Quantification of metallothionein in whole body gammarus locusta (crustacea: amphipoda) using differential pulse polarography

A polarographic method was optimised for the quantitative determination of metallothionein (MT) in whole body Gammarus locusta (Crustacea: Amphipoda). The identity of the MT-like proteins was examined using electrophoretic analysis (SDS-PAGE) and induction studies (water-borne exposures to copper). The MT-method was then used to examine changes in MT levels over the lifetime (juveniles, sub-adults and adults) of a laboratory population of G. locusta; copper (Cu) and zinc (Zn) body burden were also measured by flame atomic absorption spectrometry. MT was quantified by heat-denaturation of 30?000?g supernatants (to remove the bulk of non-MT proteins) followed by differential pulse polarography (DPP) to measure sulphydryl-group containing proteins. DPP proved to be a sensitive and reliable method to quantify heat-stable total sulphydryl-proteins. The majority of the latter was indicated to be MT from SDS-PAGE and induction studies. SDS-PAGE revealed a putative MT of 23?±?0.6?kDa app. mol. wt., consistent with the average size of dimeric MTs (10–20?kDa) found in several aquatic invertebrate species. Copper-exposure revealed parallel increases in the amounts of 23?kDa protein and total sulphydryl-proteins. Levels of whole body MT (rabbit pure MT as standard) range between 1.3 and 2.3?mg?g^?1 dry wt. over the lifetime of G. locusta. Sex-related changes in MT status were observed, the levels in females being 1.6 times higher than in males (p<0.05). Copper and Zn body content changed over the lifetime of G. locusta, the levels in juveniles (p>0.05) and sub-adults (p<0.05) being higher than in adults. It is concluded that the inclusion of MT in ecotoxicological studies with amphipod crustaceans require some caution with respect to the stages of moulting and/or sexual maturation of organisms, but this should not provide any major impediment in its use in metal pollution monitoring.     

The distribution,metabolism and toxicity of 14c-DDT in model aquarium tanks with fish and sediment simulating a tropical marine environment

Studies conducted on the distribution, fate and metabolism of DDT in a model ecosystem simulating a tropical marine environment of fish, Gobious nudiceps, Lethrinus harak, Gobious keinesis, Gobious nebulosis and white shrimp (Panaeus setiferus), show that DDT concentration in the water decreases rapidly within the first 24?h. Rapid accumulation of the pesticide in the biota also reaches a maximum level in 24?h before gradually declining. The bioaccumulation factors calculated for the fish species (G. keinesis) and white shrimp (P. Setiferus) were 270 and 351, respectively, after 24?h. There was a steady build up of DDT residues in the sediment during the first 24?h which continued to a maximum concentration of 6.66?ng/g in the seawater/fish/sediment ecosystem after 3 weeks and 5.27?ng/g in the seawater/shrimps/sediment ecosystem after 2.7 days. The depuration of the accumulated pesticide was slow with only 54% lost in G. nudiceps within 3 days of exposure in fresh sea water. By contrast, depuration was fast in the white shrimp, which lost 97% of the accumulated pesticide under the same conditions. DDT was found to be toxic to two of the fish species (G. nebulosis and L. harak) and to white shrimp, and the degree of toxicity was dependent on the particular species. The 24?h LC₅₀ at room temperature for the fish species G. nebulosis and white shrimp was found to be 0.011 and 0.116?mg/kg, respectively. These levels are comparable to the ones recorded for the temperate organisms. Degradation of DDT to its primary metabolites, DDE and DDD, was found in all the compartments of the ecosystem with DDE being the major metabolite in the fish, shrimps and sediment, while in seawater, DDD dominated as the major metabolite.     

The correlation of total and extractable heavy metals from soil and domestic sewage sludge and their transfer to maize ( Zea mays L.) plants

A study to understand the mobility and transport of heavy metals (HMs) from soil and soil amended with sewage sludge to maize plants was carried out. The total and ethylenediaminetetraaceticacid (EDTA)-extractable HMs in agricultural soil and untreated domestic sewage sludge samples, and the correlation between the total and extractable metals in soil and sewage sludge were carried out. Pot experiments were performed to study the transfer of HMs to maize grains, grown in soil (control) and in soil amended with sewage sludge (test samples). The total and extractable HMs in soil, sewage sludge, and maize grains were analysed by FAAS/ETAAS (flame atomic absorption spectrometer/electrothermal atomic absorption spectrometer) after digestion in microwave oven. Statistically significant correlations were obtained between the total contents of Cu, Cd, As and their respective extractable fractions in soil, while in domestic wastewater sludge (DWS) the better correlation was observed only for Ni and Cd. The edible part of maize plants (grains) from test samples presented high concentration of Zn, Pb, Ni, Cd, Cu, As, and Cr concentrations (80.7–85.6, 3.8–3.95, 2.35–2.5, 0.75–0.82, 3.21–3.29, 0.23–0.27, and 0.22–0.29?mg?kg^?1, respectively). Good correlations were found between metals in exchangeable fractions of both soil and DWS and total metals in control and test samples of maize grains. The transfer factor of all HMs from DWS to maize grains was also determined.     

Lead distribution in rats following respiratory exposure to lead-contaminated soils

Two preliminary experiments were performed to explore the use of an intratracheal instillation technique as a cost-effective method of determining the biokinetics of lead (Pb) following respiratory exposure to Pb-contaminated soils. A novel intratracheal instillation procedure was refined and used to deliver a defined dose of Pb-contaminated soil or PbAc to the lower respiratory tract of rats. In the first experiment, rats were sacrificed at numerous time intervals post-dosing, and liver, kidney, blood, and bone tissues were collected for Pb analysis. In the second experiment, rats were dosed with Pb-contaminated soil or PbAc via intratracheal instillation or gastric gavage. All rats were sacrificed 96 h after dose administration and tissues were collected for lead analysis. Data from these experiments indicate Pb is well-absorbed following intratracheal instillation of Pb-contaminated soil, and the intratracheal instillation technique could be used as a cost effective method for exploring the biokinetics of Pb in Pb-contaminated soils following respiratory tract exposure.     

Interaction of alcohol and kola nut on brain sodium pump activity in Wistar rats

The effect of interaction of alcohol and kola nut on sodium pump activity was studied in Wistar rats. Thirty Wistar rats were divided into six groups of five rats per group. Control received a placebo (4 mL of distilled water). Groups 2–6 were treated for a period of 21 days with 10% (v/v) alcohol, 50 mg caffeine/kg, 50 mg kola nut/kg, or a combination of 10% (v/v) alcohol + 50 mg kola nut/kg, or 10% (v/v) alcohol + 50 mg caffeine/kg, respectively. One day after the final exposure, brains were harvested and several biochemical parameters examined including activities of total ATPase, ouabain-insensitive ATPase, ouabain sensitive ATPase (Na⁺–K⁺-ATPase) and levels of nonenzymatic breakdown of ATP and inorganic phosphate (P_i) released. Results showed that the essential enzyme of the brain responsible for neuronal function, Na⁺–K⁺-ATPase, was inhibited by alcohol–kola nut co-administration relative to control, resulting in a decreased ATP production, ion transport and action potential, leading to loss of neuronal activities.     

Pharmacokinetics of [14C]-atrazine in rhesus monkeys,single-dose intravenous and oral administration

This is the first study regarding the pharmacokinetics of [¹⁴C]-atrazine conducted with rhesus monkeys. The animals received one dose (0.25 mg) intravenously (IV) or three doses (1, 10, or 100 mg) orally. Plasma, urine, and feces were collected at defined times up to 7 days post-dosing. Sample radioactivity was measured to determine the mass equivalent. IV administered [¹⁴C]-atrazine disappeared rapidly from blood, with an elimination half-life of about 5.5 ± 1.1 h. The pharmacokinetic profiles of [¹⁴C]-atrazine following oral administration at the three dose levels show that kinetic parameters such as AUC and C _max are linearly correlated with the dose. Seven days after dosing, urinary and fecal excretion of [¹⁴C]-atrazine reached 99% of total administered dose in the IV group and 91–95% in the three oral dose groups. In the IV-administered monkeys, approximately 85% of the dose was excreted in urine and 12% in feces. In three oral dose groups, urinary and fecal radioactivity recoveries approximated 57% and 21%, 58% and 25%, and 53% and 35%, respectively. More than 50% of the total urinary excreted radioactivity was found within the first 24 h after dosing. In conclusion, the principal elimination of [¹⁴C]-atrazine, IV and orally administered, is via urine. The oral bioavailability was 60% or higher. There was a significant linear correlation between administered oral dose and plasma concentration. Overall oral dose accountability ranged from 91% to 95%. Data generated may be useful in the risk assessment of human exposure to environmental atrazine contamination.     

Assessment of the state and management of the cephalopod trawl fishery resources in Greek waters

The cephalopod trawl fishery in Greek waters is reviewed in relation to fishing effort for 1964–1981. The mean annual (1964–81) catch of cephalopods amounted to 1348 tons. Total landings of cephalopods are very well described by the exponential⁹ and linear²⁹ surplus yield models, according to which optimum fishing effort, maximum sustained yield and optimum catch per unit of fishing effort range between 120000–150000HP, 1586–1630 tons and 10–13kgr/HP respectively. In addition, the analysis revealed that cephalopod resources were slightly overfished (according to the linear model) during 1975–1981. Hence, authorities must take immediate measures for the protection and rational management of cephalopod resources in Greek waters. Possible alternative measures are discussed in the text.     

Heavy metals in coastal sediments in Guipuzcoa (Spain)∗

Surficial sediment samples were wet digested (HNO₃‐HCI‐HF) and analysed by atomic absorption spectroscopy. The background values in the area, and the Pollution Index Load in the sampling points were calculated. Metal inputs of continental origin are specially important in Deva, Oria and Urola river mouths.     

Singlet molecular oxygen—mediated photooxidation of monochloro and mononitrophenols. A kinetic study

A kinetic and mechanistic study on the aerobic dye sensitized photooxidation of the mono—nitro and chlorophenols was carried out. A singlet molecular oxygen mechanism operates in the photooxidation. Solvent and substituent effects, suggest the intermediacy of a complex with partial charge transfer character, as has been postulated for other phenols.

Chemical (reactive) and physical interactions of the substrates with singlet molecular oxygen were discriminated. Quantum yields for photooxidation (higher for the chlorophenols) range from 3 × 10^‐2 to 2 × 10^‐3, as measured by substrate or oxygen consumption. These values indicate the viability of a singlet molecular oxygen photooxidation as a way for the degradation of nitro and chlorophenol environmental contaminants.     

Hydroxyl radical generation by redox cycling of some chemotherapeutic antibiotics

The anticancer drugs: adriamycin, farmorubicin and mitomycin C greatly enhance the generation of hydroxyl radicals (HO^.) from H₂O₂ in the presence of Co(II) ions (CoCl2) at pH 7.4 and 8, as measured by the deoxyribose assay. Catalase, hydroxyl radical scavengers (mannitol, cysteine, glutathione, thiourea, lactic dehydrogenase) inhibited the degradation of deoxyribose confirming that HO‐radicals are responsible for the degradation of the carbohydrate.