Toxic Compounds (toxic + compound)

Distribution by Scientific Domains

Selected Abstracts

Effect of Binary Combinations of Selected Toxic Compounds on Growth and Fermentation of Kluyveromyces marxianus

Jose M. Oliva
The inhibitory effects of various lignocellulose degradation products on glucose fermentation by the thermotolerant yeast Kluyveromycesmarxianus were studied in batch cultures. The toxicity of the aromatic alcohol catechol and two aromatic aldehydes (4-hydroxybenzaldehyde and vanillin) was investigated in binary combinations. The aldehyde furfural that usually is present in relatively high concentration in hydrolyzates from pentose degradation was also tested. Experiments were conducted by combining agents at concentrations that individually caused 25% inhibition of growth. Compared to the relative toxicity of the individual compounds, combinations of furfural with catechol and 4-hydroxybenzaldehyde were additive (50% inhibition of growth). The other binary combinations assayed (catechol with 4-hydroxybenzaldehyde, and vanillin with catechol, furfural, or 4-hydroxybenzaldehyde) showed synergistic effect on toxicity and caused a 60,90% decrease in cell mass production. The presence of aldehydes in the fermentation medium strongly inhibited cell growth and ethanol production. Kluyveromyces marxianusreduces aldehydes to their corresponding alcohols to mitigate the toxicity of these compounds. The total reduction of aldehydes was needed to start ethanol production. Vanillin, in binary combination, was dramatically toxic and was the only compound for which inhibition could not be overcome by yeast strain assimilation, causing a 90% reduction in both cell growth and fermentation. [source]

New perspectives in retinal imaging: fundus autofluorescence and age-related macular degeneration

Fundus Autofluorescence (FAF) imaging using confocal scanning laser ophthalmoscopy is a non-invasive method to to accurately record the topographic distribution of RPE lipofuscin in the human eye in vivo. Excessive lipofuscin accumkulation in the RPE is a common downstream pathogenetic pathway in various complex and monogenetic retinal diseases. Toxic compounds and molecular mechanisms of interference with normal cellular functions have been identified including the dominant fluorophore A2-E. Alterations in fundus autofluorescence (FAF) appearance in eyes with early and late age-related macular degeneration (AMD) can be striking. FAF patterns and distribution do not necessarily correlate with the features of interest in color or angiographic images of eyes with early or late AMD. In the prospective, multicenter FAM study distinct patterns of abnormal FAF were identified and classified in the junctional zone of geographic atrophy (GA). Areas of increased FAF outside GA were associated with variable degrees of loss of retinal sensitivity when tested with microperimetry which suggests a functional correlate of lipofuscin accumulation. Increased FAF preceded the development and enlargement of outer retinal atrophy associated with spread of absolute scotoma in eyes with AMD. Longitudinal examinations showed that the abnormal phenotypic FAF patterns serve as novel prognostic determinants which allows to distinguish fast vs. slow progressors. These findings are relevant and now used to design and carry out interventional trials with agents aimed at slowing down spread of atrophy, e.g. using visual cycle modulators to influence lipofuscinogenesis. Hereby FAF imaging also serves as a mean to accurately delineate and measure areas of GA over time in an automated fashion. A phenotype-genotype correlation was identified for a distinct FAF phenotype subset which was found to represent late-onset Stargardt macular dystrophy mimicking late-stage atrophic AMD. New imaging technologies were recently applied including simultaneous recordings of FAF images and high-resolution, spectral-domain optical coherence tomography (OCT) which allows to identify morphological correlates of abnormal FAF signals in optical biopsies. [source]

Bioaccumulation and biotransformation of arsenic in the Mediterranean polychaete Sabella spallanzanii experimental observations

Alessandra Notti
Abstract The Mediterranean fan worm Sabella spallanzanii is characterized by elevated basal levels of arsenic in branchial crowns (>1,000 ,g/g) and an unusual prevalence of dimethylarsinic acid (DMA), a relatively toxic compound with a possible antipredatory role. The aim of this work was to obtain further insights on the capability of this polychaete to accumulate arsenic from different compounds and to operate biotransformation reactions. Laboratory exposures to arsenate (AsV), dimethylarsinic acid (DMA), trimethylarsine (TMA), and arsenobetaine (AsB) revealed significant differences among tissues and kind of experiments. The highest increases of arsenic content were observed in branchial crowns of organisms treated with arsenate, which can enter the cell through the phosphate carrier system; lower variations were measured with DMA and TMA, while not-significant changes of total As occurred after treatments with AsB. In body tissues, exposure to AsV, DMA, and TMA confirmed a progressively lower accumulation of total arsenic, while a marked increase was caused by AsB. Obtained results suggested that accumulated arsenic could be chemically transformed, thus explaining the elevated basal levels of DMA typical of S. spallanzanii; during all the experiments, DMA was the most accumulated molecule, suggesting that this species possesses the enzymatic pathways for methylation and demethylation reactions of inorganic and trimethylated arsenicals. Only arsenobetaine was not converted into DMA, which would confirm a microbial pathway for degradation for this molecule, particularly important in body tissues of S. spallanzanii for the presence of bacteria associated to digestive tracts. Overall, the present study suggests future investigations on the biological role of arsenic and DMA in S. spallanzanii as a potential adaptive mechanism against predation in more vulnerable tissues. [source]

Pseudomonas putida KT2440 responds specifically to chlorophenoxy herbicides and their initial metabolites

Dirk Benndorf Dr.
Abstract Pseudomonas putida,KT2440 is often used as a model to investigate toxicity mechanisms and adaptation to hazardous chemicals in bacteria. The objective of this paper was to test the impact of the chlorophenoxy herbicides 2,4-dichlorophenoxyacetic acid,(2,4-D) and 2-(2,4-dichlorophenoxy)propanoic acid,(DCPP) and their metabolites 2,4-dichlorophenol,(DCP) and 3,5-dichlorocatechol,(DCC), on protein expression patterns and physiological parameters. Both approaches showed that DCC has a different mode of action and induces different responses than DCPP, 2,4-D and DCP. DCC was the most toxic compound and was active as an uncoupler of oxidative phosphorylation. It repressed the synthesis of ferric uptake regulator (Fur)-dependent proteins, e.g. fumarase,C and L -ornithine N5-oxygenase, which are involved in oxidative stress response and iron uptake. DCPP, 2,4-D and DCP were less toxic than DCC. They disturbed oxidative phosphorylation to a lesser extent by a yet unknown mechanism. Furthermore, they repressed enzymes of energy-consuming biosynthetic pathways and induced membrane transporters for organic substrates. A TolC homologue component of multidrug resistance transporters was found to be induced, which is probably involved in the removal of lipophilic compounds from membranes. [source]

Swimming characteristics of magnetic bacterium, Magnetospirillum sp.

AMB-, implications as toxicity measurement
Abstract To develop a novel toxicity measurement system using the persistent swimming property of magnetic bacteria along an externally applied magnetic field, certain characteristics of Magnetospirillum sp. AMB-1 cells were examined, including their growth pattern, motility, magnetosensitivity, swimming speed, and cell length distribution. In addition, the effect of toxic compounds on the swimming speed was assessed relative to application as a toxicity sensor. With an inoculum of 1.0 × 108 cells/mL, the cells reached the stationary phase with a concentration of about 5 × 108 cells/mL after 20 h, under both aerobic and anaerobic conditions. The distribution of the cell length did not vary significantly during the growth period, and both aerobically and anaerobically growing cells showed a similar cell length distribution. Although the cells showed similar growth patterns under both conditions, the anaerobically grown cells exhibited higher motility and magnetosensitivity. Actively growing cells under anaerobic conditions had an average swimming speed of 49 ,m/s with a standard deviation of 20 ,m/s. When the anaerobically growing cells were exposed to various concentrations of toxic compounds, such as 1-propanol and acetone, the swimming speed decreased with an increased concentration of the toxic compound. Accordingly, the relationship between swimming speed and toxicity can be used as an effective quantitative toxicity measurement; furthermore, the relative sensitivity of the proposed system was comparable to Microtox, which is commercially available. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 76: 11,16, 2001. [source]

Beauvericin Decreases Cell Viability of Wheat

Antonia, robárová
Abstract Recently, beauvericin (BEA) has been recognized as an important toxic compound synthesized by several Fusarium strains, infecting maize, wheat, and rice, worldwide. The effects of BEA on mammalian cells have been studied; however, its effects on the function of host plant cells are largely unknown. The purpose of our work was to assess whether BEA can affect the root and leaf cells of wheat cultivar (cv.) ,Arina' seedlings, using a cytotoxicity assay and fluorescence microscopy. Toxigenicity during wheat germination was higher in BEA-treated wheat seedlings than in non-treated seedlings (control). Leaf primordial, situated at the base and the tips of treated leaves, were more affected by BEA compared to the control when assayed in medium for cell viability measured by luminescent equipment. BEA-Treated plant cells secrete adenosine triphosphate (ATP) to the extracellular matrix and invoke more luminescence by luciferase than the non-treated seedlings. Our results were confirmed by fluorescence microscopy following ,4,,6-diamidino-2-phenylindole' (DAPI) staining and by confocal microscopy. In addition, the bioluminescent protein luciferase was observed in the intracellular space indicating presence of ATP. The incidence of nuclear fragmentation increased significantly in cells of seedlings treated with BEA at 40,,M concentration implying that the intracellular phytotoxin BEA plays an important role, possibly as a mediator in cell-death signalling. [source]

Life-history strategy in an oligophagous tephritid: the tomato fruit fly, Neoceratitis cyanescens

Abstract 1.,In phytophagous insects, life-history traits mainly depend on host plant range. Substantial longevity, high fecundity and larval competition are the major traits of polyphagous Tephritidae while species with a restricted host range generally exhibit a lower longevity and fecundity as well as mechanisms to avoid larval competition. Our aim in this study was to investigate the life history of an oligophagous species, the tomato fruit fly, Neoceratitis cyanescens (Bezzi). 2.,We determined life tables under laboratory conditions in order to calculate the main demographic parameters of N. cyanescens and studied the influence of larval and adult diet on life-history traits. 3.,The mean longevity of N. cyanescens females was 40 days. There was a strong synchronisation of female maturity. Oviposition showed an early peak at 9,13 days after a short pre-oviposition period (6 days). The absence of proteins in the adult diet both delayed ovarian maturation and decreased female fecundity. In addition, females originating from tomato fruits produced significantly more eggs than females originating from bugweed or black nightshade, showing that even the larval host plant may strongly affect the subsequent fecundity of adult females. 4.,The traits of N. cyanescens are then discussed in the light of those documented for polyphagous and monophagous tephritids. Neoceratitis cyanescens displayed attributes intermediate between those of polyphagous and monophagous tephritids. Its smaller clutch size compared with polyphagous species and its specialisation on the Solanaceae family whose fruits contain toxic compounds may help in reducing intra- and inter-specific competition, respectively. [source]

Determination of DNA methylation by COBRA: A comparative study of CGE with LIF detection and conventional gel electrophoresis

Simon Goedecke
Abstract DNA methylation as an epigenetic modification of the human genome is under emphatic investigation. Several studies have demonstrated a role of DNA methylation in oncogenesis. In conjunction with histone modifications, DNA methylation may cause the formation of heterochromatin and thus mediate the inactivation of gene transcription. It is important to develop methods that allow for an accurate quantification of the amount of DNA methylation in particular DNA regions, to gain information concerning the threshold of methylation levels necessary for gene inactivation. In this article, a CGE method with on-column LIF detection using SYBR Green is compared with a conventional slab-gel electrophoresis. We thus investigate the validity to analyze DNA methylation in the samples of a combined bisulfite restriction analysis. It is demonstrated that CGE is superior to gel electrophoresis in means of linearity, precision, accuracy, automatization (high throughput), and sample consumption. However, gel electrophoresis is easier to perform (simple devices, no PC usage), and the running costs are comparatively low. A further advantage of CGE is the sparse use of toxic compounds (MeOH and SYBR Green), whereas gel electrophoresis is performed in polyacrylamide gels with ethidium bromide staining. [source]

Effects of exposure to oxamyl, carbofuran, dichlorvos, and lindane on acetylcholinesterase activity in the gills of the Pacific oyster Crassostrea gigas

Gerardo A. Anguiano
Abstract Acetylcholinesterase (AChE) activity has been used to test the exposure of mollusk bivalves to pesticides and other pollutants. The Pacific oyster Crassostrea gigas is a species with a worldwide distribution, and it has a high commercial value. The use of this species as a bioindicator in the marine environment, and the use of measurements of AChE activity in tissues of C. gigas require prior evaluation of organisms exposed to several toxic compounds in the laboratory. In our study, the effects of pesticides on AChE activity in the gills and mantle tissues of C. gigas were analyzed by exposing animals to organophosphate (dichlorvos), carbamate (carbofuran and oxamyl), and organochlorine (lindane) pesticides. Adult Pacific oysters were exposed to several concentrations (0.1,200 ,M) of dichlorvos, carbofuran, and oxamyl for 96 h, and lindane (1.0 and 2.5 ,M) was applied for 12 days. In gill tissues, all pesticides analyzed caused a decrease in AChE activity when compared to the control unexposed group. The mean inhibition concentration (IC50) values were determined for dichlorvos, carbofuran, and oxamyl pesticides. Dichlorvos had the highest toxic effect, with an IC50 of 1.08 ,M; lesser effects were caused by oxamyl and carbofuran, with IC50s of 1.67 and 3.03 ,M, respectively. This study reports the effects of pesticides with several chemical structures and validates measurement of AChE activity in the gill tissues of C. gigas for use in environmental evaluations or food quality tests. © 2009 Wiley Periodicals, Inc. Environ Toxicol 25: 327,332, 2010. [source]

Assessment of sediment quality of Yangtze River estuary using zebrafish (Danio rerio) embryos

Lingling Wu
Abstract Yangtze River estuary is one of the largest estuaries worldwide. In this study, the sediment quality of Yangtze River estuary was evaluated using zebrafish (Danio rerio) embryos. Freshly fertilized zebrafish eggs (2 h after fertilization) were exposed to the whole sediment and its organic phase of extract, respectively. The parameters, including survival rate, abnormality, hatching rate, and heart rate of the zebrafish embryos, were recorded during the 96-h exposure. The results demonstrated that the concentrations of heavy metals (Zn, Cu, Cd, Ni, Cr, and As) and low-molecular weight PAHs (Fluorene) in the sediment of Yangtze River estuary exceeded their corresponding effects range low values. The maximum concentrations of Zn and Fluorene in the sediment samples were 239.6 ,g/g and 45.9 ng/g, respectively. In both whole sediment test and organic extract test, the survival rate and heart rate of zebrafish embryos were reduced, as well as abnormalities and delayed hatching were induced. For example, the highest mortality of the embryos was 39% in the whole sediment exposure. Overall, the occurrence of toxic compounds in the sediment of Yangtze River estuary may have potentially teratogenic effect on biota. The sediment from the upstream of Yangtze River estuary have more observed toxic effects on zebrafish embryos than that form the downstream. Therefore, more attention should be paid to control these pollutants, especially heavy metals in the Yangtze River estuary. © 2009 Wiley Periodicals, Inc. Environ Toxicol, 2010. [source]

Mixture and single-substance toxicity of selective serotonin reuptake inhibitors toward algae and crustaceans

Anne Munch Christensen
Abstract Selective serotonin reuptake inhibitors (SSRIs) are used as antidepressant medications, primarily in the treatment of clinical depression. They are among the pharmaceuticals most often prescribed in the industrialized countries. Selective serotonin reuptake inhibitors are compounds with an identical mechanism of action in mammals (inhibit reuptake of serotonin), and they have been found in different aqueous as well as biological samples collected in the environment. In the present study, we tested the toxicities of five SSRIs (citalopram, fluoxetine, fluvoxamine, paroxetine, and sertraline) as single substances and of citalopram, fluoxetine, and sertraline in binary mixtures in two standardized bioassays. Test organisms were the freshwater algae Pseudokirchneriella subcapitata and the freshwater crustacean Daphnia magna. In algae, test median effect concentrations (EC50s) ranged from 0.027 to 1.6 mg/L, and in daphnids, test EC50s ranged from 0.92 to 20 mg/L, with sertraline being one of the most toxic compounds. The test design and statistical analysis of results from mixture tests were based on isobole analysis. It was demonstrated that the mixture toxicity of the SSRIs in the two bioassays is predictable by the model of concentration addition. Therefore, in risk assessment based on chemical analysis of environmental samples, it is important to include the effect of all SSRIs that are present at low concentrations, and the model of concentration addition may be used to predict the combined effect of the mixture of SSRIs. [source]

Alteration of normal cellular profiles in the scleractinian coral (Pocillopora damicornis) following laboratory exposure to fuel oil

Luc Rougée
Abstract Petroleum contamination from oil spills is a continuing threat to our ocean's fragile ecosystems. Herein, we explored the effects of the water-soluble fraction of crude oil on a stony coral, Pocillopora damicornis (Linneaeus 1758). We developed methods for exposing corals to various concentrations of crude oil and for assessing the potential molecular responses of the corals. Corals were exposed to water-accommodated fraction solutions, and appropriate cellular biomarkers were quantified. When compared to the "healthy" control specimens, exposed corals exhibited shifts in biomarker concentrations that were indicative of a shift from homeostasis. Significant changes were seen in cytochrome P450 1-class, cytochrome P450 2-class, glutathione- S -transferase-pi, and cnidarian multixenobiotic resistance protein-1 biomarkers, which are involved the cellular response to, and manipulation and excretion of, toxic compounds, including polycyclic aromatic hydrocarbons. A shift in biomarkers necessary for porphyrin production (e.g., protoporphyrinogen oxidase IX and ferrochelatase) and porphyrin destruction (e.g., heme oxygenase-1 and invertebrate neuroglobin homologue) illustrates only one of the cellular protective mechanisms. The response to oxidative stress was evaluated through measurements of copper/zinc superoxide dismutase-1 and DNA glycosylase MutY homologue-1 concentrations. Likewise, changes in heat shock protein 70 and small heat shock proteins indicated an adjustment in the cellular production of proteins. Finally, the results of this laboratory study were nearly identical to what we observed previously among corals of a different species, Porites lobata, exposed to an oil spill in the field after the grounding of the Merchant Vessel Kyowa Violet. [source]

Predicting single and mixture toxicity of petrogenic polycyclic aromatic hydrocarbons to the copepod Oithona davisae

Carlos Barata
Abstract In the present study, the acute toxicity of 10 polycyclic aromatic hydrocarbons (PAH) associated with the Prestige fuel oil spill (Spain, 2002) were evaluated, either as single substances or in mixtures, in adults of the copepod Oithona davisae. All but dimethylphenanthrene had negative effects on O. davisae survival at concentrations below their water solubility, with 48-h median lethal concentrations for naphthalene and pyrene of 56.1 and 0.8 ,mol/L, respectively, making these the least and most toxic compounds. Polycyclic aromatic hydrocarbons had narcotic effects on copepods, as evidenced by the lack of motility at lower concentrations than those causing death. Naphthalene showed the greatest narcotic effects, and phenanthrene showed minor effects. Acute toxicity of the tested PAHs was inversely related (r2 = 0.9) with their octanol,water partition coefficient, thereby confirming the validity of the baseline quantitative structure,activity regression models for predicting the toxicity of PAH compounds in copepod species. When supplied in mixtures, the toxic effect of PAHs was additive. These results indicate that the many PAHs in an oil spill can be considered unambiguous baseline toxicants (class 1) acting additively as nonpolar narcotics in copepods; hence, their individual and combined toxicity can be predicted using their octanol,water partition coefficient. [source]

Bioluminescence inhibition assays for toxicity screening of wood extractives and biocides in paper mill process waters

Anna Rigol
Abstract The risk associated with wood extractives, biocides, and other additives in pulp and paper mill effluents was evaluated by performing a characterization of process waters and effluents in terms of toxicity and chemical analysis. The individual toxicity of 10 resin acids, two unsaturated fatty acids, and three biocides was estimated by measuring the bioluminescence inhibition with a ToxAlert® 100 system. Median effective concentration values (EC50) of 4.3 to 17.9, 1.2 to 1.5, and 0.022 to 0.50 mg/L were obtained, respectively. Mixtures of these three families of compounds showed antagonistic effects. Chemical analysis of process waters was performed by liquid chromatography-and gas chromatography-mass spectrometry. Biocides such as 2-(thiocyanome-thylthio)-benzotiazole (TCMTB) (EC50 = 0.022 mg/L) and 2,2-dibromo-3-nitrilpropionamide (DBNPA) (EC50 = 0.50 mg/L) were the most toxic compounds tested and were detected at concentrations of 16 and 59 ,g/L, respectively, in a closed-circuit recycling paper mill. Process waters from kraft pulp mills, printing paper mills, and packing board paper mills showed the highest concentration of resin acids (up to 400 ,g/L) and accounted for inhibition percentages up to 100%. Detergent degradation products such as nonylphenol (NP) and octylphenol (OP) and the plasticizer bisphenol A (BPA) were also detected in the waters at levels of 0.6 to 10.6, 0.3 to 1.4, and 0.7 to 187 ,g/L, respectively. However, once these waters were biologically treated, the concentration of detected organic compounds diminished and the toxicity decreased in most cases to values of inhibition lower than 20%. [source]

Discrepancy between acute and chronic toxicity induced by imidacloprid and its metabolites in Apis mellifera

Séverine Suchail
Abstract Imidaclopridi a systemic nitroguanidine insecticide that belongs to theneonicotinoid family. As an agonist of the acetylcholine receptor, it attacks the insect nervous system and is extremely effective against various sucking and mining pests. Oral acute and chronic toxicity of imidacloprid and its main metabolites (5-hydroxyimidacloprid, 4,5-dihydroxyimidacloprid, desnitroimidacloprid, 6-chloronicotinic acid, olefin, and urea derivative) were investigated in Apis mellifera. Acute intoxication by imidacloprid or its metabolites resulted in the rapid appearance of neurotoxicity symptoms, such as hyperresponsiveness, hyperactivity, and trembling and led to hyporesponsiveness and hypoactivity. For acute toxicity tests, bees were treated with doses of toxic compounds ranging from 1 to 1,000 ng/bee (10,10,000 ,g/kg). Acute toxicity (LD50) values of imidacloprid were about 60 ng/bee (600 ,g/kg) at 48 h and about 40 ng/bee (400 ,g/kg) at 72 and 96 h. Out of the six imidacloprid metabolites tested, only two (5-hydroxyimidacloprid and olefin) exhibited a toxicity close to that of imidacloprid. Olefin LD50 values were lower than those of imidacloprid. The 5-hydroxyimidacloprid showed a lower toxicity than imidacloprid with a LD50 four to six times higher than that of imidacloprid. Urea also appeared as a compound of nonnegligible toxicity by eliciting close to 40% mortality at 1,000 ng/bee (10,000 ,g/kg). However, no significant toxicity was observed with 4,5-dihydroxyimidacloprid, 6-chloronicotinic acid, and desnitroimidacloprid in the range of doses tested. To test chronic toxicity, worker bees were fed sucrose solutions containing 0.1, 1, and 10 ,g/L of imidacloprid and its metabolites for 10 d. Fifty percent mortality was reached at approximately 8 d. Hence, considering that sucrose syrup was consumed at the mean rate of 12 ,l/d and per bee, after an 8-d period the cumulated doses were approximately 0.01, 0.1, and 1 ng/bee (0.1, 1, and 10 ,g/kg). Thus, all tested compounds were toxic at doses 30 to 3,000 (olefin), 60 to 6,000 (imidacloprid), 200 to 20,000 (5-OH-imidacloprid), and >1,000 to 100,000 (remaining metabolites) times lower than those required to produce the same effect in acute intoxication studies. For all products tested, bee mortality was induced only 72 h after the onset of intoxication. [source]

In vivo production of catalase containing haem analogues

FEBS JOURNAL, Issue 12 2010
Myriam Brugna
Haem (protohaem IX) analogues are toxic compounds and have been considered for use as antibacterial agents, but the primary mechanism behind their toxicity has not been demonstrated. Using the haem protein catalase in the Gram-positive bacterium Enterococcus faecalis as an experimental system, we show that a variety of haem analogues can be taken up by bacterial cells and incorporated into haem-dependent enzymes. The resulting cofactor-substituted proteins are dysfunctional, generally resulting in arrested cell growth or death. This largely explains the cell toxicity of haem analogues. In contrast to many other organisms, E. faecalis does not depend on haem for growth, and therefore resists the toxicity of many haem analogues. We have exploited this feature to establish a bacterial in vivo system for the production of cofactor-substituted haem protein variants. As a pilot study, we produced, isolated and analysed novel catalase variants in which the iron atom of the haem prosthetic group is replaced by other metals, i.e. cobalt, gallium, tin, and zinc, and also variants containing meso-protoheme IX, ruthenium meso-protoporphyrin IX and (metal-free) protoporphyrin IX. Engineered haem proteins of this type are of potential use within basic research and the biotechnical industry. Structured digital abstract ,,MINT-7722358, MINT-7722368: katA (uniprotkb:Q834P5) and katA (uniprotkb:Q834P5) physically interact (MI:0915) by copurification (MI:0025) [source]

Engineering of a monomeric and low-glycosylated form of human butyrylcholinesterase

FEBS JOURNAL, Issue 2 2002
Expression, characterization, crystallization, purification
Human butyrylcholinesterase (BChE; EC is of particular interest because it hydrolyzes or scavenges a wide range of toxic compounds including cocaine, organophosphorus pesticides and nerve agents. The relative contribution of each N-linked glycan for the solubility, the stability and the secretion of the enzyme was investigated. A recombinant monomeric BChE lacking four out of nine N-glycosylation sites and the C-terminal oligomerization domain was stably expressed as a monomer in CHO cells. The purified recombinant BChE showed catalytic properties similar to those of the native enzyme. Tetragonal crystals suitable for X-ray crystallography studies were obtained; they were improved by recrystallization and found to diffract to 2.0 Ĺ resolution using synchrotron radiation. The crystals belong to the tetragonal space group I422 with unit cell dimensions a = b = 154.7 Ĺ, c = 124.9 Ĺ, giving a Vm of 2.73 Ĺ3 per Da (estimated 60% solvent) for a single molecule of recombinant BChE in the asymmetric unit. The crystal structure of butyrylcholinesterase will help elucidate unsolved issues concerning cholinesterase mechanisms in general. [source]

Minimizing the release of proinflammatory and toxic bacterial products within the host: A promising approach to improve outcome in life-threatening infections

Roland Nau
Abstract Various bacterial components (e.g., endotoxin, teichoic and lipoteichoic acids, peptidoglycans, DNA) induce or enhance inflammation by stimulating the innate immune system and/or are directly toxic in eukariotic cells (e.g., hemolysins). When antibiotics which inhibit bacterial protein synthesis kill bacteria, smaller quantities of proinflammatory or toxic compounds are released in vitro and in vivo than during killing of bacteria by ,-lactams and other cell-wall active drugs. In general, high antibiotic concentrations liberate lower quantities of bacterial proinflammatory or toxic compounds than concentrations close to the minimum inhibitory concentration. In animal models of Escherichia coli Pseudomonas aeruginosa and Staphylococcus aureus peritonitis/sepsis and of Streptococcus pneumoniae meningitis, a lower release of proinflammatory bacterial compounds was associated with a reduced mortality or neuronal injury. Pre-treatment with a bacterial protein synthesis inhibitor reduced the strong release of bacterial products usually observed during treatment with a ,-lactam antibiotic. Data available strongly encourage clinical trials comparing antibiotic regimens with different release of proinflammatory/toxic bacterial products. The benefit of the approach to reduce the liberation of bacterial products should be greatest in patients with a high bacterial load. [source]

RESEARCH ARTICLE: Fungicidal activity of amiodarone is tightly coupled to calcium influx

Sabina Muend
Abstract The antiarrhythmic drug amiodarone has microbicidal activity against fungi, bacteria and protozoa. In Saccharomyces cerevisiae, amiodarone triggers an immediate burst of cytosolic Ca2+, followed by cell death markers. Ca2+ transients are a common response to many forms of environmental insults and toxic compounds, including osmotic and pH shock, endoplasmic reticulum stress, and high levels of mating pheromone. Downstream signaling events involving calmodulin, calcineurin and the transcription factor Crz1 are critical in mediating cell survival in response to stress. In this study we asked whether amiodarone induced Ca2+ influx was beneficial, toxic or a bystander effect unrelated to the fungicidal effect of the drug. We show that downregulation of Ca2+ channel activity in stationary phase cells correlates with increased resistance to amiodarone. In actively growing cells, extracellular Ca2+ modulated the size and shape of the Ca2+ transient and directly influenced amiodarone toxicity. Paradoxically, protection was achieved both by removal of external Ca2+ or by adding high levels of CaCl2 (10 mM) to block the drug induced Ca2+ burst. Our results support a model in which the fungicidal activity of amiodarone is mediated by Ca2+ stress, and highlight the pathway of Ca2+ mediated cell death as a promising target for antifungal drug development. [source]

KNQ1, a Kluyveromyces lactis gene encoding a transmembrane protein, may be involved in iron homeostasis

Emmanuela Marchi
Abstract The original purpose of the experiments described in this article was to identify, in the biotechnologically important yeast Kluyveromyces lactis, gene(s) that are potentially involved in oxidative protein folding within the endoplasmic reticulum (ER), which often represents a bottleneck for heterologous protein production. Because treatment with the membrane-permeable reducing agent dithiothreitol inhibits disulfide bond formation and mimics the reducing effect that the normal transit of folding proteins has in the ER environment, the strategy was to search for genes that conferred higher levels of resistance to dithiothreitol when present in multiple copies. We identified a gene (KNQ1) encoding a drug efflux permease for several toxic compounds that in multiple copies conferred increased dithiothreitol resistance. However, the KNQ1 product is not involved in the excretion of dithiothreitol or in recombinant protein secretion. We generated a knq1 null mutant, and showed that both overexpression and deletion of the KNQ1 gene resulted in increased resistance to dithiothreitol. KNQ1 amplification and deletion resulted in enhanced transcription of iron transport genes, suggesting, for the membrane-associated protein Knq1p, a new, unexpected role in iron homeostasis on which dithiothreitol tolerance may depend. [source]

Reproductive ecology of Drosophila

T. A. Markow
Summary 1Species of the genus Drosophila reproduce in a wide range of different resources, including fruits, sap, flowers, mushrooms and cacti. Drosophila species and their resources also exhibit considerable variability in geographic distribution. 2Habitat and resource differences pose enormous challenges for Drosophila species. Host chemistry may include highly toxic compounds and breeding sites may be characterized by extreme abiotic conditions such as high and/or low temperature and humidity. 3Drosophila reproductive biology, in terms of morphology, physiology, and behaviour, is as variable among Drosophila species as is their resource use. In some species, adults are ready to reproduce upon emergence, whereas one sex or the other in other species may require weeks to become sexually mature. 4Already a robust system for transmission and population genetic studies, the sequencing of the genomes of 12 diverse Drosophila species now brings the power of genomics to investigators wishing to understand the functional aspects of Drosophila ecology [source]

Biotransformation of xenobiotics by amine oxidases

Margherita Strolin Benedetti
Although the cytochrome P450 (CYP) system ranks first in terms of catalytic versatility and the wide range of xenobiotics it detoxifies or activates to reactive intermediates, the contribution of amine oxidases and in particular of monoamine oxidases (MAOs) to the metabolism of xenobiotics is far from negligible but has been largely neglected. In this review on the involvement of amine oxidases in the metabolism of xenobiotics, the major characteristics reported for the CYP system (protein, reaction, tissue distribution, subcellular localisation, substrates, inhibitors, inducers, genetic polymorphism, impact of different physiopathological conditions on the activity, turnover) will be compared, whenever possible, with the corresponding characteristics of amine oxidases (MAOs in particular). The knowledge of the involvement of MAO-A, -B or both in the metabolism of a drug allows us to predict interactions with selective or non-selective MAO inhibitors (e.g. the metabolism of a drug deaminated by both forms of MAO is not necessarily inhibited in vivo by a selective MAO-A or -B inhibitor). If a drug is metabolized by MAOs, competitive interactions can occur with other drugs that are MAO substrates, e.g. with ,-adrenoceptor agonists and antagonists, prodrugs of dopamine, serotonin 5-HT1 -receptor agonists as well as with primaquine, flurazepam and citalopram. Moreover, the knowledge of the involvement of MAOs in the metabolism of a drug may suggest possible, although not obligatory, interactions with tyramine-containing food or drink, with over the counter medicines sold to relieve the symptoms of coughs and colds (generally containing the indirectly-acting sympathomimetic amine phenylpropanolamine) or with phenylephrine-containing preparations. Finally, biotransformation by amine oxidases, as by CYP, does not always lead to detoxication but can produce toxic compounds. [source]

Decomposition of Allelopathic Plants in Soil

T. D. Xuan
Abstract Higher plants with strong allelopathic properties are commonly incorporated into soil for weed-control purposes. To understand the phytotoxic variation in the soil, which can be utilized for weed control through the use of allelopathic plants, the decomposition of alfalfa (Medicago sativa L. cv. Rasen) and kava (Piper methysticum L.) after soil amendment were evaluated. Both alfalfa and kava strongly inhibited barnyardgrass and monochoria growth for up to 10 days (80,100 % weed control). After 20,25 days, the magnitude of inhibition was drastically reduced, but was still effective (50 % weed control). A number of phenolic acids were detected in the soil even 50 days after incorporation in low concentration, but their concentrations reached a maximum after 10,15 days and were efficacious until 20,25 days. Phenolic acids varied between alfalfa and kava. The variations in electrical conductivity (EC) and osmotic pressure (OP) were strongly related to chemicals and toxic compounds exuded into the soil during decomposition and were proportional to the magnitude of inhibition observed, whereas pH did not appear to be correlated with inhibition. The decomposition of several unknown inhibitors present in kava was also analysed and assessed. Our findings indicate that these growth inhibitors were almost disintegrated in soil after 10 days, but strong inhibition was detected until 25 days after amendment. Results from this study demonstrate that chemicals released from allelopathic plants incorporated into soil are toxic and cause inhibition of certain species and could be exploited as a biological tool for weed management. [source]

Evaluation of analogues of DRDE-07 as prophylactic agents against the lethality and toxicity of sulfur mustard administered through percutaneous route

A. S. Kulkarni
Abstract Sulfur mustard (SM), chemically bis (2-chloroethyl) sulfide is a bifunctional alkylating agent that causes serious blisters on contact with human skin. Although several antidotes have been reported for the systemic toxicity of SM in experimental animals none of them are approved so far and decontamination of SM immediately by physical or chemical means is recommended as the best protection. Two compounds amifostine [S-2(3-aminopropylamino) ethyl phosphorothioate] and DRDE-07 [S-2(2-aminoethylamino) ethyl phenyl sulfide] gave very good protection as an oral prophylactic agent against SM the in mouse model, but in the rat model the protection was only moderate. In the search for more effective and less toxic compounds, a number of analogues of DRDE-07 were synthesised and their protective efficacy was evaluated in mouse and rat models. The LD50 of S-aryl substitution was between 1 and 2 g kg,1 and S-alkyl substitution was more than 2 g kg,1. In the mouse model, DRDE-07, DRDE-10, DRDE-21, DRDE-30 and DRDE-35 gave about 20 fold protection, and DRDE-23 and DRDE-38 gave less protection of 4.8 and 9.0 fold respectively, against percutaneously administered SM. In the rat model, DRDE-07, DRDE-09, DRDE-10 and DRDE-21 gave about two fold protection. Percutaneously administered SM (19.33 mg kg,1) significantly depleted the hepatic GSH content in mice. Pretreatment with DRDE-21 significantly elevated the levels. A 4.4 fold increase in % DNA fragmentation was observed 7 days after SM administration (19.33 mg kg,1) in mice. Pretreatment with DRDE-07, DRDE-09, DRDE-10, DRDE-21, DRDE-30 and DRDE-35 significantly protected the mice from SM induced DNA damage. The histopathological lesions in liver and spleen induced by percutaneously administered SM was reduced by pretreatment with DRDE-07, DRDE-09, DRDE-10 and DRDE-21. These analogues may prove as prototypes for the designing of more effective prophylactic drug for SM. Copyright © 2006 John Wiley & Sons, Ltd. [source]

Changes in expression and activity of glutathione S -transferase in different organs of schistosoma haematobium -infected hamster

S. A. Sheweita
Abstract Schistosomiasis is a major health problem in many subtropical developing countries, causing a number of serious pathologies, including bladder cancer. Most of the toxic compounds formed as a result of these infestations are derived either exogenously or formed endogenously and can be conjugated with glutathione (GSH) via glutathione S-transferase (GST). The present study investigates the effect of Schistosma haematobium infection on the activity of GST and glutathione reductase (GR) and levels of glutathione and free radicals (measured as thiobarbituric acid reactive substances) in different organs of the male hamster. The total activity of GST was increased in several organs; in kidney by 50 and 46% at 6 and 10 weeks postinfection, respectively, and in bladder tissues by 169, 23, and 130% at 2, 4, and 6 weeks postinfection, respectively. In support of this, the expression of GST isozymes was also induced in kidney and bladder tissues at early stages (2, 4, and 6 weeks) and reduced at the later stages of infection (8 and 10 weeks). In contrast, the expression of these isozymes was decreased in the spleen and liver at 2, 4, 6, 8, and 10 weeks postinfection. Also, such activity was decreased in lungs by 74 and 78% and in bladders by 65 and 72% at 8 and 10 weeks postinfection, respectively. GSH levels increased in lungs by 95, 40, and 56% at 2, 4, and 6 weeks and in spleen by 26 and 74% at 4 and 6 weeks, respectively, but decreased at later stages of S. haematobium infection in these organs. The depletion of GSH levels also occurred in bladders by 72 and 54% at 8 and 10 weeks postinfection, respectively. The activity of GR was increased in the livers, lungs, and kidneys of the S. haematobium -infected hamster. TBARS also increased in the lung by 14, 65, 53, 828, and 624% and in the kidney by 64, 29, 87, 190, and 111%, and in the bladder by 216, 23, 1468, 528, and 1025% at 2, 4, 6, 8, and 10 weeks postinfection, respectively. This study indicates that low GST expression and high levels of free radicals could provide new evidence for damage to the bladder and other organs as a result of S. haematobium infection. © 2003 Wiley Periodicals, Inc. J Biochem Mol Toxicol 17:138,145, 2003; Published online in Wiley InterScience ( DOI 10.1002/jbt.10071 [source]

Anaerobic digestion of Aegean olive mill effluents with and without pretreatment

Gülseren Pekin
Abstract BACKGROUND: Olive oil production is an important economical activity in the Aegean region of Turkey. However, the effluents of the olive oil producing mills with their high organic loads and toxic compounds are causing serious environmental problems. The anaerobic biological treatment of olive mill wastewater (OMWW) using the treatment plants of the regional industries could be a method of choice and within the scope of this study floccular and granular sludges were investigated in batch mode for their success in the treatment of OMWW while producing biogas. The major limitation of this treatment is the inhibition of methanogenic bacteria by the phenolic compounds in OMWW. Thus an integrated solution was suggested in which a pre-treatment step (dephenolization) was also introduced before biological step. RESULTS: The effluents of 27 olive mills out of 47 were found to have total phenolics (TP) less than 3 g L,1 and could be treated anaerobically after simple dilution. The biogas production for the untreated OMWW was higher for floccular sludge than for the granular sludge (68.5 mL and 45.7 mL respectively). Combined pre-treatment experiments, first coagulation with polyaluminum chloride, followed by flocculation with cationic polyelectrolyte and finally Fenton's oxidation, could remove 80% of TP and 95% of the total suspended solids. CONCLUSION: OMWW having TP values less than 3 g L,1 can be treated anaerobically using floccular sludge after simple dilution and biogas can be produced. For OMWW samples having higher TP values pre-treatment is necessary and the pre-treatment given in this study may be used effectively. Copyright © 2010 Society of Chemical Industry [source]

Physiological effects in juvenile three-spined sticklebacks feeding on toxic cyanobacterium Nodularia spumigena -exposed zooplankton

J.-P. Pääkkönen
Feeding rate, growth and nutritional condition as well as nodularin concentration of juvenile three-spined sticklebacks Gasterosteus aculeatus were assessed in an experimental study where field-collected fish were given a diet of zooplankton fed with toxic Nodularia spumigena for 15 days. Food consumption was higher in N. spumigena bloom conditions compared with the cyanobacterium-free control, but despite this the growth rate of exposed fish did not improve. Control fish and fish fed N. spumigena -exposed zooplankton had higher RNA:DNA ratios and protein content than fish grown in cyanobacterial bloom conditions indicating good nutritional condition and recent growth of fish, whereas in bloom conditions metabolic transformation of nodularin to less toxic compounds may cause an energetic cost to the fish affecting the growth rate of the whole organism. Juvenile three-spined sticklebacks collected from the field contained higher concentrations of nodularin at the beginning of the experiment (mean 503·1 ,g kg,1). After 15 days, the lowest nodularin concentrations in fish were measured in the control treatment, suggesting that fish fed with non-toxic food are able to detoxify nodularin from their tissues more effectively than fish in continuing exposure. [source]

Characterization of Pectins and Some Secondary Compounds from Theobroma cacao Hulls

M. Arlorio
ABSTRACT This paper describes the chemical characterization of cocoa hulls, a potential source of high-methoxyl pectins (HMP). The content of some antinutritive compounds and potentially toxic compounds is also reported. Use of 2-propanol is proposed for the preliminary clean-up of the hulls and for the washing of the gel. Antinutritive and potentially toxic compounds seem not to limit the use of cocoa hulls. Lindane and ochratoxin A were easily removed together with fat using 2-propanol during preliminary clean up. The pectins (partially purified, yield: 1.29 ± 0.08%) showed a high methoxylation degree (%DE) of 60.53 ± 6.09%, and a viscosity of 16,200 cPs (5 rpm 20 °C). Washing procedures permit the decrease of the gel acidity from pH 1.97 to pH 3.76. [source]

Gastroprotective and cytotoxic effect of semisynthetic ferruginol derivatives

Carlos Areche
The gastroprotective abietane diterpene ferruginol has been shown to present high cytotoxicity. In order to obtain active compounds with less cytotoxicity, 18 semisynthetic ferruginol derivatives and totarol were assessed for their gastroprotective effects in the HCl/ethanol-induced gastric lesion model in mice, as well as for cytotoxicity in human gastric epithelial cells (AGS) and human lung fibroblasts (MRC-5). At 20 mg kg,1, the greatest gastroprotective effects were provided by abieta-8,11,13-triene (1), abieta-8,11,13-trien-12-yl-2-chloropropanoate (8), abieta-8,11,13-trien-12-yl propenoate (9), 12-(2,3,4,6-tetra- O -acetyl-,-D-glucopyranosyloxy)-abieta-8,11,13-triene (17) and 12-(,-D-galactopyranosyloxy)-abieta-8,11,13-triene (18), all of which were as active as the reference drug lansoprazole at 20 mg kg,1, reducing gastric lesions by 69, 76, 67, 72 and 61%, respectively. No relation was observed between lipophilicity and the gastroprotective effect. Compounds that showed the greatest cytotoxicity towards AGS cells were ferruginol (2), the corresponding formate (5), acetate (6), propionate (7), 8, 9, 12-(,-D-glucopyranosyloxy)-abieta-8,11,13-triene (16), 18 and totarol (20) (IC50 18,44 ,M). Ferruginol and compounds 5,9, 16, 18 and 20 were the most toxic compounds against fibroblasts (IC50 19,56 ,M), with a correlation to AGS cells. The derivative 19 was much more active against AGS cells than towards fibroblasts. The best activity/cytotoxicity ratio was found for compound 17, with a lesion index comparable with lansoprazole at 20 mg kg,1 and cytotoxicity >1000 ,M towards MRC-5 and AGS cells, respectively. In conclusion, some derivatives showed a better gastroprotective effect/cytotoxicity ratio than the parent compound ferruginol. A total of 13 new compounds are reported here for the first time. [source]


Article first published online: 24 SEP 200
Carmichael, W. W. Department of Biological Sciences, Wright State University, Dayton, Ohio 45435 USA Cyanobacteria toxins (cyanotoxins) include cytotoxins and biotoxins with cytotoxins including about 60 compounds ranging from phytoalexins to animicrobials to enzyme inhibitors to compounds that can reverse multidrug resistance. Producer organisms include marine/brackish water Cystoseira, Hormothamnin, Lyngbya, Nodularia and Synechocystis, and the freshwater/terrestrial genera Anabaena, Dichotrix, Fischerella, Hapalosiphon, Lyngbya, Microcystis, Nostoc, Oscillatoria, Planktothrix, Phormidium, Schizothrix, Scytonema, Spirulina, Stigonema and Symploca. Since many of these compounds have been identified, not during ecological studies, but during drug discovery investigations, their ecological role is only speculative. Biotoxins are responsible for acute lethal, acute, chronic and sub-chronic poisonings of wild/domestic animals and humans. They include the neurotoxins; anatoxin-a, anatoxin-a(s) and saxitoxins plus the hepatotoxins; microcystins, nodularins and cylindrospermopsin. These compounds are included when referencing harmful algal blooms (HAB's) such as the more predominate marine PSP (paralytic shellfish poisoning), DSP (diarrhetic shellfish poisoning), NSP (neurotoxic shellfish poisoning), ASP (amnesic shellfish poisoning) and EAS (estuary associated syndrome). The CTP (cyanobacteria toxin poisoning) organisms occur in freshwater lakes, ponds, rivers and reservoirs throughout the world. Organisms responsible for CTP's are Anabaena, Aphanizomenon, Cylindrosperm- opsis, Microcystis, Nodularia, Nostoc Oscillatoria (Planktothrix), Trichodesmium and certain picoplanktic genera. Concern for animal and human health impairments arises from animal poisonings, associated with cyanobacteria waterblooms, beginning with the later part of the 1800's. It was not until the 1950's that we began to understand that cyanobacteria could indeed produce highly toxic compounds. A recent 1998 compilation of all available information on toxic cyanobacteria was published by the World Health Organization. This increasing focus on the role of cyanobacteria metabolites in chemical ecology, drug discovery and toxinology has placed new importance on using correct taxonomy for communication of responsible organisms. [source]