Papers by Dorota Napierska
Comparative Biochemistry and Physiology A-molecular & Integrative Physiology, 1999
Antarctic crustaceans and South Baltic shrimps live in waters .with different temperatures. To in... more Antarctic crustaceans and South Baltic shrimps live in waters .with different temperatures. To investigate the special enzymatic adaptation of Antarctic krill Euphausia superba and shrimp Palaemonetes varians from the Vistula delta to environmental temperatures, we selected cytosol NADP-dependent malic enzyme from the abdomen muscle. Malic enzyme was purified from E. superba and P. varians. The purification procedure involved (NH&SO4 fractionation, chromatography on DEAE-Sepharose, 2',5'-ADP-Sepharose and Sepharose 6B. One band stained for
Journal of Sea Research, 2003
This paper describes the two-phase study of the environmental impact of the SwePol Link submarine... more This paper describes the two-phase study of the environmental impact of the SwePol Link submarine electrical energy transfer system between Sweden and Poland. During the first phase (1997 -1998), the potential effects of proposed technical solutions for the transmission line and different routes across the Baltic Sea were analysed. During the second phase (1999 -2000), studies on environmental and background conditions before cable installation (1999) and studies on the environmental effects after cable system installation were undertaken. During this phase, underwater TV and video inspection of the bottom, observations of the bottom habitats by scuba-divers, sampling and laboratory analysis of macrozoobenthos and measurements of the earth's magnetic field were conducted.

Ecotoxicology and Environmental Safety, 2006
The aim of the present study was to examine the activity of hepatic glutathione S-transferase (GS... more The aim of the present study was to examine the activity of hepatic glutathione S-transferase (GST) in flounder, Platichthys flesus, as a potential biomarker of exposure to xenobiotics present in the environment. The fish were collected along known pollution gradients and from areas regarded relatively free of anthropogenic input. GST was measured spectrophotometrically in each sampled specimen of flounder. The databases from the studies performed by two research institutes were compiled, and generalized linear models were used to analyze the dependence of the enzyme activity on the time and area of sampling and on selected biological parameters of the fish. Fish sex, sampling area, year, month, water temperature, and somatic indices (gonado-somatic index, hepato-somatic index) were significant in the model of GST activity. The three annual datasets showed that the biomarker activities exhibited in the Gulf of Gdansk were significantly different from those measured in the areas off the coast. r 2005 Elsevier Inc. All rights reserved.

Ecotoxicology, 2009
The objective of this study was to investigate the pattern of enzymatic activities, environmental... more The objective of this study was to investigate the pattern of enzymatic activities, environmental genotoxicity and cytotoxicity in flounder, Platichthys flesus, from the Polish coastal area of the Baltic Sea. Fish were sampled in different contaminated sites in the Gulf of Gdansk and in a reference area outside the gulf. The activities of acetylcholinesterase (AChE), glutathione s-transferase (GST), catalase (CAT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), were studied, as well as the frequency of micronuclei, nuclear buds and fragmented-apoptotic cells. A higher mean activity level of muscular AChE and a lower activity level of hepatic GST were evident in samples taken from the reference site, relative to those found in the gulf. Modeled CAT activity (in both liver and gill tissue), blood plasma LDH and CK activities were all significantly higher in flounder collected at locations within the Gulf of Gdansk than at the reference site. No statistically significant alterations were observed in the activities of ALT and AST in the blood plasma of flounder in this study. Fish collected from a location at the mouth of the Vistula River showed the highest hepatic GST and CAT, the highest gill CAT activity, and the highest frequency of blood micronuclei, nuclear buds and fragmented-apoptotic cell inductions, as well as the lowest level of blood plasma CK. The present study confirms that compared to fish from the reference area, flounder from the Gulf of Gdansk clearly demonstrate a different enzyme activity, genotoxicity and cytotoxicity biomarker response pattern.

Marine Pollution Bulletin, 2005
The results of the present study are based on enzyme biomarker measurements in flounder (Platicht... more The results of the present study are based on enzyme biomarker measurements in flounder (Platichthys flesus), a flatfish species that is widely distributed in Baltic coastal waters. The fish were collected from known pollution gradients and from reference areas regarded as relatively free of anthropogenic input. Muscular cholinesterases (ChEs), hepatic glutathione S-transferase (GST), and hepatic ethoxyresorufin-O-deethylase (EROD) activities were measured in each sampled specimen of flounder. Generalized linear models (GLM) were used to analyze the dependence of the enzyme activity on sampling year and area as well as on the biological parameters of the fish. Statistically significant differences in the activities of the measured biomarkers were observed between reference and contaminated sites. ChEs and GST activities differed with gender. The results of this study suggest that the location and year of sampling have a significant impact on the activity of all the measured biomarkers.

Pesticide Biochemistry and Physiology, 2008
The aim of this study was to investigate the impact of carbofuran, a carbamate pesticide on the e... more The aim of this study was to investigate the impact of carbofuran, a carbamate pesticide on the enzymatic activity of acetylcholinesterase (AChE) in the larvae of the herring nematode parasite Anisakis simplex. A. simplex larvae collected from herring were exposed to carbofuran in vivo at concentrations of 50, 100, 500 and 1000μg/l, for 24, 48 and 72h, at a temperature of 4°C. Generalized Linear Models (GLM) were applied to analyze the relationship between AChE activity and carbofuran concentration, the time of exposure and the biological parameters of the host. The results indicate that A. simplex larvae have a high threshold of sensitivity to carbofuran. The average enzymatic activity was higher in parasites obtained from male hosts, when compared with female hosts. These data suggest that host sex-dependent biological processes may also influence AChE enzymatic activity in parasites.

Previous studies in the Gulf of Gdańsk discussed the responses of selected enzymatic biomarkers t... more Previous studies in the Gulf of Gdańsk discussed the responses of selected enzymatic biomarkers to the contaminant gradient in fish and mussels. In the present study, flounder muscle and liver tissues were analyzed for polychlorinated biphenyls (PCB congeners: 28, 52, 101, 118, 138, 153 and 180), organochlorine pesticides (HCHs, HCB and DDTs), and trace metals (Pb, Cd, Zn, Cu, Hg, Cr). An attempt was made to identify the relationship between the measured enzymatic biomarker responses (cholinesterases, malic enzyme, isocitrate dehydrogenase, glutathione S-transferase) and contaminant concentrations in selected flounder tissues. The observed differences in enzymatic biomarker levels suggest that chronic exposure to low-concentration mixtures of contaminants may be occurring in the studied area. However, no conclusive evidence was found of a clear link between the biomarker responses and contaminant concentrations in flounder tissues.

Science of The Total Environment, 2006
The aim of the presented studies was to detect the possible effects of contaminants on the physio... more The aim of the presented studies was to detect the possible effects of contaminants on the physiological and biochemical parameters of eelpout females from the southern Baltic. Eeelpout was sampled in Polish coastal waters during November 2001, 2002 and 2003. The integrated studies included measurements of selected biomarkers in fish as well as the analysis of female reproductive capacity and fry malformation frequencies in relation to environmental conditions in examined areas. The mean values of relative fecundity (RF) and embryo somatic (ESI) indexes were the highest at the reference site. The frequency of females carrying dead and malformed fry was the highest at selected sites from the outer and inner part of the Gulf of Gdansk. The highest mean activity levels of muscular AChE were noted in fish sampled at the reference site and one site from the outer part of the gulf, whereas liver GST activity was the highest in samples from other site from the outer part of the gulf and the reference site. The results of trace metals analyses in fish muscle and liver did not indicate any substantial differences in the mean tissue concentrations between samples from contaminated sites and the reference site. The concentrations of PCBs, HCHs and DDTs in liver were markedly higher at three contaminated sites in comparison with the other sites. A similar pattern was observed in muscle tissue. Based on the preceding evidence, it cannot be concluded unequivocally that these studies on the physiology, biochemistry and contaminant concentrations of eelpout females provide evidence that the contaminants present in the coastal areas of the Polish part of the southern Baltic Sea have a harmful impact on this species.
Ices Journal of Marine Science, 2006

Marine Pollution Bulletin, 2005
The results of the present study are based on enzyme biomarker measurements in flounder (Platicht... more The results of the present study are based on enzyme biomarker measurements in flounder (Platichthys flesus), a flatfish species that is widely distributed in Baltic coastal waters. The fish were collected from known pollution gradients and from reference areas regarded as relatively free of anthropogenic input. Muscular cholinesterases (ChEs), hepatic glutathione S-transferase (GST), and hepatic ethoxyresorufin-O-deethylase (EROD) activities were measured in each sampled specimen of flounder. Generalized linear models (GLM) were used to analyze the dependence of the enzyme activity on sampling year and area as well as on the biological parameters of the fish. Statistically significant differences in the activities of the measured biomarkers were observed between reference and contaminated sites. ChEs and GST activities differed with gender. The results of this study suggest that the location and year of sampling have a significant impact on the activity of all the measured biomarkers.

Particle and Fibre Toxicology, 2010
Silica nanoparticles (SNPs) are produced on an industrial scale and are an addition to a growing ... more Silica nanoparticles (SNPs) are produced on an industrial scale and are an addition to a growing number of commercial products. SNPs also have great potential for a variety of diagnostic and therapeutic applications in medicine. Contrary to the well-studied crystalline micron-sized silica, relatively little information exists on the toxicity of its amorphous and nano-size forms. Because nanoparticles possess novel properties, kinetics and unusual bioactivity, their potential biological effects may differ greatly from those of micron-size bulk materials. In this review, we summarize the physico-chemical properties of the different nano-sized silica materials that can affect their interaction with biological systems, with a specific emphasis on inhalation exposure. We discuss recent in vitro and in vivo investigations into the toxicity of nanosilica, both crystalline and amorphous. Most of the in vitro studies of SNPs report results of cellular uptake, size-and dose-dependent cytotoxicity, increased reactive oxygen species levels and pro-inflammatory stimulation. Evidence from a limited number of in vivo studies demonstrates largely reversible lung inflammation, granuloma formation and focal emphysema, with no progressive lung fibrosis. Clearly, more research with standardized materials is needed to enable comparison of experimental data for the different forms of nanosilicas and to establish which physico-chemical properties are responsible for the observed toxicity of SNPs.

Langmuir, 2010
For the investigation of the interaction of nanoparticles with biomolecules, cells, organs, and a... more For the investigation of the interaction of nanoparticles with biomolecules, cells, organs, and animal models there is a need for well-characterized nanoparticle suspensions. In this paper we report the preparation of monodisperse dense amorphous silica nanoparticles (SNP) suspended in physiological media that are sterile and sufficiently stable against aggregation. SNP sols with various particle sizes (2-335 nm) were prepared via base-catalyzed hydrolysis and polymerization of tetraethyl orthosilicate under sterile conditions using either ammonia (Stober process (1) ) or lysine catalyst (Lys-Sil process (2) ). The series was complemented with commercial silica sols (Ludox). Silica nanoparticle suspensions were purified by dialysis and dispersed without using any dispersing agent into cell culture media (Dulbecco's Modified Eagle's medium) containing antibiotics. Particle sizes were determined by dynamic light scattering. SNP morphology, surface area, and porosity were characterized using electron microscopy and nitrogen adsorption. The SNP sols in cell culture medium were stable for several days. The catalytic activity of the SNP in the conversion of hydrogen peroxide into hydroxyl radicals was investigated using electron paramagnetic resonance. The catalytic activity per square meter of exposed silica surface area was found to be independent of particle size and preparation method. Using this unique series of nanoparticle suspensions, the relationship between cytotoxicity and particle size was investigated using human endothelial and mouse monocyte-macrophage cells. The cytotoxicity of the SNP was strongly dependent on particle size and cell type. This unique methodology and the collection of well-characterized SNP will be useful for further in vitro studies exploring the physicochemical determinants of nanoparticle toxicity.

Small, 2009
The effect that monodisperse amorphous spherical silica particles of different sizes have on the ... more The effect that monodisperse amorphous spherical silica particles of different sizes have on the viability of endothelial cells (EAHY926 cell line) is investigated. The results indicate that exposure to silica nanoparticles causes cytotoxic damage (as indicated by lactate dehydrogenase (LDH) release) and a decrease in cell survival (as determined by the tetrazolium reduction, MTT, assay) in the EAHY926 cell line in a dose-related manner. Concentrations leading to a 50% reduction in cell viability (TC50) for the smallest particles tested (14-, 15-, and 16-nm diameter) ranging from 33 to 47 µg cm−2 of cell culture differ significantly from values assessed for the bigger nanoparticles: 89 and 254 µg cm−2 (diameter of 19 and 60 nm, respectively). Two fine silica particles with diameters of 104 and 335 nm show very low cytotoxic response compared to nanometer-sized particles with TC50 values of 1095 and 1087 µg cm−2, respectively. The smaller particles also appear to affect the exposed cells faster with cell death (by necrosis) being observed within just a few hours. The surface area of the tested particles is an important parameter in determining the toxicity of monodisperse amorphous silica nanoparticles.

Nanotoxicology, 2010
Identifying the physico-chemical characteristics of nanoparticles (NPs) that drive their toxic ac... more Identifying the physico-chemical characteristics of nanoparticles (NPs) that drive their toxic activity is the key to conducting hazard assessment and guiding the design of safer nanomaterials. Here we used a set of 17 stable suspensions of monodisperse amorphous silica nanoparticles (SNPs) with selected variations in size (diameter, 2-335 nm), surface area (BET, 16-422 m(2)/g) and microporosity (micropore volume, 0-71 microl/g) to assess with multiple regression analysis the physico-chemical determinants of the cytotoxic activity in four different cell types (J774 macrophages, EAHY926 endothelial cells, 3T3 fibroblasts and human erythrocytes). We found that the response to these SNPs is governed by different physico-chemical parameters which vary with cell type: In J774 macrophages, the cytotoxic activity (WST1 assay) increased with external surface area (alphas method) and decreased with micropore volume (r(2) of the model, 0.797); in EAHY926 and 3T3 cells, the cytotoxic activity of the SNPs (MTT and WST1 assay, respectively) increased with surface roughness and small diameter (r(2), 0.740 and 0.872, respectively); in erythrocytes, the hemolytic activity increased with the diameter of the SNP (r(2), 0.860). We conclude that it is possible to predict with good accuracy the in vitro cytotoxic potential of SNPs on the basis of their physico-chemical characteristics. These determinants are, however, complex and vary with cell type, reflecting the pleiotropic interactions of nanoparticles with biological systems.

Toxicological Sciences, 2008
Because of their small size and large specific surface area (SA), insoluble nanoparticles are alm... more Because of their small size and large specific surface area (SA), insoluble nanoparticles are almost not affected by the gravitational force and are generally formulated in stable suspensions or sols. This raises, however, a potential difficulty in in vitro assay systems in which cells adhering to the bottom of a culture vessel may not be exposed to the majority of nanoparticles in suspension. J. G. Teeguarden et al., 2007, Toxicol. Sci. 95, 300-312 have recently addressed this issue theoretically, emphasizing the need to characterize the effective dose (mass or number or SA dose of particles that affect the cells) which, according to their model based on sedimentation and gravitation forces, might only represent a very small fraction of the nominal dose. We hypothesized, in contrast, that because of convection forces that usually develop in sols, the majority of the particles may reach the target cells and exert their potential toxicity. To address this issue, we exposed three different cell lines (A549 epithelial cells, EAHY926 endothelial cells, and J774 monocyte-macrophages) to a monodisperse suspension of Stö ber silica nanoparticles (SNP) in three different laboratories. Four different end points (lacticodehydrogenase [LDH] release, LDH cell content, tetrazolium salt (MTT), and crystal violet staining) were used to assess the cell response to nanoparticles. We found, in all cell lines and for all end points, that the cellular response was determined by the total mass/number/SA of particles as well as their concentration. Practically, for a given volume of dispersion, both parameters are of course intimately interdependent. We conclude that the nominal dose remains the most appropriate metric for in vitro toxicity testing of insoluble SNP dispersed in aqueous medium. This observation has important bearings on the experimental design and the interpretation of in vitro toxicological studies with nanoparticles.

Nanotoxicology, 2010
We explored how to assess the genotoxic potential of nanosize particles with a well validated ass... more We explored how to assess the genotoxic potential of nanosize particles with a well validated assay, the in vitro cytochalasin-B micronucleus assay, detecting both clastogens and aneugens. Monodisperse Stöber amorphous silica nanoparticles (SNPs) of three different sizes (16, 60 and 104 nm) and A549 lung carcinoma cells were selected as models. Cellular uptake of silica was monitored by ICP-MS. At non-cytotoxic doses the smallest particles showed a slightly higher fold induction of micronuclei (MNBN). When considering the three SNPs together, particle number and total surface area appeared to account for MNBN induction as they both correlated significantly with the amplitude of the effect. Using nominal or cellular dose did not show statistically significant differences. Likewise, alkaline comet assay and FISH-centromeric probing of MNBN indicated a weak and not statistically significant induction of oxidative DNA damage, chromosome breakage and chromosome loss. This line of investigation will contribute to adequately design and interpret nanogenotoxicity assays.
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Papers by Dorota Napierska