Bivalves Transmissible Neoplasia: Biochemical Aspects of Contagious Cancer in a Clam Macoma Balthica.

BACKGROUND/AIMS: Occurring in marine invertebrates infectious haemic neoplasia (bivalves transmissible neoplasia, BTN) arises from genome instabilities leading to multilevel malfunctions and unregulated cell division of presumably haemocyte precursors. As its biochemical characterisation remains unknown, we here present the first data describing selected aspects of the physiology and biochemistry of the disease a in model clam Macoma balthica. We chose free amino acids (FAA) composition, mitochondrial respiration and enzymatic activity, oxidative stress enzymes activities and corticosteroids profile as markers of this contagious cancer. METHODS: Selected markers were measured in neoplastic and healthy clams and two tissue types, haemolymph and solid tissue. FAA composition was assessed in the haemolymph samples using high performance liquid chromatography-mass spectrometry (LC/MS). Mitochondrial respiration analysis was performed on haemocytes using oxygen electrodes integrated system Seahorse XFp. Mitochondrial enzymes activities were measured using spectrophotometry (cytochrome oxidase, COX) and commercial kit (succinate dehydrogenase, SDH). Total Antioxidant Capacity (TAC), Acetylocholinesterase (AChE), Protein Carbonyl Content (CBO) and Malondialdehyde (MDA) levels were measured in the solid tissue using analytical kits, and glutathione (GSH) was measured spectrophotometrically. Corticosteroids profile, measured in the solid tissue, was obtained with Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (HPLC-ESI-MS/MS) technique. RESULTS: In both clam groups nine FAAs were detected with Asp, Glu, Pro, Ser constituting over 90% of total FAA content. Significantly higher Gln level was detected in BTN positive clams. In neoplastic clams, an impairment of mitochondrial metabolism was observed as a decrease in mitochondrial oxygen consumption and lower cytochrome c oxidase activity. In the neoplastic clams significantly higher concentration of low molecular weight antioxidants was found. Finally, we report high level of corticosterone and lower levels of dehydrocorticosterone, cortisol and cortisone in healthy clams and elevated cortisol level in BTN individuals. CONCLUSION: Neoplastic clams are characterized by altered mitochondrial metabolism, with a potential key role of glutamine (Gln) in cancer cells energy production. Despite low aerobic respiration, BTN cells have efficient antioxidative response to elevated concentration of ROS. Elevated cortisol level in BTN-positive clams may indicate an important role of this corticosteroid in cancer biochemistry. Thus, we here provide the first results of selected physiological and biochemical aspects of BTN, making an important step in studying cancer epidemiology in wildlife.

Horizontal transmission of disseminated neoplasia in the widespread clam Macoma balthica from the Southern Baltic Sea.

Disseminated neoplasia (DN) is one of the most challenging and unrecognised diseases occurring in aquatic fauna. It has been diagnosed in four bivalve species from the Gulf of Gdansk (Southern Baltic Sea) with the highest frequency in Macoma balthica (formerly Limecola balthica), reaching up to 94% in some populations. The aetiology of DN in the Baltic Sea has not yet been identified, with earlier studies trying to link its occurrence with environmental pollution. Taking into account recent research providing evidence that DN is horizontally transmitted as clonal cells between individuals in some bivalve species, we aimed to test whether DN is a bivalve transmissible neoplasia (BTN) in the population of M. balthica from the Gulf of Gdansk highly affected with cancer. We examined mitochondrial cytochrome c oxidase I (mtCOI) and elongation factor 1α (EF1α) sequences of genomes obtained from haemolymph and tissues of neoplastic and healthy individuals. Sequence analysis resulted in detection of an independent transmissible cancer lineage occurring in four neoplastic clams that is not present in healthy animals. This study describes the first case of BTN in the clam M. balthica (MbaBTN), providing further insights for studies on this disease.

Rapid and efficient generation of oligodendrocytes from human induced pluripotent stem cells using transcription factors.

Rapid and efficient protocols to generate oligodendrocytes (OL) from human induced pluripotent stem cells (iPSC) are currently lacking, but may be a key technology to understand the biology of myelin diseases and to develop treatments for such disorders. Here, we demonstrate that the induction of three transcription factors (SOX10, OLIG2, NKX6.2) in iPSC-derived neural progenitor cells is sufficient to rapidly generate O4+ OL with an efficiency of up to 70% in 28 d and a global gene-expression profile comparable to primary human OL. We further demonstrate that iPSC-derived OL disperse and myelinate the CNS of Mbpshi/shiRag-/- mice during development and after demyelination, are suitable for in vitro myelination assays, disease modeling, and screening of pharmacological compounds potentially promoting oligodendroglial differentiation. Thus, the strategy presented here to generate OL from iPSC may facilitate the studying of human myelin diseases and the development of high-throughput screening platforms for drug discovery.

Changes in steroid profiles of the blue mussel Mytilus trossulus as a function of season, stage of gametogenesis, sex, tissue and mussel bed depth.

This paper describes changes in the content of free steroid hormones e.g. testosterone (T), estradiol-17ß (E2), estrone (E1) and estriol (E3) of Mytilus trossulus from the southern Baltic Sea as a function of season, stage of gametogenesis, sex, tissue (gonadal and somatic) and depth. The highest levels of T, E2, E1 and E3 were found in mussels sampled in spring and summer while the lowest levels were found in winter. This pattern was stable and was seen in both sexes and tissues in mussels from both mussel beds. The spring and summer peaks in steroid levels (SL) coincided with advanced levels of gametogenesis (the highest gonadal index, GI) of our model species. But, the lowest GI (autumn) and the lowest steroids content (winter) did not overlap. Instead, water temperature increase was followed by increase of SL and vice versa. This suggests that steroids may not be actively involved in the early stages of gamete development and does not preclude them from potentially being involved as endogenous modulators in the final stages of reproduction (e.g. spawning). Hence, observed fluctuations in SL in our model species are unlikely to be caused by reproductive cycle but are rather of unknown nature, likely linked with environmental conditions. Sex-related differences in steroid content included estrogen domination in females and androgen domination in males. A trend towards higher level of steroids in gills than in gonads was found, supporting the hypothesis about an exogenous origin of steroids in bivalves. However, based on the present results, we cannot exclude the possibility that these steroids have both an endogenous and exogenous origin.

Acute hydrogen peroxide (H2O2) exposure does not cause oxidative stress in late-copepodite stage of Calanus finmarchicus.

Use of hydrogen peroxide (H2O2) for removal of salmon lice in the aquaculture industry has created concern that non-target organisms might be affected during treatment scenarios. The aim of the present study was to examine the potential for H2O2 to produce oxidative stress and reduce survival in one of the most abundant zooplankton species in Norwegian coastal areas, the copepod Calanus finmarchicus. Copepods were subjected to two 96-hr tests: (1) acute toxicity test where mortality was determined and (2) treated copepods were exposed to concentrations below the No Observed Effect Concentration (0.75 mg/L) H2O2 and analyzed for antioxidant enzyme activities, as well as levels of glutathione (GSH) and malondialdehyde (MDA). Compared to available and comparable LC50 values from the literature, our results suggest that C. finmarchicus is highly sensitive to H2O2. However, 96-hr exposure of C. finmarchicus to 0.75 mg H2O2/L did not significantly affect the antioxidant systems even though the concentration is just below the level where mortality is expected. Data suggest that aqueous H2O2 exposure did not cause cellular accumulation with associated oxidative stress, but rather produced acute effects on copepod surface (carapace). Further investigation is required to ensure that aqueous exposure during H2O2 treatment in salmon fish farms does not exert adverse effects on local non-target crustacean species and populations. In particular, studies on copepod developmental stages with a more permeable carapace are warranted.

Sex-related differences in steroid concentrations in the blue mussel (Mytilus edulis trossulus) from the southern Baltic Sea.

This paper reports on sex-related differences in free steroid hormone concentrations including the concentrations of three naturally occurring estrogens (17ß-estradiol E2, estrone E1, and estriol E3) and one androgen (testosterone T) in the tissues (gills and gonads) of the blue mussel Mytilus edulis trossulus sampled from the Gulf of Gdansk (Baltic Sea, Poland). The dissimilarity in steroid concentrations between tissues was particularly evident in the T concentration with a level in gills almost three times higher compared to gonads (on average, 15.38 ng/g w.w. and 5.31 ng/g w.w., respectively, p=0.00008), suggesting its exogenous origin. In general, a tendency towards a skewed steroid profile related to sex, with E2 more abundant for males and T for females, was observed. Female gonads were characterized by a higher level of T than testis (4.61 ng/g w.w. for females and 0.70 ng/g w.w. for males, p=0.0121). At the same time, the level of E2 found in the testis was higher than in the ovary (4.81 ng/g w.w. and 3.86 ng/g w.w., respectively); however, the difference was not statistically significant. As for gills, similar trend with T and E2 being more abundant in males was observed. At the same time, no disturbances in the sex ratio and gametogenesis process were observed which suggests i) efficient deactivation of free forms of steroids, and/or ii) their little or no physiological role.

Human stem cell models of neurodegeneration: a novel approach to study mechanisms of disease development.

The number of patients with neurodegenerative diseases is increasing significantly worldwide. Thus, intense research is being pursued to uncover mechanisms of disease development in an effort to identify molecular targets for therapeutic intervention. Analysis of postmortem tissue from patients has yielded important histological and biochemical markers of disease progression. However, this approach is inherently limited because it is not possible to study patient neurons prior to degeneration. As such, transgenic and knockout models of neurodegenerative diseases are commonly employed. While these animal models have yielded important insights into some molecular mechanisms of disease development, they do not provide the opportunity to study mechanisms of neurodegeneration in human neurons at risk and thus, it is often difficult or even impossible to replicate human pathogenesis with this approach. The generation of patient-specific induced pluripotent stem (iPS) cells offers a unique opportunity to overcome these obstacles. By expanding and differentiating iPS cells, it is possible to generate large numbers of functional neurons in vitro, which can then be used to study the disease of the donating patient. Here, we provide an overview of human stem cell models of neurodegeneration using iPS cells from patients with Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, frontotemporal dementia, Huntington's disease, spinal muscular atrophy and other neurodegenerative diseases. In addition, we describe how further refinements of reprogramming technology resulted in the generation of patient-specific induced neurons, which have also been used to model neurodegenerative changes in vitro.

Antidepressants as new endocrine disruptors? - transcriptomic profiling in gonads of Mytilus trossulus exposed to norfluoxetine.

In this study an impact of norfluoxetine (NFLU) on Mytilus trossulus gonads was investigated focusing on sex-related differences in hormonal changes, gene expression, and transcriptomic profiling. Sex-specific differences in gonadal serotonin levels were found. NFLU stimulates serotonin synthesis and/or transport in female gonads, potentially accelerating oocyte maturation and gamete release. In males, NFLU decreases serotonin level what likely leads to impeding sperm maturation and thus spawning delay. Transcriptomic analyses highlighted the presence of NFLU-induced changes in gene expression related to gametogenesis and neurotransmission. In females, NFLU upregulated genes associated with oocyte development and downregulated those involved in sperm maturation. NFLU-treated males exhibited mixed effects in their genes in relation to spermatogenesis. Additionally, sex-related differences in the expression of the CYP450 genes responsible for detoxification were found. Overall, norfluoxetine acts as an endocrine-disrupting chemical and impacts gonadal serotonin levels and gene expression, potentially disrupting reproductive success of M. trossulus.

Recent advances in the use of SPME for drug analysis in clinical, toxicological, and forensic medicine studies.

Solid-phase microextraction (SPME) has gained attention as a simple, fast, and non-exhaustive extraction technique, as its unique features enable its use for the extraction of many classes of drugs from biological matrices. This sample-preparation approach consolidates sampling and sample preparation into a single step, in addition to providing analyte preconcentration and sample clean-up. These features have helped SPME become an integral part of several analytical protocols for monitoring drug concentrations in human matrices in clinical, toxicological, and forensic medicine studies. Over the years, researchers have continued to develop the SPME technique, resulting in the introduction of novel sorbents and geometries, which have resulted in improved extraction efficiencies. This review summarizes developments and applications of SPME published between 2016 and 2022, specifically in relation to the analysis of central nervous system drugs, drugs used to treat cardiovascular disorders and bacterial infections, and drugs used in immunosuppressive and anticancer therapies.

Persistence of norfluoxetine in marine mussels.

The concentration of pharmaceuticals in coastal waters is tending towards increasing due to a shift of the human population into coastal zones. In parallel, the number of prescriptions of antidepressants, mainly selective serotonin reuptake inhibitors (SSRI), is constantly growing. Most of the SSRI is metabolised into active compounds; for instance, norfluoxetine (NFLU) is the main active metabolite of fluoxetine. In this study, we tested the bioaccumulation and depuration of NFLU in Mytilus trossulus at two environmentally relevant concentrations (100 and 500 ng/L, after six days of exposure and five days of depuration at 10 °C). The concentration of NFLU in the mussels' tissue seems not to be directly proportional to the exposure concentration. The levels of NFLU in the mussels' tissues after the depuration period were comparable to the levels detected at the end of exposure. This indicates that NFLU is not efficiently removed by the mussels and points to a potential risk for consumers of such marine organisms.

Oxytetracycline-induced inflammatory process without oxidative stress in blue mussels Mytilus trossulus.

Potentially harmful compounds including pharmaceuticals are commonly found in marine waters and sediments. Amongst those, antibiotics and their metabolites are detected worldwide in various abiotic (at concentrations as high as µg/L) and biotic matrices at ng/gram of tissue, posing a risk to non-target species exposed to them such as blue mussels. Amongst those, oxytetracycline (OTC) belongs to the most detected antibiotics in the marine environment. In this work, we concentrated on studying the potential induction of oxidative stress, activation of cellular detoxification processes (including Phase I and Phase II xenobiotic biotransformation enzymes) and multixenobiotic resistance pumps (Phase III) as well as changes in the aromatisation efficiency in Mytilus trossulus exposed to 100 µg/L OTC. Our results show that 100 µg/L OTC concentration did not provoke cellular oxidative stress and did not affect the expression of genes involved in detoxification processes in our model. Moreover, no effect of OTC on aromatisation efficiency was found. Instead, phenoloxidase activity measured in haemolymph was significantly higher in OTC exposed mussels than in those from the control (30.95 ± 3.33 U/L and 17.95 ± 2.75 U/L, respectively). OTC exposed mussels were also characterised by a tissue-dependant activation of major vault protein (MVP) gene expression (1.5 times higher in gills and 2.4 times higher in the digestive system) and a decreased expression of the nuclear factor kappa B-a (NF-κB) gene (3.4 times lower in the digestive system) when compared to those from the control. Additionally, an elevated number of regressive changes and inflammatory responses in tissues such as gills, digestive system and mantle (gonads) was observed underlining the worsening of bivalves' general health. Therefore, instead of a free-radical effect of OTC, we for the first time describe the occurrence of typical changes resulting from antibiotic therapy in non-target organisms like M. trossulus exposed to antibiotics such as OTC.

The impact of various carbon nanomaterials on the morphological, behavioural, and biochemical parameters of rainbow trout in the early life stages.

With the increasing production and the number of potential applications of carbon nanomaterials, mainly from the graphene family, their release into the natural environment, especially to aquatic ecosystems, is inevitable. The aim of the study was to determine the effects of various carbon nanomaterials (graphene nanoflakes (GNF), graphene oxide (GO), reduced graphene oxide (RGO) and silicon carbide nanofibers (NFSiC) in the concentration of 4 mg L-1 on the early life stages of the rainbow trout Oncorhynchus mykiss. The survival rates of O. mykiss were not affected after 36 days of exposure to studied materials, except for RGO, which caused significant mortality of both embryos and larvae compared to the control conditions. Larvae exposed to GO and NFSiC were characterized by a smaller standard body length at hatch, whereas at the end of the experiment, the growth of fish exposed to all materials was accelerated, especially in GO and RGO treatment, in which higher body weight and length were accompanied by lower volume of the yolk sac. Neither the markers of the oxidative damage nor the antioxidant enzymes activities were significantly affected in embryos, newly hatched larvae and larvae after 26-day exposure to studied carbon nanomaterials. Also, no neurotoxic effect expressed by the activity of the whole-body acetylcholinesterase was observed. Nevertheless, the significant increase in the velocity and the overall activity of larvae exposed to GNF (not investigated after exposure to other materials) must be highlighted. The most pronounced effect of RGO might be connected with its large particle size, sharp edges, and the presence of TiO2 nanoparticles. The results indicate for the first time that various carbon nanomaterials potentially released into aquatic ecosystems may have serious developmental implications for the early life stages of salmonid fish.

Microplastics cause neurotoxicity and decline of enzymatic activities in important bioturbator Hediste diversicolor.

Microplastics (MPs) tend to accumulate in marine sediments thus benthic fauna is particularly vulnerable to microplastic pollution. Hediste diversicolor is a widespread species in coastal marine sediments. It plays key ecological functions mostly related to bioturbation process which means sediment reworking due to the worm burrowing activity and building a network of galleries. Herein, we show that commercial plastic microspheres of two sizes (63-75 and 300-355 µm) have the potential to cause neurotoxicity in H. diversicolor. The whole-body acetylcholinesterase (AChE) activity - a common indicator of neurotoxic effect - was on average 60% lower in polychaetes exposed for 28 days to MPs served at environmentally relevant concentrations (0.08% sediment d. wt.), than in unexposed ones. Significantly reduced activities of antioxidant enzymes (SOD, CAT, GST) indicated suppression of the cellular antioxidative system in worms exposed to MPs. No changes were, however, observed in tGSH, lipid or protein oxidation measures (CBO, MDA), and in the energetic value of these polychaetes. The response was generally similar with no regard to MPs size. Only very few microspheres were found in polychaetes exposed to MPs spiked sediment. The potential role of MPs-associated pollutants as a factor responsible for observed biochemical effects, is discussed.

Do magnetic fields related to submarine power cables affect the functioning of a common bivalve?

The aim of the study was to determine the effect of static magnetic field (SMF) and electromagnetic field (EMF), of values usually recorded near submarine cables, on the bioenergetics, oxidative stress, and neurotoxicity in the cockle Cerastoderma glaucum. Bivalves maintained a positive energy balance, but the filtration rate and energy available for individual production were significantly lower in SMF-exposed animals compared to the control treatment. No changes in the respiration were noted but ammonia excretion rate was significantly lower after exposure to EMF. Changes in the activities of antioxidant enzymes and the lipid peroxidation were not observed however, exposure to both fields resulted in increased protein carbonylation. After exposure to EMF a significant inhibition of acetylcholinesterase activity was observed. As the present study for the first time revealed the oxidative damage and neurotoxicity in marine invertebrate after exposure to artificial magnetic fields, the need for further research is highlighted.

Effects of different types of primary microplastics on early life stages of rainbow trout (Oncorhynchus mykiss).

Plastic pollution is recognized as serious threat to aquatic organisms. The aim of this research was to determine the effect of environmentally realistic concentrations of various microplastics (MPs) on survival, growth, development and induction of endocrine, geno- and cytotoxic responses in the early life stages of rainbow trout Oncorhynchus mykiss. Fish were exposed for 69-days, from embryos at eyed-stage to mobile yolk-sac larvae, to pre-production pellets (3000 µm; polystyrene - PS and polyethylene terephthalate - PET). Additionally, since salmonid larvae are particularly exposed to light polymers after swimming up from the bottom, fish were also treated with PE microspheres (150-180 µm; polyethylene - PE) for both long (69-days, from embryos at eyed-stage) and short period (29 days, from larvae 3 weeks after hatching) to test the development stage-related effect on the growth parameters and fitness. Hatching success, rate and the survival of larvae did not differ among treatments. Although some alterations were found in the length gain after the long-term exposure and in the yolk-sac exhaustion rate in all PE treatments, the final size of larvae did not differ from the respective controls. PE-treated larvae have shown elevated corticosterone concentrations being significantly higher in fish exposed from the embryo stage. It was indicated for the first time that mobile yolk-sac larvae ingested MPs (up to 24% of larvae contained microspheres). No changes were recorded in cytotoxicity endpoints in any of the treatments, but exposure to PS pellets resulted in significantly higher frequencies of genotoxicity endpoints compared to the control treatment. This effect and aforementioned alterations in PE-treated larvae might result from the exposure to toxic MPs leaches. The fact that selected PAHs' levels reached the highest values in PS pellets and PE microspheres must be underlined.

Oil-mediated oxidative-stress responses in a keystone zooplanktonic species, Calanus finmarchicus.

The copepod Calanus finmarchicus is an ecologically important species in the North Atlantic, Norwegian and Barents seas. Accidental or continuous petroleum pollution from oil and gas production in these seas may pose a significant threat to this low trophic level keystone species. Responses related to oxidative stress, protein damage and lipid peroxidation were investigated in C. finmarchicus exposed to a water-accommodated fraction (WAF) of a naphthenic North Atlantic crude oil. The exposure concentration corresponded to 50% of the 96 h LC50, and samples were obtained at 0, 24, 48, 72 and 96 h after exposure initiation. Gene expressions (superoxide dismutase, catalase, glutathione S-transferase, glutathione synthetase, heat shock protein 70 and 90, ubiquitin and cytochrome P-450 330A1), enzyme activities (superoxide dismutase, catalase, glutathione S-transferase) and concentrations of total glutathione and malondialdehyde were analyzed. Gene expression analyses showed no differences between controls and the exposed animals, however significantly higher glutathione S-transferase activity and malondialdehyde concentrations were found in the exposed group, suggests lipid peroxidation as main toxic effect.

Do the graphene nanoflakes pose a potential threat to the polychaete Hediste diversicolor?

Graphene is a promising material with a wide range of future applications that could potentially lead to its transfer from numerous water and terrestrial sources to the sea, thus fate and effects of graphene in the marine ecosystem deserve attention. Within this work, the impact of the short- and long-term exposure (36 h and 24 days) of the marine benthic polychaete Hediste diversicolor to various concentrations (36 h: 0.4, 4, 40 and 400 mg L-1; 24 days: 4 and 40 mg L-1) of the pristine graphene multilayer nanoflakes (of thickness 8-12 nm) was investigated. Experiments revealed a limited toxic effect of graphene on H. diversicolor. Although the polychaetes ingested graphene, no impact on their total energy content was found. The toxic effect expressed by significant elevation of catalase activity indicating activation of defence mechanisms was recorded but only at the early stage of exposure. Activities of other antioxidant and cellular damage biomarkers (SOD, GST, GSH, MDA, CBO) remained unaffected. Moreover, no neurotoxic effect expressed by inhibition of acetylcholinesterase (AChE) activity was observed. Substantial inter-individual variability in the activities of some biomarkers at the end of the long-term experiment was found. Polychaetes were buried deeper in the sediment with graphene than in the controls indicating an escape reaction and avoidance behaviour. The latter may lead to the transfer of graphene from the sediment surface to deeper sediment layers with unknown consequences for the benthic ecosystem.

Cellular level response of the bivalve Limecola balthica to seawater acidification due to potential CO2 leakage from a sub-seabed storage site in the southern Baltic Sea: TiTank experiment at representative hydrostatic pressure.

Understanding of biological responses of marine fauna to seawater acidification due to potential CO2 leakage from sub-seabed storage sites has improved recently, providing support to CCS environmental risk assessment. Physiological responses of benthic organisms to ambient hypercapnia have been previously investigated but rarely at the cellular level, particularly in areas of less common geochemical and ecological conditions such as brackish water and/or reduced oxygen levels. In this study, CO2-related responses of oxygen-dependent, antioxidant and detoxification systems as well as markers of neurotoxicity and acid-base balance in the Baltic clam Limecola balthica from the Baltic Sea were quantified in 50-day experiments. Experimental conditions included CO2 addition producing pH levels of 7.7, 7.0 and 6.3, respectively and hydrostatic pressure 900 kPa, simulating realistic seawater acidities following a CO2 seepage accident at the potential CO2-storage site in the Baltic. Reduced pH interfered with most biomarkers studied, and modifications to lactate dehydrogenase and malate dehydrogenase indicate that aerobiosis was a dominant energy production pathway. Hypercapnic stress was most evident in bivalves exposed to moderately acidic seawater environment (pH 7.0), showing a decrease of glutathione peroxidase activity, activation of catalase and suppression of glutathione S-transferase activity likely in response to enhanced free radical production. The clams subjected to pH 7.0 also demonstrated acetylcholinesterase activation that might be linked to prolonged impact of contaminants released from sediment. The most acidified conditions (pH 6.3) stimulated glutathione and malondialdehyde concentration in the bivalve tissue suggesting potential cell damage. Temporal variations of most biomarkers imply that after a 10-to-15-day initial phase of an acute disturbance, the metabolic and antioxidant defence systems recovered their capacities.

Melatonin enhances antioxidant action of alpha-tocopherol and ascorbate against NADPH- and iron-dependent lipid peroxidation in human placental mitochondria.

Human placental mitochondria might be a significant source of NADPH- and iron-dependent production of reactive oxygen species (ROS). Preeclampsia is believed to be a consequence of overproduction of ROS in human placenta. The experimental results presented here show that melatonin inhibits NADPH- and iron-dependent lipid peroxidation of human placental mitochondria in a concentration-dependent manner. At 1.5 mm concentration, melatonin suppressed this process nearly completely. Melatonin does not influence significantly the iron oxidation at this conditions, indicating that free radical scavenging rather than metal-chelating phenomenon is the basis of its antioxidant action. The fact of inhibition of lipid peroxidation by melatonin at conditions excluding iron participation also supports this hypothesis. Elucidation of the nature of common interaction among melatonin, ascorbate, and alpha-tocopherol in human placental mitochondria was the main aim of this study. In presence of 90 mum ascorbate, the inhibition of lipid peroxidation by melatonin was strong and had a feature of synergistic interaction. At presence of 30 mum ascorbate, which stimulated lipid peroxidation, melatonin caused a loss of pro-oxidant effect of ascorbate. While the interaction of melatonin with ascorbate indicated synergism, the joint action of melatonin and alpha-tocopherol was additive. When all three antioxidants were applied together, the strongest inhibition of lipid peroxidation was observed. The experimental results presented here indicated that melatonin could be considered as an effective component of antioxidant treatment of preeclampsia, allowing the use of reduced doses of vitamin C and E owing to elevated efficiency of their antioxidant activity in placenta when used in combination.

Effects of chronic exposure to microplastics of different polymer types on early life stages of sea trout Salmo trutta.

The aim of the present study was to determine the effect of a long-term (113 days) exposure to microplastics on the development and induction of endocrine, geno- and cytotoxic responses in early life stages of sea trout Salmo trutta. Microplastic particles (3000 µm) of three most commonly mass-produced polymers (polystyrene - PS, polyethylene terephthalate - PET and polyethylene - PE) were applied in environmentally realistic concentrations (0.1% of sediment dry weight) in a laboratory experiment imitating the natural environment, typical for sea trout spawning grounds. The exposure of the sea trout, from fertilized eggs to mobile yolk-sac larvae, to microplastics did not affect the hatching success (the survival of embryos), hatching rate and the incubation period. Microplastics of any tested polymer type also had no adverse effect on the larvae survival, growth rate and the rate of yolk sack absorption. Similarly, no changes in frequencies of detected cytotoxicity endpoints compared to the control group were recorded. Exposure to polymer particles induced however the formation of genotoxicity endpoints (nuclear buds, micronuclei and blebbed nuclei cells). The level of total genotoxicity (ΣGentox) in fish larvae erythrocytes increased significantly in the following sequence: PS > PET > PE. No significant changes in the whole body corticosterone, dehydrocorticosterone and cortisone concentrations due to exposure to microplastics were recorded, while cortisol was detected in larvae exposed to PS. Our results show that long-term, non-ingestion related exposure to microplastics does not affect development of S. trutta early life stages but may lead to genotoxic responses. PS seems to be the most hazardous among all polymers studied. This is the first study demonstrating non-ingestion related toxicity of microplastics to the early life stages of fish.