Structure-Dependent Effects of Phthalates on Intercellular and Intracellular Communication in Liver Oval Cells.

Humans are exposed to phthalates released from plastics, cosmetics, or food on a daily basis. Phthalates have low acute liver toxicity, but their chronic exposures could induce molecular and cellular effects linked to adverse health outcomes, such as liver tumor promotion or chronic liver diseases. The alternation of gap junctional intercellular communication (GJIC) and MAPK-Erk1/2 pathways in liver progenitor or oval cells can disrupt liver tissue homeostatic mechanisms and affect the development and severity of these adverse outcomes. Our study with 20 different phthalates revealed their structurally dependent effects on liver GJIC and MAPK-Erk1/2 signaling in rat liver WB-F344 cell line with characteristics of liver oval cells. The phthalates with a medium-length side chain (3-6 C) were the most potent dysregulators of GJIC and activators of MAPK-Erk1/2. The effects occurred rapidly, suggesting the activation of non-genomic (non-transcriptional) mechanisms directly by the parental compounds. Short-chain phthalates (1-2 C) did not dysregulate GJIC even after longer exposures and did not activate MAPK-Erk1/2. Longer chain (≥7 C) phthalates, such as DEHP or DINP, moderately activated MAPK-Erk1/2, but inhibited GJIC only after prolonged exposures (>12 h), suggesting that GJIC dysregulation occurs via genomic mechanisms, or (bio)transformation. Overall, medium-chain phthalates rapidly affected the key tissue homeostatic mechanisms in the liver oval cell population via non-genomic pathways, which might contribute to the development of chronic liver toxicity and diseases.

High-valent iron (Fe(VI), Fe(V), and Fe(IV)) species in water: characterization and oxidative transformation of estrogenic hormones.

This paper presents solid state synthesis and characterization of tetra-oxy iron(iv) and iron(v) species in their salt forms (Na4FeO4-Fe(IV) and K3FeO4-Fe(V)). Stability of the synthesized salts, commonly called ferrates, in water was determined by applying the (57)Fe Mössbauer spectroscopy technique. Within 2 s in water, Fe(IV) converted into Fe(III) while Fe(V) transformed into Fe(VI) and Fe(III) at pH = 8.2. Comparatively, Fe(VI) (bought as K2FeO4) remained stable in aqueous solution during the short time period. The oxidative removal efficiency of the high-valent iron species was then tested against five environmentally important estrogenic hormones (estron (E1), 17-ß-estradiol (E2), estriol (E3), 17-α-ethinylestradiol (EE2), and diethylstibestrol (DES)) in effluent water of a wastewater treatment plant. Three dosages of iron species (1, 10, and 100 mg L(-1)) were applied to the effluent water. An increase in the concentration of dosages enhanced the removal of estrogens. Both Fe(V) and Fe(VI) were effective in degrading estrogens, but Fe(IV) showed limited oxidation capacity to transform estrogens. The oxidized products of the estrogens were analyzed using Raman spectroscopy and high-performance liquid chromatography-mass spectrometry (HPLC-MS) techniques. Results demonstrated the transformation of estrogens into low molecular weight oxygenated compounds such as quinone-like and opened-aromatic ring species. A detailed study on E1 by using excess Fe(VI) showed the mineralization of the parent compound. The results demonstrate great potential of high-valent iron species in the degradation of endocrine disruptor chemicals like estrogens with several superior aspects including fast reactions, complete degradation and/or formation of benign organic species, and environmentally-acceptable iron oxide by-products.

Evaluation of antibacterial properties of novel phthalocyanines against Escherichia coli--comparison of analytical methods.

We analyzed antibacterial effects of several novel phthalocyanines against Escherichia coli and evaluated the suitability of flow cytometry for the detection of antibacterial effects of phthalocyanines in comparison with routinely used cultivation. After 3h of exposure under cool white light eight cationic phthalocyanines showed very high antibacterial activity in the concentration of 2.00 mg L(-1) and four of them were even efficient in the concentration of 0.20 mg L(-1). Antibacterial activity of neutral and anionic compounds was considerably lower or even negligible. No antibacterial effect was detected when bacteria were exposed without illumination. Binding affinity to bacterial cells was found to represent an important parameter influencing phthalocyanine antibacterial activity that can be modified by total charge of peripheral substituents and by the presence of suitable functional groups inside them. Agglomeration of cells observed in suspensions treated with a higher concentration of certain cationic phthalocyanines (the strongest binders to bacterial membrane) affected cytometric measurements of total cell counts, thus without appropriate pretreatment of the sample before analysis this parameter seems not to be fully valid in the evaluation of phthalocyanine antibacterial activity. Cytometric measurement of cell membrane integrity appears to be a suitable and even more sensitive parameter than cultivation.

Survey of cyanobacterial toxins in Czech water reservoirs--the first observation of neurotoxic saxitoxins.

The environmental occurrence and concentrations of cyanobacterial toxins (cyanotoxins) were investigated in the Czech Republic. Concentrations of microcystins (MCs), cylindrospermopsin (CYN) or saxitoxins (STXs) were determined immunochemically by ELISA assays in 30 water samples collected from the surface layers of 19 reservoirs during the summer season of 2010. MCs were detected in 18 reservoirs and 83 % of samples, with median and maximal concentration being 1.5 and 18.6 µg/L, respectively. The high frequency of MC occurrence coincided with prevalence of cyanobacterium Microcystis sp., which was detected in 87 % samples, followed by Dolichospermum (Anabaena) sp. observed in 33 % samples. CYN was detected by ELISA only in one sample at a concentration of 1.2 µg/L. STXs presence was indicated for the first time in Czech water reservoirs when the toxins were found at low concentrations (0.03-0.04 µg/L) in two samples (7 %) collected from two different reservoirs, where STXs co-occurred with MCs and eventually also with CYN. In both STX-positive samples, the phytoplankton community was dominated by Microcystis sp., but Dolichospermum sp. and/or Aphanizomenon sp. were also present as putative producers of STX and/or CYN. Cyanotoxins commonly occurred in Czech water reservoirs, and MCs frequently at concentrations possibly associated with human health risks. MCs were the most prevalent and abundant cyanotoxins, but also other cyanotoxins were detected, though sporadically. Further research and regulatory monitoring of cyanotoxins other than MCs is therefore required.

Photodynamic effects of 31 different phthalocyanines on a human keratinocyte cell line.

Phthalocyanines (Pcs, colored macromolecular compounds with the ability to generate singlet oxygen) represent a promising group of photosensitizers due to their intense absorption in the red and UV portion of the spectrum which leads to their excitation. In order to characterize possible toxic effects associated with eventual practical use and application of these chemicals, we employed an in vitro cell culture model to evaluate cytotoxic effects of 31 different phthalocyanines using neutral red uptake assay. An immortalized human keratinocyte cell line HaCaT was exposed to the tested chemicals for 2 or 24h, either with or without illumination in the last 60 min of the exposure period. After 2- or 24-h exposure without illumination, no cytotoxic effects or weak cytotoxic effects were induced by any Pc under the study and EC50 values could not be obtained within the tested concentration ranges (1.25-20 mg L(-1) or 0.625-10 mg L(-1)). On the other hand, exposure to phthalocyanines under illumination induced a significant cytotoxic effect. The most pronounced cytotoxicity was elicited by Pcs previously shown to have high positive charge densities at peripheral parts of substituent groups, which is most likely the factor responsible for the binding of Pc to negatively charged membranes on the cell surface and thus guaranteeing the tight connection necessary for the singlet oxygen attack on the cell surface.

The toxicity of phthalocyanines to the aquatic plant Lemna minor (duckweed) - testing of 31 compounds.

Phthalocyanines are prospective chemicals that have applications in industry, medicine and biology due especially to their architectural flexibility and production of reactive oxygen species. Although they are used in so many areas of human activities nowadays, there is still little knowledge of their ecotoxicity. Here we present the first observation of their toxic effects on representatives of the aquatic plants Lemna minor. The tested phthalocyanines possess a wide spectrum of phytotoxicity ranging from seldom (>50 mg L(-1)) to highly toxic 0.11 mg L(-1). Moreover, the potential of phthalocyanines to be used as selective cyanocides or herbicides is discussed as well.

Multimodal action and selective toxicity of zerovalent iron nanoparticles against cyanobacteria.

Cyanobacteria pose a serious threat to water resources around the world. This is compounded by the fact that they are extremely resilient, having evolved numerous protective mechanisms to ensure their dominant position in their ecosystem. We show that treatment with nanoparticles of zerovalent iron (nZVI) is an effective and environmentally benign method for destroying and preventing the formation of cyanobacterial water blooms. The nanoparticles have multiple modes of action, including the removal of bioavailable phosphorus, the destruction of cyanobacterial cells, and the immobilization of microcystins, preventing their release into the water column. Ecotoxicological experiments showed that nZVI is a highly selective agent, having an EC(50) of 50 mg/L against cyanobacteria; this is 20-100 times lower than its EC(50) for algae, daphnids, water plants, and fishes. The primary product of nZVI treatment is nontoxic and highly aggregated Fe(OH)(3), which promotes flocculation and gradual settling of the decomposed cyanobacterial biomass.

Critical review of actually available chemical compounds for prevention and management of cyanobacterial blooms.

Cyanobacteria proliferation is among the most threatening consequences of freshwater pollution. Health risks from human and other-organism exposure to cyanobacteria have led to an effort to find practical methods for cyanobacterial water-bloom reduction. Hence, methods and techniques have been developed in order to reduce the amount of phosphorus or to decrease the abundance of nuisance phytoplankton species directly in the water bodies (in-lake measures). Although these "acute" methods do not solve the problem of catchment area eutrophication, they are cheaper, easier to manage, and for some areas they are the only way to protect human and environmental health against massive cyanobacterial proliferation. This review summarizes the extent of knowledge and published data about the management using metals (Al, Fe, Cu, Ag, Ca), photosensitizers (hydrogen peroxide, phthalocyanines, TiO(2)), herbicides and chemicals derived from natural compounds as fast and efficient removal agents of cyanobacteria. This review concludes that some compounds, when non-persistent and ecotoxicologically acceptable may help to manage cyanobacterial blooms in an efficient way compared to previous methods (e.g. copper sulfate).

Degradation of natural toxins by phthalocyanines--example of cyanobacterial toxin, microcystin.

Phthalocyanines (Pcs) are promising photosensitizers for use in various branches of science and industry. In the presence of visible light and diatomic oxygen, phthalocyanines can react to produce singlet oxygen, a member of reactive oxygen species able to damage different molecules and tissues. The aim of this study was to investigate the ability of phthalocyanines to degrade natural toxins in the presence of visible light. As the representative of hardly degradable toxins, a group of cyanobacterial peptide toxins--microcystin-LR--was chosen for this study. According to our results, phthalocyanines are able to degrade 61.5% of microcystins within a 48-hour incubation (38% of microcystins was degraded after 24 h and 24% after 12 h of incubation). Although other oxidants like hydrogen peroxide or ozone are able to degrade microcystins within several hours, we assume that by optimizing the spectrum emitted by light source and by changing the absorption characteristics of Pcs, microcystins degradation by phthalocyanines could be more effective in the near future.

In search of the main properties of phthalocyanines participating in toxicity against cyanobacteria.

Phthalocyanines are promising photosensitizers for use in various branches of science including nanotechnology. In the presence of visible light and diatomic oxygen, phthalocyanines can react to produce singlet oxygen (1O2*), which has known inhibitory effects on cellular growth and metabolic activity, although other mechanisms may be involved. The present work focuses on the properties of phthalocyanines (atom charge densities, singlet oxygen production, inhibition effects at various irradiances) contributing to toxicity against the cyanobacteria, Synechococcus nidulans. Our results indicate that positive charge densities at peripheral parts of substituents exhibit greater inhibitory effects against S. nidulans than the amount of singlet oxygen produced, potentially by binding to negatively charged membranes on the cell surface. The weak effect of 1O2* was further demonstrated by a 10% increase in phthalocyanine toxicity (the maximal inhibition detected) when the irradiance increased 3-fold from 1200 to 4000 lux.

Algicidal activity of phthalocyanines--screening of 31 compounds.

Phthalocyanines and their analogues show great potential as photodynamic agents producing reactive oxygen species (ROS), especially in medicine. However, their biocidal effects may also be employed to inhibit various undesirable organisms. This study explores their potential algicidal effects. The laboratory tests concern the effects of various phthalocyanine derivatives on the green alga Pseudokirchneriella subcapitata and cyanobacterium Synechococcus nidulans. Their effects on one example of the sensitive nontarget aquatic organism-crustacean Daphnia magna were also screened. Among 31 tested compounds, the cationic phthalocyanines substituted with heterocycle exhibited the strongest effects on phytoplankton species, some of them even below the level of 1 mg/L, while effects on crustaceans ranged from 3.6 to more than 50 mg/L. These results show that some phthalocyanine derivatives can act as potent algicides.

Growth assays with mixed cultures of cyanobacteria and algae assessed by in vivo fluorescence: One step closer to real ecosystems?

A growth toxicity assay with mixed cultures of cyanobacteria and algae using in vivo fluorescence is presented. Test organisms (the green alga Pseudokirchneriella subcapitata and the cyanobacterium Aphanothece clathrata) growing alone and in a mixture were exposed to selected chemicals. P. subcapitata featured a higher sensitivity to toxicants in the presence of A. clathrata compared to the single species assay. On the other hand, growth of a cyanobacterium was not affected by the presence or absence of the green alga. The proposed method seems to be suitable for pre-screening studies of toxicants (algistatic agents, herbicides) applied into the aquatic environment and for the assessment of their impact on natural phytoplankton communities.

Effects of aqueous extracts from five species of the family Papaveraceae on selected aquatic organisms.

The effects of aqueous root extracts from five species of the family Papaveraceae on the growth of cyanobacteria, algae, and other non-target aquatic organisms were investigated to evaluate their potential use as algicides or cyanocides in the aquatic environment. Dicranostigma lactucoides and Sanguinaria canadensis featured the highest toxicity while Macleaya microcarpa was found to be the least toxic to all aquatic organisms tested. The Chelidonium majus extract had the best properties as a potential algicide or cyanocide because of its significant toxicity to phytoplankton and lower toxicity to non-target aquatic organisms as compared with the other Papaveraceae family members.