Publications by authors named "Anna Wójtowicz"

58 Publications

Probing menstrual bloodstain aging with fluorescence spectroscopy.

Spectrochim Acta A Mol Biomol Spectrosc 2021 Mar 13;248:119172. Epub 2020 Nov 13.

Department of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, NY 12222, USA. Electronic address:

Menstrual blood (MB) is a common and important type of forensic evidence, especially in sexual assault cases. MB is composed of peripheral blood (PB), vaginal fluid, and endometrial cells of the uterine wall. In forensic investigations, the differentiation of MB and PB can determine whether the blood present is a result of tissue damage from an assault or a natural cause and thus help to reconstruct the event. Understanding how menstrual blood changes is necessary to develop a method for bloodstain aging. Fluorescence spectroscopy, a promising spectroscopic method for bloodstain analysis, was used to probe the biochemical changes that occur over time in menstrual bloodstains. It was found that steady-state fluorescence spectra underwent significant changes over first nine hours post deposition. The underlying mechanism of fluorescence changes was proposed to involve the kinetic transformation of three fluorophores: tryptophan, nicotinamide adenine dinucleotide and flavins.
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http://dx.doi.org/10.1016/j.saa.2020.119172DOI Listing
March 2021

Fetal upper mediastinum - normal and abnormal findings in obstetric ultrasound screening.

Ginekol Pol 2020 ;91(10):620-628

Department of Obstetrics and Perinatology, Jagiellonian University Medical College, Cracow, Poland.

Fetal cardiac assessment is an integral part of the obstetric ultrasound. The inclusion of the outflow tracts and the three-vessel and tracheal view into the ultrasound screening enhances the detection rate for cardiovascular anomalies. Both, international and Polish guidelines recommend routine evaluation of the upper mediastinum. The aim of the study was to present the principles for assessing the structures of the upper mediastinum in normal conditions and to draw attention to the pathologies which may be visible in this plane.
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http://dx.doi.org/10.5603/GP.a2020.0092DOI Listing
January 2020

Autophagy-related neurotoxicity is mediated via AHR and CAR in mouse neurons exposed to DDE.

Sci Total Environ 2020 Nov 29;742:140599. Epub 2020 Jun 29.

Maj Institute of Pharmacology, Polish Academy of Sciences, Department of Experimental Neuroendocrinology, Laboratory of Molecular Neuroendocrinology, Smetna street 12, 31-343 Krakow, Poland. Electronic address:

DDE (dichlorodiphenyldichloroethylene) is an environmental metabolite of the pesticide DDT, which is still present in the environment, and its insecticidal properties are used to fight malaria and the Zika virus disease. We showed for the first time that the neurotoxic effects of DDE involve autophagy, as demonstrated by elevated levels of Becn1, Map1lc3a/MAP1LC3A, Map1lc3b, and Nup62/NUP62 and an increase in autophagosome formation. The suggestion that the aryl hydrocarbon receptor (AHR) and the constitutive androstane receptor (CAR) are involved in the neurotoxic effect of DDE was supported by increases in the mRNA and protein expression of these receptors, as detected by qPCR, ELISA, immunofluorescence labeling and confocal microscopy. Selective antagonists of the receptors, including alpha-naphthoflavone, CH223191, and CINPA 1, inhibited p,p'-DDE- and o,p'-DDE-induced LDH release and caspase-3 activity, while specific siRNAs (Ahr and Car siRNA) reduced the levels of p,p'-DDE- and o,p'-DDE-induced autophagosome formation. Although the neurotoxic effects of DDE were isomer independent, the mechanisms of p,p'- and o,p'-DDE were isomer specific. Therefore, we identified previously unknown mechanisms of the neurotoxic actions of DDE that, in addition to inducing apoptosis, stimulate autophagy in mouse neocortical cultures and induce AHR and CAR signaling.
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http://dx.doi.org/10.1016/j.scitotenv.2020.140599DOI Listing
November 2020

The interference of alpha- and beta-naphthoflavone with triclosan effects on viability, apoptosis and reactive oxygen species production in mouse neocortical neurons.

Pestic Biochem Physiol 2020 Sep 1;168:104638. Epub 2020 Jul 1.

Department of Animal Nutrition, Biotechnology, and Fisheries, Agricultural University of Krakow, Al. Mickiewicza 24/28, 30-059 Krakow, Poland.

Triclosan (TCS) is commonly used worldwide in a range of personal care and sanitizing products. A number of studies have revealed the presence of TCS in human tissues. It has recently been shown that TCS can interact with AhR in mouse neurons and the one of its effects is the stimulation of reactive oxygen species (ROS) production. Reactive oxygen species perform a wide spectrum of functions in neuronal cells, where they are generated as by-products of cellular metabolism. Therefore the aim of the study was to investigate effects of two synthetic naphthoflavones, the beta-naphthoflavone (βNF) and alpha-naphthoflavone (αNF), well known agonist and antagonist of AhR on TCS-stimulated cytotoxicity, apoptosis and ROS production in mouse primary cortical neurons in vitro cultures. The results showed that both agonist (βNF) and antagonist (αNF) of AhR enhanced the LDH release and caspase-3 activity stimulated by TCS. Interestingly, both naphthoflavones decreased the TCS-stimulated ROS production, however, they showed no scavenging properties as revealed by ABTS and DPPH methods. What's more, both βNF as well as αNF inhibited the activity of xanthine oxidase (XO) stimulated by TCS. Thus, we can assume that αNF or βNF act in a competitive way over TCS and inhibit its effect on antioxidant enzyme activity.
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http://dx.doi.org/10.1016/j.pestbp.2020.104638DOI Listing
September 2020

Early- and Late-Onset Preeclampsia: A Comprehensive Cohort Study of Laboratory and Clinical Findings according to the New ISHHP Criteria.

Int J Hypertens 2019 17;2019:4108271. Epub 2019 Sep 17.

Department of Obstetrics & Perinatology, Jagiellonian University Medical College, Ul. Kopernika 23, 31-501 Kraków, Poland.

Recently, the diagnostic criteria of preeclampsia have been changed. No studies are available in the literature that analyzed in detail the differences between early-onset preeclampsia (EOP) and late-onset preeclampsia (LOP), taking into account the International Society for the Study of Hypertension in Pregnancy (ISSHP) criteria. Thus, we sought to retrospectively investigate in detail the differences in clinical and laboratory outcomes between EOP and LOP diagnosed according to the ISSHP criteria. A retrospective cohort study was conducted in 214 women with singleton pregnancies and preeclampsia admitted to the Department of Obstetrics and Perinatology of the University Hospital in Kraków, Poland, from 2013 to 2017 (113 (52.8%) women with EOP and 101 (47.2%) women with LOP). Electronic medical records were reviewed for demographics and medical history, laboratory tests, and delivery and neonatal data. Patients with preeclampsia accounted for 1.7% of the women who delivered during the study period. The EOP and LOP groups did not differ in the distribution of risk factors for preeclampsia. The most common risk factor was primiparity, which was observed in 72.0% of cases. Regarding the ISSHP diagnostic criteria, the two groups differed in the incidence of fetal growth restriction (=0.0009), hemolysis (=0.0416), and neurological complications (=00342), which were found more often in the EOP group. In addition, the EOP group had more frequent occurrence of severe cardiorespiratory ( < 0.0001) and hematological (=0.0127) complications, adverse fetoplacental conditions ( < 0.0001), and severe fetoplacental complications (=0.0003). Children born to women with EOP had lower Apgar scores ( < 0.001) and higher rates of intraventricular hemorrhage ( < 0.0001), respiratory disorders requiring mechanical ventilation ( < 0.0001), and early (=0.0004) and late sepsis (=0.002). EOP differed from LOP in terms of maternal and perinatal adverse outcomes. The observed higher rates of fetoplacental adverse conditions and severe complications indicate a significant contribution of impaired placentation to the etiopathogenesis of EOP.
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http://dx.doi.org/10.1155/2019/4108271DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6766116PMC
September 2019

Direct analysis from dried blood spot card surfaces with direct probe mass spectrometry - Evaluation study.

Rapid Commun Mass Spectrom 2019 Jul;33(13):1148-1152

Faculty of Chemistry, Jagiellonian University, Gronostajowa St. 2, 30-387, Kraków, Poland.

Rationale: The Direct Probe Mass Spectrometry (DIPMS) method allows successful analysis of powders, solid and liquid samples. The potential of direct surface analysis could find further application in the examination of surfaces with good absorption properties such as Dried Blood Spot (DBS) cards that constitute a great alternative to the classical blood collection method directly from veins.

Methods: DIPMS was performed with the ionization carried out under atmospheric pressure in an Atmospheric Pressure Chemical Ionization source. Direct analysis of diazepam solutions in methanol and after their deposition onto a DBS card was conducted. Subsequently, images of the DBS cards with and without blood samples were acquired using Scanning Electron Microscopy (SEM).

Results: Direct quantitative analysis of diazepam liquid samples by DIPMS was successfully performed. Linear correlation between the concentration of diazepam and the peak intensity with a R coefficient of 0.937 was obtained. However, the method failed when the analysis was conducted directly from the surface of the DBS cards and no diazepam peak was observed in the mass spectrum. The SEM images confirmed the good absorption properties of DBS cards and the absence of blood components on the surface.

Conclusions: DIPMS is an excellent technique for the rapid, direct analysis of powders, solid and liquid samples; however, the potential of the method is limited when samples are deposited on surfaces with good absorption properties such as DBS cards.
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http://dx.doi.org/10.1002/rcm.8447DOI Listing
July 2019

The Action of Di-(2-Ethylhexyl) Phthalate (DEHP) in Mouse Cerebral Cells Involves an Impairment in Aryl Hydrocarbon Receptor (AhR) Signaling.

Neurotox Res 2019 Jan 18;35(1):183-195. Epub 2018 Aug 18.

Department of Clinical Biochemistry, University of Opole, kard. B. Kominka 6a, 45-032, Opole, Poland.

Di-(2-ethylhexyl) phthalate (DEHP) is used as a plasticizer in various plastic compounds, such as polyvinyl chloride (PVC), and products including baby toys, packaging films and sheets, medical tubing, and blood storage bags. Epidemiological data suggest that phthalates increase the risk of the nervous system disorders; however, the impact of DEHP on the brain cells and the mechanisms of its action have not been clarified. The aim of the present study was to investigate the effects of DEHP on production of reactive oxygen species (ROS) and aryl hydrocarbon receptor (AhR), as well as Cyp1a1 and Cyp1b1 mRNA and protein expression in primary mouse cortical neurons and glial cells in the in vitro mono-cultures. Our experiments showed that DEHP stimulated ROS production in both types of mouse neocortical cells. Moreover, the results strongly support involvement of the AhR/Cyp1A1 signaling pathway in the action of DEHP in neurons and glial cells. However, the effects of DEHP acting on the AhR signaling pathways in these two types of neocortical cells were different. In neurons, AhR mRNA expression did not change, but AhR protein expression decreased in response to DEHP. A similar trend was observed for Cyp1a1 and Cyp1b1 mRNA and protein expression. Failure to induce Cyp1a1 in neurons was confirmed by EROD assay. In primary glial cells, a decrease in AhR protein level was accompanied by a decrease in AhR mRNA expression. In glial cells, mRNA and protein expression of Cyp1a1 as well as Cyp1a1-related EROD activity were significantly increased. As for Cyp1b1, both in neurons and glial cells Cyp1b1 mRNA expression did not significantly change, whereas Cyp1b1 protein level were decreased. We postulate that developmental exposure to DEHP which dysregulates AhR/Cyp1a1 may disrupt defense processes in brain neocortical cells that could increase their susceptibility to environmental toxins.
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http://dx.doi.org/10.1007/s12640-018-9946-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6313375PMC
January 2019

Impact of Elastin-Derived Peptide VGVAPG on Matrix Metalloprotease-2 and -9 and the Tissue Inhibitor of Metalloproteinase-1, -2, -3 and -4 mRNA Expression in Mouse Cortical Glial Cells In Vitro.

Neurotox Res 2019 Jan 30;35(1):100-110. Epub 2018 Jul 30.

Department of Public Health, Dietetics and Lifestyle Disorders, Faculty of Medicine, University of Information Technology and Management in Rzeszow, Sucharskiego 2, 35-225, Rzeszow, Poland.

Degradation products of elastin, i.e. elastin-derived peptides (EDPs), are involved in various physiological and pathological processes. EDPs are detectable in cerebrospinal fluid in healthy people and in patients after ischemic stroke. However, to date, no studies concerning the role of EDP in the nervous system were conducted. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) play important roles during the repair phases of cerebral ischemia, particularly during angiogenesis and reestablishment of cerebral blood flow. Therefore, the aim of this study was to investigate the impact of the specific elastin-derived peptide VGVAPG on Mmp-2, -9 and Timp-1, -2, -3 and -4 mRNA expression in mouse cortical glial cells in vitro. Primary glial cells were maintained in DMEM/F12 without phenol red supplemented with 10% fetal bovine serum and the cells were exposed to 50 nM, 1 and 50 μM of the VGVAPG peptide. After 3 and 6 h of exposition to the peptide, expression of Mmp-2, -9 and Timp-1, -2, -3 and -4 mRNA was measured. Moreover, siRNA gene knockdown, cytotoxicity and apoptosis measurement were included in our experiments, which showed that VGVAPG in a wide range of concentrations exhibited neither proapoptotic nor cytotoxic properties in mouse glial cells in vitro. The peptides enhanced mRNA expression of Timp-2 and Timp-3 genes in an elastin-binding protein (EBP)-dependent manner. However, changes in mRNA expression of Mmp-2, Mmp-9 and Timp-4 were partially EBP-dependent. The decrease in mRNA expression of Timp-1 was EBP-independent. However, further studies underlying the VGVAPG peptide's mechanism of action in the nervous system are necessary.
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http://dx.doi.org/10.1007/s12640-018-9935-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6313372PMC
January 2019

Triclosan-Evoked Neurotoxicity Involves NMDAR Subunits with the Specific Role of GluN2A in Caspase-3-Dependent Apoptosis.

Mol Neurobiol 2019 Jan 19;56(1):1-12. Epub 2018 Apr 19.

Department of Animal Biotechnology, Faculty of Animal Sciences, University of Agriculture, Redzina 1B, 30-248, Krakow, Poland.

Triclosan (TCS) is an antimicrobial agent that is used extensively in personal care and in sanitising products. A number of studies have shown the presence of TCS in different human tissues such as blood, adipose tissue, the liver, brain as well as in breast milk and urine. N-Methyl-D-aspartate receptors (NMDARs) are glutamate-gated ion channels that are widely expressed in the central nervous system and which play key roles in excitatory synaptic transmission. There is, however, no data on the involvement of NMDAR subunits in the apoptotic and neurotoxic effects of TCS. Our experiments are the first to show that TCS used at environmentally relevant concentrations evoked NMDA-dependent effects in neocortical neurons in primary cultures, as MK-801, an uncompetitive NMDA receptor antagonist, reduced the levels of TCS-induced ROS production as well as caspase-3 activity and LDH release. TCS caused a decrease in protein expression of all the studied NMDA receptor subunits (GluN1, GluN2A, GluN2B) that were measured at 3, 6 and 24 h post-treatment. However, at 48 h of the experiment, the level of the GluN1 subunit returned to the control level, and the levels of the other subunits showed a tendency to increase. In TCS-treated neocortical cells, protein profiles of NMDAR subunits measured up to 24 h were similar to mRNA expression of GluN1 and GluN2A, but not to GluN2B mRNA. In this study, cells transiently transfected with GluN1, GluN2A or GluN2B siRNA exhibited reduced levels of LDH release, which suggests the involvement of all of the studied NMDAR subunits in the neurotoxic action of TCS. According to our data, GluN1 and GluN2A were mainly responsible for neuronal cell death as evidenced by neutral red uptake, whereas GluN2A was involved in TCS-induced caspase-3-dependent apoptosis. We suggest that TCS-evoked apoptosis and neurotoxicity could be related to transient degradation of NMDAR subunits in mouse neurons. Furthermore, recycling of NMDAR subunits in response to TCS is possible. Because transfections with specific siRNA did not completely abolish the effects of TCS as compared to cells transfected with negative siRNA in this study, other NMDAR-independent mechanisms of TCS action are also possible.
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http://dx.doi.org/10.1007/s12035-018-1083-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6334736PMC
January 2019

Evaluation of the fetal palate at 11 to 13 (+6) weeks of gestation based on an analysis of static ultrasound images using modern IT techniques.

Prenat Diagn 2018 05 26;38(6):414-421. Epub 2018 Apr 26.

Department of Obstetrics & Perinatology, Jagiellonian University Medical College, Kraków, Poland.

Objective: To describe a new computer-based technique to isolate the shape of the fetal palate visible in the midsagittal plane from static ultrasound images routinely used to measure nuchal translucency.

Method: This is a retrospective interpretation of images of the midsagittal view of the fetal face at 11 to 13 (+6) weeks of gestation in 7 cases of cleft lip and palate (CLP) and 7 normal controls. The images were subjected to pattern analysis.

Results: Proprietary software was applied and for each CLP case, and palatine bones with different forms from those of fetuses in the control group were recorded. In 4 cases, an image of a continuous palatine bone was observed at a threshold of 180, whereas the remaining 3 images were obtained at 128. A continuous palatine bone structure was not observed in any fetus from the CLP group, even at a level of 128, when the surrounding structures were visible.

Conclusion: The application of pattern analysis to a 2D frozen image is a new approach for prenatal diagnostics. This technique may be a helpful tool for physicians and could assist in the diagnosis of cleft palate.
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http://dx.doi.org/10.1002/pd.5251DOI Listing
May 2018

Comparison of FTIR-ATR and Raman spectroscopy in determination of VLDL triglycerides in blood serum with PLS regression.

Spectrochim Acta A Mol Biomol Spectrosc 2017 Aug 18;183:239-246. Epub 2017 Apr 18.

Wrocław University of Science and Technology, Faculty of Fundamental Problems of Technology, Department of Biomedical Engineering, 27 Stanisława Wyspiańskiego St., 50-370 Wrocław, Poland.

Hypertriglyceridemia, related with triglyceride (TG) in plasma above 1.7mmol/L is one of the cardiovascular risk factors. Very low density lipoproteins (VLDL) are the main TG carriers. Despite being time consuming, demanding well-qualified staff and expensive instrumentation, ultracentrifugation technique still remains the gold standard for the VLDL isolation. Therefore faster and simpler method of VLDL-TG determination is needed. Vibrational spectroscopy, including FT-IR and Raman, is widely used technique in lipid and protein research. The aim of this study was assessment of Raman and FT-IR spectroscopy in determination of VLDL-TG directly in serum with the isolation step omitted. TG concentration in serum and in ultracentrifugated VLDL fractions from 32 patients were measured with reference colorimetric method. FT-IR and Raman spectra of VLDL and serum samples were acquired. Partial least square (PLS) regression was used for calibration and leave-one-out cross validation. Our results confirmed possibility of reagent-free determination of VLDL-TG directly in serum with both Raman and FT-IR spectroscopy. Quantitative VLDL testing by FT-IR and/or Raman spectroscopy applied directly to maternal serum seems to be promising screening test to identify women with increased risk of adverse pregnancy outcomes and patient friendly method of choice based on ease of performance, accuracy and efficiency.
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http://dx.doi.org/10.1016/j.saa.2017.04.020DOI Listing
August 2017

Depressive-like effect of prenatal exposure to DDT involves global DNA hypomethylation and impairment of GPER1/ESR1 protein levels but not ESR2 and AHR/ARNT signaling.

J Steroid Biochem Mol Biol 2017 07 2;171:94-109. Epub 2017 Mar 2.

Department of Animal Biotechnology, Faculty of Animal Sciences, University of Agriculture, Redzina Street 1B, 30-248 Krakow, Poland.

Several lines of evidence suggest that exposures to Endocrine Disrupting Chemicals (EDCs) such as pesticides increase the risks of neuropsychiatric disorders. Despite extended residual persistence of dichlorodiphenyltrichloroethane (DDT) in the environment, the mechanisms of perinatal actions of DDT that could account for adult-onset of depression are largely unknown. This study demonstrated the isomer-specific induction of depressive-like behavior and impairment of Htr1a/serotonin signaling in one-month-old mice that were prenatally exposed to DDT. The effects were reversed by the antidepressant citalopram as evidenced in the forced swimming (FST) and tail suspension (TST) tests in the male and female mice. Prenatally administered DDT accumulated in mouse brain as determined with gas chromatography and tandem mass spectrometry, led to global DNA hypomethylation, and altered the levels of methylated DNA in specific genes. The induction of depressive-like behavior and impairment of Htr1a/serotonin signaling were accompanied by p,p'-DDT-specific decrease in the levels of estrogen receptors i.e. ESR1 and/or GPER1 depending on sex. In contrast, o,p'-DDT did not induce depressive-like effects and exhibited quite distinct pattern of biochemical alterations that was related to aryl hydrocarbon receptor (AHR), its nuclear translocator ARNT, and ESR2. Exposure to o,p'-DDT increased AHR expression in male and female brains, and reduced expression levels of ARNT and ESR2 in the female brains. The evolution of p,p'-DDT-induced depressive-like behavior was preceded by attenuation of Htr1a and Gper1/GPER1 expression as observed in the 7-day-old mouse pups. Because p,p'-DDT caused sex- and age-independent attenuation of GPER1, we suggest that impairment of GPER1 signaling plays a key role in the propagation of DDT-induced depressive-like symptoms.
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http://dx.doi.org/10.1016/j.jsbmb.2017.03.001DOI Listing
July 2017

The significance of a prenatal diagnosis of right aortic arch.

Prenat Diagn 2017 Apr 13;37(4):365-374. Epub 2017 Mar 13.

Department of Obstetrics and Perinatology, Jagiellonian University Medical College, Kraków, Poland.

Objectives: To analyze a population of fetuses with prenatally diagnosed right aortic arch (RAA).

Methods: Retrospective study of fetuses with RAA diagnosed prenatally between 2011 and 2015 in two referral centers.

Results: Right aortic arch was found in 4.4% (46/1036) of fetuses with cardiovascular abnormalities (CVA). As an isolated anomaly, RAA was present in 30.4% of cases; in 32.6%, other CVA were detected; in 23.9%, CVA and extracardiac anomalies; and in 13.1%, only extracardiac malformations. The most common noncardiac abnormalities were thymus hypoplasia/aplasia (7/17), of which six had deletion 22q.11.2. In another three fetuses, trisomy 21 was present. One intrauterine fetal death occurred at 41 weeks of pregnancy, and two fetuses died after birth. In six of 18 infants with known follow-up, symptoms of dysphagia were reported, of which four infants underwent surgical intervention. In 12 infants, an isolated RAA was clinically silent.

Conclusions: The diagnosis of RAA is an indication for a detailed examination of cardiac and noncardiac structures, including the thymus. It is advisable to consider genetic testing, together with the assessment of deletion 22q11.2, especially in the case of accompanying defects. The prognosis depends on underlying cardiac and extracardiac anomalies and possibly coexisting genetic defects. Isolated anomalies are asymptomatic. © 2017 John Wiley & Sons, Ltd.
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http://dx.doi.org/10.1002/pd.5020DOI Listing
April 2017

Neonatal Marfan syndrome diagnosed prenatally.

Ginekol Pol 2017 ;88(1):45

Department of Obstetrics and Perinatology, Jagiellonian University Medical College, Krakow, Poland.

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http://dx.doi.org/10.5603/GP.a2017.0009DOI Listing
July 2018

Systemic application of AAV vectors targeting GFAP-expressing astrocytes in Z-Q175-KI Huntington's disease mice.

Mol Cell Neurosci 2016 12 27;77:76-86. Epub 2016 Oct 27.

Cluster of Excellence NeuroCure, University Medicine Charitè, Berlin, Germany; Department of Experimental Neurology, University Medicine Charité, Berlin, Germany.

Huntington's disease (HD) affects both neurons and astrocytes. To target the latter and to ensure brain-wide transgene expression, adeno-associated viral (AAV) vectors can be administered intravenously, as AAV vectors cross the blood-brain barrier (BBB) and enable preferential transduction of astrocytes due to their close association with blood vessels. However, there is a possibility that the subclass of GFAP-expressing astrocytes performs a distinct role in HD and reacts differently to therapeutic measures than the rest of the astrocytes. The gfaABC1D promoter allows specific targeting of the GFAP-expressing astrocytes (~25% of S100β-expressing astrocytes). We have examined the expression of three different transgenes (GCaMP6f, Kir4.1 and GLT1) and tested the effects of the AAV serotypes 9 and rh8. The AAV vectors were injected into the tail vein of 1-year-old homozygous Z-Q175-KI HD mice and their wild-type (WT) littermates. At this age, HD mice exhibit motor symptoms, including pronounced hypokinesia and circling behaviour. The expression times ranged from 3 to 6weeks. The target cell population was defined as the cells expressing S100β in addition to GFAP. Viewfields in the dorsal striatum and the overlaying cortex were evaluated and the transduction rate was defined as the percentage of target cells that expressed the reporter transgene (enhanced green fluorescent protein, EGFP, or Tomato). In all cases, the transduction rate was higher in the cortex than in the striatum. AAV9 was more efficient than AAVrh8. One of the injected constructs (AAV9-gfaABC1D-GLT1-Tomato) was tested for the first time. GLT1, the principal astrocytic glutamate transporter, is deficient in HD and therefore considered as a potential target for gene therapy. At a dose of 1.86×10 vector genome (vg) per animal, the fraction of GLT1-Tomato+ cells in the striatum and the cortex amounted to 30% and 49%, respectively. In individual Tomato+ HD astrocytes, treatment with the GLT1 vector increased the level of GLT1 immunofluorescence by 21% compared to the HD control. The described approach offers new and interesting opportunities to examine the pathophysiological consequences of brain-wide transgene expression in a specific astrocyte subpopulation.
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http://dx.doi.org/10.1016/j.mcn.2016.10.007DOI Listing
December 2016

Triclosan activates aryl hydrocarbon receptor (AhR)-dependent apoptosis and affects Cyp1a1 and Cyp1b1 expression in mouse neocortical neurons.

Environ Res 2016 Nov 27;151:106-114. Epub 2016 Jul 27.

Department of Animal Biotechnology, Animal Sciences Faculty, University of Agriculture, Redzina 1B, 30-248 Krakow, Poland. Electronic address:

Triclosan (TCS) is an antimicrobial agent that is used extensively in personal care and in sanitizing products, such as soaps, toothpastes, and hair products. A number of studies have revealed the presence of TCS in human tissues, such as fat, liver and brain, in addition to blood and breast milk. The aim of the present study was to investigate the impact of TCS on AhR and Cyp1a1/Cyp1b1 signaling in mouse neocortical neurons in primary cultures. In addition to the use of selective ligands and siRNAs, expression levels of mRNA and proteins as well as caspase-3 activity, reactive oxygen species (ROS) formation, and lactate dehydrogenase (LDH) release have been measured. We also studied the involvement of the AhR in TCS-induced LDH release and caspase-3 activation as well as the effect of TCS on ROS generation. Cultures of neocortical neurons were prepared from Swiss mouse embryos on day 15/16 of gestation. The cells were cultured in phenol red-free Neurobasal medium with B27 and glutamine, and the neurons were exposed to 1 and 10µM TCS. Our experiments showed that the expression of AhR and Cyp1a1 mRNA decreased in cells exposed to 10µM TCS for 3 or 6h. In the case of Cyp1b1, mRNA expression remained unchanged compared with the control group following 3h of exposure to TCS, but after 6h, the mRNA expression of Cyp1b1 was decreased. Our results confirmed that the AhR is involved in the TCS mechanism of action, and our data demonstrated that after the cells were transfected with AhR siRNA, the cytotoxic and pro-apoptotic properties of TCS were decreased. The decrease in Cyp1a1 mRNA and protein expression levels accompanied by a decrease in its activity. The stimulation of Cyp1a1 activity produced by the application of an AhR agonist (βNF) was attenuated by TCS, whereas the addition of AhR antagonist (αNF) reversed the inhibitory effects of TCS. In our experiments, TCS diminished Cyp1b1 mRNA and enhanced its protein expression. In case of Cyp1a1 we observed paradoxical effect of TCS action, which caused the decrease in activity and protein expression of Cyp1a1 and the increase in protein level of AhR. Therefore, we determined the effects of TCS on the production of ROS. Our results revealed that TCS increased the production of ROS and that this effect of TCS was reversed by 10µM N-acetyl-L-cysteine (NAC), the ROS scavenger. To confirm an involvement of ROS in TCS-induced neurotoxicity we measured AhR, Cyp1a1, and Cyp1b1 mRNA expression levels in cells co-treated with TCS and NAC. In the presence of NAC, TCS enhanced mRNA expression of the cytochromes and AhR at 3 and 6h, respectively. We postulate that TCS exhibits primary and secondary effects. The primary effects such as impairment of Cyp1a1 signaling are mediated by TCS-induced ROS production, whereas secondary effects of TCS are due to transcriptional activity of AhR and estrogenic properties of TCS.
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http://dx.doi.org/10.1016/j.envres.2016.07.019DOI Listing
November 2016

Dibutyl Phthalate (DBP)-Induced Apoptosis and Neurotoxicity are Mediated via the Aryl Hydrocarbon Receptor (AhR) but not by Estrogen Receptor Alpha (ERα), Estrogen Receptor Beta (ERβ), or Peroxisome Proliferator-Activated Receptor Gamma (PPARγ) in Mouse Cortical Neurons.

Neurotox Res 2017 01 31;31(1):77-89. Epub 2016 Aug 31.

Department of Experimental Neuroendocrinology, Institute of Pharmacology, Polish Academy of Sciences, Smetna 12, 31-343, Kraków, Poland.

Dibutyl phthalate (di-n-butyl phthalate, DBP) is one of the most commonly used phthalate esters. DBP is widely used as a plasticizer in a variety of household industries and consumer products. Because phthalates are not chemically bound to products, they can easily leak out to enter the environment. DBP can pass through the placental and blood-brain barriers due to its chemical structure, but little is known about its mechanism of action in neuronal cells. This study demonstrated the toxic and apoptotic effects of DBP in mouse neocortical neurons in primary cultures. DBP stimulated caspase-3 and LDH activities as well as ROS formation in a concentration (10 nM-100 µM) and time-dependent (3-48 h) manner. DBP induced ROS formation at nanomolar concentrations, while it activated caspase-3 and LDH activities at micromolar concentrations. The biochemical effects of DBP were accompanied by decreased cell viability and induction of apoptotic bodies. Exposure to DBP reduced Erα and Pparγ mRNA expression levels, which were inversely correlated with protein expression of the receptors. Treatment with DBP enhanced Ahr mRNA expression, which was reflected by the increased AhR protein level observed at 3 h after exposure. ERα, ERβ, and PPARγ antagonists stimulated DBP-induced caspase-3 and LDH activities. AhR silencing demonstrated that DBP-induced apoptosis and neurotoxicity are mediated by AhR, which is consistent with the results from DBP-induced enhancement of AhR mRNA and protein expression. Our study showed that AhR is involved in DBP-induced apoptosis and neurotoxicity, while the ERs and PPARγ signaling pathways are impaired by the phthalate.
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http://dx.doi.org/10.1007/s12640-016-9665-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5209414PMC
January 2017

Tetrabromobisphenol A (TBBPA)-stimulated reactive oxygen species (ROS) production in cell-free model using the 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) assay-limitations of method.

Environ Sci Pollut Res Int 2016 Jun 15;23(12):12246-52. Epub 2016 Mar 15.

Department of Public Health, Dietetics and Lifestyle Disorders, University of Information Technology and Management in Rzeszow, Sucharskiego 2, 35-225, Rzeszow, Poland.

Tetrabromobisphenol A (TBBPA) is a widely used brominated flame retardant, applied in a variety of commercial and household products, mainly electronic ones. Since the production of reactive oxygen species (ROS) is considered one of the principal cytotoxicity mechanisms, numerous studies undertake that aspect of TBBPA's mechanism of action. The present study verifies if the fluorogenic substrate 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) should be used to detect ROS production induced by TBBPA. To determine the ability of TBBPA alone to stimulate the conversion of H2DCFDA to its fluorescent product 2',7'-dichlorofluorescein (DCF), we used a cell-free model. In the experiments we check different cultured media also in combination with free radical scavenger N-acetyl-l-cysteine (NAC). Additionally, experiments with stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH·) have been made. The presented data showed that TBBPA in all tested concentrations interacts with H2DCFDA in phosphate-buffered saline (PBS) buffer while in micromolar concentrations in the DMEM/F12 medium with and without serum. The addition of NAC inhibited the interaction of TBBPA with H2DCFDA. Experiments with DPPH· showed that, in the presence of NAC, TBBPA acts like a free radical. TBBPA has similar properties to free radical and is susceptible to free radical scavenging properties of NAC. Our results indicated that H2DCFDA assay cannot be used to evaluate cellular ROS production in TBBPA studies. The study connected with TBBPA-stimulated ROS production in cell culture models using the H2DCFDA assay should be revised using a different method. However, due to the free radical-like nature of TBBPA, it can be very difficult. Therefore, further investigation of the nature of TBBPA as a compound with similar properties to free radical is required.
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http://dx.doi.org/10.1007/s11356-016-6450-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4893053PMC
June 2016

TBBPA causes neurotoxic and the apoptotic responses in cultured mouse hippocampal neurons in vitro.

Pharmacol Rep 2016 Feb 25;68(1):20-6. Epub 2015 Jun 25.

Department of Animal Biotechnology, Animal Sciences Faculty, University of Agriculture, Kraków, Poland. Electronic address:

Background: Tetrabromobisphenol A (TBBPA) is a brominated flame retardant widely used in a variety of commercial and household products. TBBPA can become bioaccumulated in human body fluids, and also in different brain regions. The aim of the present study was to determine the viability and apoptosis of cultured mouse hippocampal neurons in vitro after exposure to TBBPA. Additionally, we examined the involvement of ROS generation in the effect of TBBPA.

Methods: Primary hippocampal neuron cultures were prepared from Swiss mouse embryos on day 17/18 of gestation. The cultures were treated with TBBPA at concentrations ranging from 1nM to 100μM for 30min or 3, 6 or 24h. To study apoptosis, the activity of caspase-3 was measured, and apoptotic body formation was evaluated. To investigate the cytotoxic effect of TBBPA, the level of lactate dehydrogenase (LDH) was measured in the culture medium.

Results: Our results demonstrated that TBBPA concentrations ranging from 100nM to 100μM caused caspase-3 activation and apoptotic body formation. The cytotoxic effects of TBBPA were observed at concentrations ranging from 50nM to 100μM. To detect intracellular ROS, the fluorogenic dye H2DCFDA was used. We did not observe any significant increase in the level of cellular ROS in cultured cells after TBBPA treatment. However, in a cell-free model, TBBPA at concentrations ranging from 10 to 100μM interacted with H2DCFDA and enhanced the fluorescence signal.

Conclusion: We suggest that the H2DCFDA assay cannot be used to measure TBBPA-stimulated cell-mediated ROS production.
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http://dx.doi.org/10.1016/j.pharep.2015.06.005DOI Listing
February 2016

The role of PPARγ in TBBPA-mediated endocrine disrupting effects in human choriocarcinoma JEG-3 cells.

Mol Cell Biochem 2015 Nov 8;409(1-2):81-91. Epub 2015 Aug 8.

Department of Animal Biotechnology, University of Agriculture, Rędzina 1B, 30-248, Kraków, Poland.

The goal of the present study was to investigate the action of TBBPA on PPARγ protein expression in vitro in human choriocarcinoma-derived placental JEG-3 cells. We also analyzed TBBPA for its action on placental secretion of progesterone and β-hCG, cell viability, and apoptosis. Our results showed that after TBBPA treatment at 10 nM and 10 µM, PPARγ protein expression increased in a time-dependent manner until 48 h and then slightly decreased at 72 h but was still above the control level. This alteration in PPARγ protein expression was accompanied by a decreased β-hCG level. Interestingly, co-treatment with the PPARγ antagonist GW9662 reversed the TBBPA-mediated changes in PPARγ protein expression but, according to β-hCG secretion, potentiated an inhibitory effect of TBBPA. Additionally, in our study, we assessed the ability of TBBPA to increase progesterone levels in JEG-3 cells compared with those of controls. Finally, in the present study, we demonstrated that TBBPA at all of the tested doses significantly increased caspase-3 activity compared with that of the vehicle control. The apoptotic action of TBBPA was also confirmed by Hoechst 33342 staining. These results showed the up-regulation of PPARγ protein expression after TBBPA exposure in human placental cells. Although co-treatment with antagonist of PPARγ reversed the TBBPA-mediated increase in this protein expression and restored it to the control level, it did not reverse the effect on β-hCG secretion. This indicated that the mechanism of TBBPA-induced changes in β-hCG secretion is PPARγ-independent.
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http://dx.doi.org/10.1007/s11010-015-2514-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4589557PMC
November 2015

Modulation of estradiol synthesis and aromatase activity in human choriocarcinoma JEG-3 cells exposed to tetrabromobisphenol A.

Toxicol In Vitro 2015 Feb 16;29(1):44-50. Epub 2014 Sep 16.

Department of Animal Biotechnology, University of Agriculture, Rędzina 1B, 30-248 Cracow, Poland. Electronic address:

The goal of the present study was to investigate the impact of tetrabromobisphenol A (TBBPA) on human choriocarcinoma-derived placental JEG-3 cells in vitro. We determined the effect of this compound on estradiol secretion, aromatase protein expression and activity in vitro in the JEG-3 cell line. We assessed the ability of TBBPA to increase intracellular levels of cAMP as well as its effect on cell viability and proliferation. Our results indicated that TBBPA, at a wide range of concentrations (1×10(-8)-5×10(-5)M), significantly induced estradiol secretion by JEG-3 cells compared to that of controls after 24, 48 or 72 h of exposure. This effect was accompanied by an increase in the aromatase protein expression in JEG-3 cells treated with 100 nM and 10 μM of TBBPA for 24 h. Additionally, in our study, we confirmed that TBBPA-induced changes in aromatase protein expression were associated w ith the up-regulation of aromatase activity and cAMP levels. No tested doses of TBBPA inhibited JEG-3 cell proliferation, except for the highest dose of 100 μM, which had a toxic effect on cell viability at all time points. The present study clearly indicates that TBBPA alters JEG-3 cells estrogen synthesis due to its action on CYP19 protein expression and thus this compound may interfere with normal placental development during early pregnancy.
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http://dx.doi.org/10.1016/j.tiv.2014.09.003DOI Listing
February 2015

Impact of endocrine-disrupting chemicals on neural development and the onset of neurological disorders.

Pharmacol Rep 2013 ;65(6):1632-9

Department of Experimental Neuroendocrinology, Institute of Pharmacology, Polish Academy of Sciences, Smętna 12, PL 31-343 Kraków, Poland.

Even though high doses of organic pollutants are toxic, relatively low concentrations have been reported to cause long-term alterations in functioning of individual organisms, populations and even next generations. Among these pollutants are dioxins, polychlorinated biphenyls, pesticides, brominated flame retardants, plasticizers (bisphenol A, nonylphenol, and phthalates) as well as personal care products and drugs. In addition to toxic effects, they are able to interfere with hormone receptors, hormone synthesis or hormone conversion. Because these chemicals alter hormone-dependent processes and disrupt functioning of the endocrine glands, they have been classified as endocrine-disrupting chemicals (EDCs). Because certain EDCs are able to alter neural transmission and the formation of neural networks, the term neural-disrupting chemicals has been introduced, thus implicating EDCs in the etiology of neurological disorders. Recently, public concern has been focused on the effects of EDCs on brain function, concomitantly with an increase in neuropsychiatric disorders, including autism, attention deficit and hyperactivity disorder as well as learning disabilities and aggressiveness. Several lines of evidence suggest that exposure to EDCs is associated with depression and could result in neural degeneration. EDCs act via several classes of receptors with the best documented mechanisms being reported for nuclear steroid and xenobiotic receptors. Low doses of EDCs have been postulated to cause incomplete methylation of specific gene regions in the young brain and to impair neural development and brain functions across generations. Efforts are needed to develop systematic epidemiological studies and to investigate the mechanisms of action of EDCs in order to fully understand their effects on wildlife and humans.
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http://dx.doi.org/10.1016/s1734-1140(13)71524-xDOI Listing
October 2014

Reduced tonic inhibition in striatal output neurons from Huntington mice due to loss of astrocytic GABA release through GAT-3.

Front Neural Circuits 2013 26;7:188. Epub 2013 Nov 26.

Cluster of Excellence NeuroCure, University Medicine Charité Berlin, Germany ; Department of Experimental Neurology, University Medicine Charité Berlin, Germany.

The extracellular concentration of the two main neurotransmitters glutamate and GABA is low but not negligible which enables a number of tonic actions. The effects of ambient GABA vary in a region-, cell-type, and age-dependent manner and can serve as indicators of disease-related alterations. Here we explored the tonic inhibitory actions of GABA in Huntington's disease (HD). HD is a devastating neurodegenerative disorder caused by a mutation in the huntingtin gene. Whole cell patch clamp recordings from striatal output neurons (SONs) in slices from adult wild type mice and two mouse models of HD (Z_Q175_KI homozygotes or R6/2 heterozygotes) revealed an HD-related reduction of the GABA(A) receptor-mediated tonic chloride current (I(Tonic(GABA))) along with signs of reduced GABA(B) receptor-mediated presynaptic depression of synaptic GABA release. About half of I(Tonic(GABA)) depended on tetrodotoxin-sensitive synaptic GABA release, but the remaining current was still lower in HD. Both in WT and HD, I(Tonic(GABA)) was more prominent during the first 4 h after preparing the slices, when astrocytes but not neurons exhibited a transient depolarization. All further tests were performed within 1-4 h in vitro. Experiments with SNAP5114, a blocker of the astrocytic GABA transporter GAT-3, suggest that in WT but not HD GAT-3 operated in the releasing mode. Application of a transportable substrate for glutamate transporters (D-aspartate 0.1-1 mM) restored the non-synaptic GABA release in slices from HD mice. I(Tonic(GABA)) was also rescued by applying the hyperagonist gaboxadol (0.33 μM). The results lead to the hypothesis that lesion-induced astrocyte depolarization facilitates non-synaptic release of GABA through GAT-3. However, the capacity of depolarized astrocytes to provide GABA for tonic inhibition is strongly reduced in HD.
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http://dx.doi.org/10.3389/fncir.2013.00188DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3840359PMC
April 2014

PPAR-γ agonist GW1929 but not antagonist GW9662 reduces TBBPA-induced neurotoxicity in primary neocortical cells.

Neurotox Res 2014 Apr 17;25(3):311-22. Epub 2013 Oct 17.

Laboratory of Genomics and Biotechnology, Animal Sciences Faculty, University of Agriculture, Redzina 1B, 30-248, Krakow, Poland,

Tetrabromobisphenol A (2,2-bis(4-hydroxy-3,5-dibromophenyl)propane; TBBPA) is a widely used brominated flame retardant. TBBPA induces neuronal damage, but the mechanism by which this occurs is largely unknown. We studied the possible involvement of peroxisome proliferator-activated receptor gamma (PPAR-γ) in TBBPA-induced apoptosis and toxicity in mouse primary neuronal cell cultures. TBBPA enhanced both, caspase-3 activity and lactate dehydrogenase (LDH) release in neocortical cells after 6 and 24 h of exposition. These data were supported at the cellular level with Hoechst 33342 staining. Immunoblot analyses showed that, compared with control cells, 10 μM TBBPA decreased the expression of PPAR-γ protein in neocortical neurons after 1-24 h of exposure. Co-treatment with TBBPA and GW1929 inhibited the TBBPA-induced caspase-3 activity, apoptotic body formation, and LDH release as well as TBBPA-induced decrease in PPAR-γ protein expression. Thus, our data support neuroprotective potential of PPAR-γ agonists. The PPAR-γ antagonist GW9662 prevented the TBBPA-induced decrease in PPAR-γ protein level, but it potentiated TBBPA-induced apoptotic and neurotoxic effects, which suggest that the mechanism of TBBPA action in neuronal cells is not only PPAR-γ-dependent. Therefore, further studies of the mechanism of TBBPA action in the nervous system are needed.
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http://dx.doi.org/10.1007/s12640-013-9434-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3936120PMC
April 2014

[Components of plastic disrupt the function of the nervous system].

Postepy Hig Med Dosw (Online) 2013 May 27;67:499-506. Epub 2013 May 27.

Pracownia Biotechnologii i Genomiki, Katedra Trzody Chlewnej i Małych Przeżuwaczy, Wydziału Hodowli i Biologii Zwierząt, Uniwersytet Rolniczy w Krakowie, Krakow.

Development of the chemical industry leads to the development of new chemical compounds, which naturally do not exist in the environment. These chemicals are used to reduce flammability, increase plasticity, or improve solubility of other substances. Many of these compounds, which are components of plastic, the new generation of cosmetics, medical devices, food packaging and other everyday products, are easily released into the environment. Many studies have shown that a major lipophilicity characterizes substances such as phthalates, BPA, TBBPA and PCBs. This feature allows them to easily penetrate into living cells, accumulate in the tissues and the organs, and affect human and animal health. Due to the chemical structures, these compounds are able to mimic some endogenous hormones such as estradiol and to disrupt the hormone homeostasis. They can also easily pass the placental barrier and the blood-brain barrier. As numerous studies have shown, these chemicals disturb the proper functions of the nervous system from the earliest moments of life. It has been proven that these compounds affect neurogenesis as well as the synaptic transmission process. As a consequence, they interfere with the formation of the sex of the brain, as well as with the learning processes, memory and behavior. Additionally, the cytotoxic and pro-apoptotic effect may cause neurodegenerative diseases. This article presents the current state of knowledge about the effects of phthalates, BPA, TBBPA, and PCBs on the nervous system.
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http://dx.doi.org/10.5604/17322693.1051001DOI Listing
May 2013

[Congenital cytomegaly in one twin - a case report].

Med Wieku Rozwoj 2012 Jul-Sep;16(3):252-60

Klinika Chorób Dzieci Katedry Pediatrii, Uniwersytet Jagielloński, Collegium Medium w Krakowie, ul. Wielicka 265, 30-663 Kraków.

A report on dichorionic/diamniotic pregnancy in which only one, female, fetus was infected with cytomegalovirus and presented with severe congenital diseases at birth. Infection of the fetus occurred after recurrent maternal infection. The second, male, fetus did not have CMV infection. The cesarean section was performed at the 38th week of gestation. The birth weight of the infected girl was 1680g, the main symptoms, beside dystrophy, concerned the central nervous system: microcephaly, brain atrophy, hydrocephalus, corpus callosum agenesis. She also had Turner syndrome symptoms. The viral load was highest in the urine 81.2 x10^6/ml, in the cerebro-spinal fluid 15.4x10^6/ml and lower in blood 0.38 x10^5/ml. The concentration of specific IgG was 308 U/ml. Specific IgM was not detected. Throughout hospitalization, the infection maintained only one viral genotype gB2. Despite treatment with ganciclovir (10 weeks) and foscarnet (2 weeks), the girl died at the age of 8 months. Novel molecular diagnostic techniques (nested and real time PCR) confirmed the congenital infection and were helpful in the monitoring of the infection and treatment efficacy.
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April 2013

Energy demand of synaptic transmission at the hippocampal Schaffer-collateral synapse.

J Cereb Blood Flow Metab 2012 Nov 29;32(11):2076-83. Epub 2012 Aug 29.

Institute for Neurophysiology, Charité-Universitätsmedizin, Berlin, Germany.

Neuroenergetic models of synaptic transmission predicted that energy demand is highest for action potentials (APs) and postsynaptic ion fluxes, whereas the presynaptic contribution is rather small. Here, we addressed the question of energy consumption at Schaffer-collateral synapses. We monitored stimulus-induced changes in extracellular potassium, sodium, and calcium concentration while recording partial oxygen pressure (pO(2)) and NAD(P)H fluorescence. Blockade of postsynaptic receptors reduced ion fluxes as well as pO(2) and NAD(P)H transients by ∼50%. Additional blockade of transmitter release further reduced Na(+), K(+), and pO(2) transients by ∼30% without altering presynaptic APs, indicating considerable contribution of Ca(2+)-removal, transmitter and vesicle turnover to energy consumption.
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http://dx.doi.org/10.1038/jcbfm.2012.116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3493999PMC
November 2012

The effect of triclosan on hormone secretion and viability of human choriocarcinoma JEG-3 cells.

Reprod Toxicol 2012 Nov 4;34(3):385-92. Epub 2012 Jun 4.

Laboratory of Genomics and Biotechnology, University of Agriculture, Redzina 1B, 30-248 Krakow, Poland.

Triclosan is an antimicrobial agent frequently used in pharmaceuticals and personal care products. We analyzed triclosan for its action on placental secretion of progesterone, estradiol and human chorionic gonadotropin in vitro in the JEG-3 cells. We also investigated its action on cell viability, proliferation and apoptosis. The JEG-3 cells were cultured with increasing doses of triclosan (1×10(-9)-1×10(-4) M) for 24, 48 and 72 h. Triclosan was found to increase estradiol and progesterone secretion after short- and long-term exposure. The stimulatory effect was observed up to 10 μM after short- and long-term exposure to triclosan. In addition, triclosan caused an adverse effect on β-hCG secretion. The highest doses of triclosan (50 and 100 μM) showed a strong cytotoxic effect. Anti proliferative and pro-apoptotic effects were also observed. Overall, this study demonstrates that triclosan may indirectly disrupt steroidogenesis which may, in turn, affect placental development and consequently fetal growth.
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http://dx.doi.org/10.1016/j.reprotox.2012.05.094DOI Listing
November 2012