.021, 12, 243-247, 2015.06.
95. Q. Zhu, T. H. Bang, K. Ohnuki, T. Sawai, K. Sawai, Shimizu K, Inhibition of neuraminidase by Ganoderma triterpenoids and implications for neuraminidase inhibitor design, Scientific Reports, DOI: 10.1038/srep13194., 13194, 2015.06, Neuraminidase (NA) inhibitors are the dominant antiviral drugs for treating influenza in the clinic. Increasing prevalence of drug resistance makes the discovery of new NA inhibitors a high priority. Thirty-one triterpenoids from the medicinal mushroom Ga.
96. O. Matsuda, M. Hara, K. Yazaki, T. Nakagawa, Shimizu K, A. Uemura, H. Utsugi, Determination of Seed Soundness in Conifers Cryptomeria japonica and Chamaecyparis obtusa Using Narrow-Multiband Spectral Imaging in the Short-Wavelength Infrared Range, PLOS ONE, e0128358, 10, 6, 2015.06.
97. A. Ashour, Q. Zhu, M. Yasuda, H. Ishikawa, Shimizu K, Multiple biological effects of olive oil by-products such as leaves, stems, flowers, olive milled waste, fruit pulp, and seeds of the olive plant on skin., Phytotherapy Research, DOI: 10.1002/ptr.5326, 26, 9, 721-725, 2015.06.
98. Tran Hai-Bang, Kuniyoshi Shimizu, Structure-activity relationship and inhibition pattern of reishi-derived (Ganoderma lingzhi) triterpenoids against angiotensin-converting enzyme, Phytochemistry Letters, 10.1016/j.phytol.2015.04.021, 12, 243-247, 2015.06, Abstract Many triterpenoids have shown ability to inhibit hydrolyzing activity of angiotensin-converting enzyme; however, there has been no report about the structure-activity relationship and inhibition patterns of these compounds. In this study, 32 lanostane-type triterpenoids derived from Ganoderma lingzhi were assayed to determine the structural features involved in the observed inhibition effects. In silico and in vitro experiments were also used to determine the kinetics of the reaction. Fifteen compounds showed measurable in vitro IC50 values ranging from 0.194 to 0.941 mM. It was shown that carboxyl groups play an important role in inhibiting the enzyme; further, a hydroxyl group or carbonyl group at either C7 or C15 increases the inhibition rate, and a double bond at C24,25 decreases the activity. Based on the docking data we speculated that triterpenoids could not fit into the active site of the enzyme; therefore, the inhibition mode could not be competitive. Dixon plotting showed that the inhibition patterns should be uncompetitive and non-competitive instead..
99. Hui Tan, Ahmed Ashour, Yoshinori Katakura, Kuniyoshi Shimizu, A structure-activity relationship study on antiosteoclastogenesis effect of triterpenoids from the leaves of loquat (Eriobotrya japonica), Phytomedicine, 10.1016/j.phymed.2015.03.002, 22, 4, 498-503, 2015.04, Our previous results elucidated that the leaves of Eriobotrya japonica possessed the potential to suppress ovariectomy-induced bone mineral density deterioration, and ursolic acid, the major bioactive component in these leaves, suppressed the osteoclast differentiation. The aim of this study was to discover more candidates for development of novel antiosteoclastogenesis agents from the leaves of E. japonica. Phytochemical analysis following a cell-based osteoclastic tartrate-resistant acid phosphatase (TRAP) activity assay revealed 11 more compounds with a potent antiosteoclastogenesis effect. The potency of ursane-type triterpenoids from the leaves of E. japonica prompted us to investigate the structure-activity relationships underlying their antiosteoclastogenesis. The results revealed that both the hydroxyl group at C-3 and the carboxylic group at C-17 played indispensable roles in the antiosteoclastogenesis activity of ursane-type triterpenoids. The configuration at C-3 (a beta-form of the hydroxyl group) was found to be important for this activity. While introducing a hydroxyl group at C-19 increased the inhibitory activity of ursane-type triterpenoids carrying an alpha-form hydroxyl group at C-3. The bioactivity analyses of ursolic acid and oleanolic acid demonstrated that the antiosteoclastogenesis effect of ursolic acid may be related to different positions of the C-29 and C-30 methyl groups on the E-ring, since oleanolic acid showed limited activity. The addition of a hydroxyl group at C-2 would dramatically improve the inhibition of oleanane-type triterpenoids. Collectively, these findings could provide important clues for the improvement of multi-targeted antiosteoclastogenesis agents from the leaves of E. japonica..
100. Q. Zhu, T. Nakagawa, A. Kishikawa, K. Ohnuki, Shimizu, K., In vitro bioactivities and phytochemical profile of various parts of the strawberry (Fragaria×ananassa var. Amaou), Journal of Functional Foods, 13, 1, 38-49, 2015.03.
101. S. Fatmawat, T. Ersama, Shimizu, K., The inhibitory activity of aldose reductase in vitro by constituents of Garcinia mangostana Linn., Phytomedicine, 22, 1, 49-51, 2015.02.
102. A. Mira, S. Yamashita, Y. Katakura, Shimizu, K., In Vitro Neuroprotective Activities of Compounds from Angelica shikokiana Makino, Molecules, 10.3390/molecules20034813, 20, 3, 4813-4832, 2015.02.
103. Ahmed Ashour, Ryuichiro Kondo, Shimizu K, Anti-proliferative Activity of Some Oleanolic acid derivatives with Potent TopoisomeraseInhibitory Activity on B16 Melanoma Cells, Wood Research Journal, 2015.01.
104. Enos Tangke Arung, Wiwin Suwinarti, Medi Hendra, Supomo, Irawan Wijaya Kusuma, Dwi Caesarina Nanda Puteri, Huseyin Avni Eroglu, Yongung Kim, Kuniyoshi Shimizu, Hiroya Ishikawa, Determination of antioxidant and anti-melanogenesis activities of Indonesian Lai, Durio kutejensis [Bombacaceae (Hassk) Becc] fruit extract, Tropical Journal of Pharmaceutical Research, 14, 1, 41-46, 2015.01.
105. L. D. Khuong, R. Kondo, R. D. Leon, T. K. Anh, K. Shimizu, I. Kamei, Effect of chemical factors on integrated fungal fermentation of sugarcane bagasse for ethanol production by a white-rot fungus, Phlebia sp. MG-60, Bioresource Technology, https://doi.org/10.1016/j.biortech.2014.05.064, 30-40, 2014.10, Bioethanol production through integrated fungal fermentation (IFF), involving a unified process for biological delignification with consolidated biological processing by the white-rot fungus Phlebia sp. MG-60, was applied to sugarcane bagasse. Initial moisture content of the bagasse was found to affect biological delignification by MG-60, and 75% moisture content was suitable for selective lignin degradation and subsequent ethanol production. Additives, such as basal media, organic compounds, or minerals, also affected biological delignification of bagasse by MG-60. Basal medium addition improved both delignification and ethanol production. Some inorganic chemical factors, such as Fe2+, Mn2+, or Cu2+, reduced bagasse carbohydrate degradation by MG-60 during delignifying incubations and resulted in increased ethanol production. The present results indicated that suitable culture conditions could significantly improve IFF efficiency.

.
106. T. H. Bang, Shimizu, K., Potent Angiotensin-Converting Enzyme Inhibitory Tripeptides identified by a Computer-based Approach, Journal of Molecular Graphics and Modeling, 53, 206-211, 2014.09.
107. Shimizu, K., S. Fatmawati, H. Yu, C. Zhang, F. Jin, 20(S)-Ginsenoside Rh2 as aldose reductase inhibitor from Panax ginseng, Bioorganic & Medicinal Chemistry Letters, 10.1016/j.bmcl.2014.08.009, 24, 18, 4407-4409, 2014.09.
108. Ha, T.J., Shimizu, K., Kubo, I., Lipoxygenase inhibitory activity of alkyl protocatechuates, Food Chemistry, 159, 15, 471-476, 2014.09.
109. Bang, T.H., Yamamoto, A., Matsumoto, S., Ito, H., Igami, K., Miyazaki, T., Kondo, R., Shimizu, K., Hypotensive effects of Reishi (Ganoderma lingzhi)’s auto-digested extract and its potent peptides for Angiotensin-Converting Enzyme inhibition, Molecules, doi:10.3390/molecules190913473, 19, 13473-13485, 2014.08.
110. Hai-Bang Tran, Atsushi Yamamoto, Sayaka Matsumoto, Hisatomi Ito, Kentaro Igami, Toshitsugu Miyazaki, Ryuichiro Kondo,Kuniyoshi Shimizu, Hypotensive effects and angiotensin-converting enzyme inhibitory peptides of reishi (Ganoderma lingzhi) auto-digested extract, Molecules, 19, 9, 13473-13485, 2014.08.
111. Tanaka A, Zhu Q, Tan H, Horiba H, Ohnuki K, Yamauchi R, Mori Y, Yamauchi R, Ishikawa H, Iwamoto A, Kawahara H, Shimizu K, Biological activities and phytochemical profiles of extracts from different parts of bamboo (Phyllostachys pubescens), Molecules 2014, 19(6), 8238-8260, 10.3390/molecules19068238, 2014.06.
112. Ashour, A., EI-Sharkawy, S., Amer, M., Marzouk, A., Kishikawa, A., Ohzono, M., Kondo, R., Shimizu, K., Production of citric acid from corncobs with its biological evaluation
, Journal of Cosmetics, Dermatological Sciences and Applications, 4, 3, 141-149, 2014.06.
113. Purnomo, A.S., Putri, E.M.M., Nawfa, R., Putra, S.R., Shimizu, K., Kondo, R., Biodegradation of heptachlor-contaminated soil by white-rot fungal inocula, Environmental Science and Pollution Research, 2014.05.
114. L. D. Khuong, R. Kondo, R. D. Leon, T. K. Anh, K. Shimizu, I. Kamei, Bioethanol production from alkaline-pretreated sugarcane bagasse by consolidated bioprocessing using Phlebia sp. MG-60, International Biodeterioration & Biodegradation, 88, 3, 62-68, 2014.03.
115. T. Hui, S. Furuta, T. Nagata, K. Ohnuki, T. Akasaka, B. Shirouchi, M. Sato, R. Kondo, K. Shimizu, The Inhibitory Effect of Loquat Leaves (Eriobotrya japonica) on Ovariectomy-induced Bone Loss and Osteoclast Differentiation, Journal of Agricultural and Food Chemistry, doi: 10.1021/jf402735u, 62, 4, 836-41, 2014.01.
116. A. Ashour, S. El-Sharkawy, M. Amer, B. F. Abdel, Y. Katakura, T. Miyamoto, N. Toyota, T. H. Bang, R. Kondo, K. Shimizu, Rational design and synthesis of topoisomerase I and II inhibitors based on oleanolic acid moiety for new anti-cancer drugs., Bioorg Med Chem., doi: 10.1016/j.bmc.2013.11.034., 22, 1, 211-20, 2014.01.
117. Hai Bang, T., Suhara, H., Doi, K., Ishikawa, H., Fukami, K., Parajuli, G.P., Katakura, Y., Yamashita, S., Watanabe, K., Adhikari, M.K., Manandhar, H.K., Kondo, R., Shimizu, K., Wild Mushrooms in Nepal: Some Potential Candidates as Antioxidant and ACE-Inhibition Sources, Evidence-based Complementary and Alternative Medicine, http://dx.doi.org/10.1155/2014/195305, 2014.01, Twenty-nine mushrooms collected in the mountainous areas of Nepal were analyzed for antioxidant activity by different methods, including Folin-Ciocalteu, ORAC, ABTS, and DPPH assays. Intracellular H2O2-scavenging activity was also performed on HaCaT cells. The results showed that phenolic compounds are the main antioxidant of the mushrooms. Among studied samples, Inonotus andersonii, and Phellinus gilvus exhibited very high antioxidant activity with the phenolic contents up to 310.8 and 258.7 mg GAE/g extracts, respectively. The H2O2-scavenging assay on cells also revealed the potential of these mushrooms in the prevention of oxidative stress. In term of ACE-inhibition, results showed that Phlebia tremellosa would be a novel and promising candidate for antihypertensive studies. This mushroom exhibited even higher in vitro ACE-inhibition activity than Ganoderma lingzhi, with the IC50 values of the two mushrooms being 32 μg/mL and 2 μg/mL, respectively. This is the first time biological activities of mushrooms collected in Nepal were reported. Information from this study should be a valuable reference for future studies on antioxidant and ACE-inhibitory activities of mushrooms..
118. Tan, H., Furuta, S., Nagata, T., Ohnuki, K., Akasaka, T., Shirouchi, B., Sato, M., Kondo, R., Shimizu, K., Inhibitory Effects of the Leaves of Loquat (Eriobotrya japonica) on Bone Mineral Density Loss in Ovariectomized Mice and Osteoclast Differentiation, Journal of Agricultural and Food Chemistry, 62, (4), 836-841, 2014.01, The loquat, Eriobotrya japonica Lindl. (Rosaceae), is a small tree native to Japan and China that is widely cultivated for its succulent fruit. Its leaves are used as an ingredient of a tasty tea called “Biwa cha” in Japanese. The anti-osteoporosis effects of the leaves of loquat in vitro and in vivo have been investigated. After 15 days of feeding normal diet or diet supplemented with 5% loquat leaves, the body weight, viscera weights, and bone mineral density (BMD) of both groups of eight ovariectomized (OVX) mice were compared. The result showed that the loss of BMD in loquat-fed mice was significantly prevented in three parts of the body, especially in the trabecular bone of the head (P < 0.05), abdomen (P < 0.01), and lumbar (P < 0.05) compared to the control group. No hypertrophy in the uterus by the loquat leaves diet was observed. The effect of the extract (447.25 g) prepared from the dried leaves of loquat (2.36 kg) was further studied on RANKL-induced osteoclast differentiation and cell viability. The extract suppressed the differentiation of osteoclasts under 50, 125, 250, and 500 μg/mL. Through bioactivity-guided fractionation, ursolic acid (1) was isolated and inhibited osteoclast differentiation under 4 and 10 μg/mL. It was concluded that loquat leaves possess the potential to suppress ovariectomy-induced bone mineral density deterioration..
119. E. T. Arung, H. Kuspradini, I. W. Kusuma, T. H. Bang, S. Yamashita, Y. Katakura, K. Shimizu, Effects of isolated compound from Sonneratia caseolaris leaf : a validation of traditional utilization by melanin biosynthesis and antioxidant assays, submitted, 2013.12.
120. S. Fatmawati, RYUICHIRO KONDO, Shimizu K, Structure-activity relationships of lanostane-type triterpenoids from Ganoderma lingzhi as α-glucosidase inhibitors, 10.1016/j.bmcl.2013.08.084, 23, 21, 5900-5903, 2013.11.
121. A. Ashour, M. Amer, A. Marzouk, K. Shimizu, R. Kondo, S. El-Sharkawy, Corncobs as a potential source of functional chemicals., Molecules, doi:10.3390/molecules181113823, 18, 11, 13823-13830, 2013.11.
122. T. H. Bang, A. Yamamoto, S. Matsumoto, H. Ito, K. Igami, T. Miyazaki, R. Kondo, K. Shimizu, Hypotensive effects of Reishi (Ganoderma lingzhi)’s auto-digested extract and its potent peptides for Angiotensin-Converting Enzyme inhibition., submitted, 2013.09.
123. T. H. Bang, H. Suhara, K. Doi, H. Ishikawa, K. Fukami, G. P. Parajuli, Y. Katakura, S. Yamashita, K. Watanabe, M. K. Adhikari, H. K. Manandhar, R. Kondo, K. Shimizu, Wild Mushroom in Nepal: Some potential Candidates as Antioxidant and ACE Inhibition Sources, Evidence-Based Complementary and Alternative Medicine, Volume 2014 (2014), Article ID 195305, 11 pages http://dx.doi.org/10.1155/2014/195305, 2014, 2013.08.
124. A. Mira, A. Tanaka, Y. Tateyama, R. Kondo, K. Shimizu, Comparative Biological Study of Roots, Stems, Leaves, and Seeds of Angelica shikokiana Makino, J Ethnopharmacol, 148, 980-987, 2013.08.
125. A. Ashour, S. El-Sharkawy, M. Amer, F.A. Bar, A. Mera, T. Nagata, R. Kondo, K. Shimizu, Biotransformation of Khellin to Khellol by Aspergillus niger and the Evaluation of their Biological Activities., The Open Bioactive Compounds Journal, 4, 1-3, 2013.08.
126. A. Ashour, M. Amer, A. Marzouk, K. Shimizu, R. Kondo, S. El-Sharkawy, A new lignan from corncobs, a potential source of functional chemicals, 2013.08.
127. J. Liu, K. Shimizu, A. Tanaka, W. Shinobu, K. Ohnuki, T. Nakamura, R. Kondo, A protocol for isolation of target proteins of natural compound, Protocol Exchange, doi:10.1038/protex.2013.051, 2013.05.
128. K. Shimizu, Antibacterial compounds from shoot skins of moso bamboo (Phyllostachys pubescens), Journal of Wood Science, 10.1007/s10086-012-1310-6, 59, 2, 155-159, 2013.04.
129. Kubo, I., Fujita, K.-I., Shimizu, K. , Anti-salmonella agents from the Brazilian medicinal plant tanacetum balsamita and their applications, Natural Antioxidants and Biocides from Wild Medicinal Plants, 15, 239-253, 2013.04.
130. A. Ashour, S. El-Sharkawy, M. Amer, F. A.l Bar, R. Kondo, K. Shimizu, Melanin Biosynthesis Inhibitory Activity of Compounds Isolated from unused parts of Ammi visinaga, Journal of Cosmetics, Dermatological Sciences and Applications, 3, 40-43, 2013.03.
131. J. Liu, K. Shimizu, A. Tanaka, W. Shinobu, K. Ohnuki, T. Nakamura, R. Kondo., Target proteins of ganoderic acid DM provides clues to various pharmacological mechanisms, Sci. Rep. 2, 905, 2012.11, 霊芝には、古来よりガンを初め、様々な疾病に関する効能が伝承されている。その効能を裏付けようと多くの基礎研究が実施されており、霊芝の薬理活性に関する研究は、近年、劇的に増大している。その薬理活性は、含有されるβ-グルカンに代表される多糖類と、特徴的な構造を有するラノスタン型トリテルペノイド類に由来すると言われている。候補者は、霊芝の前立腺肥大症や骨粗鬆症に関する改善効果を見いだし、その活性成分の一つとして、ラノスタン型トリテルペノイド類に属するganoderic acid DMを単離した。本化合物は、5α-リダクターゼ阻害活性、男性ホルモン受容体結合活性、破骨細胞分化抑制効果、ガン細胞増殖抑制効果を示した。本化合物の作用機構の本質に迫るべく、細胞培養系での挙動並びに、構造活性相関を検討し、本知見を元に、ganoderic acid DM結合磁性プローブを用いて標的タンパク質の探索を行ったところ、微小管形成において重要な役割を担うタンパク質であるチューブリンと特異的に相互作用することを見いだした。さらに、同化合物は、チューブリンの重合を促進させ、微小管を不安定化させることにより、細胞の増殖・機能に影響を与えることを明らかにした。以上、霊芝由来トリテルペノイドの生理活性発現に関する新規メカニズムを提案した。.
132. Tukiran, S. S. Cahyasari, K. Shimizu, An ester of 4-methoxy cynnamic acid isolated from Xylocarpus moluccencis (Lamk) M. Roem (Meliaceae), Indonesian Journal of chemistry, 12, 2, 184-188, 2012.07.
133. T.X. Meng, C.-F. Zhang, T. Miyamoto, H. Ishikawa, K. Shimizu, S Ohga, R. Kondo, The melanin biosynthesis stimulating compounds isolated from the fruiting bodies of Pleurotus citrinopileatus, Journal of Cosmetics, Dermatological Sciences and Applications, 2, 151-157, 2012.07.
134. S. Yokoyama, T. H. Bang, K. Shimizu, R. Kondo, Osteoclastogenesis inhibitory effect of ergosterol peroxide isolated from Pleurotus eryngii, Natural Products Communications, 7, 9, 1163-1164, 2012.07.
135. E. T. Arung, S. Furuta, K. Sugamoto, K. Shimizu, H. Ishikawa, Y. Matsushita, R. Kondo, The inhibitory effects of representative chalcones contained in Angelica keiskei on melanin biosynthesis in B16 melanoma cells, Natural Products Communications, 7, 8, 1007-1010, 2012.07.
136. E. T. Arung, H. Kuspradini I. W. Kusuma, K. Shimizu, R. Kondo, Validation of Eupatorium triplinerve Vahl leaves, a skin care herbal from East Kalimantan in melanin biosynthesis assay, Journal of Acupuncture and Meridian Studies, 5, 2, 87-92, 2012.07.
137. E. Matsubara, K. Shimizu, M. Fukagawa, Y. Ishizi, C. Kakoi, T. Hatayama, J. Nagano, T. Okamoto, K. Ohnuki, R. Kondo, Volatiles emitted from the roots of Vetiveria zizanioides suppress the decline in attention during a visual display terminal task, Biomedical Research, 35, 5, 299-308, 2012.07.
138. E. T. Arung, I. W. Kusuma, Y.-U. Kim, K. Shimizu, R. Kondo, Antioxidantive compounds from leaves of Tahongai (Klienhovia hospita), Journal of Wood Science,58(1)77-80, 58, 1, 77-80, 2012.04.
139. T. X. Meng, H. Ishikawa, K. Shimizu, S. Ohga, R. Kondo, A glucosylceramide with antimicrobial activity from the edible mushroom Pleurotus citrinopileatus, Journal of Wood Science, 58, 1, 81-86, 2012.02.
140. J. Liu, K.i Kurashiki, A. Fukuta, S.i Kaneko, Y. Suimi, K. Shimizu, R. Kondo, Quantitative determination of the representative triterpenoids in the extracts of Ganoderma lucidum with different growth stages using high-performance liquid chromatography for evaluation of their 5α-reductase inhibitory properties, Food Chemistry, 133, 1034-1038, 2012.02.
141. Kubo, I., Shimizu, K., Nihei, K.-I., Maniçoba, a quercetin rich amazonian dish: Antioxidative prospects
, Quercetin: Dietary Sources, Functions and Health Benefits, 26, 129-154, 2012.01, A large quantity of quercetin was characterized from a local dish, known as maniçoba in Pará, Brazil. The fresh leaf of Manihot esculenta Crantz (Euphorbiaceae), a principal ingredient for maniçoba, was found to contain a large amount of rutin or quercetin 3-O-rutinoside which was completely hydrolyzed to quercetin during cooking processes. Antioxidant activity of quercetin and its microfloral metabolites was examined. Quercetin inhibited the oxidation of linoleic acid catalyzed by soybean lipoxygenase-1 (EC 1.13.11.12, Type 1) with an IC50 of 4.5 μM, while rutin did not inhibit this oxidation up to 200 μM. The progress curves for the enzyme reaction indicate that quercetin shows slow binding kinetics. In the pseudoperoxidase activity, hydroperoxides slowly oxidize quercetin to a rather stable intermediate, 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone. A microfloral metabolite of quercetin and rutin after ingestion, 3,4-dihydroxyphenylacetic acid, also scavenged superoxide anion generated by xanthine oxidase and inhibited uric acid formation. The antioxidant activity observed with the methanol extract of maniçoba is due mainly to multifunctional quercetin's response..
142. S. Fatmawati, K. Shimizu, R. Kondo, Structure-activity relationships of ganoderma acids from Ganoderma lucidum as aldose reductase inhibitors, Bioorganic & Medicinal Chemistry Letters, 21(24), 7295-7297, 2011.12.
143. I. Kubo, S. H. Lee, K. Shimizu, Combination effect of miconazole with polygodial against Candida albicans, Open Journal of Medical Microbiology, 1, 7-11, 2011.12.
144. A. Tanaka, H. J. Kim, S. Oda, K. Shimizu, R. Kondo, Antibacterial activity of moso bamboo shoot skin (Phyllostachys pubescens) against Staphylococcus aureus., Journal of Wood Science, 57, 6, 542-544, 2011.10.
145. E. T. Arung, K. Shimizu, Antioxidant activity of compounds from wood of Artocarpus heterophyllus, Indonesian Journal of Pharmaceutical Sciences, 9, 2, 72-77, 2011.09.
146. Arung, E.T.,Shimizu, K., Kondo, R., Artocarpus Plants as a Potential Source of Skin Whitening Agents, Natural Product Communications, https://doi.org/10.1177/1934578X1100600943, 6, 9, 1397-1402, 2011.08, Artocarpus plants have been a focus of constant attention due to the potential for skin whitening agents. In the in vitro experiment, compounds from the Artocarpus plants, such as artocarpanone, norartocarpetin, artocarpesin, artogomezianol, andalasin, artocarbene, and chlorophorin showed tyrosinase inhibitory activity. Structure-activity investigations revealed that the 4-substituted resorcinol moiety in these compounds was responsible for their potent inhibitory activities on tyrosinase. In the in vitro assay, using B16 melanoma cells, the prenylated polyphenols isolated from Artocarpus plants, such as artocarpin, cudraflavone C, 6-prenylapigenin, kuwanon C, norartocarpin, albanin A, cudraflavone B, and brosimone I showed potent inhibitory activity on melanin formation. Structure-activity investigations revealed that the introduction of an isoprenoid moiety to a non-isoprenoid-substituted polyphenol enhanced the inhibitory activity of melanin production in B16 melanoma cells. In the in vivo investigation, the extract of the wood of Artocarpus incisus and a representative isolated compound from it, artocarpin had a lightening effect on the skin of guinea pigs’ backs. Other in vivo experiments using human volunteers have shown that water extract of Artocarpus lakoocha reduced the melanin formation in the skin of volunteers. These results indicate that the extracts of Artocarpus plants are potential sources for skin whitening agents..
147. T.X. Meng, S. Furuta, S. Fukamizu, R. Yamamoto, H. Ishikawa, E.T. Arung, K. Shimizu, S. Ohga, R. Kondo, Evaluation of biological activities of extracts from the fruiting body of Pleurotus citrinopileatus for skin cosmetics, Journal of Wood Science, 57, 5, 452-458, 2011.05.
148. E. T. Arung, S. Furuta, H. Ishikawa, H. Tanaka, K. Shimizu, R. Kondo, Melanin Biosynthesis Inhibitory and Antioxidant Activities of Quercetin-3'-O-b-D-glucoside Isolated from Allium cepa, Zeitschrift für Naturforschung C, 66, 5-6, 209-214, 2011.04.
149. S. Fatmawati, K. Shimizu, R. Kondo , Ganoderol B: a potent alpha-glucosidase inhibitor isolated from the fruiting body of Ganoderma lucidum, Phytomedicine, 18, 12, 1053-1055, 2011.02.
150. E. T. Arung, K. Shimizu, I. W. Kusuma, R. Kondo., Tyrosinase inhibitory effect of quercetin 4'-O-b-glucopyranoside from dried skin of Red Onion (Allium cepa)
, Natural Product Research, 25, 3, 256-263, 2011.02.
151. E. T. Arung, E. Matsubara, I. W. Kusuma, E. Sukaton, K. Shimizu, R. Kondo, Inhibitory components from the buds of clove (Syzygium aromaticum) on melanin formation in B16 melanoma cells, Fitoterapia, 82, 2, 198-202(2011), 2011.02.
152. E. Matsubara, M. Fukagawa, T. Okamoto, K. Ohnuki, K. Shimizu, R. Kondo, (-)-Bornyl acetate induces autonomic relaxation and reduces arousal level after visual display terminal work without any influences of task performance in low-dose condition, Biomedical Research, 32, 2, 151-157, 2011.02.
153. E. Matsubara, M. Fukagawa, T. Okamoto, K. Ohnuki, K. Shimizu, R. Kondo, The essential oil of Abies sibirica (Pinaceae) reduces arousal levels after visual display terminal work, Flavour and Fragrance Journal, 26, 3, 204-210, 2011.02.
154. E. Matsubara, M. Fukagawa, T. Okamoto, A. Fukuda, C. Hayashi, K. Ohnuki, K. Shimizu, R. Kondo, Volatiles emitted from the leaves of Laurus nobilis L. improve vigilance performance in visual discrimination task, Biomedical Research, 32, 1, 19-28, in press, 2011.02, 月桂樹(Laurus nobilis L.)の葉を乾燥させたものは、香辛料として広く使用されている。また、葉から水蒸気蒸留により得られた精油も、食品の香り付けや薬、化粧品などにも使われる。月桂樹葉は、抗けいれん作用や抗炎症作用、抗菌活性、抗酸化活性など多岐にわたる生理活性を有することが知られている。さらにリウマチや神経痛、疥癬に効くとされ、ゴキブリ忌避作用や鎮静、麻酔作用なども知られている。本研究では、月桂樹葉揮発成分によるビジランス作業効率への影響について検討した。月桂樹葉の供試量を二段階(低濃度群と高濃度群)に設定し、清浄空気を対照とした三条件で、作業効率と生理心理学的な効果について比較した。低濃度の月桂樹葉揮発成分を吸入することによって、正答率の低下抑制に寄与することが分かった。さらに高濃度群では、吸入によりネガティブ感情を引き起こす可能性が示唆された。低濃度と高濃度群ではともに、心血管系循環促進が示唆され、覚醒状態は維持されていたことが分かった。これらの知見から、低濃度の月桂樹葉揮発成分には、感情への負の影響を引き起こすことなく、生理学的な覚醒状態の維持に寄与し、正答率の低下抑制というビジランス維持の機能を持つことが示唆された。.
155. Koyama, F., Koyama, F.,Shimizu, K., Matsubara, E., Yoshida, E., Kondo, R., Reduction of Ammonia Emission during Composting of Poultry Manure by Covering with Cedar Bark, Mokuzai Gakkaishi/Journal of the Japan Wood Research Society, 57, 6, 370-376, 2011.01, The aim was to investigate the extent of reduction of ammonia emissions from poultry manure by utilizing cedar bark, which occurs plentifully from timber trimming facilities and has an ability to adsorb ammonia. In a laboratory scale study, cedar bark meal was contacted with an ammonia concentration of 620 ppm in Tedlar bags for 5 minutes. The result showed that the bark reduced ammonia by 99.5%. This study compiled data on ammonia above poultry manure covered with cedar saw dust or cedar bark during a period of 3 weeks. The results indicated that both types of covering agents achieved significant reductions in ammonia concentrations. The adsorption of ammonia was 11 mg-g-1 and 33 mg-g-1 for saw dust and bark respectively. Water-soluble nitrogen, ionic nitrogen, and organic nitrogen accounted for 22%, 23% and 47%, respectively, of increased nitrogen in the bark during composting. It suggested that the increase in ionic nitrogen was due to the higher cation exchange capacity. The increase in organic nitrogen may be attributed to covalent bonds. The results showed that the reduction in the emission of ammonia appears to be caused by the bark cover acting as an inexpensive and simple deodorizer..
156. S. Fatmawati, K. Shimizu, R. Kondo, Inhibition of aldose reductase in vitro by constituents of Ganoderma lucidum, Planta Med, 76, 15, 1691-1693, 2010.11.
157. E. T. Arung, B. D. Wicaksono, Y. A. Handoko, I. W. Kusuma, K. Shimizu, D. Yulia, F. Sandra, Cytotoxic effect of artocarpin on T47D cells, J. Nat. Med. , 64, 4, 423-429, 2010.10.
158. S. Fatmawati, K. Shimizu, R. Kondo, Ganoderic acid Df, a new triterpenoid with aldose reductase activity from the fruiting body of Ganoderma lucidum , Fitoterapia., 81, 8, 1033-1036, 2010.09.
159. J. Liu, K. Shimizu, R. Kondo, The effects of ganoderma alcohols isolated from Ganoderma lucidum on the androgen receptor binding and the growth of LNCaP cells, Fitoterapia., 81, 8, 1067-1072, 2010.09.
160. J. Liu, K. Shimizu, H. Yu,C. Zhang, F. Jin, R. Kondo, Stereospecificity of hydroxyl group at C-20 in antiproliferative action of ginsenoside Rh2 on prostate cancer cells, Fitoterapia., 81, 7, 902-905, 2010.09.
161. E. T. Arung, S. Furuta, H. Ishikawa, I. W. Kusuma, K. Shimizu, R. Kondo, Anti-melanogenesis properties of quercetin- and its derivative-rich extract from Allium cepa, Food Chemistry, 124, 3, 1024-1028, 2010.09.
162. T.J. Ha, K. Shimizu, T. Ogura, I. Kubo, Inhibition mode of soybean lipoxygenase-1 by quercetin, Chem. Biodiversity, 7, 8, 1893-1903, 2010.08.
163. M. Nagano, K. Shimizu, R. Kondo, C. Hayashi, D. Sato, K. Kitagawa, K. Ohnuki, Reduction of depression and anxiety by 4 weeks Hericium erinaceus intake, Biomedical Research, 31, 4, 231-237, 2010.08, Hericium erinaceumはサンゴハリタケ科のキノコであり、日本では、その形状から、ヤマブシタケ(山伏茸)と呼ばれている。本キノコは、抗ガン活性、脂質低減活性、細胞毒性活性、抗菌活性、抗酸化活性、小胞体ストレス抑制活性、神経成長因子(NGF)合成促進活性など多岐にわたる生理活性を有することが知られている。我々は、ヤマブシタケの新たな機能性の探索のために、中年女性における不定愁訴改善効果の検証を、無作為化二重盲検比較試験にて、クッパーマン更年期障害指数、CES-Dうつ病自己評価尺度、ピッツバーグ睡眠質問票、不定愁訴アンケートを用いて行った。その結果、4週間のヤマブシタケ含有食品摂取(1日ヤマブシタケ乾燥粉末4グラム)によって、様々な不定愁訴の有意な改善が認められた。.
164. I. W. Kusuma, E. T. Arung, E. Rosamah, S. Purwatiningsih, H. Kuspradini, Syafrizal, J. Astuti, Y-U, Kim, K. Shimizu, Antidermatophyte and antimelanogenesis compound from Eleutherine americana growing in Indonesia, J. Nat. Med., 64, 2, 223-226, 2010.07.
165. E. T. Arung, K. Shimizu, H. Tanaka, R. Kondo, 3-Prenyl luteolin, a new prenylated flavone with melanin biosynthesis inhibitory activity from wood of Artocarpus heterophyllus, Fitoterapia., 81, 6, 640-643, 2010.06.
166. S. Yokoyama, S. Takeno, T. Nagata, K. Shimizu, R. Kondo, Screening of extracts of Japanese medicinal plants for HMG-CoA reductase inhibition, Bulletin of the Kagoshima University Forest, 37, 37-41, 2010.05.
167. J. Liu, J. Shiono, K. Shimizu, R. Kondo, Ganoderic acids from Ganoderma lucidum. Inhibitory activity of osteoclastic differentiation and structural criteria, Planta Med., 76, 137-139, 2010.05.
168. I. Kubo, T.J. Ha, K. Shimizu, Lipoxygenase inhibitory activity of 6-pentqadecanylsalicylic acid without prooxidant effect, Natural Product Communications , 5, 1, 85-90, 2010.05.
169. E. T. Arung, K. Shimizu, H. Tanaka, R. Kondo, Melanin biosynthesis inhibitors from wood of Artocarpus heterophyllus: The effect of isoprenoid substituent of flavone with 4-substituted resorcinol moiety at B ring, Lett. Drug Des. Discovery, 7, 8, 602-605, 2010.05.
170. E. T. Arung, K. Yoshikawa, K. Shimizu, R. Kondo, Isoprenoid-substituted flavonoids from wood of Artocarpus heterophyllus on B16 melanoma cell -Cytotoxicity and structural criteria-, Fitoterapia , 81, 120-123, 2010.04.
171. Kusuma, I.W., Arung, E.T., Rosamah, E., Purwatiningsih, S., Kuspradini, H., Syafrizal, Astuti, J., Kim, Y.-U.,Shimizu, K., Antidermatophyte and antimelanogenesis compound from Eleutherine americana grown in Indonesia, Open Bioactive Compounds Journal, 64, 2, 223-226, 2010.04, An active compound from the bulb of Eleutherine americana L. Merr. (Iridaceae) collected from East Kalimantan, Indonesia, was tested for its antidermatophyte and antimelanogenesis activity. Antifungal assay-directed fractionation of the n-hexane-soluble fraction of the methanolic extract of the bulb of E. americana led to the isolation of 1 as an active compound. The compound was identified as the naphthoquinone eleutherin by EI-MS and 1H-, 13C-, and two-dimensional NMR analyses. Antidermatophyte assay of 1 at concentrations of 10, 20, 40, 60, and 80 μg/disk and myconazole, a commercial antidermatophyte, at 10 μg/disk displayed 7, 8, 13, 16, 17, and 14 mm of inhibition zone against Trichophyton mentagrophytes, respectively. In a melanin formation inhibition assay, compound 1 displayed potent antimelanogenesis activity at 5 ppm with low toxicity compared with arbutin, a commercial skin-whitening agent. The results showed the high potential of 1, an active compound from E. americana, to be applied as an antidermatophyte and antimelanogenesis agent..
172. Arung, E.T., Yoshikawa, K., Shimizu, K., Kondo, R., Isoprenoid-substituted flavonoids from wood of Artocarpus heterophyllus on B16 melanoma cells: Cytotoxicity and structural criteria, Fitoterapia, 81, 2, 120-123, 2010.03, As a result of cytotoxicity-guided fractionation, nine flavonoids, artocarpin (1), cudraflavone C (2), 6-prenylapigenin (3), kuwanon C (4), norartocarpin (5), albanin A (6), cudraflavone B (7), brosimone I (8) and artocarpanone (9) were identified from the methanol extract of the wood of Artocarpus heterophyllus, known commonly as Nangka in Indonesia. A structure–activity investigation of the effect of these isolated compounds (1–9) and structurally related compounds on B16 melanoma cells indicated that isoprenoid moiety substitutions in flavonoids enhance their cytotoxicity, and that the position of attachment and the number of isoprenoid-substituent moieties per molecule influence flavonoid cytotoxicity..
173. Kubo, I., Masuoka, N., Ha, T.J., Shimizu, K., Nihei, K., Multifunctional antioxidant activities of alkyl gallates, Open Bioactive Compounds Journal, 3, 1-11, 2010.01, A series (C1 to C16) of alkyl gallates was tested for their antioxidant activity for food protection and human health. One molecule of alkyl gallate, regardless of alkyl chain length, scavenges six molecules of 1,1-diphenyl-2-picrylhydrazyl (DPPH). Alkyl gallates inhibited the linoleic acid peroxidation catalyzed by soybean lipoxygenase-1 (EC 1.13.11.12, Type 1) without being oxidized. The progress curves for enzyme reactions were recorded by both spectrophotometric and polarographic methods and the inhibitory activity was a parabolic function of their lipophilicity (log P) and maximized with alkyl chain length between C12 and C16. Tetradecanyl (C14) gallate exhibited the most potent inhibition with an IC50 of 0.06 µM. The inhibition kinetics of dodecyl gallate (C12) revealed competitive and slow-binding inhibition. Alkyl gallates chelate transition metal ions and this chelation ability should be of considerable advantage as antioxidants. Additionally, gallic acid was found to inhibit superoxide anion generated by xanthine oxidase (EC 1.1.3.22) but did not inhibit enzymatically catalyzed uric acid formation..
174. Liu, J., Shiono, J., Shimizu, K., Kondo, R., Ganoderic acids from ganoderma lucidum: Inhibitory activity of osteoclastic differentiation and structural criteria, Planta Medica, 10.1055/s-0029-1186035, 76, 2, 137-139, 2010.01, Increased osteoclastic bone resorption plays a central role in the pathogenesis of many bone diseases. Ganoderma lucidum is known for medicinal effects such as anti-inflammatory and anti-bone resorption activities. In this study, we investigated the inhibitory effects of ganoderic acids isolated from G. lucidum on osteoclastic differentiation using RAW264 cells in vitro. A carbonyl in C7 is essential to elicit the inhibition of osteoclast differentiation..
175. E. T. Arung, I. W. Kusuma, E. O. Christy, K. Shimizu, R. Kondo, Evaluation of medicinal plants from Central Kalimantan for anti melanogenesis, J. Nat. Med. , 63, 473-480, 2009.12.
176. Z. Huang, K. Hashida, R. Makino, F. Kawamura, K. Shimizu, R. Kondo, S. Ohara, Evaluation of biological activities of extracts from 22 African tropical wood species, J. Wood Sci, 55, 3, 225-229, 2009.05.
177. M. Hasegawa, R. Wakimoto, E. Yoshida, K. Shimizu, R. Kondo, A. Widyatmoko, A. Nirsatmanto, S. Shiraishi , Provenance variation in growth and wood properties of A. mangium and A. auriculiformis in Central Java, Indonesia : selecting potential hybrid parents for good performance, Bull. Kyushu Univ. For. , 90, 25-37, 2009.05.
178. J. Liu, J. Shiono, Y. Tsuji, K. Shimizu, R. Kondo, Methyl ganoderic acid DM: a selective potent osteoclastgenesis inhibitor, The Open Bioactive Compounds Journal, 2, 37-42, 2009.05.
179. J. Liu, J. Shiono, K. Shimizu, H. Yu,C. Zhang, F. Jin, R. Kondo, 20(R)-Ginsenoside Rh2, not 20(S), is a selective osteoclastgenesis inhibitor without any cytotoxicity, Bioorg. Med. Chem. Lett., 19, 3320-3323, 2009.05.
180. J. Liu, J. Shiono, K. Shimizu, A. Kukita, T. Kukita, R. Kondo, Ganoderic acid DM: anti-androgenic osteoclastogenesis inhibitor, Bioorg. Med. Chem. Lett., 19, 2154-2157, 2009.05.
181. E.T. Arung, I.W. Kusuma, S. Purwatiningsih, S.-S. Roh, C.H. Yang, S. Jeon, Y.-U. Kim, E. Sukaton, J. Susilo, Y. Astuti, B.D. Wicaksono, F. Sandra, K. Shimizu, R. Kondo, Antioxidant Activity and Cytotoxicity of the Traditional Indonesian Medicine Tahongai (Kleinhovia hospita L.) Extract, JAMS Journal of Acupuncture and Meridian Studies, 2, 4, 306-308 , 2009.05.
182. K. Shimizu, I. Miyamoto, J. Liu, F. Konishi, S. Kumamoto, R. Kondo, Estrogen-like activity of ethanol extract of Ganoderma lucidum, J. Wood Sci., 55, 53-39 , 2009.04.
183. J. Liu, S. Tamura, K. Kurashiki, K. Shimizu, K. Noda, F. Konishi, S. Kumamoto, R. Kondo, Anti-androgen effects of extracts and compounds from Ganoderma lucidum, Chem. Biodiversity, 6, 231-243, 2009.04.
184. J. Liu, K. Shimizu, R. Kondo, Anti-androgenic activity of fatty acids, Chemistry&Biodiversity, 6, 4, 503-512, 2009.04.
185. I. Miyamoto, J. Liu, K. Shimizu, A. Kukita, T. Kukita, M. Sato, R. Kondo, Regulation of osteoclastogenesis by ganoderic acid DM isolated from Ganoderma lucidum, Eur. J. Pharmacol., 602, 1, 1-7, 2009.04.
186. S. Fatmawatia, K. Kurashiki, S. Takeno, Y. Kim, K. Shimizu, M. Sato, K. Imaizumi, K. Takahashi, S. Kamiya, S. Kaneko, R. Kondo , The inhibitory effect on aldose reductase by an extract of Ganoderma lucidum , Phytotherapy Research, 23, 28-32, 2009.03.
187. Noguchi, M., Kakuma, T., Tomiyasu, K., Yamada, A., Itoh, K., Konishi, F., Kumamoto, S., Shimizu, K., Kondo, R., Matsuoka, K., Randomized clinical trial of an ethanol extract of Ganoderma lucidum in men with lower urinary tract symptoms, Asian Journal of Andrology, https://doi.org/10.1111/j.1745-7262.2008.00361.x, 10, 5, 777-785, 2008.08, Aim: To evaluate the safety and efficacy of an extract of Ganoderma lucidum that shows the strongest 5α‐reductase inhibitory activity among the extracts of 19 edible and medicinal mushrooms by a double‐blind, placebo‐controlled, randomized and dose‐ranging study in men with lower urinary tract symptoms (LUTS). Methods: In this trial, we randomly assigned 88 men over the age of 49 years who had slight‐to‐moderate LUTS to 12 weeks of treatment with G. lucidum extract (6 mg once a day) or placebo. The primary outcome measures were changes in the International Prostate Symptom Score (IPSS) and variables of uroflowmetry. Secondary outcome measures included changes in prostate size, residual urinary volume after voiding, laboratory values and the reported adverse effects. Results:G. lucidum was effective and significantly superior to placebo for improving total IPSS with 2.1 points decreasing at the end of treatment (mean difference, −1.18 points; 95% confidence interval, −1.74 to −0.62; P < 0.0001). No changes were observed with respect to quality of life scores, peak urinary flow, mean urinary flow, residual urine, prostate volume, serum prostate‐specific antigen or testosterone levels. Overall treatment was well tolerated with no severe adverse effects. Conclusion: The extract of G. lucidum was well tolerated and improved IPSS scores. These results encouraged a further, large‐scale evaluation of phytotherapy for a long duration using the extract of G. lucidum on men with LUTS..
188. Masanori Noguchi, Tatsuyuki Kakuma, Katsuro Tomiyasu, Yoshiko Kurita, Hiroko Kukihara, Fumiko Konishi, Shoichiro Kumamoto,Kuniyoshi Shimizu, Ryuichiro Kondo , Kei Matsuoka, Effect of an extract of Ganoderma lucidum in men with lower urinary tract symptoms: A double-blind, placebo-controlled randomized and dose-ranging study, Asian Journal of Andrology, 10, 4, 651-658, 2008.07.
189. M. Noguchi, K. Tomiyasu, M, Nakiri, K. Matsuoka, T. Kakuma, F. Konishi, S. Kumamoto, K. Shimizu, R. Kondo, An extract of ganoderma lucidum in men with lower urinary tract symptoms: A double-blind, placebo-controlled randomized and dose-ranging study, Nishinihon Journal of Urology, 70, 9, 466-472, 2008.04.
190. K. Shimizu, M. Gyokusen, S. Kitamura,T. Kawabe, T. Kozaki, K. Ishibashi, R. Izumi, W. Mizunoya, K. Ohnuki, R. Kondo, Essential Oil of Lavender Inhibited the Decrement of Attention in the Long-term Task in Human, Bioscience, Biotechnology, and Biochemistry, 72, 7, 1944-1947, 72(7), 1944-1947 , 2008.04.
191. E. T. Arung, S. Muladi, E. Sukaton, K. Shimizu, R. Kondo, Artocarpin, a promising compound as whitening agent and skin cancer, Journal of Wood Science and Technology, 6, 33-36, 2008.04.
192. M. Noguchi, T. Kakuma, K. Tomiyasu, Y. Kurita, H. Kukihara, F. Konishi, S. Kumamoto, K. Shimizu, R. Kondo, K. Matsuoka , The effect of an extract of Ganoderma lucidum in men with lower urinary tract symptoms: A double-blaind, placebo-controlled randomized and dose-ranging study, Asian Journal of Andrology, 10, 4, 651-658, 2008.03.
193. M. Noguchi, T. Kakuma, K. Tomiyasu, A. Yamada, K. Itoh, F. Konishi, S. Kumamoto, K. Shimizu, R. Kondo, K. Matsuoka , A randamized clinical trial of an ethanol extract of Ganoderma lucidum in men with lower urinary tract symptoms, Asian Journal of Andrology, 10, 5, 777-785, 2008.03.
194. J. Liu, R. Ando, K. Shimizu, K. Hashida, R. Makino, S. Ohara, R. Kondo, Steroid 5α-reductase inhibitory activity of condensed tannins from woody plants , J. Wood Sci , 54, 1, 68-75, 2008.03.
195. K. Shimizu, S. Fukunaga, K. Yoshikawa, R. Kondo, Screening of extracts of Japanese woods for melanin biosynthesis inhibition, J. Wood Sci., 53, 153-160, 2007.10.
196. Arung, E.T., Shimizu, K., Kondo, R., Structure-activity relationship of prenyl-substituted polyphenols from Artocarpus heterophyllus as inhibitors of melanin biosynthesis in cultured melanoma cells, Chemistry and Biodiversity, https://doi.org/10.1002/cbdv.200790173, 4, 9, 2166-2171, 2007.09, A series of prenylated, flavone‐based polyphenols, compounds 1–8, were isolated from the wood of Artocarpus heterophyllus. These compounds, which have previously been shown not to inhibit tyrosinase activity, were found to be active inhibitors of the in vivo melanin biosynthesis in B16 melanoma cells, with little or no cytotoxicity. To clarify the structural requirement for inhibition, some structure–activity relationships were studied, in comparison with related compounds lacking prenyl side chains. Our experiments indicate that both prenyl and OH groups, as well as the type of substitution pattern, are crucial for the inhibition of melanin production in B16 melanoma cells..
197. Y. Kim, J. Han, S. S. Lee, K. Shimizu, Y. Tsutsumi, R. Kondo , Steroid 9a-hydroxylation during testosterone degradation by resting Rhodococcus equi Cells, Arch. Pharm. Chem. Life Sci.., 340, 209-214, 2007.07.
198. J. Liu, K. Shimizu, F. Konishi, K. Noda, S. Kumamoto, K. Kurashiki, R. Kondo, Anti-androgenic activities of the triterpenoids fraction of Ganoderma lucidum , Food Chem. , 100, 1691-1696, 2007.06.
199. J. Liu, K. Shimizu, F. Konishi, S. Kumamoto, R. Kondo, The anti-androgen effect of ganoderol B isolated from the fruiting body of Ganoderma lucidum, Bioorg. Med. Chem. , 15, 14, 4966-4972, 2007.03.
200. Enos T. Arung, K. Shimizu, R. Kondo , Isoprenoid-substituted polyphenols from Artocarpus heterophyllus -Melanin biosynthesis inhibitory activity and structural criteria-, Chemistry & Biodiversity , 4, 2166-2171, 2007.03.
201. Y. Kim, J. Han, S. S. Lee, K. Shimizu, Y. Tsutsumi, R. Kondo, Steroid 9a-hydroxylation during testosterone degradation by resting Rhodococcus equi Cells, Arch. Pharm. Chem. Life Sci, 340, 209-214 (2007), 2007.01.
202. J. Liu, K. Kurashiki, K. Shimizu,R. Kondo, Structure activity relationship for inhibition of 5α-reductase by triterpenoids isolated from Ganoderma lucidum, Bioorg. Med. Chem., 14, 24, 8654-60, 2006.12.
203. Tukiran, Saidah, Suyatno, Nurul Hidayati, K Shimizu, Briononic acid from the hexane extract of Sandoricum koetjape Merr. stem bark (Meliaceae), Indo. J. Chem. , 6, 3, 304-306, in press, 2006.11.
204. Tukiran, K. Sakai, K. Shimizu, S.A. Achmad, Hemsleyanoside F, an oligoresveratrol glucoside from the stem bark of Shorea parvistipulata F. Heim., Bull. Soc. Nat. Prod. Chem. (Indonesia), 6, 7-12, 2006.11.
205. Masuoka, N., Shimizu, K., Kubo, I., Antioxidant activity of anacardic acids, Food Chemistry, https://doi.org/10.1016/j.foodchem.2005.08.023, 99, 3, 555-562, 2006.08, Anacardic acids, 6-pentadec(en)ylsalicylic acids isolated from the cashew Anacardium occidentale L. (Anacardiaceae) nut and apple, were found to possess preventive antioxidant activity while salicylic acid did not show this activity. These anacardic acids prevent generation of superoxide radicals by inhibiting xanthine oxidase (EC 1.1.3.22, Grade IV) without radical-scavenging activity. Notably, the inhibition kinetics of anacardic acids do not follow hyperbolic dependence of enzyme inhibition on inhibitor contents (Michaelis–Menten equation) but follow the Hill equation instead. Anacardic acid (C15:1) inhibited the soybean lipoxygenase-1 (EC 1.13.11.12, Type 1) catalyzed oxidation of linoleic acid with an IC50 of 6.8 μM. The inhibition is a slow and reversible reaction without residual enzyme activity. The inhibition kinetics indicate that anacardic acid (C15:1) is a competitive inhibitor and the inhibition constant, KI, was 2.8 μM. Anacardic acids act as antioxidants in a variety ways, including inhibition of various prooxidant enzymes involved in the production of the reactive oxygen species and chelate divalent metal ions such as Fe2+ or Cu2+, but do not quench reactive oxygen species. The C15-alkenyl side chain is largely associated with the activity.
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206. K. Shimizu, M. Yamanaka, M. Gyokusen, S. Kaneko, M. Tsutsui, J. Sato, I. Sato, M. Sato, R. Kondo, Estrogen-like activity and prevention effect of bone loss in calcium deficient ovariectomized rats by the extract of Pleurotus eryngii., Phytotherapy Research, 20, 659-664, 2006.01.
207. J. Liu, K. Shimizu, R. Kondo, Ganoderic acid TR, a new lanostanoid with 5a-reductase inhibitory activity from the fruiting body of Ganodema lucidum., Natural Product Communications, 1, 5, 345-350, 2006.01.
208. J. Liu, K. Kurashiki, K. Shimizu, R. Kondo, 5a-reductase inhibitory effects of triterpenoids isolated from Ganoderma lucidum., Biol. Pharm. Bull., 29, 392-395, 2006.01.
209. Enos T. Arung, K. Shimizu, R. Kondo, Inhibitory Effect of Isoprenoid-substituted Flavonoids Isolated from Artocarpus heterophyllus on Melanin Biosynthesis, Planta Med, 72, 847-850, 2006.01.
210. Enos T. Arung, K. Shimizu, R. Kondo, Inhibitory effect of Artocarpanone from Artocarpus heterophyllus on melanin biosynthesis, Biol Pharm. Bull., 29, 1966-1969, 2006.01.
211. Tukiran, S.A. Achmad, E.H. Hakim, Y.M. Syah, L. Makmur, L.D. Juliawaty, K. Shimizu, and K. Sakai., Oligostilbenoids from the stem bark of Shorea selanica Blume (Dipterocarpaceae), Malaysian Journal of Science (MJS),, 24, 1, 27-35, 2005.01.
212. Tukiran, S.A. Achmad, E.H. Hakim, L. Makmur, K. Sakai, K. Shimizu, and Y.M. Syah, Oligostilbenoids from Shorea balangeran., Biochemical systematic and Ecology (BSE),, 10.1016/j.bse.2004.10.016, 33, 6, 631-634, 2005.01.
213. J. Liu, R. Fujita, M. Sato, K. Shimizu, F. Konishi, K. Noda, S. Kumamoto, C. Ueda, H. Tajiri, S. Kaneko, Y. Suimi, R. Kondo, The effect of strain, growth stage and cultivationg condition of Ganoderma lucidum on 5a-reductase inhibition, J. Wood Sci, 10.1007/s10086-004-0645-z, 51, 2, 189-192, 2005.01.
214. Enos T. Arung, K. Yoshikawa., K. Shimizu, R. Kondo, The effect of chlorophorin and its derivatives on melanin biosynthesis., Holzforschung, 10.1515/HF.2005.085, 59, 5, 514-518, 2005.01.
215. Enos T. Arung, Irawan W. Kusuma, Yetti M. Iskandar, S. Yasutake, K. Shimizu, R. Kondo, Screening of Indonesian plants for tyrosinase inhibitory activity, J. Wood Sci.., 10.1007/s10086-004-0690-7, 51, 5, 520-525, 2005.01.
216. Tukiran, Sjamsul A. Achmad, Euis H. Hakim, Yana M. Syah, Lukman Makmur, Lia D. Juliawaty, K. Shimizu, K. Sakai, Resveratrol oligomers from the stem bark of Hopea bancana (Dipterocarpaceae), Bull. Soc. Nat. Prod. Chem. (Indonesia), 4, 17-22, 2004.01.
217. K. Fukuhara, K. Shimizu, I. Kubo, Arudonine, an allelopathic steroidal glycoalkaloid from the root bark of Solanum arundo Mattei, Phytochemistry, 65, 9, 1283-1286, 2004.01.
218. I. Kubo, Y Xu, K Shimizu, Antibacterial activity of ent-kaurene diterpenoids from Rabdosia rosthornii., Phytother Res., 18, 2, 180-183, 2004.01.
219. I kubo, K. Nihei,K. Shimizu, Oxidation products of quercetin catalyzed by mushroom tyrosinase, Bioorg. Med. Chem., 12, 5343-5347, 2004.01.
220. Shimizu, K., Fujita, R., Kondo, R., Sakai, K., Kaneko, S., Morphological features and dietary functional components in fruit bodies of two strains of Pholiota adiposa grown on artificial beds, Journal of Wood Science, 49, 2, 193-196, 2003.04, The morphological features and dietary functional components of two strains (FPF-13 and Oninome-B) of Pholiota adiposa (numerisugitake mushroom) grown on artificial bed blocks were examined. The components examined were chitin, mannitol, trehalose, guanosine 5′-monophosphate, ergosterol, and β-glucan. There was a significant difference in the external shape of the two strains. However, the differences in the contents of functional components between the two strains, as well as between the pilei and stipes of the strains, were small. In both strains the trehalose content was much higher than the mannitol content, in contrast to those of Lentinula edodes. From a commercial point of view, Oninome-B has a clear advantage over other strains of P. adiposa owing to its less removable scale..
221. I. Kubo, K. Shimizu, Y.L. Xu, entIsopimarane diterpenoids of the leaves from Rabdosia forrestii, Fitoterapia, 74, 7-8, 649-9, 2003.01.
222. K. Towatari, K. Yoshida, N. Mori, K. Shimizu, R. Kondo, K. Sakai, Polyphenols from the heartwood of Cercidiphyllum japonicum and their effects on proliferation of mouse hair epithelial cells, Planta Med., 10.1055/s-2002-35657, 68, 11, 995-998, 2002.03.
223. K. Shimizu, R. Kondo, K. Sakai, N. Takeda, T. Nagahata, The skin-lightening effects of artocarpin on UVB -induced pigmentation., Planta Med. , 10.1055/s-2002-20057, 68, 1, 79-81, 2002.03.
224. K. Shimizu, R. Kondo, K. Sakai, Antioxidant activity of heartwood extracts of Papua New Guinean woods., J. Wood. Sci., 10.1007/BF00770708, 48, 5, 446-450, 2002.03.
225. K. Shimizu, R. Kondo, K. Sakai, N. Takeda, T. Nagahata, T. Oniki, Novel vitamin E derivative with 4-substituted resorcinol moiety has both antioxidant and tyrosinase inhibitory properties., Lipids, 10.1007/s11745-001-0847-9, 36, 12, 1321-1326, 2001.03.
226. K. Shimizu, R. Kondo, K. Sakai, A geranylated chalcone with 5α-reductase inhibitory properties from Artocarpus incisus., Phytochemistry, 10.1016/S0031-9422(00)00187-4, 54, 8, 737-739, 2001.03.
227. @Kuniyoshi Shimizu, Masayuki Fukuda, Ryuichiro Kondo*, and, Kokki Sakai, The 5α-reductase inhibitory components from heartwood of Artocarpus incisus: Structure-activity investigations, Planta Medica, 66, 1, 16-19, 2000.11.
228. K. Shimizu, R. Kondo, K. Sakai, Y. Shoyama, H. Sato, T. Ueno, The 5α-reductase inhibitory activity and hair regrowth effects of extract from Boehmeria nipononivea., Biosci. Biotechnol. Biochem., 64, 875-877, 2000.03.
229. @Kuniyoshi Shimizu, Ryuichiro KondoEmail author, Kokki Sakai, Supanida Buabarn and Uraiwan Dilokkunanant, 5α-Reductase inhibitory component from leaves of Artocarpus altilis, Journal of Wood Science, 46, 5, 385-389, 2000.10.
230. K. Shimizu, R. Kondo, K. Sakai , S. Buabarn, U. Dilokkunanant, The 5α-reductase inhibitory component from leaves of Artocarpus altilis., J. Wood. Sci., 46, 385-389, 2000.03.
231. K. Shimizu, R. Kondo and K. Sakai, Inhibition of tyrosinase by flavonoids, stilbenes and related 4-substituted resorcinols: Structure-activity investigations., Planta Med., 10.1055/s-2000-11113, 66, 1, 11-15, 2000.03.
232. K. Shimizu, M. Fukuda, R. Kondo, K. Sakai, The 5α-reductase inhibitiory components from heartwood of Artocarpus incisus: Structure-activity investigations., Planta Med., 10.1055/s-2000-11114, 66, 1, 16-19, 2000.03.
233. K. Shimizu, R. Kondo, K. Sakai, S.-H. Lee, H. Sato, The Inhibitory Components from Artocarpus incisus on Melanin Biosynthesis., Planta Med., 10.1055/s-2006-957470, 64, 5, 408-412, 1998.03.
234. K. Shimizu, R. Kondo, K. Sakai, A Stilbene Derivative from Artocarpus incisus., Phytochemistry, 10.1016/S0031-9422(97)00143-X, 45, 6, 1297-1298, 1997.03.
235. C. Pilotti, R. Kondo, @K. Shimizu, K. Sakai, An Examination of the Anti-fungal Components in the Heartwood Extracts of Pterocarpus indicus., Mokuzai gakkaishi, 41, 6, 593-597, 1995.03.