Effect of <Emphasis Type="Italic">Pinus radiate</Emphasis> bark extracts with different molecular weight distributions on cell growth of NIH/3T3 fibroblasts and dendrite retraction of B16 melanoma cells

Hot water extract (HWE) of Pinus radiata bark was separated into monomeric polyphenol (MPP), oligomeric proanthocyanidin (OPA), and polymeric proanthocyanidin (PPA) fractions by monitored chromatography using a Sephadex LH 20 column and an UV detector at 250 nm. The effects of these fractions on NIH/3T3 fibroblasts and B16 melanoma cells were examined by evaluating cell viability, melanogenesis (melanin content), morphological changes, and tyrosinase inhibitory activity. The polyphenolic fractions had a proliferation effect on fibroblasts, with cell growth increasing significantly (P < 0.01) even at the high concentration of 1250 μg/ml. At 125 μg/ml, HWE, MPP, and OPA had no effect on melanogenesis, whereas PPA significantly upregulated melanogenesis (P < 0.05). Melanogenesis was significantly upregulated in melanoma cells treated with these fractions at the high concentration of 600 μg/ml (P < 0.01). B16 melanoma cells cultured with the proanthocyanidin (PA)-rich fractions (HWE, OPA, and PPA) showed marked dendrite retraction, leading to significant morphological transformation. OPA in particular showed colored adhesion on the surface of melanoma cells. All four fractions significantly inhibited mushroom tyrosinase activity when compared to arbutin and ascorbic acid 2-glucoside (P < 0.05). The tyrosinase inhibitory activity of OPA and PPA did not differ significantly (P = 0.11), indicating that the inhibitory effects had a low correlation with molecular weight distribution. The inhibition kinetics of PPA determined using a Lineweaver-Burk plot indicated that PPA is a noncompetitive inhibitor of l-3,4-dihydroxyphenylalanine oxidation by mushroom tyrosinase.     

Antimicrobial activity against <Emphasis Type="Italic">Streptococcus sobrinus</Emphasis> and glucosyltransferase inhibitory activity of taxifolin and some flavanonol rhamnosides from kempas (<Emphasis Type="Italic">Koompassia malaccensis</Emphasis>) extracts

Twenty plant materials collected from the islands of Java and Kalimantan in Indonesia were extracted with 50% aqueous ethanol (crude extract). The crude extracts were assayed for antimicrobial activities against Streptococcus sobrinus and for glucosyltransferase (GTase) inhibition. Fourteen extracts inhibited the growth of S. sobrinus by more than 50% and six extracts inhibited GTase activity by more than 50% at a concentration of 100 μg/ml. Koompassia malaccensis (kempas) extracts showed 90% depression of S. sobrinus growth and 80% inhibition of GTase activity at a concentration of 100 μg/ml. Kempas crude extracts were subjected to column chromatography using Sephadex LH-20 and then preparative high-performance liquid chromatography to isolate four compounds A, B, C, and D. These compounds were identified as taxifolin and the flavanonol rhamnoside isomers neoastilbin, astilbin, and isoastilbin, respectively, from ¹H and ¹³C nuclear magnetic resonance (NMR) spectra and other two-dimensional NMR techniques (COSY, HMBC, and HMQC). Each compound depressed the growth of S. sobrinus over a concentration range of 9.3242.7 μg/ml and showed GTase inhibitory activity with IC₅₀ values in the range 27.4–57.3 μg/ml. Taxifolin and flavanonol rhamnoside isomers isolated for the first time from kempas could be potent compounds for preventing dental caries. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society Conference, Hiroshima, 2007     

Evaluation of biological activities of extracts from 22 African tropical wood species

Heartwoods of 22 African tropical wood species were extracted with methanol and the contents of total phenolic compounds in these extracts were measured. Three bioassays were conducted to evaluate the antioxidant activity, tyrosinase inhibitory activity, and antifungal activity of the methanol extracts. The results indicated that the extracts from 13 species exhibited high antioxidant potential, and their inhibitory concentrations that caused 50% scavenging of the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (IC₅₀) were less than 10 μg/ml. The crude extract from Cylicodiscus gabunensis showed the highest antioxidant activity, and was even higher than that of (+)-catechin, which is known as a potent antioxidant. There was a good correlation between the antioxidant activity and the content of total phenolic compounds, indicating that phenolic compounds played a predominant role in the antioxidant property of the wood extracts. Among all 22 species, only 1 species, Milicia excelsa, contained extractives that showed very high tyrosinase inhibitory activity. The extracts from 9 species showed high antifungal activity. No consistent relationship was observed between the tyrosinase inhibitory activity or antifungal activity and the content of total phenolics in the extracts. Part of this article was presented at the 56th Annual Meeting of the Japan Wood Research Society, Akita, August 2006     

Screening antiacne potency of Indonesian medicinal plants: antibacterial,lipase inhibition,and antioxidant activities

Indonesian medicinal plants were screened as potential sources of antiacne agents. The screening methods were performed using antibacterial assay against Propionibacterium acnes, lipase inhibitor assay, and antioxidant assay. The results showed that from 40 plant materials extracted with methanol and 50% ethanol in water, Caesalpinia sappan was the best extract based on the combined activities: antibacterial (minimum inhibitory concentration 0.13 mg/ml; minimum bactericidal concentration 0.25 mg/ml), lipase inhibitory [50% inhibitory concentration (IC₅₀) 120.0 μg/ml], and antioxidative (IC₅₀ 6.47 μg/ml). Another prospective extract is Intsia palembanica based on its lipase inhibitory activity (IC₅₀ 4.1 μg/ml) and antioxidant activity (IC₅₀ 3.87 μg/ml). Part of this report was presented at the 58th Annual Meeting of the Japan Wood Research Society, Tsukuba, March 2008     

Inhibitory components from the buds of clove (Syzygium aromaticum) on melanin formation in B16 melanoma cells

In the course to find a new whitening agent, we evaluated the methanol extract from bud of clove (Syzygium aromaticum) on melanin formation in B16 melanoma cells. Eugenol and eugenol acetate were isolated as the active compounds and showed melanin inhibition of 60% and 40% in B16 melanoma cell with less cytotoxicity at the concentration of 100 and 200 μg/mL, respectively. Furthermore, an essential oil prepared from the bud of clove, which contain eugenol and eugenol acetate as dominant components, showed melanin inhibition of 50% and 80% in B16 melanoma cells at the concentration of 100 and 200 μg/mL, respectively.     

Screening of Indonesian plants for tyrosinase inhibitory activity

In our efforts to find new tyrosinase inhibitory materials, we investigated 44 Indonesian plants belonging to 24 families for tyrosinase inhibitory activity. The extracts of 5 Artocarpus woods showed potent tyrosinase inhibitory activity (over 80% at 100 μg/ml) similar to a positive control, kojic acid. In Artocarpus woods, the extracts of the sapwoods showed stronger inhibitory activity than those of the heartwoods. Chlorophorin was isolated as one of the active compounds in the sapwood of Artocarpus heterophyllus. The content of chlorophorin in sapwood was higher than that in heartwood. Part of this paper was presented at the 53rd (Fukuoka, March 2003) and 54th (Hokkaido, August 2004) Annual Meetings of the Japan Wood Research Society, and the 5th International Wood Science Symposium (Kyoto, September 2004)     

Evaluation of biological activities of extracts from the fruiting body of <Emphasis Type="Italic">Pleurotus citrinopileatus</Emphasis> for skin cosmetics

Pleurotus citrinopileatus Singer has recently become a popular delicacy in East Asian countries. We prepared a methanol extract, soluble fractions from the methanol extract, and a hot water extract of the fruiting bodies of P. citrinopileatus. The biological activities such as melanin biosynthesis inhibition, tyrosinase inhibition, antioxidant activities [1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, oxygen radical absorbance capacity (ORAC), and superoxide dismutase (SOD)-like activity], antibacterial activities, and antihyaluronidase activities of these extracts were evaluated. We found that the n-hexanesoluble, diethyl ether-soluble, and ethyl acetate-soluble fractions exhibited melanin biosynthesis inhibition in B16 melanoma cells, as well as antioxidant (ORAC) and antibacterial activities. However, the n-butanol-soluble and water-soluble fractions and the methanol and hot water extracts exhibited antioxidant (DPPH radical scavenging, SOD-like activity) and antihyaluronidase activities. These results indicate that the fruiting bodies of P. citrinopileatus have the potential to be used as an ingredient in skin cosmetics.     

In vitro screening of angiotensin I-converting enzyme inhibitors from Japanese cedar (Cryptomeria japonica)

Screening and isolation of angiotensin I-converting enzyme (ACE) inhibitors from Japanese cedar (Cryptomeria japonica) based on the in vitro ACE inhibitory assay were attempted. The ethanol extract from outer bark showed the highest inhibitory activity (IC₅₀ is 16g/ml) among 24 extracts prepared from roots, leaves, heartwood, sapwood, inner bark, and outer bark by successive extraction with four solvents. The fractionation of the outer bark ethanol extract followed by the bioassay resulted in the isolation of two strong ACE inhibitors, catechin and dimeric procyanidin B3. The bioassay of three flavan-3-ols including (+)-catechin and six flavones revealed that most of these compounds have high ACE inhibitory activity. The results suggest that the phenolic hydroxyl group at the C7 position and heterocyclic oxygen atom of these compounds are important for expressing the inhibitory activity.     

Fumigant and repellent activities of essential oil from <Emphasis Type="Italic">Coriandrum sativum</Emphasis> (L.) (Apiaceae) against red flour beetle <Emphasis Type="Italic">Tribolium castaneum</Emphasis> (Herbst) (Coleoptera: Tenebrionidae)

The biological activity of essential oil extracted from coriander, Coriandrum sativum L. (Apiaceae) against eggs, larvae and adults of Tribolium castaneum (Herbst) was investigated in a series of laboratory experiments. Bioassays conducted in air-tight glass chambers showed vapour toxicity and strong repellency on filter paper arena test towards all the stages used. Responses varied significantly (P < 0.001) within insect stages, dosages and exposures. At 12.0 μg/ml dosage, mortality of adults reached 95%, but for 12-, 14- and 16-days larvae, mortalities were 65, 74 and 82%, respectively. On the developmental inhibition, individuals fumigated at the larval stage confirmed that the percentage of larvae reaching to pupal stage and pupae to adult stage, decreased significantly (P < 0.001) with increasing dosage concentration. The oil fumigation yielded 100% mortality for T. castaneum larvae, pupae and adults at 0.08 μg/ml dosage. The oil had fumigant activity against eggs and the toxicity progressively increased with increased exposure times and concentrations. One hundred per cent T. castaneum egg mortalities were obtained with oil fumigation at 20 μg/ml and 96 h exposure period. C. sativum oil also showed highly repellent activity to the adults of T. castaneum, with overall repellency was 90% and at a dosage of 12 μg/ml repellency was 100% in a filter-paper arena test.     

Possibility of improvement in fundamental properties of wood of acacia hybrids by artificial hybridization

Growth, specific gravity, and wood fiber length of Acacia mangium, Acacia auriculiformis, artificial acacia hybrid clones, and combinations, which were planted in a trial forest in Bavi, Vietnam, in July 2001, were examined. The radial variations from pith to bark were investigated to clarify the effect of genetic factors on these traits. Superiority of hybrids over their parents ranged from 36.3% to 41.6% for diameter, from 20.0% to 25.3% for height, from 6.9% to 20.7% for specific gravity, and from 6.1% to 12.8% for wood fiber length. The hybrid possessed heterosis in diameter, height, specific gravity, and wood fiber length regardless of whether the female parent was A. mangium or A. auriculiformis. The profiles of wood fiber length and specific gravity in the radial direction were similar for all the trees investigated. Wood fiber length was initially 0.5–0.6 mm near the pith and then increased slowly, finally reaching 1.0–1.2 mm near the bark. The specific gravity of acacia increased from 0.49–0.58 near the pith to 0.63–0.74 near the bark. From a relative distance of 30% from the pith, the specific gravity increased slightly and seemed to be stable. The relations among tree diameter, specific gravity, and wood fiber length were fair and could be represented by positive linear regression formulas. Hybrids for which A. auriculiformis was the female parent and A. mangium was the male parent had a faster growth rate and longer wood fibers than the inverse hybrids. Part of this report was presented at the 6th Pacific Regional Wood Anatomy Conference, Kyoto, Japan, December 2005     

3-Prenyl luteolin,a new prenylated flavone with melanin biosynthesis inhibitory activity from wood of Artocarpus heterophyllus

In our efforts to find new whitening agent from natural resources, we focused on wood of Artocarpus heterophyllus which shows anti-melanogenesis activity. By activity-guided fractionation of A. heterophyllus wood extract, a new prenylated flavonoid, 3-prenyl luteolin (1) was isolated. The IC₅₀ of mushroom tyrosinase inhibitotry activity of 1 was 76.3 µM. The results of the comparison with that of luteolin showed the prenyl substituent at C-3 position of 1 play an important role for revealing tyrosinase inhibition. In melanin formation inhibition on B16 melanoma cells, IC₅₀ of 1 was 56.7 µM with less cytotoxicity.     

Effects of Ca^2＋concentrations on accumulations of mineral elements in the components of Pteroceltis tatarinowii

The bark ofPteroceltis tatarinowii is a raw material for manufacturing Xuan Paper. The effects of Ca²⁺ concentrations on the accumulation of mineral elements in the bark, leaf and root ofPteroceltis tatarinowii were studied under controlled conditions. The types of Hoagland nutrient solution with three Ca²⁺ concentrations levels (200, 400 and 600 μg·g⁻¹) and a control (without Ca²⁺ were designed to culturePteroceltis tatarinowii. After 6 months, contents of seven mineral elements including Ca, K, Mg, Mn, Zn, Cu and Na in the root, leaf and bark were analyzed. The results indicated that Ca accumulations content in the root, leaf and bark had positively relation with Ca²⁺ concentrations (200, 400, 600 μg·g⁻¹), and the order of the Ca content in the three components was root>leaf>bark. Ca content in the root treated with 600 μg·g⁻¹ Ca²⁺ concentrations was 5.5 times as high as that of the control, and about 1.4 times as high as that of the root treated in 200 and 400 μg/g Ca²⁺ concentrations respectively. On the contrary, K and Mg contents in the root, leaf and bark were negatively related to Ca²⁺ concentrations, especially in the bark, and their accumulation trend followed the order of leaf>root>bark. K content in the bark treated with 600 μg·g⁻¹ Ca²⁺ concentrations was 39.3% of that of the control, and was 79.0% and 91.8% of that of the bark treated with 200μg·g⁻¹ and 400μg·g⁻¹ Ca²⁺ concentrations respectively; Mg content in the bark treated with 600μg·g⁻¹ Ca²⁺ concentrations was 23.4% of that of the control, and was 27.1% and 35.4% of that of the bark treated with 200 and 400 μg·g⁻¹ Ca²⁺ concentrations respectively. Compared with the control, the general tendency of Mn, Zn and Cu content decreased with increasing of Ca²⁺ concentrations and their contents were in the order: root>leaf>bark. Based on the results of this study, the experiment has been useful for providing academic bases in improving the bark quality ofPteroceltis tatarinowii on non-limestone soil. Foundation item: This paper is supported by National Natural Science Foundation of China (No. 39970608). Biography: Fang Shengzun (1963), male, Professor in Stiiviculture, Nanjing Forestry University, Nanjing 210027, P.R. China. Responsible editor: Zhu Hong     

Inhibitory effect of garlic, clove and carrot on growth ofAspergillus flavus and aflatoxin production

The inhibitory effect of crude extracts of garlic, clove and carrot at concentrations of 20,000, 40,000, 60,000, 80,000 and 100,000 μg/mL on growth ofAspergillus flavus and aflatoxin production in rice was investigated. The results showed that garlic, clove and carrot could inhibit the growth ofA. flavus and aflatoxin production. Garlic and clove at 100,000 μg/mL highly reduced the level of aflatoxin from 5.94 to 0.15 and 0.06 μg/g respectively whereas carrot at 20,000 μg/mL reduced the most level of aflatoxin from 5.94 to 0.03 μg/g. Garlic, clove and carrot at 100,000μg/mL also inhibited the mould growth. The most effective herb for inhibiting the growth ofA. flavus and aflatoxin production was garlic. Responsible Editor: Sun Yueqi     

Antioxidative and antiradical activities of <Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis> leaf oils from Thailand

The leaf essential oils (six samples) from three clones of Eucalyptus camaldulensis Dehnh. were characterized by gas chromatography-mass spectrometry. Radical scavenging and antioxidant properties were investigated by the 1,1-diphenyl-2-picylhydrazyl (DPPH) assay and the β-carotene bleaching test. It was found that the whole essential oil and its fractions had significant antioxidant effects when they were tested by each method. In the DPPH assay, the E. camaldulensis leaf oils showed IC₅₀ inhibitory concentrations in the range of 1.75–12.62 mg/ml. In the β-carotene bleaching test, the IC₅₀ valued were in the range of 14.30–118.55 μg/ml. Part of this report was presented at the 18th meeting of the Chugoku-Shikoku Branch of the Japan Wood Research Society, November 2006     

Brazilin from Caesalpinia sappan wood as an antiacne agent

In screening experiments for antiacne activity, methanolic and 50% ethanolic extracts of Caesalpinia sappan wood showed the most potent activity out of 28 species of plants extracts. These extracts showed inhibition of Propionibacterium acnes growth, lipase inhibitory activity, and antioxidant activity. In order to isolate the active compound from C. sappan, separation of the extract components was performed by column chromatography and preparative high-performance liquid chromatography (HPLC). Brazilin, protosappanin A, and sappanone B were isolated from methanolic extracts. Brazilin showed better antibacterial activity [minimum inhibitory concentration (MIC) = minimum bactericidal concentration (MBC) = 0.50 mg/ml] than protosappanin A (MIC = MBC = 1.00 mg/ml) and sappanone B (MIC = MBC > 2.00 mg/ml). The 50% inhibitory concentration (IC₅₀) for lipase inhibition was lowest for brazilin (6 μM), which showed strong inhibition compared with protosappanin A (100 μM) and chloramphenicol (677 μM, positive control). The antioxidant activity of brazilin (IC₅₀ 8.8 μM) was not significantly different from protosappanin A (9.1 μM) and (+)-catechin (10.2 μM). The antioxidant activity of brazilin and protosappanin A were higher than sappanone B (IC₅₀ 14.5 μM). Brazilin is considered to have suffi ciently potent activity for use as an antiacne agent.     

Chemical changes in terpenes of sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) wood during steam drying in kiln at high temperature

Sawdusts of sugi (Cryptomeria japonica) wood prepared before and after steam drying at 120°C in a kiln were extracted with n-hexane and ethyl acetate to give n-hexane extracts and ethyl acetate extracts. From gas chromatography-mass spectrometry analysis of the ethyl acetate extracts from woods before and after steam drying, the components of 4-epi-cubebol, cubebol, and 2,7(14),10-bis-abolatrien-1-ol-4-one, which existed in the raw sugi wood, were proved to disappear in the steam-dried wood. These components were also absent in the ethyl acetate extract of the steam-condensed solution of waste steam from the kiln outlet. When these three components were treated with 0.2% (v/v) acetic acid solution at 120°C, δ-cadinene was produced as a major product from both 4-epi-cubebol and cubebol by dehydration and cleavage of the cyclopropane ring, and cryptomerone from 2,7(14),10-bisabolatrien-1-ol-4-one by hydration. The chemical changes of the three components presumably occur during steam drying of the sugi wood. This study was presented in part at the 85th Spring Meeting of the Chemical Society of Japan, Kanagawa, Japan, March 26–29, 2005     

A new norlignan from Taxodium ascendens

A new norlignan, (2R,3R,4S,5S)-2,4-bis(4-hydroxyphenyl)-3,5-dihydroxy-tetrahydropyran (1), together with 9 known compounds were isolated from the branches and leaves of Taxodium ascendens. Their structures were mainly determined on the basis of MS, IR, 1D and 2D NMR spectral evidences. Methanol extract showed inhibitory activity on carbonic anhydrase II with an IC₅₀ value of 4.27 µg/ml, acetone extract and methanol extract inhibited activity of cathepsin B with IC₅₀ values of 2.12 and 3.71 µg/ml, respectively.     

Effects of bark stripping by sika deer, Cervus nippon, on wind damage to coniferous trees in subalpine forest of central Japan

We evaluated the effect of bark stripping by sika deer (Cervus nippon), and subsequent wood decay, on tree fall in a coniferous forest on Mt Ohdaigahara in central Japan from July to September 2006. This valuable primeval coniferous forest is declining because of bark stripping. Broken trunks of Abies homolepis and Picea jezoensis var. hondoensis had more serious decay and larger bark-stripping wounds on the trunk than standing trees, suggesting that bark stripping causes trunk decay and results in broken trunks and uprooting by typhoons.     

Extraction of <Emphasis Type="Italic">Trigonella foenum-graecum</Emphasis> L. by supercritical fluid CO<Subscript>2</Subscript> and its contact toxicity to <Emphasis Type="Italic">Rhyzopertha dominica</Emphasis> (Fabricius) (Coleoptera: Bostrichidae)

Rhyzopertha dominica (Fabricius) has developed extensive pesticide resistance in the last several decades. We have developed a supercritical fluid extraction method for Trigonella foenum-graecum L. (TFG) and studied the contact toxicities of the extracts to R. dominica. The extraction method was designed with orthogonal experiments to preserve and collect all the possible active components. Contact toxicity and efficiency of extraction were used as standard values to optimize extraction conditions, which were achieved at 55°C under 25 Mpa of pressure. The extraction efficiency for 200 g of dry sample reached 6.21% with 30 ml of 95% alcohol. Extracts loaded on filter paper showed dose and time dependent toxicities to adult R. dominica with a LC₅₀ value of 65.02 μg/cm² after 3 days post treatment. Our extensive in vivo studies indicated the extracts from Trigonella foenum-graecum seeds have high efficacy against pesticide resistant R. dominica. The active ingredient(s) from the extract shows promise as a novel pesticide candidate.     

寄生小蜂对小青×黑杨树皮及杨圆蚧挥发油的行为反应

Four-armed airflow olfactometer was used to determining the foraging behavior ofPteroptrix longgiclava (Girault) (Hymenoptera: Aphelinidae) andEncarsia gigas (Tshumakova) (Hymenoptera: Aphelinidae) to the essential oils which emitted from the healthy bark ofPopulus pseudo-simonii×P. nigra, the infested bark injured byQuadraspidiotus gigas (Thiem & Gerneck), the body and scale of fixed 1st-instar-nymph ofQ. gigas. The results from these experiments showed that the volatile oils produced from the injured bark and from the scale of fixed 1st-instar-nymph had a higher attractive ability to female adults of the two species of wasps. The essential oil produced from the scale of the pest at dosages of 3–7 μL and the essential oil emitted by injured bark at dosages of 5–9 μL had a stronger alluring effect on the host searching behavior ofPteroptrix longgiclava. The essential oil from the body of fixed 1st-instar-nymph ofQ. gigas also had certain effect on the host locating effort ofPteroptrix longgiclava andEncarsia gigas. Those two wasps did not shown any reaction to the essential oil produced by the healthy bark of poplar. Foundation item: This paper was supported by National Natural Science Foundation of China (39970620) and the “TRAPOYT” Biography: CHI De-fu (1962-), male, Ph.D., Professor in Northeast Forestry University, Harbin 150040, P.R. China Responsible editor: Chai Ruihai