Epicatechin and catechin in cocoa inhibit amyloid beta protein induced apoptosis

To elucidate additional health benefits of cocoa phytochemicals on the neurotoxicity induced by amyloid beta protein (Abeta), PC12 cells were treated with toxic peptide (Abeta(25)(-)(35)) and the effects of epicatechin, catechin, and cocoa were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction, lactate dehydrogenase (LDH) release, and trypan blue exclusion methods. Significant increase in neuronal cell death was observed on PC12 cells treated with Abeta(25)(-)(35) (25 microM), while epicatechin and catechin and their mixture prevented the Abeta-induced neuronal cell death. Abeta treatment also led to the increased membrane instability of PC12 cells. The membrane protective effects of the phenolics determined by LDH release and trypan blue exclusion assays demonstrated that epicatechin, catechin, and their mixture protect cellular membrane from Abeta-induced cytotoxicity. In these three different cell viability assays, the mixture of epicatechin and catechin showed the highest protective effect and synergistic activity. The present results showed that the major flavonoids of cocoa, epicatechin and catechin, protect PC12 cells from Abeta-induced neurotoxicity, and suggest that cocoa may have anti-neurodegenerative effect in addition to other known chemopreventive effects.     

Nutrient availability and response of sago palm (Metroxylon sago Rottb.) to controlled release N fertilizer on coastal lowland peat in the tropics

Sago palms (Metroxylon sagu Rottb.) growing on peat soils were found to grow more slowly and to show a lower production than palms growing on mineral soils. This difference was related to the physical and chemical constraints of peat soils, which include low bulk density, high acidity, and low N, P, K, Ca, Zn, and Cu levels. In coastal lowland peat soils, the distance from the sea has been found to be an important determinant of soil elemental composition. We predicted that a sufficient supply of N at the rosette stage would improve sago palm growth and that the availability of N in soil to which controlled release N fertilizer was applied might be higher than that in soil treated with soluble fertilizer. To investigate the changes in the nutrient composition of peat soils at various distances from the sea and the effect on sago palm growth, we studied sago palm areas in Indonesia and Malaysia. To observe the influence of N on the growth performance, we also conducted a fertilizer experiment on coastal lowland peat soil in Indonesia. Distance from the sea had no significant effect on the cation concentration in the soil solution (with the exception of Mg) or on the levels of soil-exchangeable cations. No significant differences were observed between the concentrations of exchangeable cations in surface peat soils and those in mature leaves. However, the concentrations of K, Na, and Ca in mature leaves increased significantly with their concentrations in the soil solution. This finding implies that the concentrations of cations in sago palm leaves depend directly on the concentrations of cations in the soil solution. No significant effect of N fertilizers on plant height and leaf formation was observed. N fertilizers applied twice a year did not affect appreciably the foliar concentration of N determined in December 1998 (5 months after the initial application) and December 1999. In June 2000, we detected a significantly higher concentration of N (p < 0.01) in young leaves of the palms treated with LP-100 or urea than in control leaves. However, no significant difference was detected between the LP-100 and urea treatments in the concentration of N in both mature and young leaves. This finding indicated that the concentration of N in sago palm leaves increased with the level of soil-applied N, regardless of whether N was applied as controlled release fertilizer or in the soluble form. We anticipate that a significant difference in the effects of these N fertilizers may occur during the next rainy season, when there should be a considerable loss of soluble N.     

Induction of apoptosis by garcinol and curcumin through cytochrome c release and activation of caspases in human leukemia HL-60 cells

Garcinol, a polyisoprenylated benzophenone, was purified from Garcinia indica fruit rind. The effects of garcinol and curcumin on cell viability in human leukemia HL-60 cells were investigated. Garcinol and curcumin displayed strong growth inhibitory effects against human leukemia HL-60 cells, with estimated IC(50) values of 9.42 and 19.5 microM, respectively. Garcinol was able to induce apoptosis in a concentration- and time-dependent manner; however, curcumin was less effective. Treatment with garcinol caused induction of caspase-3/CPP32 activity in a dose- and time-dependent manner, but not caspase-1 activity, and induced the degradation of poly(ADP-ribose) polymerase (PARP). Pretreatment with caspase-3 inhibitor inhibited garcinol-induced DNA fragmentation. Treatment with garcinol (20 microM) caused a rapid loss of mitochondrial transmembrane potential, release of mitochondrial cytochrome c into cytosol, and subsequent induction of procaspase-9 processing. The cleavage of D4-GDI, an abundant hematopoietic cell GDP dissociation inhibitor for the Ras-related Rho family GTPases, occurred simultaneously with the activation of caspase-3 but preceded DNA fragmentation and the morphological changes associated with apoptotic cell death. Of these, Bcl-2, Bad, and Bax were studied. The level of expression of Bcl-2 slightly decreased, while the levels of Bad and Bax were dramatically increased in cells treated with garcinol. These results indicate that garcinol allows caspase-activated deoxyribonuclease to enter the nucleus and degrade chromosomal DNA and induces DFF-45 (DNA fragmentation factor) degradation. It is suggested that garcinol-induced apoptosis is triggered by the release of cytochrome c into the cytosol, procaspase-9 processing, activation of caspase-3 and caspase-2, degradation of PARP, and DNA fragmentation caused by the caspase-activated deoxyribonuclease through the digestion of DFF-45. The induction of apoptosis by garcinol may provide a pivotal mechanism for its cancer chemopreventive action.     

Novel polyphenol molecule isolated from licorice root (Glycrrhiza glabra) induces apoptosis, G2/M cell cycle arrest, and Bcl-2 phosphorylation in tumor cell lines

Herbal therapies are commonly used by patients with cancer, despite little understanding about biologically active chemical derivatives. We recently demonstrated that the herbal combination PC-SPES, which contains licorice root, had anti-prostate cancer activity attributable to estrogen(s) that produced a chemical castration. A recent study also demonstrated that licorice root alone decreased circulating testosterone in men. Other studies demonstrated antitumor activity of PC-SPES in vitro associated with decreased expression of the anti-apoptotic protein Bcl-2 and in patients independent of chemical castration, suggesting that other mechanisms of antitumor activity exist separate from chemical castration. In the present study, we assessed licorice root extract for effects on Bcl-2 to identify novel cytotoxic derivatives. Licorice root extract induced Bcl-2 phosphorylation as demonstrated by immunoblot and G2/M cell cycle arrest, similarly to clinically used antimicrotubule agents such as paclitaxel. Bioassay-directed fractionations resulted in a biologically active fraction for Bcl-2 phosphorylation. HPLC separation followed by mass spectrometry and NMR identified 6 compounds. Only one molecule was responsible for Bcl-2 phosphorylation; it was identified as 1-(2,4-dihydroxyphenyl)-3-hydroxy-3-(4'-hydroxyphenyl) 1-propanone (beta-hydroxy-DHP). The effect on Bcl-2 was structure specific, because alpha-hydroxy-DHP, 1-(2,4-dihydroxyphenyl)-2-hydroxy-3-(4'-hydroxyphenyl) 1-propanone, in contrast to beta-hydroxy-DHP, was not capable of Bcl-2 phosphorylation. Pure beta-hydroxy-DHP induced Bcl-2 phosphorylation in breast and prostate tumor cells, G2/M cell cycle arrest, apoptosis demonstrated by Annexin V and TUNEL assay, decreased cell viability demonstrated by a tetrazolium (MTT) assay, and altered microtubule structure. Therefore, these data demonstrate that licorice root contains beta-hydroxy-DHP, which induced Bcl-2 phosphorylation, apoptosis, and G2/M cell cycle arrest, in breast and prostate tumor cells, similarly to the action of more complex (MW >800) antimicrotubule agents used clinically.     

Midbody targeting of the ESCRT machinery by a noncanonical coiled coil in CEP55

The ESCRT (endosomal sorting complex required for transport) machinery is required for the scission of membrane necks in processes including the budding of HIV-1 and cytokinesis. An essential step in cytokinesis is recruitment of the ESCRT-I complex and the ESCRT-associated protein ALIX to the midbody (the structure that tethers two daughter cells) by the protein CEP55. Biochemical experiments show that peptides from ALIX and the ESCRT-I subunit TSG101 compete for binding to the ESCRT and ALIX-binding region (EABR) of CEP55. We solved the crystal structure of EABR bound to an ALIX peptide at a resolution of 2.0 angstroms. The structure shows that EABR forms an aberrant dimeric parallel coiled coil. Bulky and charged residues at the interface of the two central heptad repeats create asymmetry and a single binding site for an ALIX or TSG101 peptide. Both ALIX and ESCRT-I are required for cytokinesis, which suggests that multiple CEP55 dimers are required for function.     

Flowering estimation in apple orchards by image analysis

Tree-specific management practice related to individual tree physiological condition is necessary for higher quality and quantity in apple fruit production. Detection of apple flowering abundance based on analysis of HSL (hue, saturation, luminance) images was used to estimate the number of flower clusters (FC) of individual trees in a high density apple orchard. The image acquisition was performed with a still camera and an industrial color camera during the day and night. The FC estimation algorithm included HSL thresholding with parameter optimization. Three hypothetical, tree-specific management practices (sprayings) were assumed, using >25, >50 and >100 FC thresholds to carry out the practice. When an industrial camera was used for image acquisition during the daytime and hypothetical spraying was done by on/off criterion >100 FC per tree, 10 % incorrect executions were identified. Comparable FC counting performance was achieved by using a still camera or an industrial camera.     

埋地管道交流腐蚀的实验室评价方法

针对长期以来埋地管道交流腐蚀评价只能依赖现场测取的交直流参数且准确性欠佳的问题,提出了一种埋地管道交流干扰腐蚀的实验室评价方法.通过现场测取实际管道的交流干扰信号,在实验室使用取自管道沿线的土样进行大范围交流腐蚀模拟实验,评价埋地管道在现场交流干扰条件下的真实腐蚀风险.研究结果表明:采用对称电路和数字信号处理技术,可以实现在实验室内精确模拟实际管道在交流腐蚀和阴极保护条件下的腐蚀速率;通过覆盖现场测量数据的交流干扰腐蚀失重实验,可以最大程度地在实验室内实现埋地管道交流干扰的腐蚀风险评价与表征.     

Preparation and characterization of high molecular weight poly(trimethylene terephthalate) by solid-state polymerization

Solid-state polymerization of poly(trimethylene terephthalate)(PTT) was carried out to obtain high molecular weight polymers. Two kinds of commercial PTT chips were polymerized in the solid state by the heat treatment at 190∼220°C for various times and they were characterized by end group content, molecular weight, thermal analysis, and X-ray diffraction. In the solid-state polymerization of PTT, the overall reaction rate was governed by the solid-state polymerization temperature and time, and pellet size. The content of carboxyl end groups decreased during the solid-state polymerization with increasing solid-state polymerization temperature and time. The melting temperature and crystallinity of the PTT were higher for the ones treated at higher temperature and longer time. The activation energy for the solid-state polymerization of PTT was in the range of 24∼25 kcal/mol for both chips. Through the solid-state polymerization of commercial PTT chips, high molecular weight polymers up to an intrinsic viscosity of 1.63 dl/g was obtained, which corresponded to about a 117,000 weight-average molecular weight.