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.     

Effect of white African mineral hair dye on the activities of phosphatases and malondialdehyde level in selected tissues of albino rats

The effect of white African mineral dye Yombofita (YF) on the activities of alkaline phosphatase (ALP), acid phosphatase (ACP) and malondialdehyde (MDA) levels in the skin, liver, kidney and serum of albino rats was investigated. The chemical analysis of the dye was first carried out using solubility test, pH determination and X-ray fluorescence (XRF) elemental analysis. Six different concentrations (0.05, 0.15, 0.25, 0.5, 0.75 and 1.00%) of the dye were prepared using hydrogen peroxide (30 volume) as solvent. A total of 80 albino rats (Rattus norvegicus) were used for the study. The rats were divided into 8 groups of 10 each and were maintained on commercial feed for the period of the experiment i.e. 30 days. In group 1, the control group, the animals were applied distilled water on their heads, whereas in group 2 the vehicle i.e. hydrogen peroxide was applied. In groups 3 to 8 various concentrations of YF (white) dye ranging from 0.05, 0.15, 0.25, 0.5, 0.75 to 1.00% was applied respectively. At the end of the experiment, blood samples were collected and portions of the selected tissues were excised for the determination of ALP and ACP activities. The MDA level was also determined in the skin of experimental animals. The results revealed a significant decrease (p?p?相似文献   

Determination of types of interactions exhibited by binary mixtures of heavy metals tested against the hermit crab,Clibanarius africanus

Single and joint action toxicity of heavy metal compounds (ZnCO₃, CuSO₄ and CdSO₄) were carried out against the hermit crab, Clibanarius africanus in laboratory bioassays. On the basis of 96?h LC₅₀ values, CuSO₄ (96?h LC₅₀?=?0.03?mM) was found to be the most toxic followed by CdSO₄ and ZnCO₃ (96?h LC₅₀?=?0.24?mM) in decreasing order of toxicity. The evaluation of the metal compounds in mixtures of ZnCO₃–CuSO₄, ZnCO₃–CdSO₄ and CdSO₄–CuSO₄ in pre-defined molar ratios revealed that interactions between Zn–Cu and Zn–Cd test mixtures were in conformity with the models of antagonism (reduction in toxicity of constituent metals). The synergistic ratios (S.R.s) and relationship between observed/predicted 96?h LC₅₀ values (relative toxic unit, RTU) were consistently below 1, indicating antagonistic interactions. Analysis using isobolograms also indicated the interactions between the mixture components to be antagonistic. However, with regards to CdSO₄–CuSO₄ test mixtures, the interactions between these metals were mainly in conformity with the model of synergism. The S.R. and RTU values were largely above 1, indicating synergistic interactions. The importance of the joint action evaluations for the management and protection of aquatic ecosystems, particularly in the setting up of water quality criteria was discussed.     

A study of biochemical parameters among individuals residing in a fluoride endemic area in India

Fluoride (Fl) exerts an inhibitory effect on many metabolic enzymes of various tissues. A study was carried out among individuals residing in a Fl-affected area in India. The biochemical parameters including serum enzyme activities of alkaline phosphatase (ALKP), glutamic pyruvic transaminase (SGPT), lactate dehydrogenase (LDH), and serum creatinine (CRTN) were estimated in 259 subjects compared to 233 controls. The results showed that among the Fl-exposed population, ALKP and SGPT were significantly increased compared to control. The CRTN levels were also significantly increased compared to the control subjects. LDH levels were not markedly altered. Data suggest that Fl exposure affects renal and hepatic functions in endemic areas of India.     

Fatty acid composition of marine macroalgae from the Black Sea and Dardanelles

The fatty acid (FA) composition was determined in 14 species of marine macroalgae belonging to the Chlorophyta, Phaeophyta and Rhodophyta, which were collected from ?ile in the Black Sea and Kepez in the Dardanelles. Generally, polyunsaturated FAs and monounsaturated FAs were major components (50–77%). Total saturated FAs ranged from 22% to 50%, with 16?:?0 as the most abundant saturate (32–38%). Two samples of Cystoseira barbata collected from a different station had some differences from each other in their contents of 18?:?2n-6 and 18?:?3n-3 and in the 18?:?2n-6/20?:?4n-6 ratios. Green algal species had a significantly higher proportion of unsaturated FAs and a significantly lower proportion of saturated FAs than the red and brown algae. The amount of n-3 FAs was significantly higher in Ulva rigita, Chaetomorpha linum, Enteromorpha linza and Gracilaria verrucosa (8.88, 6.44, 5.31 and 5.24, respectively).     

In vitro genotoxicity of fly ash leachate in earthworm coelomocytes

Fly ash is the major coal combustion byproduct from thermal power plants. Considering its plant–nourishing constituents, its soil amendment in farmland is one of its promoted disposal methods. A substantial amount of heavy metals present in fly ash, which may leach out due to rainwater or irrigation water, may cause serious problem with long term use, especially to soil organisms. These metals may cause DNA damage through Reactive Oxygen Species (ROS) generation. In the present study, single cell gel electrophoresis [(SCGE) i.e., comet assay] was used to detect DNA damage in earthworm (Dichogaster curgensis) coelomocytes, following an in vitro exposure. Significant DNA damage was observed at the lowest concentration of fly ash leachate (6.25%) examined. DNA damage by all the tested concentrations (6.25%, 12.5%, 25%, 50%) differed significantly (p?<?0.001) from that of the negative control. Hence, long-term application of fly ash might prove harmful for earthworm populations.     

Adult respiratory distress syndrome in human organophosphate poisoning cases

Adult respiratory distress syndrome (ARDS) is a clinical syndrome which occurs due to various direct or indirect noxious agents and pyhsical mechanisms. One of the etiological factors is ingested chemicals (e.g. paraquat, ethylene glycol). The organophosphorous (OP) insecticides are the most toxic and frequently encountered chemicals that produce poisoning in humans. Studies were undertaken to investigate whether OP insecticides may produce ARDS. Fifteen autopsy cases due to suicidal OP insecticide poisoning were investigated. Toxicological analyses were performed with a gas chromatography-nitrogen phosphorous detector (GC-NPD). Methamidophos, dimefox and dichlorvos were the detected OP. Lungs were examined macroscopically. Two cases showed features compatible with ARDS in which the only cause of death was OP poisoning. Due to the severity of ARDS it is important to assess insecticidal exposure in an attempt to enhance survival patterns following OP poisoning in patients in order to initiate therapy.     

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.     

Feasibility of metabolic parameter estimation in pharmacokinetic models of carbon tetrachloride exposure in rats1

Carbon tetrachloride (CCl₄) is a toxic chemical that was once used in degreasers and detergents, and some remnants of the chemical may be present in the water supply. Physiologically-based pharmacokinetic (PBPK) modeling can assist in understanding resulting internal doses of CCl₄ after exposure, but the pharmacokinetic parameters describing the metabolism of CCl₄ are not well characterized. The goal of this study was to provide insights into how to more accurately estimate these values in rats using PBPK modeling and data from previous studies. Three different PBPK models were constructed to describe CCl₄ exposure in rats via inhalation, oral ingestion, and venous injection. Each of these models was compared to data, and sensitivity analysis was performed for each model to determine whether the available data could be used to accurately determine the metabolic parameters of interest. These parameter sensitivities were so low that optimization to the available data yielded physiologically unrealistic results. Model sensitivities were analyzed for different doses and routes of exposure in order to find experimental conditions that would allow for greater identifiability of the metabolic parameters. Data were simulated from these models at optimal conditions with varying levels of noise from a normal distribution. Optimizations were then performed to confirm that the original values could be obtained. The experiments developed are left as suggestions for investigators who wish to further pursue estimating these metabolic parameters.