Paeonol promotes arylamine N‐acetyltransferase activity in human bladder tumor cells

Paeonol was used to determine effects of arylamine N‐acetyltransferase (NAT) activity in human bladder tumor cells. The NAT activity was measured by high performance liquid chromatography assaying for the amounts of N‐acetyl‐2‐aminofluorene (2‐AAF) and N‐acetyl‐p‐amino‐benzoic acid (N‐Ac‐PABA) and remaining 2‐aminofluorene (2‐AF) and p‐aminobenzoic acid (PABA). The NAT activity in human bladder tumor cells was promoted by paeonol in a dose‐dependent manner, i.e. the higher the concentration of paeonol, the higher the promotion of NAT activity. The data also indicated that paeonol increases the apparent values of K_m and K_max from human bladder tumor cells in cytosols and intact cells. This report is the first demonstration to show paeonol did promote human bladder tumor cell NAT activity.     

Gene expression profiling of human liver carcinoma (HepG2) cells exposed to the marine toxin okadaic acid

The marine toxin, okadaic acid (OA) is produced by dinoflagellates of the genera Prorocentrum and Dinophysis and is the causative agent of the syndrome known as diarrheic shellfish poisoning. In addition, OA acts as both a tumor promoter, attributed to OA-induced inhibition of protein phosphatases as well as an inducer of apoptosis. To better understand the potentially divergent toxicological profile of OA, the concentration-dependent cytotoxicity and alterations in gene expression on the human liver tumor cell line HepG2 upon OA exposure were determined using RNA microarrays, DNA fragmentation, and cell proliferation assays as well as determinations of cell detachment and cell death in different concentrations of OA. mRNA expression was quantified for approximately 15,000 genes. Cell attachment and proliferation were both negatively correlated with OA concentration. Detached cells displayed necrotic DNA signatures but apoptosis also was broadly observed. Data suggest that OA has a concentration dependent effect on cell cycle, which might explain the divergent effects that at low concentration OA stimulates genes involved in the cell cycle and at high concentrations it stimulates apoptosis.     

Flavin-containing monooxygenase activity in the gumboot chitonCryptochiton stelleri

Flavin-containing monooxygenase (FMO) activity was observed in microsomes from the digestive gland of the gumboot chitonCryptochiton stelleri. Digestive-gland microsomal oxygen uptake was initiated after the addition of N,N-dimethylaniline (DMA), methimazole, phenylhydrazine (PH), 2-aminofluorene (2-AF), or 1,1-dimethylhydrazine (DMH) to the incubations. The pH optimum for 2-AF dependent oxygen uptake was 8.4 and required NADPH. The formation of DMA N-oxide and the oxidation of methimazole was measured and confirmed the presence of FMO activity in this tissue.     

Effects of ellagic acid on arylamine N‐acetyltransferase activity in human colon tumor cells

The inhibition of arylamine N‐acetyltransferase (NAT) activity by ellagic acid was determined in a human colon tumor (adenocarcinoma) cell line. Two assay systems were performed, one with cellular cytosols (9000g supernatant), the other with intact colon tumor cell suspensions. The NAT activity in a human colon tumor cell line was inhibited by ellagic acid in a dose‐dependent manner in both types of examined systems: i.e. the greater the concentration of ellagic acid in the reaction, the greater the inhibition of NAT activities in both systems. The data also indicated that ellagic acid decreased the apparent K _m and V _max of NAT enzymes from human colon tumor cells in both the systems examined. This report is the first demonstration which showed ellagic acid affect human colon tumor cell NAT activity.     

Epigenetic effects of N-methyl-N′-nitro-N-nitrosoguanidine on Kazakh esophageal epithelial cells

Epigenetic change is a key factor for esophageal cancer progression. This study shows that upon exposure of primary cultures of human esophageal epithelial cells obtained from people of the Kazakh nationality in China to N-methyl-N′-nitro-N-nitrosoguanidine at concentrations of 0.75, 1.50, and 3.00 mg/L, (1) cell proliferation was inhibited, the percentages of cells in the G₀ and G₁ phases declined, and those in the S and G₂ + M phases increased at all concentrations, while apoptosis rates increased at medium and high concentrations; (2) the content of DNA-methyltransferases 3a and 3b and the activities of DNA-methyltransferase 1 and DNA-methyltransferase 3a increased at all concentrations while the content of DNA-methyltransferase 1 and the activity of DNA-methyltransferase 3b were increased only at the high concentration; (3) messenger RNA and protein levels of DNA-methyltransferases 1, 3a, and 3b were increased at all concentrations. Obviously, N-methyl-N′-nitro-N-nitrosoguanidine in the low mg/L range can inhibit the proliferation of normal Kazakh esophageal epithelial cells, can cause cell cycle arrest in the S and G₂ + M phases, promote apoptosis, and increase the activities of DNA-methyltransferases 1, 3a, and 3b.     

Cytotoxicity of single-walled carbon nanotubes,multi-walled carbon nanotubes,and chrysotile to human lung epithelial cells

Carbon nanotubes (CNTs) have found numerous applications in various industries. Recently, adverse effects of these materials on human and animal cells in vitro have been reported. In the present study, the cytotoxicity of single-walled carbon nanotubes (SWCNTs), multi-walled carbon nanotubes (MWCNTs), and chrysotile asbestos in human lung epithelial cells has been studied using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The cells were exposed for 6 h and 24 h to between 0.97 and 1500 μg mL^?1 of CNTs and chrysotile fibers prepared in two culture media containing 5% serum and 0.5% dimethylsulfoxide. Dose–response curves were obtained to determine the nonobservable adverse effect concentration and the half-maximum inhibitory concentration (IC₅₀). The way of dispersion affects the cytotoxicity of CNTs. For MWCNT, the toxicological indexes were lower than for SWCNT. Chrysotile fibers were even less cytotoxic than CNTs. Therefore, workplace control measures are recommended as priority for occupational and environmental conditions.     

Nitrogen fixation (acetylene reduction) in the phyllosphere of Thalassia testudinum

N₂ fixation (C₂H₂ reduction) associated with the leaves of the sea grass Thalassia testudinum was investigated at 5 sites in South Florida (Biscayne Bay) and one site in the Bahamas (Bimini Harbor). Significant activities were correlated with the occurrence of a heterocystous blue-green alga (Calothrix sp.) on the leaves. C₂H₂ reduction was not stimulated by organic compounds, either aerobically or anaerobically in the light or dark. Therefore, other physiological types of microbes were not important in N₂ fixation. Diurnal and seasonal variations in N₂ fixation occurred, with maximal rates during the daytime and in the late spring and early summer. N₂ fixation was negligible at four stations in Biscayne Bay. At the fifth station, near Fowey Rock, about 5 kg N ha^-1 year^-1 was fixed. In the summer, the N₂ fixed per day (4–5 mg N m^-2) could provide 4 to 23% of the foliar productivity demands of T. testudinum at this site and the station in Bimini Harbor. N₂ fixation at the periphery of a sea-grass patch, near Fowey Rock, could provide 8 to 38% of the daily nitrogen requirement for leaf production, and thereby might compensate for a less effective trapping and recycling of nitrogen from dead leaves in such regions.     

Toxicological effects of N,N,N′,N′-tetramethylethylenediamine on electric eel (Electrophorus electricus) acetylcholinesterase and human serum butyrylcholinesterase

This study examined the adverse effects of N,N,N′,N′-tetramethylethylenediamine (TEMED) on electric eel (Electrophorus electricus) acetylcholinesterase (AChE) and human serum butyrylcholinesterase (BChE). Data show that TEMED inhibited AChE in electric eel as well as human serum BChE. Kinetic studies indicated that the inhibition produced by TEMED in both sources was of mixed type, i.e. K_m increased and V _max decreased in a concentration-dependent manner. K_I (constant of ChE–substrate–TEMED complex into ChE–substrate complex and TEMED) was estimated to be 0.674 mM for electric eel and 0.024 mM for human serum BChE. The γK_m (dissociation constant of ChE–substrate–TEMED complex into ChE–TEMED complex and substrate) was 0.083 and 0.2 mM for electric eel AChE and human serum BChE, respectively. The IC₅₀ for electric eel and for human serum ChE was 1.57 and 0.043 mM, respectively. The present results suggest that TEMED produced adverse effects on electric eel and human serum via inhibition of ChE.     

Epigenetic effect of long-term bisphenol A exposure on human breast adenocarcinoma cells

In this research, epigenetic effects of bisphenol A (BPA) on human breast cancer MCF-7 cells were analyzed. Genome-wide DNA methylation and gene expression were analyzed in MCF-7 cells exposed to BPA (10^?5 and 10^?6 mol/L for 5 weeks). No significant changes in the global level of 5-methyl-2′-deoxycytidine and 5-hydroxymethyl-2′-deoxycytidine were observed. DNA methylation profiling analysis indicated that BPA exposure resulted in the hypermethylation of FOXK2, LKB1, LMX1A and CUGBP2 and the hypomethylation of PTPRN2, TRIM27, BCAS3 and ZNF423. Decreased expression of apoptosis genes (P38 and BCL2L1) and increased expression of chemokine (Cxcl2 and ccl20) were detected. Changes of these genes were speculated to affect the ERα-related cell growth as well as cell apoptosis.     

Bioavailability and metabolism of hexavalent chromium compounds †

Intratracheal instillation of ⁵¹CrCl₃ in anaesthetized rabbits resulted in partial absorption. In blood, the absorbed material was entirely confined to the plasma compartment. Only trace amounts were deposited in liver and kidney. By contrast, after similar application of Na, ⁵¹CrO₄ the bulk of blood radioactivity was present in red blood cells (RBC). Substantial deposition occurred in liver and kidneys. It is concluded that Cr(VI) may enter the body unreduced via the lung and is partially deposited in cells over a prolonged period of time.

Since chromium was accumulated in liver after administration of Cr(VI) we investigated the intracellular disposition of Cr(VI) in the isolated perfused liver. No significant sex differences in chromium distribution were observed. At the end of the experiments (1 h), 60% of the applied dose (312μg Cr/liver) was located in the cytosol, whilst 14% was in the mitochondria, 9% in the microsomal pellet and 2% was associated with the nuclei. Gel chromatography of the cytosolic compartment showed that the overwhelming part of chromium was eluted in fractions with an apparent molecular weight of 6,000 dalton. These fractions exhibited absorption maxima at 410nm and 548nm. It is concluded, that cytosolic reduction might be the main intracellular redox pathway for chromates. This view was confirmed by monitoring the reaction of Cr(VI) with GSH in vitro. GSH reduced Cr(VI) without further cofactors under formation of GSH‐chromium complexes, which possibly represent major intermediates in the metabolism of Cr(VI).     

Effect of UV-B radiation on the marine diatoms Lauderia annulata and Thalassiorsira rotula grown in different salinities

The marine diatoms Lauderia annulata Cleve and Thalassiosira rotula Meunier were grown at different salinities (20, 35 and 45) and exposed to different levels of midultraviolet, UV-B) 439, 717 and 1230 J m^-2 d^-1, weighted) for 2 d. A low UV-B dose (439 J m^-2 d^-1) usually caused a slight increase in biomass production (dry weight) compared to non-UV-B irradiated cells. Enhanced UV-B radiation (717 J m^-2 d^-1) depressed protein and pigment content (chlorophyll a, chlorophyll c ₁+c₂ and carotenoids), especially in algae grown at 20 or 35 salt concentration of the nutrient solution. The effect of UV-B radiation (717 J m^-2 d^-1) on the pattern and concentration of amino acids was species-dependent. Aspartic acid was reduced in all tested diatoms. A drastic increase in glutamine and a reduction in glutamic acid pools could be observed in L. annulata samples, but no significant variation of the impact of UV-B was found in dependence on the salt concentration of the nutrient medium. T. rotula cells grown at 35 S showed an increase of glutamic acid and a decrease of glutamine levels after UV-B radiation. The results are discussed in relation to the impact of UV-B upon carbon and nitrogen metabolism.     

DNA damage in mouse organs and in human sperm cells by bisphenol A

Abstract

The in vivo genotoxic potential of bisphenol A using the comet assay in mice and in human sperm cells in vitro without metabolizing enzymes was studied. Male mice were exposed by oral gavage to the following doses of bisphenol A (0 125, 250 and 500?mg/kg body weight). DNA damage was investigated in liver, kidney, testes, urinary bladder, colon and lungs cells. In testicular cells, a significant increase in DNA strand breaks was observed in the lowest, but not in the medium or highest dose groups. Histopathological investigation of the testicular samples did not show any treatment dose-related effects. No DNA strand breaks were observed in any of the other investigated tissues. In human sperm cells in vitro, bisphenol A did not induce DNA strand breaks.     

溴阻燃剂对肝细胞的毒性

为评价四溴联苯醚(2,2,4,4-tetrabromodipheny lether,PBDE-47)、五溴联苯醚(2,2',4,4',5-pentabromodiphenylether,PBDE-99)、九溴联苯醚(polybrominated dipheny lethers206,PBDE-206)、六溴环十二烷(hexabromocyclododecane,HBCD)、四溴双酚A(tetrabromobisphenol A,TBBPA)对人肝细胞(LO2)的体外细胞毒性。将5种溴阻燃剂与人肝细胞LO2共同培养,采用四甲基偶氮噻唑蓝比色法(MTT法)检测5种溴阻燃剂对LO2细胞的相对增殖率,并按照GB/T16886.5-2003/ISO10993-5:1999标准评价5种溴阻燃剂对LO2的细胞毒性。体外实验表明,5种溴阻燃剂对LO2细胞均具有抑制作用,其中PBDE-47、PBDE-99和HBCD呈现剂量依赖关系。     

Effect of nitrification inhibitor on plant biomass and N2O emission rates in alpine meadows on the Tibetan Plateau

ABSTRACT

Treatment with nitrification inhibitors, such as dicyandiamide (DCD) and 3,4-dimethylpyrazole phosphate (DMPP) have been strongly indicated to increase grassland biomass and mitigate soil N₂O emission rates. However, the responses of both alpine meadow aboveground biomass and N₂O emission rates to nitrification inhibitors remains unclear. We separately applied three doses of DCD and DMPP to alpine grassland soils with three duplicates. The biomass and N₂O emission rates were subsequently measured by a clear-cut method and in-situ static chamber gas chromatography during the growing season. Our findings indicated that aboveground biomass increased significantly, and N₂O emission rate decreased significantly at 6.8?kg?ha^?1 DCD and DMPP. Furthermore, the biomass increase effect was more significant than the N₂O emission rate mitigation effect (p?<?0.05). The highest ratios of DCD treatments on meadow production increase and N₂O emission rate decrease were 27.2% and 36.3%, respectively. Our findings provide insight into the enhanced grassland primary production and decreased N₂O flux by nitrification inhibitor treatment in alpine meadows, which may be beneficial to help mitigate global warming.     

Wateriness of white muscle: A comparison between cod (Gadus morhua) and jelly cat (Lycichthys denticulatus)

Histological sections of starved cod (Gadus morhua Linnaeus, 1758) and jelly cats (Lycichthys denticulatus Kröyer, 1844) with similar water contents are compared. Starvation causes the white muscle fibres of cod to shrink into irregular shapes inside their connective tissue matrix, while those of jelly cats of similar water content are seen to be rounded. It appears to be less important for jelly cat muscle to be packed tightly with contractile fibres than for the muscle of cod, so the former may be adapted to a permanent state of unusually high hydration.     

Effects of environmental hypercapnia on hemato-immunological parameters,carbonic anhydrase,and Na+, K+-ATPase enzyme activities in rainbow trout (Oncorhynchus mykiss) tissues

In this study, the effects of environmental hypercapnia on hemato-immunological parameters and the activities of respiratory enzymes such as carbonic anhydrase (CA) and Na⁺, K⁺-ATPase were investigated in rainbow trout (Oncorhynchus mykiss) tissues (gill, liver and kidney). Batches of 12 fish were exposed to 4.5 mg L^?1 (control) and 14 mg L^?1 CO₂. No mortalities occurred during the 14 days of the experimental period. Red blood cell (RBC), hemoglobin (Hb), and hematocrit (Ht) levels, and innate immune parameters such as nitro blue tetrazolium (NBT), lysozyme, and myeloperoxidase activities, and the melano-macrophage frequency were negatively affected by elevated CO₂ levels. Patterns of change in CA activity differed among the gill, liver, and kidney. Compared with the activities of CA in the control group, the CA enzyme was significantly stimulated at day 7 in the gill tissue, whereas it was stimulated at day 14 of the experiment in the liver tissue of fish exposed to 14 mg L^?1 CO₂ (P < 0.05). In contrast to the pattern of CA enzyme activities, the Na⁺, K⁺-ATPase enzymes were stimulated significantly in the liver after day 7 but inhibited in the kidney and gill (P < 0.05). These results suggest that a subchronic exposure to hypercapnia of rainbow trout tissues may lead to adaptive changes in the respiratory enzymes and negatively affects hemato-immunological parameters.     

铜污染草地对放牧乌蒙半细毛羊矿物质元素代谢的影响

为了评价草地铜污染对放牧乌蒙半细毛羊的影响,探讨铜污染的治理方法,在乌蒙山区的妈姑镇和凉水沟开展草地放牧试验和矿物质元素补充研究。应用原子发射光谱分析土壤、牧草和动物组织的重金属含量,应用全自动血液分析仪分析血液指标。放牧试验的结果显示:铜污染草地放牧明显增加了动物组织铜和锌的含量(P 0.01),降低了动物组织钼和铁的含量(P 0.01),试验结束时,试验组动物的血红蛋白(Hb)、红细胞压积(PCV)、红细胞数(RBC)和平均红细胞体积(MCV)显著低于对照组(P 0.01),羊毛产量和长度显著低于对照组(P 0.01)。矿物质元素补充试验的结果显示:补充硫酸钠明显降低了铜污染草地放牧动物血液和肝脏铜元素含量,补充试验结束时,处理Ⅰ组动物血液的Hb、PCV、RBC和MCV显著高于处理Ⅱ组(P 0.01),血液指标达到正常范围。处理Ⅱ组动物血液和肝脏铜元素的含量继续升高,动物组织钼的含量继续降低(P 0.01),Hb、PCV和RBC继续下降,试验结束时出现溶血性贫血,但补充硫对羊毛品质的相关指标没有显著的影响。研究表明,铜污染草地严重影响了乌蒙半细毛羊的矿物质代谢,补充硫酸钠可以部分拮抗铜的毒性。     

The influence of bacterial quorum-sensing inhibitors against the formation of the diatom-biofilm

Quorum-sensing inhibitor (QSI) is one of the most promising and environmentally friendly agents for marine antifouling. In this study, the activities of three kinds of QSIs 3,4-dibromo-2(5)H-furanone, 4-nitropyridine-N-oxide and indole were evaluated on the growth of two marine diatoms Cylinthrotheca sp. and Nitzschia closterium. At the same time, the effects of QSIs on the formation of the diatom-biofilm were also discussed. All the results showed that QSIs significantly inhibited the growth, and the effects depended on the dose and diatom species. The extracellular polymeric substance contents in the diatom-biofilm were significantly reduced by QSIs. However, the contents of polysaccharide in culture mediums were increased, which might result in the destruction of diatom cells. Combined with the results of crystal violet staining-biofilm and images of scanning electron microscopy, it was further demonstrated that QSIs inhibited the biofilm formation of Cylindrotheca sp., and the inhibitory effect of 4-nitropyridine-N-oxide was superior to that of 3,4-dibromo-2(5)H-furanone and indole.     

Spatial variations in the N2O emissions and denitrification potential of riparian buffer strips in a contaminated urban river

Spatial variations in the N₂O emissions and denitrification potential of riparian buffer strips (RBS) in a polluted river were examined. The river received large pollutant inputs from urban runoff and wastewater discharge, resulting in impaired water quality in the river and downstream reservoir. The potential for nitrogen removal by RBS was evaluated by measuring in situ N₂O emission fluxes in static closed chambers and sediment denitrification potentials with acetylene inhibition techniques. The results showed that N₂O emission fluxes decreased from the upstream (16.39 μg/(m²·h)) to downstream (0.30 μg/(m²·h)) sites and from the water body to upland sites. The trend in decreasing N₂O emission fluxes in the downstream direction was mainly associated with sediment/soil textures (clay loam→sandy soil) and sediment/soil water contents and was also related to the vegetation along the RBS and nutrients in the sediments/soils. The correlation coefficient was highest (r=0.769) between the N₂O emission flux and sediment/soil water content. Sediment/soil denitrification potentials under N-amended and ambient conditions were higher (highest 32.86 mg/(kg·h)) for the upstream sites, which were consistent with in situ N₂O flux rates.     

Physiologische Anpassungen eines marinen insekts. II. Die Eigenschaften von schwimmenden und absinkenden Eigelegen

The gelatinous egg-masses of Clunio marinus (Diptera; Chironomidae) consist of a tube of jelly containing up to 175 eggs. The swimming egg-mass of the Atlantic populations (Race A) and the sinking egg-mass of the Baltic populations (Race B) have a higher density than the sea water, on which the female spawns; salinity is not important to the swimming or sinking of the egg-masses. Density and sinking velocity of gelatinous egg-masses from Atlantic midges are significantly higher than those from Baltic midges if the eggs are laid on water of the same salinity. The density of the eggs and the quantitative relation of eggs to jelly are the same in both races. The jelly of the egg-masses from the Baltic Sea race swells more than that from the Atlantic Sea race (increase of volume by absorption of water = 3-fold and 2.4-fold, respectively). During swelling the jelly alters from a trough-like shape into a tube-like from which closes around the eggs. If the surface tension of the spawning medium is lowered by use of detergents, the A-spawn will sink. A- and B-spawns occupy different positions at an oil-water interface. Interference contrast microscopy revealed that the A-jelly forms a well-defined outer line in relationship to the surrounding aqueous medium; this line is missing in the B-jelly. The morphological fine surface structures, as seen by the scanning electron microscope, do not explain the different properties of the A- and B-spawn. Extracts and hydrolysates of jelly material were separated by thin-layer chromatography. The results indicate that the spawn jelly of C. marinus consists of a polysaccharide-protein-complex, similar to the jellies of other invertebrates. It is rather probable that the chemical structure or the components of the jellies from A- and B-populations are not exactly the same; this would explain the different properties described. The ecological significance of swimming and sinking gelatinous egg-masses and the taxonomical position of both midge populations are discussed.