邻苯二甲酸二甲酯在离体人红细胞内的分布和含量研究

邻苯二甲酸酯类污染物(PAEs)在环境中普遍存在,可沿食物链富集,危害人体健康。本文利用荧光光谱法和高效液相色谱法(HPLC)探究了邻苯二甲酸二甲酯(DMP)在离体人红细胞内的分布情况。结果表明,红细胞在310 nm、490 nm和609nm处各具有一个荧光特征峰,其来源分别为:红细胞内的蛋白;还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)、黄素腺嘌呤二核苷酸(FAD)和膜脂;锌卟啉和原卟啉。DMP染毒后,310 nm处的荧光峰出现明显下降,其原因为进入红细胞内的DMP与蛋白发生了结合;490 nm和609 nm处的荧光峰变化很小。高效液相色谱(HPLC)实验结果表明,DMP能透过红细胞膜进入细胞内部,其进入量随暴露量增加而增加,进入量和暴露量的比值随暴露量增加而减少。上述研究成果能加深对PAEs在血液运输过程中与红细胞毒性作用的理解,可为PAEs的危险性评估和相关疾病预防提供数据参考。     

Ameliorated effects of garlic (Allium sativum) on biomarkers of subchronic acrylamide hepatotoxicity and brain toxicity in rats

Acrylamide (ACR) exerts its toxicity through stimulation of the oxidative stress; yet, its effect on neurotransmitter catabolic enzymes has not been elucidated. We investigated the effects of ACR exposure on brain and hepatic tissues antioxidant enzymes activities and different markers such as, acetylcholinesterase (AChE), nitric oxide (NO), monoamine oxidase (MAO), and lipid profile, and to evaluate the protective effects of garlic against ACR toxicity. Male Sprague-Dawley rats were exposed to ACR (1 mg kg^?1 body weight) with or without diet containing 1.5% of garlic powder for 40 days. ACR administration showed a decrease in AChE activity associated with an increase in MAO activity in both brain and hepatic tissues. In addition, ACR administration increased the lipid peroxidation and NO levels of both tissues while decreased the activities of glutathione (GSH), superoxide dismutase, and glutathione-S-transferase (GST). On the other hand, the activities of glutathione peroxidase (GPx) and catalase activities increased as a consequence of GSH depletion after ACR exposure. Finally, ACR exposure increased the brain and liver lipid profile of cholesterol, triglycerides and total lipid, while phospholipids level was decreased. Coadministration of garlic powder with ACR significantly attenuated oxidative stress, MAO activity, and inflammation in brain and hepatic tissues but did not ameliorate AChE activity. In conclusion, our results emphasized the role of garlic as a potential adjuvant therapy to prevent ACR neurotoxicity and hepatotoxicity.     

Use of Mytilus galloprovincialis and Tapes philippinarum as sentinel organisms for the development of a biosurveillance program in the Pialassa Baiona coastal lagoon (Ravenna, Italy)

The Pialassa Baiona is an intertidal lagoon located north from the town of Ravenna (Italy). It shows a certain degree of contamination caused by past uncontrolled discharges, and we aim at developing a bioassessment program to be used for future monitoring and conservation plans. A battery of six biomarkers (lysosome membrane stability, lipofuscin and neutral lipid accumulation, hsp70 expression, metallothionein levels and acetylcholinesterase (AChE) activity) was investigated in Mytilus galloprovincialis transplanted for 30 days in different sites. Tapes philippinarum are widely distributed within the lagoon, and we explored the possibility to use resident clams for future biosurveillance programs. Adenylyl cyclase (ACase) activity was also assessed in tissues of both bivalves as a future possible biomarker. Taken together, our results indicate that mussels are suitable sentinel organisms also within such a peculiar environment; alteration of four biomarkers was observed in tissues of mussels transplanted in two sites out of the five analyzed, indicating their low water quality. T. philippinarum showed a barely detectable AChE activity and absence of neutral lipids in digestive glands, therefore appeared unsuitable for our purpose, at least using the selected battery of biomarkers. ACase activity was not different among animals collected from the different sites.     

Structural and molecular alterations in arsenic-induced hepatic oxidative stress in rats: a FTIR study

Arsenic (As) is an ubiquitously distributed environmental toxicant predominantly contaminating drinking water. A number of studies indicated that oral exposure of humans to inorganic As produced damage to various body tissues including liver. Oxidative stress is thought to play a major role in As-induced hepatotoxicity. In this study, Fourier transforms infrared (FTIR) spectroscopy approach was applied to determine whether chronic As exposure at 25 ppm, intragastrically for 12 weeks, affected oxidative stress status in rat liver. Data demonstrated that chronic As administration exacerbated oxidative stress as was evidenced by suppressed antioxidant defense system and increased lipid peroxidation and protein oxidation. The FTIR study showed that peak area value of amide A decreased significantly followed by reduced amide I and amide II peak area in an As-treated rat liver suggesting altered protein profile. The change of Olefinic?CH stretching band and C?O stretching of triglycerides band indicated the altered lipid levels due to metallic exposure. The fall in the peak area of PO₂^? asymmetric stretching in the As-treated group might be due to compositional changes of nucleic acids. Hence, the results of this study indicate that As-induced oxidative stress was associated with structural and molecular modifications in proteins, lipids, and glycogen in a rat liver that may help to elucidate molecular mechanisms underlying metal-mediated hepatic damage.     

Impaired phospholipid metabolism in Saccharomyces cerevisiae by increased phospholipase B activity induced by 4-(methyl nitrosamino)-1-(3-pyridyl)-1-butanone

Tobacco products and cigarette smoke cause many respiratory diseases including cancer. 4-(Methyl nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a potent carcinogen in cigarette smoke, but its effect on lipid metabolism remains enigmatic. Hence, Saccharomyces cerevisiae was exposed to different concentrations of NNK (0–400?µmol?L^?1) to elucidate its role in lipid metabolism. Exposure to NNK substantially decreases (about 60%) of the phospholipid content with a concomitant increase in lysophospholipids. Significant reduction was observed in the phosphatidylcholine followed by phosphatidylethanolamine with NNK-treated cells. On the contrary, cells accumulated significant amount of neutral lipids and free fatty acids. Exposure of yeast cells (wild-type cells and three plbΔ mutant strains) to NNK greatly enhances the hydrolysis of phospholipid in the presence of calcium. We are the first to report that exposure to NNK enhances phospholipase B (PLB), particularly plb1p activity. Furthermore, NNK also promotes the alteration of phospholipid fatty acid (FA) content. These results suggest that NNK aids in the degradation of phospholipids by enhanced PLB activity and is accompanied with FA alteration. Understanding the altered phospholipid metabolism in the presence of NNK remains a worthy pursuit.     

The effects of anthracene on biochemical responses of Mediterranean mussels Mytilus galloprovincialis

Mediterranean mussels Mytilus galloprovincialis is among the most widely used bioindicators, and anthracene (AN) is one of the most commonly found hydrocarbons in the aquatic environment. M. galloprovincialis were exposed to nominal concentrations of 0.05, 0.15 and 0.4?μg/L AN. Chemical analysis using high-performance liquid chromatography revealed the uptake of AN in the whole soft body and digestive gland at different amounts. After a short exposure (2, 4 and 8 days), the results revealed that AN induced malondialdehyde (MDA), glutathione (GSH) and acetylcholinesterase (AChE) activity in digestive gland. Our findings demonstrated also that AN reduced the filtration rates in a concentration-dependent manner. Increase in lipid peroxidation (MDA content) in digestive gland (p?M. galloprovincialis, where digestive gland constitutes a valuable organ for investigating AN biotransformation and toxicity.     

Purification of metallothionein from thymus and spleen,induced by zinc administration

Zinc is an essential trace element to maintain human and generally mammal homeostasis; its biological signification could be explained through the participation or as structural element of more than 200 protein molecules in mammals.

The synthesis of proteins in spleen and thymus of rats is induced by zinc, a fact which moved us to purify and isolate such proteins, before proceeding to its identification or characterization. Rats are treated with zinc during 4 weeks and then sacrificed; blood is extracted under light ether anesthesia. The extracts of thymus and spleen were prepared by homogenization with Tris‐HC1 buffer 1.0 × 10^‐2M pH 7.4 and submitted to Sephadex G‐75 column (40 × 1.6 cm) after centrifugation and filtration through 0.45 μ filters. The elution profile of proteins were established at 253 and 280 nm, and zinc concentration was measured by atomic absorption spectrophotometry. The peak containing zinc was submitted to a second column of Sephadex G‐25 (40×1.6 cm) obtaining a peak of purified protein with physico‐chemical characteristics similar to metallothionein.     

The effect of copper and zinc on the metabolism of the mussel Mytilus edulis

The effects of copper and zinc on the metabolism of the mussel Mytilus edulis (L.) and its component tissues were studied. 500 ppm copper sodium citrate inhibited oxygen consumption of the whole animal and gill tissue, but no similar effect was observed on digestive gland tissue. 500 ppm zinc sodium citrate exerted no effect upon gill or digestive gland respiration, and neither metal salt affected the respiration of homogenates of gill, digestive gland or gonad. Direct observation of gill tissues during exposure to the metals revealed that 500 ppm copper sodium citrate caused inhibition of ciliary activity; exposure of tissues to 2 ppm Cu for 24 h resulted in only partial inhibition of the cilía. It is suggested that metabolic suppression noted in whole animals and gill tissues is due to the inhibition of an energy-consuming process such as ciliary activity rather than interference with respiratory enzyme systems.     

Effects of potassium concentration and osmotic pressure of sea water on the cell-turgor pressure of Chaetomorpha linum

The unicellular littoral alga Chaetomorpha linum is especially capable of maintaining its cell-turgor constant by regulation of the internal osmotic pressure, when the salinity of the sea water is altered. The decrease or increase of the external potassium concentration is seen to be an important cause of this turgor regulation, as well as the alteration of the external osmotic pressure, which was already known to be an important factor. This has been shown by experiments in artificial sea water with reduced osmolality and variable potassium concentration (1 to 50 mMol/l).     

Response of ultraviolet-B induced antioxidant defense system in a medicinal plant, Acorus calamus

Ultraviolet-B (UV-B) radiation generates an oxidative stress in plant cells due to excessive generation of reactive oxygen species (ROS). ROS can denature enzymes and damage important cellular components. In the present study, an important medicinal plant Acorus calamus (Sweet flag) was subjected to two doses of supplemental UV-B radiation (sUV-B): sUV1 (+ 1.8 kJ m(-2) d(-1)) and sUV2 (+3.6 kJ m(-2) d(-1)) to evaluate the relative response of antioxidant defense potential. Stimulation of activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) was observed at initial growth period while the activities of CAT and SOD decreased at later age of sampling. sUV-B induced lipid peroxidation (LPO) was observed showing alteration of membrane properties. No definite trend of change was observed for ascorbic acid (AsA), while increments in thiol, proline, phenol and protein contents were observed due to sUV-B. Results suggested that sUV-B radiation may stimulate the enzymatic and non-enzymatic defense system of Acorus plants, showing its better adaptation at lower dose of sUV-B.     

Fatty acid profiles of juvenile salmon indicate prey selection strategies in coastal marine waters

Juvenile salmon exhibit high growth rates upon their arrival into the marine environment. Dietary changes from freshwater and estuarine habitats to those derived from the marine environment may play an important role in ultimate adult survival. We measured the total lipid and fatty acid (FA) composition of juvenile Chinook salmon (Oncorhynchus tshawytscha), coho salmon (O. kisutch), and 18 of their potential prey items sampled from coastal waters during their first few months at sea. Coho salmon had significant reductions in their lipid content (% wet weight) between May and June, likely due to early marine growth. We did not find a significant drop between May and June Chinook salmon lipid content, which may indicate an earlier ontogenetic selection to marine prey that are higher in lipids and essential fatty acids (EFAs). Juvenile salmon ate prey of both high and low lipids. Significant FA compositional changes occurred for both coho and Chinook salmon between May and June. In May, the FA profile of juvenile salmon, especially coho salmon, did not resemble their prey items; however, in June, there was a strong correlation between salmon and their common fish prey as determined by gut content analysis. Significant increases in the level of EFAs, especially docosahexaenoic acid (DHA, 22:6n-3) accounted for the majority of the monthly differences in salmon tissue FA composition. In order for juvenile salmon to adequately meet their physiological requirements, they may have adapted to select advantageous prey with higher levels of EFAs, especially DHA, in order to rapidly increase their growth and ultimate survival.     

In situ variation in heart rate of the shore crab Carcinus maenas in relation to environmental factors and physiological condition

Heart rate of Carcinus maenas was recorded continuously for two days in situ, together with water temperature, salinity, depth and light intensity. Of each sex, 25 crabs were used and assigned to groups on the basis of size and colour (the carapace changes colour with prolongation of the intermoult phase). Wet weight:dry weight ratios of midgut gland, muscle, gonads and whole body were examined together with midgut gland lipid content and haemolymph protein concentration. Heart rates and wet wt:dry wt ratios for all the tissues examined were higher in early intermoult than in late intermoult stages of the adult C. maenas (P<0.05). Heart rate, lipid content and haemolymph protein concentrations were higher and wet wt:dry wt ratios of midgut gland, gonads and whole body were lower in juvenile crabs than in adults (P<0.05). The relationship between heart rate and wet wt:dry wt of whole body differed among C. maenas in early and late intermoutl. Heart rate was positively correlated with midgut gland lipid content (a Darwinian fitness parameter) in crabs that were in late intermoult. Physico-chemical environmental parameters, sex and colour accounted for 12% of the variation in heart rate of C. maenas recorded in situ. Temperature and prolonged intermoult were the most important factors influencing heart rate. The findings are discussed in relation to laboratory studies of cardiac activity and observed behaviour of C. maenas in the field.     

Effect of sexual maturation on the tissue biochemical composition of<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> and<Emphasis Type="Italic"> O</Emphasis>.<Emphasis Type="Italic"> defilippi</Emphasis> (Mollusca: Cephalopoda)

Changes in the protein, lipid, glycogen, cholesterol and energy contents, total amino acid and fatty acid profiles of Octopus vulgaris and O. defilippi tissues (gonad, digestive gland and muscle) during sexual maturation (spermatogenesis and oogenesis) were investigated. Both species showed an increase of amino acids and protein content in the gonad throughout sexual maturation (namely in oogenesis), but allocation of these nitrogen compounds from the digestive gland and muscle was not evident. The major essential amino acids in the three tissues were leucine, lysine and arginine. The major non-essential amino acids were glutamic acid, aspartic acid and alanine. With respect to carbon compounds, a significant increasing trend (P<0.05) in the lipid and fatty acid contents in the three tissues was observed, and, consequently, there was also little evidence of accumulated lipid storage reserves being used for egg production. It seems that for egg production both Octopus species use energy directly from food, rather than from stored products. This direct acquisition model contrasts with the previous model for Octopus vulgaris proposed by ODor and Wells (1978: J Exp Biol 77:15–31). Most of saturated fatty acid content of the three tissues was presented as 16:0 and 18:0, monounsaturated fatty acid content as 18:1 and 20:1 and polyunsaturated fatty acid content as arachidonic acid (20:4n-6), eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3). Though cholesterol is an important precursor of steroid hormones, this sterol content exhibited variations that do not seem to be related with the maturation process. Moreover, significant differences (P<0.05) were obtained between genders, suggesting that perhaps there is a greater physiological demand for cholesterol during spermatogenesis than oogenesis. If the component sterols of octopus are of a dietary origin, considerable variation in the cholesterol content between species might be expected on the basis of the sterol composition of their prey. The glycogen reserves increased significantly in the gonad and decreased significantly (P<0.05) in the digestive gland and muscle of O. vulgaris (these trends were not evident in O. defilippi). Glycogen may play an important role in the maturation process and embryogenesis of these organisms, because carbohydrates are precursors of metabolic intermediates in the production of energy. It was evident that sexual maturation had a significant effect upon the gonad energy content, but the non-significant energy variation (P>0.05) in the digestive gland and muscle revealed no evidence that storage reserves are transferred from tissue to tissue. The biochemical composition of digestive gland and muscle may not be influenced by sexual maturation, but rather by other biotic factors, such as feeding activity, food availability, spawning and brooding.Communicated by S.A. Poulet, Roscoff     

Impaired behavior and changes in some biochemical markers of bivalve (Ruditapes decussatus) due to zinc toxicity

Susceptibility and response of bivalves, the clam Ruditapes decussatus, to zinc (Zn) were studied by monitoring behavior using valve movement and some biochemical parameters. The LC₅₀ was 4.1 mg L^?1 at 7 days. Depuration of Zn from their tissues was also followed. Accumulation of Zn increased with concentration and duration of exposure. After 20 days, Zn caused impairment of valve movement as well as the antioxidant system, decreasing glutathione and protein levels and increasing the lipid peroxidation which is indicative of oxidative damage. Results suggested that behavior and biochemical parameters of clam R. decussatus were sensitive and suitable responses for assessing the effects of Zn on the aquatic ecosystems. It could be concluded that there is a time–dose–response relationship of Zn with behavior and oxidative stress of R. decussatus.     

Effects of experimental feeding and starvation on the proximate composition of the European bass Dicentrarchus labrax

Juvenile bass, Dicentrarchus labrax (L.), fed ad libitum on trout pellets, the shore crab Carcinus maenas and filleted saithe, Gadus virens, did not differ significantly in proximate composition although the diets were absorbed with high efficiencies of 94 to 99% and differed greatly in composition. Compared with wild fish, the white muscle, dark muscle, liver and mesenteric fat bodies of laboratory-reared bass were lower in moisture and higher in lipid. Starvation caused a rapid initial decrease in carbohydrate and a progressive decline in lipid in all tissues, coupled with an increase in moisture and ash contents. The liver and fat body somatic indices also declined, indicating that these organs are the major lipid storage depots in bass, in contrast with the skeletal muscle which has this role in other active fishes such as salmonids and clupeids. Lipid and protein are inversely related to moisture content in whole specimens.     

Histochemical and ultrastructural characterisation of mantle storage cells in the hydrothermal-vent bivalve Bathymodiolus azoricus

This study reports on two types of storage cells that are present in the mantle connective tissue of the deep-sea mussel Bathymodiolus azoricus. One type corresponds to the adipogranular cells, a kind of storage cell previously described in other bivalves. In these cells extensive regions of the cytoplasm are filled with glycogen deposits and these zones became strongly stained after histochemical (PAS) or ultrastructural detection of polysaccharides. Several lipid droplets and membrane bound granules containing homogeneous electron-dense material are also present in adipogranular cells. A second type of cell contains large lysosomes in addition to numerous lipid droplets, but lacking cytoplasmic glycogen deposits. Due to these characteristics we named them adipolysosomal cells. They can be identified in semi-thin sections stained with PAS reaction because the lysosomes are the only positively stained structures. In the connective tissue of the mantle, some cells containing many lysosomes and a few lipid droplets were also observed. These cells differ from the adipolysosomal cells mainly because they have a reduced amount of lipid reserves, and could be an initial stage in the development of adipolysosomal cells. The vesicular connective tissue cells that in other Mytilidae are specialised in glycogen storage were not detected in B. azoricus. The reserves accumulated in the two types of storage cells described in B. azoricus may be important for the survival of these hydrothermal-vent bivalves if their nutrition is affected by a temporary loss or reduction of endosymbiotic bacteria due to sulphide and/or methane shortage caused by oscillations in vent activity.     

Barnacles: Possible indicators of zinc pollution?

Barnacles [Balanus balanoides (L.), Elminius modestus Darwin, Lepas anatifera (L.)] from several different sites were found to accumulate the heavy metal zinc. The majority of the zinc was deposited in the tissues associated with the gut, and the level of zinc in soft body tissue generally reflected well the level of zinc in the immediate sea-water environment. The zinc accumulated in the gut tissues was in the form of discrete granules, mainly within the parenchyma cells which surround the gut. These granules probably exist as an insoluble zinc salt. A comparison of the zinc level in barnacles with those recorded for some other marine organisms indicates the possible usefulness of barnacles as indicators of zinc pollution.     

Changes in phospholipase A2 activity and lipid content during early development of Atlantic halibut (Hippoglossus hippoglossus)

During early development in fish, phospholipase A₂ (EC 3.1.1.4) regulates membrane lipid modifications, which relates to changes in environmental conditions and provision of fatty acids required for metabolic energy substrates and prostaglandin biosynthesis. A method to analyze phospholipase A₂ in rat tissues has been modified to measure its activity in embryonic Atlantic halibut (Hippoglossus hippoglossus L.). Egg and embryo samples were collected during the 1994 spawning season. Enzyme activity was undetectable at fertilization but in 10-d embryos was 230 pmol mg⁻¹ h⁻¹ (at 20 °C) and increased by ∼120% at hatch (17-d). Significant alterations in the fatty acid composition of important phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), were also observed. The content of some critical polyunsaturated fatty acids, and the ratio of unsaturated/saturated fatty acids, declined significantly over development. Acyl-chain restructuring mediated through the activity of phospholipase A₂, coupled with other observed lipid changes (significant increases in the PC/PE ratio and cholesterol content), would produce a decreased fluidity of membranes during embryonic development, coinciding with the predicted upward movement of larvae in the water column. Arachidonic acid (20:4n-6) removed from PE could serve as a precursor for biosynthesis of 2-series prostaglandins, and eicosapentaenoic acid (20:5n-3) from PC is a likely source for other prostaglandin types. Despite removal of polyunsaturated fatty acids, there was an overall increase in lipid and fatty acid concentration, which can be attributed to amino acid catabolism during early developmental stages. Received: 9 September 1996 / Accepted: 8 September 1997     

Biochemical and histopathological changes of Biomphalaria alexandrina snails exposed to bisphenol A

The effects of sub-lethal concentrations of bisphenol A on some biochemical parameters in Biomphalaria alexandrina snail's hemolymph and tissues as well as on histological changes of the hermaphrodite gland were evaluated. Adult snails were exposed to sub-lethal concentrations for four weeks and samples of snails were investigated after two and four weeks. Total protein and albumin levels in hemolymph, as well as lipid peroxidation and glutathione contents of snails’ tissues, were measured. After two weeks of exposure, total protein content and albumin levels decreased in all groups. Elevation of lipid peroxidation levels was correlated with decreasing glutathione content in the treated snails. Histological examination of the hermaphrodite gland revealed deformations in eggs and affected sperm production.