Natalia Pipova

Scopus Publications

Lateralization at the individual and population levels of European green lizard in Slovak Karst
Mário Pikalík, Natália Pipová, Viktória Majláthová, Vincent A. Connors, and Igor Majláth
Springer Science and Business Media LLC

Prevalence of relapsing fever spirochete Borrelia miyamotoi in Ixodes ricinus ticks from eastern Slovakia
Dominika Bubanová, Igor Majláth, Blažena Vargová, Natália Pipová, Sándor Szekeres, and Viktória Majláthová
Wiley
Borrelia miyamotoi, recurrent fever borrelia, pathogenic to humans, has been found recently in Ixodes ricinus ticks. There is still little information about its circulation in nature and potential local impact on human health as well as the occurrence in natural localities. In our study, a total of 1609 ticks (745 nymphs, 358 females and 506 males) collected from different localities in eastern Slovakia were analysed using a polymerase chain reaction targeting a gene encoding glycerophosphodiester phosphodiesterase (glpQ), which is specific to the Borrelia species in the relapsing fever group. B. miyamotoi was found in 3 nymphs, 6 females and 9 males of quested I. ricinus ticks. Although no human case of infection with B. miyamotoi has been reported in Slovakia yet, its presence needs to be studied to obtain knowledge regarding the prevalence and the distribution of this human pathogen in questing ticks and to increase medical healthcare awareness.

SEX-DEPENDENT DIFFERENCES IN STRESS-INDUCED DEPRESSION IN WISTAR RATS ARE ACCOMPANIED PREDOMINANTLY BY CHANGES IN PHOSPHATIDYLCHOLINES AND SPHINGOMYELINS
E. Leskanicova, M. Babinčák, F. Mochnacký, N. Pipová Kokošová, D. Kukelova, N. Urbańska, M. Kolesárová, D. Macekova, J. Kostolný and T. Kisková

With a high annual and lifetime prevalence, depression is becoming the leading contributor to the global disease burden. During the COVID-19 crisis, the depression and mood disorders accelerated significantly. Despite the growing evidence, the precise underlying mechanisms of depression disorders (DD) remain unknown. When studying DD in humans, there are many uncontrollable factors such as medication history, age of the patient or living conditions. In this regard, animal models provide an essential step for examining neural circuitry or molecular and cellular pathways in a controlled environment. As far as we know, women have a consistently higher prevalence of DD than men. Thus, the aim of our study was to evaluate sex-related changes in blood metabolites in a model of stress-induced depression in Wistar rats. Pregnant females were stressed using restriction of mobility in the final week of the pregnancy three times a day for 45 minutes each, three following days. After the birth, the progeny aged 60 days was stressed repeatedly. The perturbation in overall energy metabolism as well as in lipid metabolism was found. While in males, phosphatidylcholines (the most phosphatidylcholine with acyl-alkyl residue sum C40:4 - PC ae C40:4), sphingomyelins, and acylcarnitines were changed, in females, lipid metabolism perturbation was seen with the most critical alteration in hydroxysphingomyelin with acyl residue sum C16:1 (SM OH C16:1). Our results confirm that the animal model may be used further in the research of depression. Our results may provide an essential insight into the sex-dependent pathogenesis of depression and contribute to the search for effective treatment and prevention of depression with respect to sex.

Morphometric analysis – effect of the radiofrequency interface of electromagnetic field on the size of hatched dermacentor reticulatus larvae
Blažena Vargová, Igor Majlath, Juraj Kurimský, Roman Cimbala, Natalia Pipova, Jozef Živčák, Piotr Tryjanowski, Branislav Peťko, Jaroslav Džmura, Gabriela Ižariková,et al.
Institute of Rural Health
INTRODUCTION AND OBJECTIVE Electromagnetic radiation interactions with living systems have been one of determining factors in biological evolution. This study investigates the effect of 900 MHz radiofrequency (RF) electromagnetics field (EMF) exposure of eggs on the development of Dermacentor reticulatus larvae. The basic objective was to determine whether the 900 MHz RF-EMF has the potential to influence the size of the body of the hatched larvae of D. reticulatus ticks. MATERIAL AND METHODS To this aim, eggs from 3 fully engorged females of D. reticulatus were included in the test procedure. Altogether four groups of eggs were designated which included eggs from each female. We used RF-EMF frequency of 900 MHz. Eggs were exposed to EMF for different time periods (30, 60 and 90 minutes) in dark, electromagnetically shielded anechoic chamber. After the irradiation eggs were allowed to hatch in climatic chamber. Randomly selected 200 larval individuals were measured to get basic morphological records. Four body traits including the total body length (TBL), length of gnathosoma with scutum (GSL), the total body width (TBW), and the width of basis capituli (BCW) were measured. RESULTS The D. reticulatus larvae hatched from eggs exposed for 60 minutes, had demonstrably larger dimensions of all measured body traits not only as a control unexposed group but also as other experimental groups. CONCLUSIONS The study shows, particularly, that artificial EMF that is used in smartphone technology impacts seriously D. reticulatus larvae development.

Enlisting the ixodes scapularis embryonic ISE6 cell line to investigate the neuronal basis of tick—pathogen interactions
Lourdes Mateos-Hernández, Natália Pipová, Eléonore Allain, Céline Henry, Clotilde Rouxel, Anne-Claire Lagrée, Nadia Haddad, Henri-Jean Boulouis, James J. Valdés, Pilar Alberdi,et al.
MDPI AG
Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like Ixodes scapularis embryonic cell line, ISE6, is an effective tool frequently used for examining tick–pathogen interactions. We detected 37 neuropeptide transcripts in the I. scapularis ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, Anaplasma phagocytophilum. The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in A. phagocytophilum-infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of I. scapularis synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete Ixodes neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick–pathogen interactions.

Melatonin mitigates hippocampal and cognitive impairments caused by prenatal irradiation
Natália Pipová Kokošová, Terézia Kisková, Katarína Vilhanová, Andrea Štafuriková, Rastislav Jendželovský, Enikő Račeková, and Beňadik Šmajda
Wiley
Formation of new neurons and glial cells in the brain is taking place in mammals not only during prenatal embryogenesis but also during adult life. As an enhancer of oxidative stress, ionizing radiation represents a potent inhibitor of neurogenesis and gliogenesis in the brain. It is known that the pineal hormone melatonin is a potent free radical scavenger and counteracts inflammation and apoptosis in brain injuries. The aim of our study was to establish the effects of melatonin on cells in the hippocampus and selected forms of behaviour in prenatally irradiated rats. The male progeny of irradiated (1 Gy of gamma rays; n = 38) and sham‐irradiated mothers (n = 19), aged 3 weeks or 2 months, were used in the experiment. Melatonin was administered daily in drinking water (4 mg/kg b. w.) to a subset of animals from each age group. Prenatal irradiation markedly suppressed proliferative activity in the dentate gyrus in both age groups. Melatonin significantly increased the number of proliferative BrdU‐positive cells in hilus of young irradiated animals, and the number of mature NeuN‐positive neurons in hilus and granular cell layer of the dentate gyrus in these rats and in CA1 region of adult irradiated rats. Moreover, melatonin significantly improved the spatial memory impaired by irradiation, assessed in Morris water maze. A significant correlation between the number of proliferative cells and cognitive performances was found, too. Our study indicates that melatonin may decrease the loss of hippocampal neurons in the CA1 region and improve cognitive abilities after irradiation.

Ticks and radio-frequency signals: Behavioural response of ticks (Dermacentor reticulatus) in a 900 MHz electromagnetic field
Blažena Vargová, Juraj Kurimský, Roman Cimbala, Michal Kosterec, Igor Majláth, Natália Pipová, Piotr Tryjanowski, Łukasz Jankowiak, and Viktoria Majlathova
Systematic and Applied Acarology Society
The electromagnetic field (EMF) is present in the environment throughout the world and encompasses both natural and human-made sources of electromagnetic fields. It has been shown that EMF influences a variety of biological systems, including the behavioural responses of both vertebrates and invertebrates. As such, determining the effects of the EMF on the ecosystem in detail may be important for understanding the ecology and biology of organisms, particularly those, such as ticks, that are important in disease transmission. Our main goal was to determine if the tick, Dermacentor reticulatus, interacts with the EMF. An experimental behavioural test of tick sensitivity to radio frequency power radiation was performed under laboratory conditions. Tests were performed in an electromagnetic compatibility laboratory in a radio frequency electromagnetic field (RF-EMF) anechoic chamber. Ticks were irradiated using a Double-Ridged Waveguide Horn Antenna with 900 MHz RF-EMF. The applied radio-frequency power was tuned below the proposed limit for public exposure to mobile phone base stations. We found that exposure induces an immediate tick locomotor response manifested either in a previously unreported jerking movement of the whole body or in jerking of the first pair of legs. Overall, ticks exhibited significantly greater movement in the presence of the RF-EMF. Significant sex differences relative to RF-EMF exposure were observed in both response variables. In the presence of RF-EMF, body jerking by females was greater than in males and vice versa for leg jerks. This study represents the first experimental evidence of a behavioural response of D. reticulatus ticks to exposure to RF-EMF.

Stress exposure during the preimplantation period affects blastocyst lineages and offspring development
Ján BURKUŠ, Martina KAČMAROVÁ, Janka KUBANDOVÁ, Natália KOKOŠOVÁ, Kamila FABIANOVÁ, Dušan FABIAN, Juraj KOPPEL, and Štefan ČIKOŠ
Japanese Society of Animal Reproduction
We found retardation of preimplantation embryo growth after exposure to maternal restraint stress during the preimplantation period in our previous study. In the present study, we evaluated the impact of preimplantation maternal restraint stress on the distribution of inner cell mass (ICM) and trophectoderm (TE) cells in mouse blastocysts, and its possible effect on physiological development of offspring. We exposed spontaneously ovulating female mice to restraint stress for 30 min three times a day during the preimplantation period, and this treatment caused a significant increase in blood serum corticosterone concentration. Microscopic evaluation of embryos showed that restraint stress significantly decreased cell counts per blastocyst. Comparing the effect of restraint stress on the two blastocyst cell lineages, we found that the reduction in TE cells was more substantial than the reduction in ICM cells, which resulted in an increased ICM/TE ratio in blastocysts isolated from stressed dams compared with controls. Restraint stress reduced the number of implantation sites in uteri, significantly delayed eye opening in delivered mice, and altered their behavior in terms of two parameters (scratching on the base of an open field test apparatus, time spent in central zone) as well. Moreover, prenatally stressed offspring had significantly lower body weights and in 5-week old females delivered from stressed dams, fat deposits were significantly lower. Our results indicate that exposure to stress during very early pregnancy can have a negative impact on embryonic development with consequences reaching into postnatal life.

Neuronal Analysis and Behaviour in Prenatally Gamma-Irradiated Rats
Natália Kokošová, Lenka Tomášová, Terézia Kisková, and Beňadik Šmajda
Springer Science and Business Media LLC
The intrauterinal development in mammals represents a very sensitive period of life in relation to many environmental factors, including ionizing radiation (IR). The developing nervous system is particularly vulnerable to IR, and the consequences of exposure are of importance because of its potential health risks. The aim of our work was to assess whether prenatal irradiation of rats on the 17th day of embryonic development with a dose of 1 Gy would affect the formation of new cells and the number of mature neurons in the hippocampus and the selected forms of behaviour in the postnatal period. Male progeny of irradiated and control females was tested at ages of 3 weeks, 2 and 3 months. The number of mitotically active cells in the gyrus dentatus (GD) of the hippocampus was significantly reduced in irradiated rats aged 3 weeks. In irradiated rats aged 2 months, a significant reduction of mature neurons in CA1 area and in GD of the hippocampus was observed. The IR negatively influenced the spatial memory in Morris water maze, significantly decreased the exploratory behaviour and increased the anxiety-like behaviour in elevated plus-maze in rats aged 2 months. No significant differences were observed in animals aged 3 months compared with controls of the same age. A significant correlation between the number of mature neurons in the hilus and of the cognitive performances was found. Our results show that a low dose of radiation applied during the sensitive phase of brain development can influence the level of neurogenesis in the subgranular zone of GD and cause an impairment of the postnatal development of mental functions.

Resveratrol enhances the chemopreventive effect of celecoxib in chemically induced breast cancer in rats
Terézia Kisková, Rastislav Jendželovský, Erdenetsetsek Rentsen, Alexandra Maier-Salamon, Natália Kokošová, Zuzana Papčová, Jaromír Mikeš, Peter Orendáš, Bianka Bojková, Peter Kubatka,et al.
Ovid Technologies (Wolters Kluwer Health)
Resveratrol and celecoxib were used as chemopreventive agents in animal models of carcinogenesis, and exert antiproliferative and proapoptotic effects on cancer cells. Therefore, the aim of this study was to evaluate whether combining resveratrol with celecoxib may exert more potent anticarcinogenic effects than the single agents. Mammary carcinogenesis was initiated in 70 female Sprague–Dawley rats with N-methyl-N-nitrosourea (NMU). The chemoprevention with resveratrol, celecoxib, and their combination started 2 weeks before the first carcinogen dose and lasted until the end of the experiment. Tumor incidence and frequency, latency period, tumor volume, the expression of cyclooxygenase 2 (COX2) and growth differentiation factor 15 (GDF15), and also the formation of reactive oxygen species were analyzed using different methods. In addition, the levels of resveratrol and its metabolites in blood and selected tumor tissues were determined by high-performance liquid chromatography. Finally, the anticancer effects of the reagents were studied in the human breast cancer cell line MCF-7. Celecoxib as a single agent significantly decreased tumor frequency, prolonged tumor latency, and decreased the total number of malignant tumors compared with the NMU conditions. Tumor volume was nonsignificantly reduced (0.68±0.25 vs. 0.93±0.28 cm3). Importantly, the addition of resveratrol to celecoxib reduced tumor volume by 60% compared with celecoxib alone (from 0.68±0.25 to 0.27±0.07 cm3, P<0.05). Furthermore, the combination of resveratrol and celecoxib reduced tumor frequency by 29% compared with celecoxib alone (P=0.53). Tumor latency was not influenced by this combination compared with celecoxib alone (126.56±3.45 vs. 120.71±4.08 days). In addition, COX2 mRNA and immunoreactive protein stained on tumor sections were reduced and GDF15 protein increased significantly by the combination studied compared with the NMU conditions. In agreement with these data, a significant reduction in reactive oxygen species in blood lymphocytes of the combination was detected, which may have contributed toward the cancer-preventive effects of this application. This study showed that in NMU-induced mammary cancer in rats, the combination of resveratrol and celecoxib led to a significant reduction in all tumor parameters. In addition, in terms of tumor volume, the combination was more efficient than celecoxib as a single agent.

Ionizing radiation as preconditioning against transient cerebral ischemia in rats
Natália Kokošová, Viera Danielisová, Beňadik Šmajda, and Jozef Burda
AEPress, s.r.o.
Induction of ischemic tolerance (IT), the ability of an organism to survive an otherwise lethal ischemia, is the most effective known approach to preventing postischemic damage. IT can be induced by exposing animals to a broad range of stimuli. In this study we tried to induce IT of brain neurons using ionizing radiation (IR). A preconditioning (pre-C) dose of 10, 20, 30 or 50 Gy of gamma rays was used 2 days before an 8 min ischemia in adult male rats. Ischemia alone caused the degeneration of almost one half of neurons in CA1 region of hippocampus. However, a significant decrease of the number of degenerating neurons was observed after higher doses of radiation (30 and 50 Gy). Moreover, ischemia significantly impaired the spatial memory of rats as tested in Morris's water maze. In rats with a 50 Gy pre-C dose, the latency times were reduced to values close to the control level. Our study is the first to reveal that IR applied in sufficient doses can induce IT and thus allow pyramidal CA1 neurons to survive ischemia. In addition, we show that the beneficial effect of IR pre-C is proportional to the radiation dose.

Ionizing radiation as preconditioning against transient cerebral ischemia in rats

Morphological and molecular characterization of Karyolysus - A neglected but common parasite infecting some European lizards
Božena Haklová-Kočíková, Adriana Hižňanová, Igor Majláth, Karol Račka, David James Harris, Gábor Földvári, Piotr Tryjanowski, Natália Kokošová, Beáta Malčeková, and Viktória Majláthová
Springer Science and Business Media LLC
BackgroundBlood parasites of the genus Karyolysus Labbé, 1894 (Apicomplexa: Adeleida: Karyolysidae) represent the protozoan haemogregarines found in various genera of lizards, including Lacerta, Podarcis, Darevskia (Lacertidae) and Mabouia (Scincidae). The vectors of parasites are gamasid mites from the genus Ophionyssus.MethodsA total of 557 individuals of lacertid lizards were captured in four different localities in Europe (Hungary, Poland, Romania and Slovakia) and blood was collected. Samples were examined using both microscopic and molecular methods, and phylogenetic relationships of all isolates of Karyolysus sp. were assessed for the first time. Karyolysus sp. 18S rRNA isolates were evaluated using Bayesian and Maximum Likelihood analyses.ResultsA total of 520 blood smears were examined microscopically and unicellular protozoan parasites were found in 116 samples (22.3% prevalence). The presence of two Karyolysus species, K. latus and K. lacazei was identified. In total, of 210 samples tested by polymerase chain reaction (PCR), the presence of parasites was observed in 64 individuals (prevalence 30.5%). Results of phylogenetic analyses revealed the existence of four haplotypes, all part of the same lineage, with other parasites identified as belonging to the genus Hepatozoon.ConclusionsClassification of these parasites using current taxonomy is complex - they were identified in both mites and ticks that typically are considered to host Karyolysus and Hepatozoon respectively. Furthermore although distortions to the intermediate host erythrocyte nuclei were observed, the defining characteristic of Karyolysus, the haplotypes were nearly identical to those reported from lizards in the Iberian Peninsula, where such distortions were not reported and which were thus identified as Hepatozoon. Based on the phylogenetic analyses, neither vertebrate host, nor geographical patterns of the studied blood parasites could be established.

Natalia Pipova

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