Metallobiochemistry of ultratrace levels of bismuth in the rat II. Interaction of 205+206Bi3+ with tissue,intracellular and molecular components

BackgroundKnowledge on Bi metabolism in laboratory animals refers to studies at “extreme” exposures, i.e. pharmacologically relevant high-doses (mg kg⁻¹ b.w.) in relation to its medical use, or infinitesimal doses (pg kg⁻¹b.w.) concerning radiobiology protection and radiotherapeutic purposes. There are no specific studies on metabolic patterns of environmental exposure doses (ultratrace level, μg kg⁻¹ b.w.), becoming in this context Bi a “heavy metal fallen into oblivion”. We previously reported the results of the metabolic fate of ultratrace levels of Bi in the blood of rats [1]. In reference to the same study here we report the results of the retention and tissue binding of Bi with intracellular and molecular components.MethodsAnimals were intraperitoneally injected with 0.8 μg Bi kg⁻¹ b.w. as ^205+206Bi(NO)₃, alone or in combination with ⁵⁹Fe for the radiolabeling of iron proteins. The use of ^205+206Bi radiotracer allowed the determination of Bi down to pg fg⁻¹ in biological fluids, tissues, subcellular fractions, and biochemical components isolated by differential centrifugation, size exclusion chromatography, solvent extraction, precipitation, immunoprecipitation and dialysis.Main findingsAt 24 h post injection the kidney contained by far the highest Bi concentration (10 ng g⁻¹ wt.w.) followed by the thymus, spleen, liver, thyroid, trachea, femur, lung, adrenal gland, stomach, duodenum and pancreas (0.1 to 1.3 ng g⁻¹ wt.w.). Brain and testis showed smaller but consistently significant concentrations of the element (0.03 ng g⁻¹ wt.w). Urine was the predominant route of excretion. Intracellularly, liver, kidney, spleen, testis, and brain cytosols displayed the highest percentages (35%–58%) of Bi of homogenates. Liver and testis nuclei were the organelles with the highest Bi content (24 % and 27 %). However, when the recovered Bi of the liver was recorded as percent of total recovered Bi divided by percent of total recovered protein the lysosomes showed the highest relative specific activity than in other fractions. In the brain subcellular fractions Bi was incorporated by neuro-structures with the protein and not lipidic fraction of the myelin retaining 18 % of Bi of the total homogenate. After the liver intra-subcellular fractionation: (i) 65 % of the nuclear Bi was associated with the protein fraction of the nuclear membranes and 35 % with the bulk chromatin bound to non-histone and DNA fractions; (ii) about 50 % of the mitochondrial Bi was associated with inner and outer membranes being the other half recovered in the intramitochondrial matrix; (iii) in microsomes Bi showed a high affinity (close to 90 %) for the membranous components (rough and smooth membranes); (iv) In the liver cytosol three pools of Bi-binding proteins (molecular size > 300 kDa, 70 kDa and 10 kDa) were observed with ferritin and metallothionein–like protein identified as Bi-binding biomolecules. Three similar protein pools were also observed in the kidney cytosol. However, the amount of Bi, calculated in percent of the total cytosolic Bi, were significantly different compared to the corresponding pools of the liver cytosol.ConclusionsAt the best of our knowledge the present paper represents the first in vivo study, on the basis of an environmental toxicology approach, aiming at describing retention and binding of Bi in the rat at tissue, intracellular and molecular levels.     

Toxicity of baited spinosad formulations to Ceratitis capitata: from the laboratory to the application

GF‐120, a fruit fly bait designed to attract and kill adult fruit flies, was tested in the laboratory and outdoors to determine effects of pre‐treatment diet and bait aging on mortality of Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Two spinosad‐based compounds, GF‐120 and Tracer^® Ultra, had generated two distinctive dose–mortality responds, with LC⁸⁰, LC⁹⁰, and LC⁹⁹ values of 2.4, 2.8, and 4.1 p.p.m., and 255, 479, and 1 143 p.p.m., respectively. The residues of GF‐120 drops, after feeding to the flies, generated 14.3% mortality. The droplet size of the baited spray plays an important role. The toxicity of large drops lasted more than that of small droplets. In the field, exposure to the sun further deteriorates the compound, which lost 50% of its toxicity within 6 days. Disappearance of the compound in the field, due to consumption by various insects, also played a role as 50% of the GF‐120 drops disappeared within 7 days. As mortality was directly related to the amount of insecticide eaten, the effect of GF‐120 depended on the feeding status of the flies: well‐fed flies were almost unaffected compared with starved ones.     

DNA-Binding and Photocleavage Properties of Ru(II) Polypyridyl Complexes with DNA and Their Toxicity Studies on Eukaryotic Microorganisms

Four Ru(II) polypyridyl complexes, [Ru(bpy)₂(7-NO₂-dppz)]²⁺, [Ru(bpy)₂(7-CH₃-dppz)]²⁺, [Ru(phen)₂(7-NO₂-dppz)]²⁺, and [Ru(phen)₂(7-CH₃-dppz)]²⁺ (bpy = 2,2′-bipyridine, phen = 1,10-phenanthroline), (7-Nitro-dppz = 7-Nitro dipyrido[3,2-a:2′-3′-c]phenazine, 7-CH₃-dppz = 7-Methyl dipyrido[3,2-a:2′-3′-c]phenazine), have been synthesized and characterized by IR, UV, elemental analysis, ¹H NMR, ¹³C-NMR, and mass spectroscopy. The DNA-binding properties of the four complexes were investigated by spectroscopic and viscosity measurements. The results suggest that all four complexes bind to DNA via an intercalative mode. Under irradiation at 365 nm, all four complexes were found to promote the photocleavage of plasmid pBR 322 DNA. Toxicological effects of the selected complexes were performed on industrially important yeasts (eukaryotic microorganisms).     

SHORT-TERM IMPACTS OF FORMULATIONS OF BACILLUS THURINGIENSIS VAR. ISRAELENSIS DE BARJAC AND THE ORGANOPHOSPHATE TEMEPHOS,USED IN BLACKFLY (DIPTERA: SIMULIIDAE) CONTROL,ON RHEOPHILIC BENTHIC MACROINVERTEBRATES IN THE MIDDLE ORANGE RIVER,SOUTH AFRICA

Summary

The impacts of larvicides used in the control of blackflies (Diptera: Simuliidae) on macroinvertebrates in the stones-in-current biotope were assessed during 8 field trials in the middle Orange River, South Africa. Two Bacillus thuringiensis var. israelensis (B.t.i.) products (Vectobac^R 12AS and Teknar^R HP-D) and the organophosphate temephos (Abate^R 200EC) were applied at recommended and high dosages to simulate “operational” and “worst-possible” scenario's respectively. Mortality was evaluated either by direct counting of invertebrates on stones before and after application, or by ranking invertebrates on a 4-point relative abundance scale before and after application. In addition, the re-appearance of benthic invertebrate population densities after temephos application was examined.

At the recommended dosage (1.2 ppm/10 min), B.t.i. significantly reduced blackfly larval numbers (P<0.001) and those of the chironomid Rheotanytarsus fuscus Freeman (P<0.05). At high dosage (20 ppm/10 min), numbers of the filter-feeding mayfly Tricorythus discolor (Burmeister) (P<0.01) and the chironomid Cardiocladius sp. (P<0.05) were also significantly reduced. No Simulium predators were directly affected by B.t.i., but there were indications of food shortage amongst Hydropsychidae and Hirudinea.

Temephos caused significant reductions in the relative abundance of 5 taxa at 0.05 ppm, 3 to 6 taxa at 0.1 ppm, and 9 taxa at 1.0 ppm (P<0.05). “Non-target” organisms which were most affected included the chironomid R. fuscus, the mayflies Baetis glaucus Agnew and Choroterpes elegans Barnard, and the caddisflies Cheumatopsyche thomasseti Ulmer and Amphipsyche scottae Kimmins. The mayfly T. discolor was tolerant of temephos, even at high dosage (1.0 ppm/10 min). In winter, most taxa re-appeared within 19 days, and population densities were back to pre-treatment levels within 35 days.

It is concluded that good reduction of blackfly populations may be obtained with minimal direct impact on the “non-target” fauna, provided recommended dosages of temephos are not exceeded. Overdosing with temephos may result in high mortality of “non-target” organisms, including blackfly predators, and should be avoided.     

The Effect of Formaldehyde on Tissue Lipids and on Histochemical Reactions for Carbonyl Groups

Histcchemical and chemical evidence indicates that formaldehyde combines with unsaturated lipids at the double bond. The resulting complex contains a free carbonyl group which probably originates from the formaldehyde. The reaction occurs over a wide pH range, and takes place in the absence of oxygen or moisture. The reaction product is visualized by the Schiff reagent, and by the Ashbel-Seligman procedure. In the plasmal procedure, when performed on formalin-treated material, the reaction has the same significance as the pseudo-plasmal reaction, i.e. it denotes the presence of double bonds. The Ashbel-Seligman technic seems to be more sensitive to this complex than the Schiff reagent and shows it more markedly than it does the true plasmals and the atmospherically oxidized unsaturated compounds.     

Inhibition of two enzyme systems in Euchlanis dilatata (Rotifera: Monogononta) as biomarker of effect of metals and pesticides

The inhibitory effects on esterases and phospholipase A2 (PLA2) in the freshwater rotifer Euchlanis dilatata, native to Mexico, were assessed by fluorimetry after in vivo exposure (30?min) in laboratory conditions to sublethal concentrations of metals and pesticides. EC₅₀ values for esterases ranged from 7.9?×?10^?7 for DDT to 61.9 μg l^?1 for methyl parathion, while corresponding values for PLA2 ranged from 0.96?×?10^?6 for mercury to 69.2 μg l^?1 for lead. These enzyme systems in E. dilatata are very sensitive to the tested agents and suggest they would be suitable biomarkers. However, sensitivity to other environmental contaminants should be investigated in laboratory conditions and field studies to assess their potential as environmental biomarkers.     

A strategy for modelling dynamic responses in metabolic samples characterized by GC/MS

A multivariate strategy for studying the metabolic response over time in urinary GC/MS data is presented and exemplified by a study of drug-induced liver toxicity in the rat. The strategy includes the generation of representative data through hierarchical multivariate curve resolution (H-MCR), highlighting the importance of obtaining resolved metabolite profiles for quantification and identification of exogenous (drug related) and endogenous compounds (potential biomarkers) and for allowing reliable comparisons of multiple samples through multivariate projections. Batch modelling was used to monitor and characterize the normal (control) metabolic variation over time as well as to map the dynamic response of the drug treated animals in relation to the control. In this way treatment related metabolic responses over time could be detected and classified as being drug related or being potential biomarkers. In summary the proposed strategy uses the relatively high sensitivity and reproducibility of GC/MS in combination with efficient multivariate curve resolution and data analysis to discover individual markers of drug metabolism and drug toxicity. The presented results imply that the strategy can be of great value in drug toxicity studies for classifying metabolic markers in relation to their dynamic responses as well as for biomarker identification.     

低剂量混合污染生态毒理与风险评价研究进展

环境中的化学品往往以低剂量混合形式存在.对单一化学品高剂量暴露下的生态毒性研究成果,难以适用于环境中低剂量混合物的生态毒理效应诊断及风险评价.文中概述了低剂量化学品混合污染生态毒理及风险评价方面的研究进展,主要包括低剂量化学品混合污染诊断的分子毒理研究方法、风险评价方法,并介绍了简单和复杂混合物的风险评价方案.对低剂量混合污染生态毒理与风险评价研究的发展动向提出了见解,指出低剂量化学混合物的研究需要寻找敏感终点,引入多学科手段,积累更多的数据,建立完善、统一的评价体系.     

代谢组学及其在环境毒理学中的应用

代谢组学作为系统生物学的一部分,因其具有分析速度快的特点,被广泛用于生物医学等方面的研究。目前代谢组学在环境毒理学方面的研究主要针对单一污染物,但也需要考虑到被污染地的复杂情况。通过介绍代谢组学及其发展历程,总结了目前主流代谢组学技术的各自特点,讨论了代谢组学在环境重金属、有机污染物和抗生素中的毒性评估以及环境胁迫耐受性中的评价等方面内容,综述了其在环境毒理学中的应用,并指出其应用不足,旨在为代谢组学应用于环境毒理学的研究提供参考。     

石墨烯健康风险研究现状及展望

石墨烯是一种新型的二维碳纳米材料,由于具有优异的电子、光学、机械等特性,已经被广泛应用于电子器件、复合材料、能源储存等领域.近年来,石墨烯在生物医药领域崭露头角,其在诸如生物传感器、细胞成像、药物输运、抗菌材料等方面的广泛应用,为生物医药技术带来了突破,也为人体健康带来了福音.然而,随着石墨烯以不同途径进入人们的生活,其对人体及其他生物体的安全构成潜在威胁,引发的健康风险正受到广泛关注.本文从石墨烯对生物体的影响及其同生物体的相互作用方面入手,综述了近年来石墨烯健康风险的研究进展,并且总结归纳了人体抵御石墨烯健康风险的途径及机制,最后指出了未来石墨烯健康风险方面的研究方向.