Metallothionein induction in early larval stages of tilapia (Oreochromis mossambicus)

Amounts of whole-body metallothionein (MT) in tilapia (Oreochromis mossambicus) larvae increased to a peak (1,500 ng mg(-1) protein) 1 d after hatching (H1), decreased rapidly thereafter, and was maintained at a constant level (700 ng mg(-1)) 3 d after hatching (H3). Waterborne Cd(2+) could stimulate MT expression in newly hatched (H0) larvae in dose-dependent and time-dependent patterns. H0 larvae, which were treated with 35 microg L(-1) Cd(2+) for 24 h, showed a 1.7-fold increase in the MT amount (174.0+/-64.7) and a 6. 5-fold increase in accumulated Cd(2+) but no significant change in Ca(2+) content, compared with the H0 control (MT, 102.6+/-48.1). H3 larvae with the same treatment revealed about a 10-fold increase in accumulated Cd(2+), a 10% decrease in Ca(2+) content, but no change in MT (261.2+/-120.0), compared with the H3 control (MT, 330+/-74.0). H0 larvae could synthesize more MT to bind Cd(2+) for detoxification in 35 microg L(-1) Cd(2+), a dose that would not affect normal physiology or survival of H0 larvae. On the other hand, 35 microg L(-1) Cd(2+) caused H3 larvae to experience hypocalcemia, an abnormal physiological condition, in which H3 larvae could not synthesize sufficient MT, thus causing greater than 25% mortality. These results indicate for the first time that the inducibility of MT by waterborne Cd(2+) is development dependent, being correlated with inconsistent sensitivities to Cd(2+) during larval development.     

镉对满江红（Azolla imbricata）-鱼腥藻（Anabaena azollae）共生体氮代谢的影响

在实验室条件下,研究了不同浓度(0、0.01、0.05、0.1、0.5mg/L)的Cd对满江红-鱼腥藻共生体异型胞频率,固氮酶、谷氨酰氨合成酶活性以及铵态氮、游离氨基酸、可溶性蛋白、总氮含量的影响.结果表明,在整个实验期间,0.01mg/L Cd处理对上述指标均没产生显著影响,说明满江红-鱼腥藻共生体对Cd具有较强的耐性.当培养液中Cd浓度≥0.05mg/L时,随溶液中Cd浓度的增加和处理时间的推移,异型胞频率、固氮酶活性、谷氨酰氨合成酶活性、可溶性蛋白含量和总氮含量逐渐下降,而铵态氮含量在处理初期显著降低,随后迅速增加,游离氨基酸含量则逐渐增加.研究结果表明高浓度的Cd处理导致满江红-鱼腥藻共生体氮代谢的紊乱,最终造成氮素积累量的下降.     

Renal function in the freshwater rainbow trout after dietary cadmium acclimation and waterborne cadmium challenge

Renal function was examined in adult rainbow trout (Oncorhynchus mykiss) after chronic exposure to a sublethal level of dietary Cd (500 mg/kg diet) for 52 d and during a subsequent challenge to waterborne Cd (10 microg/L) for 72 h. Dietary Cd had no major effects on UFR (urine flow rate) and GFR (glomerular filtration rate) but caused increased renal excretion of glucose, protein, and major ions (Mg(2+), Zn(2+), K(+), Na(+), Cl(-) but Ca(2+)). However, dietary Cd did not affect any plasma ions except Na(+) which was significantly elevated in the Cd-acclimated trout. Plasma glucose and ammonia levels fell by 25% and 36% respectively, but neither plasma nor urine urea was affected in Cd-acclimated fish. Dietary Cd exposure resulted in a remarkable increase of Cd load in the plasma (48-fold, approximately 22 ng/mL) and urine (60-fold, 8.9 ng/mL), but Cd excretion via the kidney was negligible on a mass-balance basis. Clearance ratio analysis indicates that all ions, Cd, and metabolites were reabsorbed strongly (58-100%) in both na?ve and dietary Cd exposed fish, except ammonia which was secreted in both groups. Mg(2+), Na(+), Cl(-) and K(+) reabsorption decreased significantly (3-15%) in the Cd-exposed fish relative to the control. Following waterborne Cd challenge, GFR and UFR were affected transiently, and only Mg(2+) and protein excretion remained elevated with no recovery with time in Cd-acclimated trout. Urinary Ca(2+) and Zn(2+) excretion rates dropped with an indication of renal compensation towards plasma declines of both ions. Cadmium challenge did not cause any notable effects on urinary excretion rates of metabolites. However, a significant decrease in Mg(2+) reabsorption but an increase in total ammonia secretion was observed in the Cd-acclimated fish. The study suggests that dietary Cd acclimation involves physiological costs in terms of renal dysfunction and elevated urinary losses.     

Effects of Cadmium on Lipid Storage and Metabolism in the Freshwater Crab Sinopotamon henanense

Since environmental effects of molecular traits are often questioned we analyze here the molecular effects of cadmium (Cd) on lipid pathways and their effects on tissues development. Lipids are an important energy source for the developing embryo, and accumulate in the ovary and hepatopancreas of decapod crustaceans. The extend of Cd affecting lipid storage and metabolism, is studied here with the freshwater crabs Sinopotamon henanense. Crabs were exposed to water-born Cd at 1.45, 2.9, 5.8 mg/l for 10, 15, and 20 days. With significantly increased Cd accumulation in exposed crabs, lipid content in hepatopancreas and ovary showed a time-dependent and concentration-dependent reduction, being at least one of the reasons for a lower ovarian index (OI) and hepatopancreatic index (HI). After 10-day exposure increased triglyceride (TG) level in hemolymph and up-regulation of pancreatic lipase (PL) activity in the hepatopancreas suggested an increased nutritional lipid uptake. However, two processes led to lower lipid levels upon Cd exposure: an increased utilization of lipids and a down-regulated lipoprotein lipase (LPL) led to insufficient lipid transport. 10-day Cd exposure also triggered the production of β-nicotinamide adenine dinucleotide 2''-phosphate reduced tetrasodium salt hydrate (NADPH), as well as to the synthesis of adenosine triphosphate (ATP) and fatty acids. With increasing exposure time, the crabs at 15 and 20-day exposure contained less lipid and TG, suggesting that more energy was consumed during the exposure time. Meanwhile, the level of NADPH, ATP and the activity of PL, LPL, fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC) activity was down-regulated suggesting an impairment of the crab metabolism by Cd in addition to causing a lower lipid level.     

Acute tolerance and metabolic responses of Chinese mitten crab (Eriocheir sinensis) juveniles to ambient nitrite

The lethal concentration of nitrite to the Chinese mitten crab Eriocheir sinensis was tested by exposing the animals to 17.78, 23.71, 31.62, 42.17, and 56.23 mg NaNO2 L(-1) at 20 degrees C for 24, 48, 72, and 96 h. The corresponding LC50 value for each time exposure was 43.87 (38.70-51.70), 40.24 (34.88-46.01), 38.87 (33.72-46.01) and 38.87 (33.72-46.01) mg NaNO2 L(-1) or 29.25 (25.80-34.47), 26.83 (23.25-30.67), 25.91(22.48-30.67), 25.91(22.48-30.67) mg NO2-N L(-1), respectively. The physiological response of the crab to nitrite toxicity was further investigated by exposing the crab to 0, 10, 20, 30 and 40 mg NaNO2 L(-1) for 2 d. The changes of nitrogenous compounds in haemolymph, oxyhemocyanin and metabolism were measured at 3, 6, 24 and 48 h upon exposure. Haemolymph nitrite was significantly enhanced by the increase of nitrite from 10 to 40 mg NaNO2 L(-1) during the 2-day exposure. The concentrations of nitrate, urea and glutamate in haemolymph increased concomitantly with the exposing time and ambient nitrite levels, suggesting that the formation of nitrate, urea and glutamine may be the possible end products of nitrite detoxification in crabs. The diffusion of nitrite caused a reduction of oxyhemocyanin, resulting to hypoxia in tissues. Under a hypoxia condition, crabs increased energy demand for metabolism as indicated by the elevated levels of glucose and lactate in haemolymph. Our data showed that ambient nitrite could affect oxygen carrying capacity through oxyhemocyanin reduction and the increase of energy catabolism in crabs. This study suggests that nitrite could be detoxified through the pathway of nitrate, urea and glutamine formation in crabs.     

铜、镉对三种豆科植物生长及氮磷钾含量的影响

为了解豆科植物在Cu、Cd单一污染土壤中的生长状况及对土壤养分的吸收利用特点,采用盆栽实验研究了Cu~(2+)、Cd~(2+)单一污染下紫花苜蓿、红三叶、沙打旺3种豆科植物的株高、根长、生物量和叶、茎部N、P、K、Cu和Cd含量的变化情况.Cu~(2+)、Cd~(2+)处理浓度分别为:0、400、800、1200 mg·kg~(-1)和0、1、10、20 mg·kg~(-1).结果表明,3种豆科植物对Cu和Cd均有较强的吸收能力,除红三叶叶片中Cu含量外,3种豆科植物根、茎、叶中Cd和Cu的含量均与土壤中重金属添加量呈显著正相关.3种豆科植物在严重Cd~(2+)污染的土壤中均能正常生长.在Cu~(2+)添加量≤1200 mg·kg~(-1)时,红三叶能正常生长,而紫花苜蓿的生长则受到显著抑制作用,沙打旺在Cu~(2+)添加量≥800 mg·kg~(-1)时生长受到抑制.土壤Cu~(2+)添加量≤1200 mg·kg~(-1)时,能促进紫花苜蓿对N、P、K的吸收;Cu~(2+)添加量≤800 mg·kg~(-1)时,对红三叶N、P、K含量没有明显影响;Cu~(2+)添加量≤400 mg·kg~(-1)时,可提高沙打旺中N、P、K含量,但当Cu~(2+)添加量≥800 mg·kg~(-1)时则显著降低.土壤Cd~(2+)添加量≤20 mg·kg~(-1)时,对紫花苜蓿和红三叶茎叶以及沙打旺茎部的N、P、K含量有促进作用,但对沙打旺叶片的N、P、K含量起抑制作用.总体来看,3种豆科植物对Cu~(2+)和Cd~(2+)均有一定的耐性,红三叶对Cu~(2+)的耐性较好,紫花苜蓿对Cd~(2+)的耐性较好.

Abstract：

Aimed to understand the growth status and nutrient uptake of leguminous plants under soil copper-or cadmium contamination, a pot experiment with Medicago sativa, Trifolium pre-tense, and Astragalus adsurgens was conducted, with their plant height, root length, plant bio-mass, and N, P, K, Cu and Cd contents in leaf and stem measured. The application amounts of Cu~(2+) and Cd~(2+) were 0,400, 800, and 1200 mg·kg~(-1), and 0, 1, 10, and 20 mg·kg~(-1), re-spectively. All the test three leguminous plants had strong capability of absorbing Cu and Cd.The Cu and Cd contents in their roots, stems, and leaves, except the Cu content in T. Pratease leaf, were significantly positively correlated with the application amounts of Cu~(2+) and Cd~(2+). Un-der the application of Cd~(2+), all the three leguminous plants grew normally. When the application amount of Cu~(2+) was≤1200 mg·kgM-1, T. Pratense grew normally, while the growth of M. Sativa was significantly inhibited. The growth of A. Adsurgens was inhibited when the application amount of Cu~(2+) was≥800 mg·kg~(-1). An apphcation amount of≤1200 mg·kg~(-1) of Cu~(2+) pro-moted the N, P and K absorption of M. Sativa, but applying≤800 mg·kg~(-1) of Cu~(2+) had little effects on the N, P and K absorption of T. Pretense. The N, P and K contents of A. Adsurgens in-creased when the application amount of Cu~(2+) was ≤400 mg·kg~(-1), but decreased significantly when the Cu~(2+) application amount was≥800 mg·kg~(-1). When the application amount of Cd~(2+)was≤20 mg·kg~(-1) , the N, P and K contents in M. Sativa and T. Pratense stems and leaves and in A. Adsurgens stem increased, while those in A. Adsurgens leaf decreased. As a whole, the three leguminous plants all had certain tolerance to Cu~(2+) and Cd~(2+) stresses, especially T. Prat-ease to Cu~(2+) and M. Sativa to Cd~(2+).     

Changes in the activity levels of glutamine synthetase, glutaminase and glycogen synthetase in rats subjected to hypoxic stress

 Exposure to high altitude causes loss of body mass and alterations in metabolic processes, especially carbohydrate and protein metabolism. The present study was conducted to elucidate the role of glutamine synthetase, glutaminase and glycogen synthetase under conditions of chronic intermittent hypoxia. Four groups, each consisting of 12 male albino rats (Wistar strain), were exposed to a simulated altitude of 7620 m in a hypobaric chamber for 6 h per day for 1, 7, 14 and 21 days, respectively. Blood haemoglobin, blood glucose, protein levels in the liver, muscle and plasma, glycogen content, and glutaminase, glutamine synthetase and glycogen synthetase activities in liver and muscle were determined in all groups of exposed and in a group of unexposed animals. Food intake and changes in body mass were also monitored. There was a significant reduction in body mass (28–30%) in hypoxia-exposed groups as compared to controls, with a corresponding decrease in food intake. There was rise in blood haemoglobin and plasma protein in response to acclimatisation. Over a three-fold increase in liver glycogen content was observed following 1 day of hypoxic exposure (4.76±0.78 mg·g⁻¹ wet tissue in normal unexposed rats; 15.82±2.30 mg·g⁻¹ wet tissue in rats exposed to hypoxia for 1 day). This returned to normal in later stages of exposure. However, there was no change in glycogen synthetase activity except for a decrease in the 21-days hypoxia-exposed group. There was a slight increase in muscle glycogen content in the 1-day exposed group which declined significantly by 56.5, 50.6 and 42% following 7, 14, and 21 days of exposure, respectively. Muscle glycogen synthetase activity was also decreased following 21 days of exposure. There was an increase in glutaminase activity in the liver and muscle in the 7-, 14- and 21-day exposed groups. Glutamine synthetase activity was higher in the liver in 7- and 14-day exposed groups; this returned to normal following 21 days of exposure. Glutamine synthetase activity in muscle was significantly higher in the 14-day exposed group (4.32 μmol γ-glutamyl hydroxamate formed·g protein⁻¹·min⁻¹) in comparison to normal (1.53 μmol γ-glutamyl hydroxamate formed·g protein⁻¹·min⁻¹); this parameter had decreased by 40% following 21 days of exposure. These results suggest that since no dramatic changes in the levels of protein were observed in the muscle and liver, there is an alteration in glutaminase and glutamine synthetase activity in order to maintain nitrogen metabolism in the initial phase of hypoxic exposure. Received: 30 March 1998 / Revised: 18 November 1998 / Accepted: 25 November 1998     

镉对尖紫蛤抗氧化酶活性及脂质过氧化的影响

为阐明镉(Cd2+)对尖紫蛤消化盲囊和鳃抗氧化酶的毒性影响程度,研究了不同浓度的Cd2+(0.005、0.05、0.5 mg/L)在不同暴露时间(24h、72h、120h)对尖紫蛤鳃和消化盲囊中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)的活性以及丙二醛(MDA)含量的影响。结果表明,在Cd2+浓度为0.005 mg/L时,在整个实验期间Cd2+对消化盲囊和鳃内的SOD、CAT、GSH-PX活性并无显著影响。在Cd2+浓度为0.05 mg/L和0.5 mg/L时,SOD、CAT、GSH-PX在鳃和消化盲囊中的活性都呈现出明显的时间剂量依赖关系。在0.05 mg/L暴露时,鳃和消化盲囊中的SOD、CAT和GSH-PX的活性随时间逐渐增强,在72h时达到最大值,但在120h时略有降低。在0.5 mg/L暴露时,消化盲囊中SOD、CAT及鳃中CAT活性在24h时上升达到最大值,但鳃中SOD直到72h才达到最大值,并均在120h下降到最低,其中消化盲囊中SOD和CAT活性在120h低于对照组,这可能与消化盲囊对Cd2+的敏感性高于鳃有关。在0.5 mg/L暴露的鳃中,GSH-PX在24h、72h活性并不上升,在120h甚至低于正常水平,0.5 mg/L暴露的消化盲囊中,24h时迅速增高,然后逐渐下降到正常值。这可能与Cd2+结合了GSH-PX的活性中心,降低了GSH-PX的活性有关。与3种酶活性随着时间延长和剂量的增加,酶活性会降低的变化趋势不同,鳃和消化盲囊中的MDA的含量随时间延长和剂量的增加而增加,并不出现下降,这表明尖紫蛤鳃和消化盲囊中的MDA含量可以灵敏的反映机体内的氧化损伤程度,但不能敏感的反映水体中Cd2+的污染情况。     

镉对克氏原螯虾肝胰腺抗氧化系统的影响

采用毒性试验方法,研究了不同浓度Cd2+(1.72、3.44、6.89、13.77和27.55mg·L-1)对克氏原螯虾肝胰腺抗氧化酶(SOD、CAT、GSH-PX)和抗氧化物质(GSH、Vc)的影响.结果表明:超氧化物歧化酶(SOD)活性受低浓度Cd2+诱导和高浓度Cd2+抑制,且受抑制程度与Cd2+浓度呈正相关.过氧化氢酶(CAT)活性总体表现为先激活后下降,且在第3天达最大值,CAT活性对低浓度Cd2+(≤6.89 mg·L-1)暴露敏感,而高浓度Cd2+对其影响较小.谷胱甘肽过氧化物酶(GSH-PX)活性对Cd2+浓度敏感,当Cd2+≤6.89 mg · L-1时,GSH-PX活性先升高后下降,更高浓度下GSH-PX活性在暴露后第1天即表现出抑制作用.还原型谷胱甘肽(GSH)含量在Cd2+≤6.89 mg·L-1时始终被诱导,且均在第1天达最大值,而高浓度Cd2+(≥13.77 mg·L-1)对GSH含量影响不明显.维生素C(Vc)对Cd2+胁迫敏感,各处理组Vc含量在第1天均显著下降,且下降程度与Cd2+浓度呈正相关,之后具有一定的合成恢复能力.抗氧化酶和非酶抗氧化物质在抵御Cd2+胁迫反应中共同发挥作用,且大多表现出明显的时间和剂量效应性.GSH-PX和Vc可以作为Cd2+污染的潜在生物指示物.     

Effects of Ce3+, Cd2+, and Hg2+ on activities and secondary structure of trypsin

The different effects of Ce³⁺, Cd²⁺, and Hg²⁺ on the activities and secondary structure of trypsin were studied. The results showed that trypsin activity was increased substantially by Ce³⁺ in 0.5–5 μmol/L concentration, but the activity was decreased significantly by Cd²⁺ or Hg²⁺ in 0.5–5 μmol/L concentration. The ultraviolet-visible spectrum of trypsin with 4 μmol/L Ce³⁺ treatment was the same as that of the control, but the 232-nm characteristic peak of trypsin with 4 μmol/L Cd²⁺ or Hg²⁺ treatment was blue-shifted and the peak intensity weakened. The circular dichroism (CD) spectrum of trypsin with 4 μmol/L Ce³⁺ treatment was similar to that of the control. The secondary structure of trypsin did not change with Ce³⁺ treatment. However, the CD spectrum of trypsin with 4 μmol/L Cd²⁺ or Hg²⁺ treatment was different from that of the control and Ce³⁺ treatment. The secondary structure of trypsin with Cd²⁺ or Hg²⁺ treatment changed greatly; for example, the α-helix and β-sheet contents were reduced significantly, the β-turn was enhanced greatly, and the random coil contents increased or decreased.     

Response of Soybean Seed Germination to Cadmium and Acid Rain

Cadmium (Cd) pollution and acid rain are the main environmental issues, and they often occur in the same agricultural region. Nevertheless, up to now, little information on the combined pollution of Cd(2+) and acid rain action on crops were presented. Here, we investigated the combined effect of Cd(2+) and acid rain on the seed germination of soybean. The results indicated that the single treatment with the low level of Cd(2+) (0.18, 1.0, 3.0?mg?L(-1)) or acid rain (pH ≥3.0) could not affect the seed germination of soybean, which was resulted in the increased activities of peroxidase and catalase. The single treatment with the high concentration of Cd(2+) (>6?mg?L(-1)) or acid rain at pH?2.5 decreased the activities of peroxidase and catalase, damaged the cell membrane and then decreased the seed germination of soybean. Meanwhile, the same toxic effect was observed in the combined treatment with Cd(2+) and acid rain, and the combined treatment had more toxic effect than the single treatment with Cd(2+) or acid rain. Thus, the combined pollution of Cd(2+) and acid rain had more potential threat to the seed germination of soybean than the single pollution of Cd(2+) or acid rain.     

Functional characterization of purified zinc transporter from renal brush border membrane of rat

Major zinc binding protein purified from renal brush border membrane (BBM) (R. Kumar, R. Prasad, Biochim. Biophys. Acta 1419 (1999) 23) was reconstituted into liposomes and its functional characteristics were investigated. Physical incorporation of the major zinc binding protein into the proteoliposomes was checked by SDS-PAGE, which showed a single band on silver staining. The structural integrity of the proteoliposomes was assessed by phase contrast microscopy, which revealed the proteoliposomes as globular structures and intact boundaries. Further structural integrity/leakiness of the proteoliposomes was checked by monitoring efflux of Zn(2+) from the pre-loaded proteoliposomes in the presence of either 2 mM Ca(2+) or Cd(2+) or Zn(2+). It was observed that even after 2 h of the initiation of efflux, 85-95% of Zn(2+) was retained in the proteoliposomes, thereby indicating that proteoliposomes were not leaky and maintained structural integrity during the uptake study. Zinc uptake into the proteoliposomes followed Michaelis-Menten kinetics with affinity constant (K(m)) of 1.03 mM and maximal velocity (V(max)) of 1333 nmol/mg protein per min. The uptake process followed first-order kinetics with a rate constant (k) of 1. 09x10(-3) s(-1). The specificity of zinc transport system was determined by studying the interaction of divalent cations viz. Ca(2+) and Cd(2+) with the zinc uptake. It was observed that Cd(2+) competitively inhibited the zinc uptake process with inhibitory concentration (K(i)) of 2.9 mM. Kinetic analysis of inhibitory effect of Cd(2+) on zinc uptake revealed an increase in K(m) to 1.74 mM without influencing V(max). Zn(2+) uptake into the proteoliposomes was found to be temperature sensitive and Arrhenius plot showed a breakpoint at 27 degrees C. The apparent energies of activation (E(a)) were found to be 7.09 and 2.74 kcal/mol below and above the breakpoint, respectively. The initial velocity of Zn(2+) uptake increased with the increase in outwardly directed proton gradient ([H](i) greater than [H](o)). The Zn(2+) uptake was inhibited by DCCD, thereby suggesting the involvement of -COOH groups in the translocation of Zn(2+) across the lipid bilayer. The ratio of acidic to basic amino acids (1.26) strongly indicates that it is an acidic protein. The cysteine content in this protein was insignificant, which further corroborates the possibility that the acidic amino acids might be prominent candidates for binding to zinc. The findings of the present study confirms that 40 kDa major zinc binding glycoprotein purified from renal BBM is a zinc transporter involved in the influx of Zn(2+) into the epithelial cells of the renal tubular system.     

The mode of action of acute and chronic concentrations of waterborne Cd in the digestive gland of the acclimated infested freshwater crab (<Emphasis Type="Italic">Potamonautes warreni</Emphasis>)

Cadmium (Cd) uptake, transport and accumulation were investigated in the digestive gland of the freshwater crab, Potamonautes warreni, acclimated in its natural habitat to stresses, such as microbial gill infestations, Cd(2+) and NH(4)(+), and subsequently exposed to increasing concentrations of Cd in the laboratory for up to 21 days. Cd exposure (0.2 mg l(-1)) for 7-14 days led to Cd permeating cell membranes in a particulate form; it was adsorbed intracellularly to endocytotic circulating amoebocytes, lipid droplets and Golgi vesicles in R-cells. Cd also caused dissociation of the fibrillar rough endoplasmic reticulum (RER) and an increase in phagocytotic activity in F- and B-cells. After 21 days, Cd accumulated as crystal deposits on the basal membranes of cells in the haemolymph space and along the microvilli of cells lining the tubular lumen. Elevated Cd concentrations were found in the cytosol, amoebocytes, Golgi vesicles and P/Ca granules in R-cells. Chronic exposure to higher concentrations of Cd (0.5 and 1.0 mg l(-1)) increased crystal deposition, whereas concentrations of Cd, copper and iron decreased in the cell membranes and in amoebocytes and increased in Golgi vesicles. Reduced lipid content, swollen nuclei with vesiculated nucleoli and enhanced activity of RER in R-cells were also noted. Cd was stored in the P/Ca and Ca granules of B-cells. Acute exposure to Cd (2.0 mg l(-1) for 48 h) caused metal granule accumulation along cells lining the tubular lumen and cellular dissociation, with acidosis and necrosis in the cytoplasm and Cd deposits in mitochondria. Cd accumulated in the cells of the digestive gland in a time-, concentration- and cell-type-specific fashion.     

Protein intensity changes in the hemoglobin and plasma electrophoretic patterns of Oreochromis niloticus in response to single and combined Zn and Cd exposure

The purpose of this study was to evaluate the effects of metals on the electrophoretic patterns of hemoglobin and blood plasma proteins of Oreochromis niloticus. Fish were exposed to 0.5 and 5.0 mg/L Zn, 0.1 and 1.0 mg/L Cd, and 0.5 mg/L Zn + 0.1 mg/L Cd, and 5.0 mg/L Zn + 1.0 mg/L Cd mixtures for 7 and 28 days. In all concentrations tested, electrophoretic pattern of hemoglobin and plasma proteins by cellulose acetate electrophoresis consist of three and eight bands, respectively. The three bands for hemoglobin are one cathodic (Hb1) and two anodic (Hb2 and Hb3) bands. The protein intensity in hemoglobins of fish following Zn, Cd, and Zn + Cd exposures decreased in Hb1, whereas it increased in Hb3. The eight bands for plasma proteins are 60, 78, 87, and 94 kDA high molecular weight proteins (HMP) for four bands and 120, 132, 176, and 273 kDA very high molecular weight proteins (VHMP) for four bands. The level of 60, 78, and 94 kDA HMP and 120, 132, and 176 kDA VHMP increased in response to single and combined Zn and Cd exposure. Also, there was increasing level of the metals in the whole blood with increasing concentrations of metals in the exposure medium and with increasing duration of exposure.     

水稻幼苗根对羧基化多壁碳纳米管复合镉胁迫的生长生理响应

为了探究羧基化多壁碳纳米管(MWCNTs-COOH)复合镉(Cd)胁迫对植物生长生理的影响,采用液体培养方法,以水稻(Oryza sativa L.)为受试作物,测定了0~12.0 mg·L^-1MWCNTs-COOH、10μmol·L-1Cd单一和复合处理水稻幼苗21天后根生长、氧化损伤、抗氧化酶活性及根中Cd含量的变化。结果表明:(1)MWCNTs-COOH单一处理,根长、根鲜重均低于对照,并表现出先升高后降低的趋势,当其浓度达到12.0 mg·L^-1时,较对照分别下降了9.3%和15.2%,且低于10μmol·L^-1Cd单一处理;而复合处理组水稻幼苗根长、根鲜重、干重皆低于对应的单一处理;(2)MWCNTs-COOH单一胁迫下,水稻根的超氧自由基(O2-·)明显积累,并伴随着超氧化物歧化酶(SOD)及过氧化物酶(POD)活性升高,3.0和6.0mg·L^-1MWCNTs-COOH处理下,SOD、POD活性最高;(3)MWCNTs-COOH复合Cd胁迫下,水稻根的SOD、POD活性大均低于单一处理组,而丙二醛(MDA)及羰基化蛋白含量均显著高于单一处理;(4)MWCNTs-COOH复合Cd后,水稻幼苗根尖细胞死亡加剧,其中,10μmol·L-1Cd与12.0 mg·L^-1MWCNTs-COOH复合处理的根尖根冠区细胞伊文斯蓝染色最深;(5)1.5~6.0 mg·L^-1MWCNTs-COOH复合Cd处理水稻幼苗后,其根中Cd含量呈上升趋势,且在6.0 mg·L^-1浓度处理时达到最大值303.30μg·g-1;高浓度MWCNTs-COOH及其与Cd的复合均对水稻根产生毒性效应。     

重金属离子对凡纳滨对虾肝胰脏、鳃丝和血液SOD活力的影响

研究了3种重金属离子(Cu²⁺、Zn²⁺、Cd²⁺)在96 h内对凡纳滨对虾(Litopenaeus vannamei)对肝胰脏、鳃丝和血液超氧化物歧化酶(SOD)活力的影响.结果表明,凡纳滨对虾SOD活力在3种重金属离子作用下随取样时间变化显著(P＜0.0),Cu²⁺在实验浓度范围内(0.1～1 mg·L^-1),肝胰脏、鳃丝和血液的SOD活力随时间延长呈一峰值变化,Zn²⁺在10 mg·L^-1时对肝胰脏表现为显著抑制作用,Cd²⁺在0. mg·L^-1时对肝胰脏和鳃丝起显著抑制作用,0.2 mg·L^-1对鳃丝SOD活力无显著变化(P＞0.0),其他浓度Zn²⁺(＜10 mg·L^-1)、Cd²⁺(＜0.2 mg·L^-1)对各组织器官SOD活力的影响随时间延长均呈现先升高后下降的趋势.3种重金属离子对凡纳滨对虾肝胰脏、鳃丝、血液SOD活力的影响呈现明显的剂量-时间效应关系.其SOD活力大小顺序为肝胰脏＞鳃丝＞血液,3种重金属离子对凡纳滨对虾伤害大小顺序为Cd²⁺＞Cu²⁺＞Zn²⁺.     

Investigations into enzymes of nitrogen metabolism of the ectomycorrhizal basidiomycete, Suillus bovinus

Axenic mycelia of the ectomycorrhizal basidiomycete, Suillus bovinus, were grown in liquid media under continuous aeration with compressed air at 25 degrees C in darkness. Provided with glucose as the only carbohydrate source, they produced similar amounts of dry weight with ammonia, with nitrate or with alanine, 60-80% more with glutamate or glutamine, but about 35% less with urea as the respectively only exogenous nitrogen source. In crude extracts of cells from NH4(+)-cultures, NADH-dependent glutamate dehydrogenase exhibited high aminating (688 nmol x mg protein(-1) x min(-1)) and low deaminating (21 nmol x mg protein(-1) x min(-1)) activities. Its Km-values for 2-oxoglutarate and for glutamate were 1.43 mM and 23.99 mM, respectively. pH-optimum for amination was about 7.2, that for deamination about 9.3. Glutamine synthetase activity was comparatively low (59 nmol x mg protein(-1) x min(-1)). Its affinity for glutamate was poor (Km = 23.7 mM), while that for the NH4+ replacing NH2OH was high (Km = 0.19 mM). pH-optimum was found at 7.0. Glutamate synthase (= GOGAT) revealed similar low activity (62 nmol x mg protein(-1) x min(-1)), Km-values for glutamine and for 2-oxoglutarate of 2.82 mM and 0.28 mM, respectively, and pH-optimum around 8.0. Aspartate transaminase (= GOT) exhibited similar affinities for aspartate (Km = 2.55 mM) and for glutamate (Km = 3.13 mM), but clearly different Km-values for 2-oxoglutarate (1.46 mM) and for oxaloacetate (0.13 mM). Activity at optimum pH of about 8.0 was 506 nmol x mg protein(-1) x min(-1) for aspartate conversion, but only 39 nmol x mg protein(-1) x min(-1) at optimum pH of about 7.0 for glutamate conversion. Activity (599 nmol x mg protein(-1) x min(-1)), substrate affinities (Km for alanine = 6.30 mM, for 2-oxoglutarate = 0.45 mM) and pH-optimum (6.5-7.5) proved alanine transaminase (= GPT) also important in distribution of intracellular nitrogen. There was comparatively low activity of the obviously constitutive enzyme, urease, (42 nmol x mg protein(-1) x min(-1)) whose substrate affinity was rather high (Km = 0.56 mM). Nitrate reductase proved substrate induced; activity could only be measured after exposure of the mycelia to exogenous nitrate. Routes of entry of exogenous nitrogen and tentative significance of the various enzymes in cell metabolism are discussed.     

A comparison of 60, 70, and 90 kDa stress protein expression in normal rat NRK-52 and human HK-2 kidney cell lines following in vitro exposure to arsenite and cadmium alone or in combination

Arsenite and cadmium are two potent nephrotoxicants and common Superfund site elements. These elements are included among the stress protein inducers, but information regarding relationships between toxicity produced by combinations of these agents to the stress protein response is lacking. In this study, the immortalized cell lines normal rat kidney NRK-52E and human kidney HK-2 were exposed in vitro to arsenite (As(3+)), cadmium (Cd(2+)), or to equimolar As(3+) plus Cd(2+) mixture combinations for 3 and 5 h over a concentration range of 0.1-100 microM. After a 12-h recovery period, cultured cells were then evaluated for expression of the 60, 70, and 90 kDa major stress protein families. Results indicated that expression of stress proteins varied depending on the species of kidney cells exposed, the exposure concentrations, and the length of exposure to each element on an individual basis and for combined mixtures. For the HK-2 kidney cell line, increased levels of the 70 kDa stress protein was observed for single and combined element exposures whereas there was no change or a decrease of stress proteins 60 and 90 kDa. Increased 70 kDa expression was observed for 10-microM doses of single elements and for a lower dose of 1 microM of the As plus Cd mixture at 3- and 5-h exposures. NRK-52 kidney cells exposed to equivalent doses of As(3+) and Cd(2+) alone or in combination showed increased levels of all stress proteins 60, 70, and 90 kDa. This increase was seen for 10 microM of the As plus Cd mixture at 3 h whereas for single element exposures, increased stress protein levels were generally observed for the 100-microM doses. At 5 h- exposure, 60 and 90 kDa levels increased for 10 microM of Cd(2+) and 60 kDa levels increased for 1 microM of As(3+). However, exposures to 10 microM of the As plus Cd mixture decreased 60 kDa protein expression to control levels at 5 h. For both kidney cell lines, there was a decrease in the stress protein expression levels for all three stress protein families for 100-microM doses of the mixture combination for 3- and 5-h exposures. These data indicate a dose- and combination-related correlation between depression of the stress protein response and the onset of overt cellular toxicity and/or cell death. The threshold for these changes was cell line specific.