Chemical Structures and Anti-proliferative Effects of Valeriana fauriei Constituents on Cancer Stem Cells.

We isolated the new sesquiterpenes, valerianaterpenes IV and V, and the new lignans valerianalignans I-III from the methanol extracts of the rhizomes and roots of Valeriana fauriei and elucidated their structures based on chemical and spectroscopic findings. The absolute configuration of valerianaterpene IV and valerianalignans I-III were established by comparing experimental and predicted electronic circular dichroism (ECD) data. Among the isolated compounds, valerianalignans I and II exerted anti-proliferative activity against human astrocytoma cells (U-251 MG) and their cancer stem cells (U-251 MG CSCs). Interestingly, valerianalignans I and II notably exerted anti-proliferative activities at lower concentrations against CSCs than non-CSCs, and the absolute configurations of these compounds affected their activities.

Comparison of the Concentration of Suspended Particles and Their Chemical Composition near the Ground Surface and Dust Extinction Coefficient by LIDAR.

In many epidemiological studies, the dust extinction coefficient measured by light detection and ranging (LIDAR) is used as an indicator of exposure to Asian dust. However, few reports on the relationship between the distribution of total suspended particles (TSPs) near the ground surface and the dust extinction coefficient exist. In this study, we examined the relationship between the concentrations of TSPs near the ground surface, substances indicative of mineral content, and air pollutants that may be transported with Asian dust and dust extinction coefficients in two regions: Imizu and Yurihama-Matsue, from March to May in 2011 and 2013. In both years, large dust extinction coefficients were observed in Imizu and Matsue on days when the concentrations of TSPs and mineral content indicators were high near the ground surface in Imizu and Yurihama, and Asian dust was expected to be highly suspended. In both regions, the concentrations of TSPs and mineral content indicators were significantly positively correlated with the dust extinction coefficient. The concentrations of all air pollutants analyzed were significantly positively correlated with the dust extinction coefficient in each region in 2013, but not in 2011. These results suggest that the dust extinction coefficient is a useful indicator of Asian dust near the ground surface; however, as harmful air pollutants occasionally move with Asian dust, it is necessary to monitor these pollutants near the ground surface when conducting an epidemiological study on the health effect of airborne particles.

Anti-Proliferative Effects of Iridoids from Valeriana fauriei on Cancer Stem Cells.

We isolated seven new iridoid glucosides (valerianairidoids I-VII; 1-3, 6, 7, 9, and 12) and six known compounds from the methanol extract of the dried rhizomes and roots of Valeriana fauriei. Chemical and spectroscopic data were used to elucidate the chemical structures of the seven new iridoid glucosides, and their absolute configurations were determined by comparing their electronic circular dichroism (ECD) spectra with those determined experimentally. Aglycones 1a, 6a, and 9a, which were obtained by enzymatic hydrolysis of the isolated iridoid glucosides, exhibited anti-proliferative activities against cancer stem cells (CSCs) established by a sphere-formation assay using human breast cancer (MDA-MB-231) and human astrocytoma (U-251MG) cells. Interestingly, these iridoids selectively showed anti-proliferative activities against CSCs from MDA-MB-231 cells. These results suggest that the iridoids obtained in this study may have potency as a breast cancer treatment and as preventive agent via exterminating CSCs.

Linderapyrone: A Wnt signal inhibitor isolated from Lindera umbellata.

Linderapyrone, a Wnt signal inhibitor was isolated from the methanolic extract of the stems and twigs of Lindera umbellata together with epi-(-)-linderol A. Linderapyrone inhibited TCF/ß-catenin transcriptional activity that was evaluated using cell-based TOPFlash luciferase assay system. To evaluate the structure-activity relationship and mechanism, we synthesized linderapyrone and its derivatives from piperitone. As the results of further bioassay for synthesized compounds, we found both of pyrone and monoterpene moieties were necessary for inhibitory effect. cDNA microarray analysis in a linderapyrone derivative treated human colorectal cancer cells showed that this compound downregulates Wnt signaling pathway. Moreover, we successes to synthesize the derivative of linderapyrone that has stronger inhibitory effect than linderapyrone and ICG-001 (positive control).

Construction of sulfur-containing compounds with anti-cancer stem cell activity using thioacrolein derived from garlic based on nature-inspired scaffolds.

Sulfur-containing compounds, such as cyclic compounds with a vinyl sulfane structure, exhibit a wide range of biological activities including anticancer activity. Therefore, the development of efficient strategies to synthesize such compounds is a remarkable achievement. We have developed a unique approach for the rapid and modular preparation of nature-inspired cyclic and acyclic sulfur-containing compounds using thioacrolein, a naturally occurring chemically unstable intermediate. We constructed thiopyranone derivatives through the regioselective sequential double Diels-Alder reaction of thioacrolein produced by allicin, a major component in garlic, and two molecules of silyl enol ether as the diene partner. The cytotoxicity toward cancer stem cells of the thiopyranones was equal to or higher than that of (Z)-ajoene (positive control) derived from garlic, and the thiopyranones had higher chemical stability than (Z)-ajoene.

Association of Protein and Endotoxin in Outdoor Air with Emergency Department Visits for Children and Adults with Asthma in Fukuoka, Japan.

We examined the association of biological components in airborne particles, i.e., proteins and endotoxins, in outdoor air with asthma exacerbation in the Fukuoka metropolitan area, Fukuoka, Japan. Data on emergency department (ED) visits for asthma in children (age, 0-14 years) and adults (age, 15-64 years) were collected at a medical center from December 2014 to November 2015. One hundred eighty-one children and 143 adults visited the ED for asthma, and the weekly number of ED visits in children increased in autumn, i.e., September (second week) to November (first week). Fine (aerodynamic diameter ≤2.5 µm) and coarse (≥2.5 µm) particles were collected for 3 or 4 weeks per month, and protein and endotoxin concentrations were analyzed. Protein was largely prevalent in fine particles (0.34-7.33 µg/m3), and concentrations were high in April, May, June, and October. In contrast, endotoxin was mainly included in coarse particles (0.0010-0.0246 EU/m3), and concentrations were high in September (third week), October (first, second, and fourth weeks), February (fourth week), and July (first week). The results of a Poisson regression analysis indicated that endotoxin (in fine and coarse particles alike) was a significant factor for ED visits related to asthma in children, even after adjusting for meteorological factors, i.e., temperature, relative humidity, and wind speed. However, there was no association between environmental factors and ED visits for asthma in adults. These results suggest that endotoxin in outdoor air is significantly associated with an increased risk of asthma exacerbation in children.

Relationship of Asian Dust Events with Atmospheric Endotoxin and Protein Levels in Sasebo and Kyoto, Japan, in Spring.

Asian dust events are caused by dust storms originating from deserts in Mongolia and northern China, and these events are observed in Japan, mainly in spring. To explore the effect of Asian dust events on atmospheric endotoxin and protein levels, we collected the total suspended particles (TSP) in the spring months (March, April, and May) of 2015 in Sasebo and Kyoto, Japan, and assessed the levels of biological elements at both locations. At both locations, the daily concentrations of TSP, water-soluble Ca2+ (an indicator mineral of soil in dust), endotoxins, and proteins were found to be high during and after Asian dust events recorded by the Japan Meteorological Agency. The concentration of Ca2+ showed a strong positive correlation with the concentrations of TSP and endotoxin, while the concentration of protein was moderately positively correlated with Ca2+ in both Sasebo and Kyoto. There were large concentrations of endotoxins, and the fluctuation ranges were higher in Sasebo than in Kyoto. In contrast, protein concentrations showed low levels of fluctuation, and no major differences were found in the concentration at each location.

Structures and Antimutagenic Effects of Onoceranoid-Type Triterpenoids from the Leaves of Lansium domesticum.

A methanol extract of the dried leaves of Lansium domesticum showed antimutagenic effects against 3-amino-1,4-dimethyl-5 H-pyrido[4,3- b]indole (Trp-P-1) and 2-amino-1-methyl-6-phenylimidazo[4,5- bI]pyridine (PhIP) using the Ames assay. Nine new onoceranoid-type triterpenoids, lansium acids I-IX (1-9), and nine known compounds (10-16) were isolated from the extract. The structures of the new compounds were elucidated on the basis of chemical and spectroscopic evidence. The absolute stereostructures of the new compounds were determined via their electronic circular dichroism spectra. Several isolated onoceranoid-type triterpeneoids showed antimutagenic effects in an in vitro Ames assay. Moreover, oral intake of a major constituent, lansionic acid (10), showed antimutagenic effects against PhIP in an in vivo micronucleus test.

Seasonal Fluctuation of Endotoxin and Protein Concentrations in Outdoor Air in Sasebo, Japan.

To determine the levels of endotoxin, which is a major component of outer membrane of Gram-negative bacteria, and protein in the atmosphere in Sasebo, Japan, we measured these biological materials in fine (aerodynamic diameter ≤2.5 µm) and coarse (≥2.5 µm) particles collected for 81 weeks (September 2014 to May 2016). The monthly concentrations (i.e., the mean value of weekly concentrations for each month) of endotoxin were higher in coarse particles than in fine particles. Fluctuations in monthly endotoxin concentrations were large in both fine (0.0005-0.0208 EU/m3) and coarse (0.0032-0.1164 EU/m3) particles. Furthermore, the endotoxin concentrations in coarse particles were highest in October 2014 and 2015 as well as September 2014 (0.0407-0.1164 EU/m3). However, the monthly protein concentrations were higher in fine particles than in coarse particles. Compared to the endotoxin concentrations, the fluctuations in the monthly protein concentrations were smaller in both coarse and fine particles. To our knowledge, this study is the first to report long-term atmospheric concentrations of endotoxin and protein in Japan. Since the endotoxin concentrations in coarse particles were positively associated with the concentrations of Na+ and Cl-, it suggests the involvement of Gram-negative bacteria from seawater to the endotoxin levels in the atmosphere. For fine particles, the protein concentrations were positively associated with the concentrations of particles, NO3- and SO42-. These results suggest that combustion of organic materials, such as biomass burning, may be a contributor to atmospheric protein during this study period.

Chemical Structures of Novel Maillard Reaction Products under Hyperglycemic Conditions.

Two novel and two known compounds, 4-quinolylaldoxime and indole-3-aldehyde, were isolated from a reaction mixture consisting of D-glucose and L-tryptophan at physiological temperature and pH. The chemical structures of the two novel compounds were elucidated by spectroscopic analysis such as X-ray crystallography. One of the novel compound and the indole-3-aldehyde showed mutagenicity toward Salmonella typhimurium YG1024 with S9 mix. Furthermore, 4-quinolylaldoxime was detected from streptozotocin-induced diabetic rat plasma by LC-MS/MS analysis; however, the isolated compounds were not detected in rat diet extracts. To our knowledge, this is the first report in which 4-quinolylaldoxime was detected in rat plasma. These results suggest that amino-carbonyl reaction products may be formed in diabetic condition and induce genetic damage.

Association of airborne particles, protein, and endotoxin with emergency department visits for asthma in Kyoto, Japan.

BACKGROUND: The health effects of biological aerosols on the respiratory system are unclear. The purpose of this study was to clarify the association of airborne particle, protein, and endotoxin with emergency department visits for asthma in Kyoto City, Japan. METHODS: We collected data on emergency department visits at a hospital in Kyoto from September 2014 to May 2016. Fine (aerodynamic diameter ≤ 2.5 µm) and coarse (≥ 2.5 µm) particles were collected in Kyoto, and protein and endotoxin levels were analyzed. The association of the levels of particles, protein, endotoxin, and meteorological factors (temperature, relative humidity, wind speed, and air pressure) with emergency department visits for asthma was estimated. RESULTS: There were 1 to 15 emergency department visits for asthma per week, and the numbers of visits increased in the autumn and spring, namely many weeks in September, October, and April. Weekly concentration of protein in fine particles was markedly higher than that in coarse particles, and protein concentration in fine particles was high in spring months. Weekly endotoxin concentrations in fine and coarse particles were high in autumn months, including September 2014 and 2015. Even after adjusting for meteorological factors, the concentrations of coarse particles and endotoxin in both particles were significant factors on emergency department visits for asthma. CONCLUSIONS: Our results suggest that atmospheric coarse particles and endotoxin are significantly associated with an increased risk of asthma exacerbation.

Comparison of Air Pollution in Metropolises in China (Beijing) and Japan (Osaka and Nagoya) on the Basis of the Levels of Contaminants and Mutagenicity.

Public concern regarding the transport of air pollutants from mainland East Asia to the leeward area by the prevailing westerlies in spring and winter monsoon has been growing in recent years. We collected total suspended particle (TSP) in Beijing, a metropolis of China located windward of Japan, in spring (late February 2011-May 2011) and in winter (November 2012-early February 2013), then analyzed metals, ions, and organic compounds and mutagenicity, and compared the pollution levels with samples collected at two Japanese metropolises (Osaka and Nagoya) during the same periods. The medians of concentration of TSP and other factors in Beijing were much larger than those in the Japanese metropolises. Especially, the concentrations of polycyclic aromatic hydrocarbons (PAHs) were remarkably high in Beijing in winter, and the median of total PAHs concentration in Beijing was 62-63 times larger than that in the Japanese sites. The mutagenicity of TSP from Beijing toward Salmonella typhimurium YG1024, with and without a mammalian metabolic system (S9 mix), was 13-25 times higher than that from the Japanese sites in winter. These results suggest that air pollution levels in Beijing are very high compared with those at the two Japanese metropolises we evaluated. The diagnostic ratios of PAHs and nitrated polycyclic aromatic hydrocarbons (NPAHs) suggest that the major sources of PAHs and NPAHs in Beijing are different from those at the two Japanese sites in winter, and that the major source in Beijing is coal/biomass combustion.

Spatial correlativity of atmospheric particulate components simultaneously collected in Japan.

The simultaneous sampling of total suspended particles was performed at 14 sites in Japan during July 2008-June 2009. The spatial correlativity of each particulate composition toward Osaka was obtained for nine selected sites to overview the chemical composition and geographical distribution of particulate components across a wide range of areas nationwide. The spatial correlatives of atmospheric particulate components were extended to an even wider range of areas up to 950 km distance (meso-alpha scale region, >200 km) for a far-reaching distance analysis unique in the literature. Overall, the spatial correlations of ionic species and both organic and elemental carbons were significant, suggesting their shared advections, including their long-range transport from East Asia. Although sulfate ions are widely dispersed across Japan, such is not necessarily correlated with organic and elemental carbon, possibly indicating that the sulfate emission source, including long-range transport, differs from that of carbonaceous particulates. By contrast, the characteristics of spatial correlatives of metallic constituents vary; for example, particulate Pb and Cd show a significantly wide range of spatial correlatives to Osaka, while Mn-though limited to cities neighboring Osaka-shows significant spatial correlations. Other metallic constituents showed no significant spatial correlatives, indicating the effects of local pollutants. Moreover, the extent of the spatial dispersion of the particulate components and the relationships among chemical components were analyzed via factor analysis to highlight the effects of long-range inflow and local original emissions. In this treatment, 13 particulate components among the 19 measured were implicated in long-range transport.

Seasonal Fluctuations in Air Pollution in Dazaifu, Japan, and Effect of Long-Range Transport from Mainland East Asia.

To clarify the seasonal fluctuations in air pollution and the effect of long-range transport, we collected airborne particles (n=118) at Dazaifu in Fukuoka, Japan, from June 2012 to May 2013 and measured Pb and SO4(2-), which are indicators of the long-range transport of anthropogenic air pollutants, as well as their mutagenicity, and other factors. The levels of airborne particles, Pb, and SO4(2-) were very high on March 4, 8, 9, and 19, and May 13, 21, and 22, 2013. The backward trajectories indicated that air masses had arrived from the Gobi Desert and northern China on those days. The mutagenicity of airborne particles was examined using the Ames test on Salmonella typhimurium YG1024. Highly mutagenic airborne particles were mostly collected in winter, and most of them showed high activity both with and without S9 mix. High levels of polycyclic aromatic hydrocarbons (PAHs) were found in many samples that showed high mutagenicity. For the samples collected on January 30, February 21, and March 4, the levels of Pb, SO4(2-), PAHs, and mutagenicity were high, and the backward trajectories indicated that air masses present on those days had passed through northern or central China. The Japan Meteorological Agency registered Asian dust events at Fukuoka on March 8, 9, and 19, 2013. The results of the present study suggest that high levels of anthropogenic air pollutants were transported with Asian dust. Similarly, long-range transport of air pollutants including mutagens occurred on days when Asian dust events were not registered.

In vivo genotoxicity of a novel heterocyclic amine, aminobenzoazepinoquinolinone-derivative (ABAQ), produced by the Maillard reaction between glucose and l-tryptophan.

We recently demonstrated that a novel heterocyclic amine, 5-amino-6-hydroxy-8H-benzo[6,7]azepino[5,4,3-de]quinolin-7-one (ABAQ), is produced from glucose and l-tryptophan by the Maillard reaction at physiological temperature and pH, and that ABAQ was strongly mutagenic for Salmonella strains in the presence of S9 mix. Here, we present the results of three in vivo genotoxicity assays of ABAQ. The comet assay revealed that DNA damage was significantly increased in the livers, kidneys, lungs, and bone marrows of ICR mice, 3h after i.p. injection of ABAQ (50mg/kg body weight (bw)). To evaluate clastogenicity, the peripheral blood micronucleus test was performed, also in ICR mice. ABAQ induced micronucleated reticulocytes (MNRETs) in a dose-dependent manner; the frequency of MNRETs was significantly elevated at all i.p. doses (12.5, 25, and 50mg/kg bw) after 48h. To investigate the mutagenicity of ABAQ in vivo, gpt delta transgenic mice were treated with five consecutive administrations of ABAQ by gavage at doses of 25 or 50mg/kg per week for 3 weeks. The frequencies of gpt mutations (MF) in the liver of mice increased significantly compared with controls, in a dose-dependent manner. No significant increase of gpt MF was detected in the kidneys. Base substitutions predominated; both G:C→A:T and A:T→C:G mutations were significantly increased by ABAQ. The Spi(-) MF was also significantly increased in the liver after ABAQ treatment. If formed in vivo, ABAQ may give rise to adverse genotoxic effects.

Cytotoxic activities of alkaloid constituents from the climbing stems and rhizomes of Sinomenium acutum against cancer stem cells.

From the methanolic extract of the climbing stems and rhizomes of Sinomenium acutum, two new aporphine analogues, acutumalkaloids I and II, were isolated together with fifteen known compounds including lysicamine. The chemical structures of the isolated new compounds were elucidated based on chemical/physicochemical evidence such as NMR and MS spectra. For acutumalkaloids I and II, the absolute configurations were established by comparison of experimental and predicted electronic circular dichroism (ECD) data. We compared anti-proliferative activities of isolated compounds with reported naturally occurring Wnt/ß-catenin pathway inhibitor, nuciferine. Among the isolated compounds, we found lysicamine have anti-proliferative activity against both of HT-29 human colon cancer cell line and its cancer stem cells (CSCs). The IC50 values of lysicamine against non-CSCs and its CSCs were lower than that of nuciferine. In addition, the results of western blotting analysis suggested that lysicamine inhibited the expression of Wnt/ß-catenin pathway target protein such as survivin. These results suggested that lysicamine show cytotoxic activity via inhibition of Wnt/ß-catenin pathway.

Linderapyrone analogue LPD-01 as a cancer treatment agent by targeting importin7.

The Wnt/ß-catenin signaling pathway plays important roles in several cancer cells, including cell proliferation and development. We previously succeeded in synthesizing a small molecule compound inhibiting the Wnt/ß-catenin signaling pathway, named LPD-01 (1), and 1 inhibited the growth of human colorectal cancer (HT-29) cells. In this study, we revealed that 1 inhibits the growth of HT-29 cells stronger than that of another human colorectal cancer (SW480) cells. Therefore, we have attempted to identify the target proteins of 1 in HT-29 cells. Firstly, we investigated the effect on the expression levels of the Wnt/ß-catenin signaling pathway-related proteins. As a result, 1 inhibited the expression of target proteins of Wnt/ß-catenin signaling pathway (c-Myc and Survivin) and their genes, whereas the amount of transcriptional co-activator (ß-catenin) was not decreased, suggesting that 1 inhibited the Wnt/ß-catenin signaling pathway without affecting ß-catenin. Next, we investigated the target proteins of 1 using magnetic FG beads. Chemical pull-down assay combined with mass spectrometry suggested that 1 directly binds to importin7. As expected, 1 inhibited the nuclear translocation of importin7 cargoes such as Smad2 and Smad3 in TGF-ß-stimulated HT-29 cells. In addition, the knockdown of importin7 by siRNA reduced the expression of target genes of Wnt/ß-catenin signaling pathway. These results suggest that importin7 is one of the target proteins of 1 for inhibition of the Wnt/ß-catenin signaling pathway.

In vivo examination of the genotoxicity of the urban air and surface soil pollutant, 3,6-dinitrobenzo[e]pyrene, with intraperitoneal and intratracheal administration.

3,6-Dinitrobenzo[e]pyrene (3,6-DNBeP) was identified as a new potent mutagen toward Salmonella strains in surface soil and airborne particles. Because data of in vivo examination of the genotoxicity of 3,6-DNBeP are limited, micronucleus test was performed in peripheral blood and bone marrow, and comet assay in the lungs of mice treated with 3,6-DNBeP. In male ICR mice intraperitoneally (i.p.) injected with 3,6-DNBeP, the frequency of micronuclated polychromatic erythrocytes (MNPCEs) was increased in the peripheral blood and bone marrow after 24 h in a dose-dependent manner. Compared to controls, the highest dose of 3,6-DNBeP (40 mg/kg B.W.) induced 7.3- and 8.7-fold increases of MNPCE frequency in the peripheral blood and bone marrow, respectively. Furthermore, when 3,6-DNBeP was intratracheally (i.t.) instilled to male ICR mice, 3,6-DNBeP at the highest dose of 0.1 mg/kg body exhibited 3.1-fold increase of DNA tail moment in the lungs at 3 h after the instillation compared to controls. The values of DNA tail moment at 9 and 24 h after the instillation were increased up to 3.5 and 4.2-fold, respectively. These data indicate that 3,6-DNBeP is genotoxic to mammalians in in vivo and suggest that 3,6-DNBeP may be a carcinogenic compound present in the human environment.

Azaphilones produced by Penicillium maximae with their cell death-inducing activity on Adriamycin-treated cancer cell.

BACKGROUND: Heat shock proteins (Hsps) are overexpressed in several tumors and contribute to cell proliferation, metastasis, and anticancer drug resistance. Therefore, Hsp inhibitors have enhanced cytotoxicity as chemotherapeutic agents and may be effective with a reduced dosage for tumor therapy to avoid side effects. RESULTS: Four new azaphilones, maximazaphilones I-IV (1-4), and three known compounds (5-7) have been isolated from the airborne-derived fungus Penicillium maximae. Inhibitory effects of isolated compounds against induction of Hsp105 were evaluated by the luciferase assay system using Hsp105 promoter. In this assay, 2-4, 6, and 7 significantly inhibited hsp105 promoter activity without cytotoxicity. In addition, all isolated compounds except for 5 significantly induced the death of Adriamycin (ADR)-treated HeLa cells. Interestingly, 1-4, 6, and 7 didn't show anti-proliferative and cell death-inducing activity without ADR. CONCLUSION: This study revealed the chemical structures of maximazaphilones I-IV (1-4) and the potency of azaphilones may be useful for cancer treatment and reducing the dose of anticancer agents. In addition, one of the mechanisms of cell death-inducing activity for 2-4, 6, and 7 was suggested to be inhibitory effects of Hsp105 expression.

Chemical structures and induction of cell death via heat shock protein inhibition of the prenylated phloroglucinol derivatives isolated from Hypericum erectum.

Four new prenylated phloroglucinol derivatives (+)-erectumol I (1a), (-)-erectumol I (1b), (-)-erectumol II (2a), and (+)-erectumol II (2b) were isolated from the methanol extracts of the whole plants of Hypericum erectum. These new compounds were isolated as a pair of enantiomers, respectively. The planar chemical structures and relative configurations of the new compounds were suggested by Cu-Kα X-ray diffraction analysis and been confirmed by high-resolution mass and 1D and 2D NMR spectroscopic data. The absolute configuration of the four new compounds were established by comparing the experimental and predicted electronic circular dichroism data. Isolated compounds 1b and 2b induced death of Adriamycin-treated HeLa cells. Their enantiomers 1a and 2a did not. In addition, the apparent mechanism of cell death of 1b was the inhibited expression of heat shock protein 105.