Inhibitory effects of (-)-epigallocatechin gallate on the mutation,DNA strand cleavage,and DNA adduct formation by heterocyclic amines

Green tea is known to be a potential chemopreventive agent against cancer. In this study, we investigated the inhibitory activities of tea extracts, and in particular the polyphenolic component (-)-epigallocatechin gallate (EGCG), against heterocyclic amine-induced genotoxicity. The tea extracts displayed inhibition of 2-hydroxyamino-6-methyldipyrido[1,2-a,3',2'-d]imidazole (Glu-P-1(NHOH))-induced mutagenicity. This inhibition can be accounted for by the presence of EGCG in the extracts. The mutagenic effect of Glu-P-1(NHOH), which induces single-strand cleavage in supercoiled circular DNA under neutral conditions, was inhibited by EGCG. Using the Drosophila repair test, a test for gross DNA damage, and DNA adduct detection by (32)P-postlabeling, we showed that EGCG prevented 2-amino-3,8-dimethylimidazo[4,5-f]quinoline-induced DNA damage and adduct formation in insect DNA. EGCG was found to accelerate the degradation of Glu-P-1(NHOH) in vitro. This observation suggested that the inhibition by EGCG is associated with an accelerated degradation of metabolically activated heterocyclic amines.     

农杆菌T-DNA 转化桑树的表型分析

用自然杂交桑种子培育的3月龄幼年桑(Moru salba L．)作材料，以野生型根癌农杆菌(Agrobacterium tunefaciens）的突变体M-21介导，将根癌农杆菌T-DNA导入桑树基因组中。将幼年桑摘芯后用细针对腋芽分生组织刺伤感染获得4个形态异常的株系。PCR检测表明：4个株系都有T-DNA插入。将当代(T0)进行扦插繁殖获得T1代，T1表现出与T0不同的性状，显然T0是由正常产生生长素及细胞分裂素的未转化部分(第10叶位以下)和产生高水平细胞分裂素的转化部分共同作用决定的嵌合体表型，而T1是T0新梢的继续发育体，扦插后，它脱离了T0的作用。T0和T1的表型差异可能与T-DNA的插入有关。     

Epigenetic dynamics of imprinted X inactivation during early mouse development

The initiation of X-chromosome inactivation is thought to be tightly correlated with early differentiation events during mouse development. Here, we show that although initially active, the paternal X chromosome undergoes imprinted inactivation from the cleavage stages, well before cellular differentiation. A reversal of the inactive state, with a loss of epigenetic marks such as histone modifications and polycomb proteins, subsequently occurs in cells of the inner cell mass (ICM), which give rise to the embryo-proper in which random X inactivation is known to occur. This reveals the remarkable plasticity of the X-inactivation process during preimplantation development and underlines the importance of the ICM in global reprogramming of epigenetic marks in the early embryo.     

Oxidative stress and antioxidant responses in juvenile Brazilian flounder <Emphasis Type="Italic">Paralichthys orbignyanus</Emphasis> exposed to sublethal levels of nitrite

This study evaluated the effects of short-term exposure to sublethal levels of nitrite on oxidative stress parameters and histology of juvenile Brazilian flounder Paralichthys orbignyanus. An assessment of fish recovery was also performed. Fish were exposed to 0.08 (control), 5.72, 10.43, and 15.27 NO₂-N mg L^?1 for 10 days followed by the same recovery time. Gill, liver, and muscle samples were collected after 1, 5, and 10 days of exposure and after recovery for the measurement of antioxidant capacity against peroxyl radicals (ACAP), glutathione-S-transferase (GST) activity, content of non-protein (NPSH) and protein thiols (PSH), and lipid peroxidation levels by thiobarbituric acid-reactive substances (TBARS) content. Nitrite exposure induced alterations which compromised the overall antioxidant system (reduced ACAP and GST activity) and enhanced oxidative damage in lipids and proteins. Increases in GST activity and NPSH and PSH contents were also demonstrated. The recovery period allowed for resumption of basal levels for all (treatment 5.72 NO₂-N mg L^?1) or some of the evaluated parameters (other treatments). In conclusion, exposure to nitrite concentrations from 5.72 to 15.27 NO₂-N mg L^?1 induced oxidative stress and antioxidant responses in juvenile Brazilian flounder. The 10-day recovery period was sufficient for a complete resumption of basal physiological condition of fish exposed to concentrations of up to 5.72 NO₂-N mg L^?1.     

Natural variation in photoperiodic flowering pathway and identification of photoperiod-insensitive accessions in wild wheat, <Emphasis Type="Italic">Aegilops tauschii</Emphasis>

The D-genome progenitor of hexaploid wheat, Aegilops tauschii Coss., has a wide natural species range in central Eurasia and possesses wide natural variation in heading and flowering time. Here, we report identification of two Ae. tauschii accessions insensitive to short day length. Similarly to a loss or reduced degree of vernalization requirement, the photoperiod-insensitive mutations were found only in the early flowering sublineage (TauL1b) of Ae. tauschii. Quantitative trait locus (QTL) analyses using two F₂ mapping populations showed that a QTL for heading time on the long arm of chromosome 5D was related to the early heading phenotype of the photoperiod-insensitive accessions under short-day conditions. In the photoperiod-insensitive accession, expression patterns of two flowering-related genes were altered under short-day conditions compared with the patterns in photoperiod-sensitive accessions. This study indicates that analysis of natural variations in the Ae. tauschii population is useful to find novel genetic loci controlling agronomically important traits.     

Interannual decrease in condition factor of the western sand lance Ammodytes japonicus in Japan in the last decade: Evidence for food‐limited decline of the catch

Annual catch of the western sand lance Ammodytes japonicus in the eastern Seto Inland Sea, Japan, has shown a decreasing trend since the 1990s. To examine whether food shortage was the main cause for the catch decrease, we investigated the condition factor of the age‐0 fish at the beginning of the estivation period (late July) in Harima‐Nada, eastern Seto Inland Sea, for 10 years from 2008 to 2017. The mean abundance of copepods as food for the age‐0 fish during the fish growth period (from February to June) around the estivation area was also determined. The condition factor showed a significant decrease, and values for 2011 and later years were mostly lower than the known minimum threshold (4.2) for maturation. In the recent 4 years from 2013 to 2016, the mean copepod abundance was much lower than values for the other years. The condition factor showed a significant positive correlation with the copepod abundance. These results indicate that decline of western sand lance catch after 1990 was caused mainly by food shortage.     

In situ growth and mortality of juvenile Acropora over 2 years following mass spawning in Sekisei Lagoon,Okinawa (24°N)

The growth of Acropora in Sekisei Lagoon was investigated in situ using ceramic coral settlement devices (CSDs) and marine blocks (MBs) with small holes on their surfaces that were deployed prior to a mass spawning event. Acropora that had settled in 10-mm holes on the upper surface of the MBs were found to have grown inwards after 348 days post-mass spawning. After 733 days, the colonies had grown outwards, encrusting the blocks and attaining a maximum diameter of 21.5 mm (3.7 mm). CSDs that had been stacked above each other on frames were separated and observed in situ on 405 and 764 days after mass spawning. After 405 days, the maximum diameter of the encrusting Acropora was 7.6 mm (±2.4 mm), which increased to 19.4 mm (±5.5 mm) after 764 days, with 30% of colonies extending short branches. The length of the branches of Acropora grown on the CSDs fixed to the MBs 3 months after the mass spawning event exceeded the size of the encrusting portion of the colony 629 days after spawning. The mortality of colonies between 1 and 2 years was slight, with the temperatures conducive to coral bleaching that occurred during the study also having only a slight effect.     

Purification and characterization of two novel cysteine protease inhibitors, Eel-CPI-2 and Eel-CPI-3, in the skin mucus of the Japanese eel Anguilla japonica

We have discovered multiple acidic cysteine protease inhibitors, in addition to the known Eel-CPI-1, in the skin mucus extract of the Japanese eel Anguilla japonica by using the two-dimensional gel system of gelatin reverse zymography. Two of the acidic inhibitors, which we have named Eel-CPI-2 and Eel-CPI-3, were purified to homogeneity by anion exchange chromatography on a column of DEAE-Sepharose CL-6B, followed by fast protein liquid chromatography on Superdex 75 10/300 GL and HiTrap Q HP columns. The amino acid compositions of Eel-CPI-2 and Eel-CPI-3 were found to be almost identical and closely similar to that of the eel galectin AJL1. The molecular masses of Eel-CPI-2 and Eel-CPI-3 were elucidated to be 16,089.080 and 16,089.137 Da, respectively, by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The apparent dissociation constant of Eel-CPI-2 and Eel-CPI-3 for cysteine protease papain was determined to be 1.79 × 10⁻⁷ and 1.05 × 10⁻⁷ M, respectively, by a quartz crystal microbalance technique.