Nucleosomal structure as probed by H3 histone thiol reactivity

Two H3 histone variants are found in equal amount in HeLa cells, and they have been characterized by two-dimensional gel electrophoresis followed by reaction with specific antibodies. These molecules are the only cysteine-containing histones, and they have been used as the target for thiol-specific reagents, in intact nuclei, isolated nucleosomes, histone complexes, and purified histones. Cysteine residues are available toN-ethylmaleimide only when histones are disassembled from the core particles. Upon reaction with these reagents, one of the H3 variants undergoes profound conformational changes, as revealed by an altered electrophoretic mobility.     

The reaction of fluorescein isothiocyanate with thiols: a method for assay of isothiocyanates

The reversible formation of dithiourethanes from fluorescein isothiocyanate and mercaptoethanol or mercaptoethylamine has been studied. It is shown that it is possible to use the reaction as the basis for a spectrophotometric titration of a number of isothiocyanates with mercaptoethanol.     

Comparative reactivity analysis of small-molecule thiol surrogates

Targeted covalent inhibitors represent an increasingly popular approach to modulate challenging drug targets. Since covalent and non-covalent interactions are both contributing to the affinity of these compounds, evaluation of their reactivity is a key-step to find feasible warheads. There are well-established HPLC- and NMR-based kinetic assays to tackle this task, however, they use a variety of cysteine-surrogates including cysteamine, cysteine or acetyl-cysteine and GSH. The diverse nature of the thiol sources often makes the results incomparable that prevents compiling a comprehensive knowledge base for the design of covalent inhibitors. To evaluate kinetic measurements from different sources we performed a comparative analysis of the different thiol surrogates against a designed set of electrophilic fragments equipped with a range of warheads. Our study included seven different thiol models and 13 warheads resulting in a reactivity matrix analysed thoroughly. We found that the reactivity profile might be significantly different for various thiol models. Comparing the different warheads, we concluded that – in addition to its human relevance - glutathione (GSH) provided the best estimate of reactivity with highest number of true positives identified.     

A Perspective on the Toxicity of Petrogenic PAHs to Developing Fish Embryos Related to Environmental Chemistry

Numerous studies demonstrate polynuclear aromatic hydrocarbons (PAHs) dissolved from weathered crude oil adversely affect fish embryos at 0.5 to 23 μg/l. This conclusion has been challenged by studies that claim (1) much lower toxicity of weathered aqueous PAHs; (2) direct contact with dispersed oil droplets plays a significant role or is required for toxicity; (3) that uncontrolled factors (oxygen, ammonia, and sulfides) contribute substantively to toxicity; (4) polar compounds produced by microbial metabolism are the major cause of observed toxicity; and (5) that based on equilibrium models and toxic potential, water contaminated with weathered oil cannot be more toxic per unit mass than effluent contaminated with fresh oil. In contrast, several studies demonstrate high toxicity of weathered oil; shifts in PAH composition were consistent with dissolution (not particle ablation), embryos accumulated dissolved PAHs at low concentrations and were damaged, and assumed confounding factors were inconsequential. Consistent with previous empirical observations of mortality and weathering, temporal shifts in PAH composition (oil weathering) indicate that PAHs dissolved in water should (and do) become more toxic per unit mass with weathering because high molecular weight PAHs are more persistent and toxic than the more abundant low molecular weight PAHs in whole oil.     

Toxicity of cadmium to Helisoma anceps and its effect on kidney ultrastructure

When specimens of Helisoma anceps were exposed to water containing 0.1, 1, and 5 ppm cadmium, mortality was proportional to concentration but tissue levels of Cd were not. The shell was the site of highest concentration and total amount of Cd due to adsorption and active accumulation. Concentrations of Cd in the kidney and mantle were equal to or greater than the remaining soft tissue. Ultrastructural changes in the kidney associated with Cd exposure were almost exclusively confined to the nephrocytes. Graded transformation of the residual body to a secondary lysosome was proportional to the Cd concentration of the medium and length of exposure. Atypical nucleoli and reticular bodies appeared in the 1-ppm Cd dose. Only after autolysis of the nephrocytes in 5 ppm did ultrastructural changes occur in the ureter cells. Ultrastructural changes in the kidney epithelia were proportional to Cd dosage but were not necessarily indicative of the tissue burden of Cd.     

Disruption of nucleobase stacking to restore reactivity

Strong intermolecular interaction can prevent an organic molecule from dissolving in a reaction solution, thereby jeopardizing its reactivity and usefulness. Nucleobases and nucleosides (especially many purines and their derivatives) are notoriously difficult to dissolve in most organic solvents, generally attributed to their strong intermolecular interactions caused by the aromaticity, polarity and hydrogen-bonding. Guided by our computational study and prediction, to address this challenge, we have found that by doping the reaction solution with toluene (an inert aromatic compound), the added solvent molecules are capable of generating the stacking interaction with the solute molecules (e.g., purine derivatives) and disrupting the intermolecular stacking of the solute molecules. Thus, this inert doping can successfully address the insoluble challenge, dissolve the poorly soluble reactants (such as purine phosphoramidites), and restore the amidite reactivity for oligonucleotide synthesis. Our research has offered a simple strategy to efficiently synthesize labile oligonucleotides, via disrupting stacking interaction with inert aromatic molecules.     

Comparison of the Acute Toxicity of Corexit 9500 and Household Cleaning Products

Surfactant formulations used in chemical dispersing agents are derived from the same functional components used in numerous household products such as dishwashing soaps and laundry detergents. During the Deep Water Horizon (DWH) oil spill response, a significant volume of chemical dispersant was deployed, causing members of the public and the media to question the role of chemical dispersant (Corexit 9500) usage in mitigating oil spill effects. Consequently, laboratory tests were conducted by regulatory agencies to further evaluate and substantiate the existing aquatic toxicity of Corexit dispersants. To help put dispersant toxicity in context, two independent accredited labs were commissioned to conduct parallel studies that compared the acute toxicity of Corexit 9500 to common household cleaning agents. The results indicate that the acute toxicity of Corexit 9500 to marine aquatic organism is either within the median range or less toxic than the household cleaning agents tested. The median LC50 value for Corexit 9500 exposures to Americamysis bahia was 42.5 mg/L (four products were less toxic and four products were more toxic); whereas, the median LC50 value for Corexit 9500 exposures to Menidia beryllina was 73.1 mg/L (one product was less toxic and seven products were more toxic).     

Fate and biodegradability of sulfonated aromatic amines

Ten sulfonated aromatic amines were tested for their aerobic and anaerobic biodegradability and toxicity potential in a variety of environmental inocula. Of all the compounds tested, only two aminobenzenesulfonic acid (ABS) isomers, 2- and 4-ABS, were degraded. The observed degradation occurred only under aerobic conditions with inocula sources that were historically polluted with sulfonated aromatic amines. Bioreactor experiments, with non-sterile synthetic wastewater, confirmed the results from the aerobic batch degradation experiments. Both ABS isomers were degraded in long-term continuous experiment by abioaugmented enrichment culture. The maximum degradation rate in the aerobic bioreactor was 1.6–1.8 gl^–1 d^–1 for 2-ABS and a somewhat lower value for 4-ABS at hydraulic retention times (HRT) of 2.8–3.3h. Evidence for extensive mineralization of 2- and 4-ABS was based on oxygen uptake and carbon dioxide production during the batch experiments and the high levels of chemical oxygen demand (COD) removal in the bioreactor. Furthermore, mineralization of the sulfonate group was demonstrated by high recovery of sulfate. The sulfonated aromatic amines did not show any toxic effects on the aerobic and anaerobic bacterial populations tested. The poor biodegradability of sulfonated aromatic amines indicated under the laboratory conditions of this study suggests that these compounds may not be adequately removed during biological wastewater treatment.     

Anthocyanins,thiols, and antioxidant scavenging enzymes are involved in Lemna gibba tolerance to arsenic

The influence of arsenic (As) on the growth and the antioxidant system of Lemna gibba L. exposed to five concentrations of As (0.0, 0.25, 0.5, 1.0, and 1.5 mg L^?1) was studied. Although As exposure reduced relative growth rate, L. gibba continued to grow even after the high accumulation of this element after five days of exposure. The concentration of the superoxide anion was unaffected by As, whereas an increase of hydrogen peroxide concentrations was observed with the increasing of As concentration. Increasing concentrations of As also increased the enzyme activity of superoxide dismutase, peroxidase, and glutathione reductase and the total and nonprotein thiols, up to 0.5 mg L^?1, whereas the anthocyanin content increased constantly with As concentration. Catalase and ascorbate peroxidase activities as well as the content of chloroplastic pigments were reduced in plants exposed to all As concentrations. These results support a major role of anthocyanins, nonprotein thiols, and antioxidant scavenging enzymes in L. gibba tolerance to toxic As concentrations.     

Acute Toxicity for some Evaporating Aromatic Hydrocarbons for Freshwater Snails and Crustaceans

Some typical and more frequent freshwater invertebrates of running waters were studied to examine the influence of styrene, xylene and benzene on their mortality. Snails Amphimelania holandri FÉR. and Lymnaea stagnalis L. and crustacenas Asellus aquaticus L. and Gammarus fossarum KOCH. , were used in the semi-static test. Compounds were added in volume concentrations of 0.005 to 0.4% v/v. For all concentrations LC₅₀ was calculated by probit method, which demonstrated that mortality depends much more on increased concentrations (depending on the initial concentration) than on the length of exposure. Styrene was the most toxic, followed by xylene and then benzene. The species G. fossarum showed marked sensitivity, followed by A. aquaticus, and the species A. holandri and L. stagnalis showed less sensitivity.     

A Perspective on the Toxicity of Low Concentrations of Petroleum-Derived Polycyclic Aromatic Hydrocarbons to Early Life Stages of Herring and Salmon

This article presents a critical review of two groups of studies that reported adverse effects to salmon and herring eggs and fry from exposure to 1 μg/L or less of aqueous total polycyclic aromatic hydrocarbons (TPAH), as weathered oil, and a more toxic aqueous extract of “very weathered oil.” Exposure media were prepared by continuously flowing water up through vertical columns containing gravel oiled at different concentrations of Prudhoe Bay crude oil. Uncontrolled variables associated with the use of the oiled gravel columns included time- and treatment-dependent variations in the PAH concentration and composition in the exposure water, unexplored toxicity from other oil constituents/degradation products, potential toxicity from bacterial and fungal activity, oil droplets as a potential contaminant source, inherent differences between control and exposed embryo populations, and water flow rate differences. Based on a review of the evidence from published project reports, peer-reviewed publications, chemistry data in a public database, and unpublished reports and laboratory records, the reviewed studies did not establish consistent dose (concentration) response or causality and thus do not demonstrate that dissolved PAH alone from the weathered oil resulted in the claimed effects on fish embryos at low μg/L TPAH concentrations. Accordingly, these studies should not be relied on for management decision-making, when assessing the risk of very low–level PAH exposures to early life stages of fish.     

Luminol dependent chemiluminescence and thiol group oxidation provoked by neutrophils is attributable to different oxidizing species

Luminol-dependent chemiluminescence and thiol group oxidation of glutathione and human serum albumin were measured in order to demonstrate whether the inhibition of polymorphonuclear leukocyte chemiluminescence by albumin was attributable to thiol group oxidation. We have shown that:

• 1 thiol groups on glutathione and albumin are oxidized by PMNL stimulated by soluble and phagocytic stimuli;
• 2 thiol group oxidation in albumin and glutathione did not correlate with the inhibitory effects of these substances on luminol-dependent chemiluminescence with respect to time course, magnitude, effects of known scavengers or extracellular activity. It was therefore concluded that thiol group oxidation was not the cause of albumin inhibition of luminol-dependent chemiluminescence;
• 3 a metastable oxidant was identified after PMNL activation which was capable of oxidizing thiol groups but unable to elicit chemiluminescence form luminol.

     

Toxicity of hydrolysis volatile products of Brassica plants to Sclerotinia sclerotiorum,in vitro

Oilseed rape stem rot disease caused by Sclerotinia sclerotiorum causes serious yield losses worldwide. Glucosinolates as specific secondary metabolites of Brassicaceae are produced in various parts of the host plants. Their enzymatic hydrolysis releases chemical components, particularly isothiocyanates, with fungitoxic activity and volatile characteristics. To investigate the effect of volatiles derived from Brassica tissues, the pathogen was exposed to hydrolysis products of Brassica shoot parts as sources of glucosinolates including oilseed rape varieties and two species, black and white mustard. The results showed significant differences in inhibition of S. sclerotiorum growth between varieties and species. All tissues of black mustard inhibited completely the exposed colonies of the pathogen and oilseed rape varieties Dunkeld, Oscar and Rainbow had significant inhibitory effect on the fungus. The genotypes demonstrated significant differences for the production of toxic volatiles, indicating that GSL contents in Brassica species and even cultivars have different potentials for toxic products.     

百里酚对刺足根螨的毒力实验

根据植物精油在抗菌、杀虫和杀螨等方面的研究,考虑其无污染、无残留、用量少、价格低以及可阻止或延缓螨类的抗药性的特点,我们采用药膜法,进行了百里酚对刺足根螨[Rhizoglyphus echinopus(Fumouze&Rob in)]的毒力实验。结果表明,24 h的LC50=2.2222 g.L-1;48 h的LC50=0.6086 g.L-1。该实验为植物油应用于害螨防治提供了参考。     

Patterns of cortisol reactivity to laboratory stress

Cortisol responses to a laboratory stress protocol were investigated in 82 male firefighters. Saliva samples were collected during an adaptation period beginning between 9 and 10 am, and then at the end of each of six 10-min trials (a mental arithmetic task, an inter-task recovery period, a speech task, and three recovery periods). Individual differences in the mean cortisol response to the stress tasks were characterized by variation in the direction of the response, as well as the size of the response. Neither pre-stress cortisol levels nor responses were correlated with cardiovascular and mood responses. Cortisol levels before stress task presentation were negatively correlated with recent stress severity. Larger mean cortisol responses were associated with lower reports of recent stress exposure, lower negative affect scores, and a coping style characterized less experience of anger, more control over anger expression, and a tendency to screen out threatening information in stressful situations. Thus, increased cortisol activity was associated with less recent stress exposure and a more adaptive behavioral style than for those whose cortisol levels fell or were largely unchanged in response to a laboratory stressor.     

Species Differences in Developmental Toxicity of Epoxiconazole and Its Relevance to Humans

Epoxiconazole, a triazole‐based fungicide, was tested in toxicokinetic, prenatal and pre‐postnatal toxicity studies in guinea pigs, following oral (gavage) administration at several dose levels (high dose: 90 mg/kg body weight per day). Maternal toxicity was evidenced by slightly increased abortion rates and by histopathological changes in adrenal glands, suggesting maternal stress. No compound‐related increase in the incidence of malformations or variations was observed in the prenatal study. In the pre‐postnatal study, epoxiconazole did not adversely affect gestation length, parturition, or postnatal growth and development. Administration of epoxiconazole did not alter circulating estradiol levels. Histopathological examination of the placentas did not reveal compound‐related effects. The results in guinea pigs are strikingly different to those observed in pregnant rats, in which maternal estrogen depletion, pathological alteration of placentas, increased gestation length, late fetal death, and dystocia were observed after administration of epoxiconazole. In the studies reported here, analysis of maternal plasma concentrations and metabolism after administration of radiolabeled epoxiconazole demonstrated that the different results in rats and guinea pigs were not due to different exposures of the animals. A comprehensive comparison of hormonal regulation of pregnancy and birth in murid rodents and primates indicates that the effects on pregnancy and parturition observed in rats are not applicable to humans. In contrast, the pregnant guinea pig shares many similarities to pregnant humans regarding hormonal regulation and is therefore considered to be a suitable species for extrapolation of related effects to humans. Birth Defects Res (Part B) 98:230–246, 2013. © 2013 Wiley Periodicals, Inc.     

蔗渣浆CEH漂白废水的毒性及毒性污染负荷的研究

采用发光细菌法对蔗渣浆CEH三段漂白废水进行了毒性研究 ,其中C段废水的毒性最大 ,其EC50 为 2 8.83% ,属于强毒级别 ;E段、H段废水的EC50 分别为 6 8.92 %、2 6 7.9% ,分别属于毒、无毒级别 ;其中C段的毒性排放负荷约占总废水毒性排放负荷的 6 7.5 %左右 ;结果表明 ,漂白废水的毒性排放负荷较大 ,约为 4 2 3.92 (TU·m3 吨浆 ) ,相当于 4 2 .93(gHgCl2 吨浆 ) ,对其治理不容忽视     

A miniaturized technique for assessing protein thermodynamics and function using fast determination of quantitative cysteine reactivity

Protein thermodynamic stability is a fundamental physical characteristic that determines biological function. Furthermore, alteration of thermodynamic stability by macromolecular interactions or biochemical modifications is a powerful tool for assessing the relationship between protein structure, stability, and biological function. High-throughput approaches for quantifying protein stability are beginning to emerge that enable thermodynamic measurements on small amounts of material, in short periods of time, and using readily accessible instrumentation. Here we present such a method, fast quantitative cysteine reactivity, which exploits the linkage between protein stability, sidechain protection by protein structure, and structural dynamics to characterize the thermodynamic and kinetic properties of proteins. In this approach, the reaction of a protected cysteine and thiol-reactive fluorogenic indicator is monitored over a gradient of temperatures after a short incubation time. These labeling data can be used to determine the midpoint of thermal unfolding, measure the temperature dependence of protein stability, quantify ligand-binding affinity, and, under certain conditions, estimate folding rate constants. Here, we demonstrate the fQCR method by characterizing these thermodynamic and kinetic properties for variants of Staphylococcal nuclease and E. coli ribose-binding protein engineered to contain single, protected cysteines. These straightforward, information-rich experiments are likely to find applications in protein engineering and functional genomics.     

Temporal discounting and heart rate reactivity to stress

Temporal discounting is the reduction of the value of a reinforcer as a function of increasing delay to its presentation. Impulsive individuals discount delayed consequences more rapidly than self-controlled individuals, and impulsivity has been related to substance abuse, gambling, and other problem behaviors. A growing body of literature has identified biological correlates of impulsivity, though little research to date has examined relations between delay discounting and markers of poor health (e.g., cardiovascular reactivity to stress). We evaluated the relation between one aspect of impulsivity, measured using a computerized temporal discounting task, and heart rate reactivity, measured as a change in heart rate from rest during a serial subtraction task. A linear regression showed that individuals who were more reactive to stress responded more impulsively (i.e., discounted delayed reinforcers more rapidly). When results were stratified by gender, the effect was observed for females, but not for males. This finding supports previous research on gender differences in cardiovascular reactivity and suggests that this type of reactivity may be an important correlate of impulsive behavior.     

Toxicity of fipronil to German and American cockroaches

Topical and oral toxicity of fipronil, compared to chlorpyrifos, was determined for the German cockroach, Blattella germanica (L.), and American cockroach, Periplaneta americana (L.). Fipronil and Combat bait matrices were evaluated for their attractancies to both species. In the topical toxicity tests, LD50's of fipronil, at 72 h after topical application, were 0.03 and 0.02 µg/g for adult B. germanica and P. americana, respectively. Fipronil was significantly more toxic than topically applied chlorpyrifos (LD50's were 0.06 and 0.16 µg/g for B. germanica and P. americana, respectively). The oral toxicity of fipronil and chlorpyrifos in Petri dish experiments, against both species, was affected by stage (for B. germanica), diet concentration, and feeding assay. Fipronil caused higher mortality of B. germanica than chlorpyrifos in two feeding assays (continuous and abbreviated). Both compounds were equally toxic to adult males of P. americana at all rates. Fipronil caused higher nymphal mortality than chlorpyrifos 48–72 h after exposure in both feeding assays. In large population chamber tests, fipronil bait was more effective and faster in killing P. americana than Raid and Combat. LT50's were 0.8, 2.4, and 7.6 d for fipronil, Raid (a.i. = chlorpyrifos), and Combat (a.i. = hydramethylnon) baits, respectively. Mortality reached 96.5, 93.4, and 84.6%, respectively, at the end of the 14 d test. In the bait attractancy tests, both strains of B. germanica were attracted similarly to fipronil and Combat bait matrices. P. americana were attracted more to fipronil than to Combat bait matrix or to other alternative foods.