Significant scientific results 2022

Trace amines: new hope in the treatment of brain disorders

Daniil Grinchii, Talah Khoury, Roman Dekhtiarenko, Reyhaneh Nejati Bervanlou, Eliyahu Dremencov

Monoamines are significantly involved in central nervous system (CNS) diseases such as depression and schizophrenia. The monoamine group includes the ‘classical’ monoamines (e.g. serotonin and dopamine) and the recently discovered ‘trace’ amines. Recent studies have shown that trace amine agonists have antidepressant and antipsychotic effects. In our study, we investigated the effect of the trace aminergic agonist RO5256390 (Hofmann le Roche LTD, Switzerland) on the activity of monoamine-releasing neurons. We found in serotonergic and dopaminergic neurons RO5256390 altered the way action potentials, called “spikes”, were generated, with a greater number of them occurring within clusters (“bursts”). It is known that “burst” activity of neurons is more efficient for neurotransmitter release. Trace aminergic agonists could therefore be a new generation of drugs that would allow “reprogramming” the activity of neurons releasing “classical” monoamines away from the “sick” to the “healthy” mode.

Trace amines modulate the excitability of serotonin (5-HT; A) and dopamine (B) neurons. Oct,octopamine; PEA, phenylethylamine; RO-90, RO5256390.

GRINCHII, Daniil – HOENER, Marius C. – KHOURY, TalahDEKHTIARENKO, RomanBERVANLOU, Reyhaneh Nejati – JEŽOVÁ, Daniela – DREMENCOV, Eliyahu**. Effects of acute and chronic administration of trace amine-associated receptor 1 (TAAR1) ligands on in vivo excitability of central monoamine-secreting neurons in rats. In Molecular Psychiatry, 2022, vol. 27, no. 12, p. 4861-4868. (2021: 13.437 – IF, Q1 – JCR, 4.390 – SJR, Q1 – SJR). ISSN 1359-4184.

Expression of integrin αV and its potential partners in reproductive tissues, germ cells and spermatozoa of the bull

Jana Antalíková, Petra Sečová, Katarína Michalková, Ľubica Horovská, Jana Jankovičová

Integrins are the most common partner of tetraspanins within the membrane multi-molecular network, essential for adhesion, intercellular interactions, and bidirectional signaling across the plasma membrane. At the same time, they are, together with them, part of the extracellular vesicles produced by different cell types involved in intercellular transport and communication. By a comprehensive analysis of the bull reproductive system, we confirmed integrin αV in the plasma membrane of bull spermatozoa already at the stage of round spermatids developing in the testis, during maturation in the epididymis, and after ejaculation until acrosomal exocytosis. The localization of αV in the sperm head region corresponded with tetraspanins CD9 and CD81. Based on the similar reaction pattern of αV, CD9 and CD81 detected in testicular tissue, we hypothesized the presence of these molecules also in extracellular vesicles produced in the testis, which we termed “testisomes”. These vesicles, involving integrins and tetraspanins, may be part of intercellular communication during sperm development and maturation.

ANTALÍKOVÁ, JanaSEČOVÁ, PetraMICHALKOVÁ, KatarínaHOROVSKÁ, Ľubica – PÁLENÍKOVÁ, Veronika – JANKOVIČOVÁ, Jana**. Expression of αV integrin and its potential partners in bull reproductive tissues, germ cells and spermatozoa. In International Journal of Biological Macromolecules, 2022, vol. 209, part A, p. 542-551. (2021: 8.025 – IF, Q1 – JCR, 1.100 – SJR, Q1 – SJR).

Glutamate may act as a signaling molecule in preimplantation embryos

Alexandra Špirková, Veronika Kovaříková, Zuzana Šefčíková, Jozef Pisko, Martina Kšiňanová, Juraj Koppel, Dušan Fabian, Štefan Čikoš

Free amino acids are found in the natural environment of the preimplantation embryo (glutamic acid/glutamate being among the most abundant), and it is generally assumed that the action of extracellular amino acids on the preimplantation embryo is conditioned by their transport into the embryonic cells. In our work, we have demonstrated the expression of several subunits/types of glutamate receptors in mouse blastocysts, showing the possibility of extracellular action of glutamate. Extracellular concentrations of glutamic acid starting at 5 mM disrupted the development of mouse blastocysts, and this fact may be of great practical significance since glutamic acid and its salts are widely used as food additives. Experiments with specific glutamate receptor ligands (combined with gene expression analysis) revealed that specific AMPA and kainate receptors and the metabotropic glutamate receptor GRM5 are involved in the above effects. Thus, glutamic acid may act as a signalling molecule in preimplantation embryos, exerting its effects by activating specific cellular receptors.

ŠPIRKOVÁ, Alexandra*KOVAŘÍKOVÁ, Veronika*ŠEFČÍKOVÁ, ZuzanaPISKO, JozefKŠIŇANOVÁ, MartinaKOPPEL, JurajFABIAN, DušanČIKOŠ, Štefan**. Glutamate can act as a signaling molecule in mouse preimplantation embryos. In Biology of Reproduction, 2022, vol. 107, no. 4, p. 916-927. (2021: 4.161 – IF, Q2 – JCR, 1.082 – SJR, Q1 – SJR, karentované – CCC). (2022 – Current Contents). ISSN 0006-3363.

Beneficial potential of the bacteriocin-producing strain Lactiplantibacillus plantarum LP17L/1 isolated from barreled sheep lump cheese

Andrea Lauková, Renáta Szabóová, Eva Bino, Jana Ščerbová, Monika Pogány Simonová

Sheep breeding has a long tradition in Slovakia. On a European scale, a number of traditional sheep’s milk products have been designated as Protected Geographical Products (PGI). Sheep lump cheese has the designation Traditional Specialty Guaranteed (TSG), while barreled sheep’s cheese is the raw material for the production of another traditional product – “bryndza”. These products are a source of beneficial indigenous microbiota. The strain Lactiplantibacillus plantarum LP17L/1, isolated from lump sheep’s cheese, produces the bacteriocin plantaricin with antilisterial and antistaphylococcal activity and is a safe strain from the point of view of the consumer. Staphylococci and listeria are unwanted contaminating microbiota in dairy products, which can lead to product spoilage but also endanger the health of the consumer, and plantaricin reduces this possibility. LP17L/1 tolerates a more acidic environment; it produces beta-galactosidase (a lactose cleaving enzyme), making it useful in the production of products for lactose intolerant people and has also shown anti-parasitic effects. It is part of a pending patent application (PP 50021-2022).

LAUKOVÁ, Andrea** – TOMÁŠKA, Martin – FRAQUEZA, Maria Joao – SZABÓOVÁ, RenátaBINO, EvaŠČERBOVÁ, JanaPOGÁNY SIMONOVÁ, Monika – DVOROŽŇÁKOVÁ, Emília. Bacteriocin-Producing Strain Lactiplantibacillus plantarum LP17L/1 Isolated from Traditional Stored Ewe’s Milk Cheese and Its Beneficial Potential. In Foods, 2022, vol. 11, no. 7, art. no. 959. (2021: 5.561 – IF, Q1 – JCR, 0.726 – SJR, Q1 – SJR, karentované – CCC). (2022 – Current Contents). ISSN 2304-8158.

Utilization of anticancer and antimicrobial effects of plant isothiocyanates

Anna Bertová, Szilvia Kontár, Zdena Sulová, Albert Breier, Denisa Imrichová

We addressed the possibility of using plant isothiocyanates in inhibiting the growth of neoplastically transformed cells or cells of microbial pathogens. Although the first knowledge of these compounds is more than 50 years old, a precise understanding of the mechanisms of their effects is still lacking. Globally, we are witnessing a renaissance of interest in these agents. Plant isothiocyanates are found in abundance in plants but not in the free state but in storage glucosinolates. Of these, they are released by the enzyme myrosinase (EC when the plant tissue is damaged. We have published results on the cytotoxic effect of sulfurafane (a degradation product of glucoraphanin in the myrosinase reaction) on leukaemic cells, which was mainly through the initiation of autophagy. We collaborated in the development of a procedure for the isolation of electrophoretically homogeneous myrosinase from Lepidium sativum seeds, as well as in the development of a procedure for the extraction of glucoraphanin from Lepidium draba and the possibility of its cleavage to sulforaphane, which is the subject of utility model number 9579. These findings point to the possibility of preparing a two-component preparation consisting of plant glucoraphanin (which does not exhibit any anticancer or antimicrobial effects) and plant myrosinase. With a suitable method of application, an effective amount of sulforaphane could be released at the site where the effect is to occur.

Sulfurafane, when released from glucoraphanin, induces autophagy. Under this condition, apoptosis is blocked in the early stages and cellular repair mechanisms are rescued. If these are successful, the leukemic cell continues to proliferate, otherwise progression to cell death occurs. We also demonstrated the need for repeated administration of sulforaphane, as a large proportion of leukemic cells were able to survive at a single dose.

BERTOVÁ, AnnaKONTÁR, Szilvia – POLOZSANYI, Zoltán – ŠIMKOVIČ, Martin – ROSENBERGEROVÁ, Zuzana – REBROŠ, Martin – SULOVÁ, ZdenaBREIER, Albert**IMRICHOVÁ, Denisa**. Effects of Sulforaphane-Induced Cell Death upon Repeated Passage of Either P-Glycoprotein-Negative or P-Glycoprotein-Positive L1210 Cell Variants. In International Journal of Molecular Sciences, 2022, vol. 23, no. 18, art. no. 10818. (2021: 6.208 – IF, Q1 – JCR, 1.176 – SJR, Q1 – SJR, karentované – CCC).(2022 – Current Contents). ISSN 1422-0067.

GALÁDOVÁ, Helena – POLOZSÁNYI, Zoltán – BREIER, Albert – ŠIMKOVIČ, M. Sulphoraphane Affinity-Based Chromatography for the Purification of Myrosinase from Lepidium sativum Seeds. In Biomolecules : Open Access Journal, 2022, vol. 12, no. 3, art. no. 406. (2021: 6.064 – IF, Q2 – JCR, 1.019 – SJR, Q2 – SJR). ISSN 2218-273X.

The dopaminergic system behind judgment formation processes in poultry

Katarína Pichová, Ľubica Niederová, Ľubor Košťál

The cognitive bias principle used to study affective states tracks changes in processes leading to reward or punishment avoidance. In this context, changes in the neuronal pathways of the brain involved in the aforementioned processes are expected, and it has been shown that the pathways involved in the appraisal of reward-related stimuli are concentrated around dopaminergic (DA) neurons in different brain regions. We tested the involvement of the DA system in the mechanisms responsible for judgment bias by applying DA receptor agonists and antagonists, hypothesizing that blockade of the system induces negative judgment bias and, conversely, activation of the system induces positive judgment bias in Japanese quail. Our findings confirmed only a negative shift in the bag of judgment after application of DA receptor antagonists, which prolonged latency and also reduced the proportion of animals’ responses in the test. However, the present results demonstrate the involvement of DA signaling in the underlying mechanisms of judgment bias in Japanese quail, giving a more comprehensive view on the issue of livestock welfarism.

PICHOVÁ, Katarína**NIEDEROVÁ, ĽubicaKOŠŤÁL, Ľubor. The acute pharmacological Manipulation of dopamine receptors modulates judgment bias in Japanese quail. In Frontiers in Pharmacology, 2022, vol. 13, art. no. 883021. (2021: 4.755 – IF, Q1 – JCR, 1.126 – SJR, Q1 – SJR).

Different mechanisms of drug resistance to hypomethylating agents in the treatment of myelodysplastic syndrome and acute myeloid leukemia

Kristína Šimoničová, Ľuboš Janotka, Helena Kavcová, Zdena Sulová, Albert Breier, Lucia Messingerová

Stratagem – New diagnostic and therapeutic tools against multidrug resistant tumours (CA 17104),

As part of the involvement in the COST project Stratagem, the group at the Institute of Molecular Physiology and Genetics CBs SAS was asked to prepare a review article on the development of resistance to hypomethylating agents in myeloid blasts of hematological malignancies in the prestigious journal Drug Resistance Updates. Multidrug resistance of cancer transformed cells is a special phenotype in which these cells lose sensitivity to several drugs used in cancer therapy. The development of this phenotype leads to a situation in which the cancer cells no longer respond sufficiently to treatment with these drugs and thus fail to effectively suppress their proliferation. Demethylating drugs (vidaza and decitabine) are approved (both FDA and EMA) for the treatment of high-risk MDS and AML in patients unsuitable or resistant to intensive chemotherapy. In some cases, such patients are resistant to demethylating therapy from the outset, and some others may develop resistance during treatment. Given the fact that there is no alternative treatment protocol for these patients, their prognosis is poor. Therefore, it is essential to understand the basic principles of resistance to demethylating agents and to design options to reverse it. In the present review article, the latest knowledge on this topic is described in detail, and our team’s results are put in context with the literature.

Summary of various possible mechanisms of resistance to chemotherapeutic drugs, especially in hematological malignancies; HSC – hematopoietic stem cell, LSC – leukemic stem cell, ECM – extracellular matrix, ATP – adenosine triphosphate

ŠIMONIČOVÁ, Kristína*JANOTKA, Ľuboš*KAVCOVÁ, HelenaSULOVÁ, ZdenaBREIER, Albert**MESSINGEROVÁ, Lucia**. Different mechanisms of drug resistance to hypomethylating agents in the treatment of myelodysplastic syndromes and acute myeloid leukemia. In Drug Resistance Updates, 2022, vol. 61, art. no. 100805. (2021: 22.841 – IF, Q1 – JCR, 3.845 – SJR, Q1 – SJR). ISSN 1368-7646.

Phosphatidylglycerol and its role in the manifestations of Barth syndrome

Mária Balážová, Paulína Káňovičová, Lenka Bábelová, Dominika Kubalová, Martin Valachovič

In collaboration with the Institute of Experimental Medicine of the CAS and Tunghai University in Taiwan, we focused on the role of the mitochondrial lipid phosphatidylglycerol (PG) in the manifestation of symptoms of Barth syndrome (BTHS). Together, we described the pgc1Δtaz1Δ double mutant of Saccharomyces cerevisiae as a novel yeast model of BTHS. In our results, we showed that increased PG leads to more pronounced mitochondrial respiratory defects and an increased incidence of aberrant mitochondria compared to the taz1Δ mutant. Overall, our findings point to an important role for PG in the bioenergetics of both yeast and human cell lines.

PGC1 gene deletion increases the incidence of morphologically aberrant mitochondria in taz1Δ cells. (I) Mitochondrial morphology was visualized by Mitotracker Red staining in standard strain, pgc1Δ, taz1Δ, and pgc1Δtaz1Δ cells grown in SMDGE I- medium for 24 hours. (II) Mitochondria of the pgc1Δtaz1Δ mutant represent two populations with different structures. Ultrathin sections of pgc1Δtaz1Δ cells imaged by transmission electron microscopy are presented.

KÁŇOVIČOVÁ, Paulína – ČERMÁKOVÁ, Petra – KUBALOVÁ, DominikaBÁBELOVÁ, Lenka – VESELÁ, Petra – VALACHOVIČ, Martin – ZAHUMENSKÝ, Jakub – HORVÁTH, Anton – MALÍNSKÝ, Ján** – BALÁŽOVÁ, Mária**. Blocking phosphatidylglycerol degradation in yeast defective in cardiolipin remodeling results in a new model of the Barth syndrome cellular phenotype. In Journal of Biological Chemistry, 2022, vol. 298, no. 1, art. no. 101462. (2021: 5.485 – IF, Q2 – JCR, 1.871 – SJR, Q1 – SJR).

CHU, I* – CHEN, Ying-Chih* – LAI, Ruo-Yun – CHAN, Jui-Fen – LEE, Ya-Hui – BALÁŽOVÁ, Mária – HSU, Yuan- Hao Howard**. Phosphatidylglycerol Supplementation Alters Mitochondrial Morphology and Cardiolipin Composition. In Membranes : Open Access Journal, 2022, vol. 12, no. 4, art. no. 383. (2021: 4.562 – IF, Q1 – JCR, 0.517 – SJR, Q2 – SJR).

The form of fibre and zinc in pig diets after weaning may affect the absorption of trace elements from the digestive tract and influence the biomechanical properties of bones

Grešáková Ľ., Čobanová, K., Tokarčíková, K., Takácsová, M.

Zinc (Zn) is essential for the proper functioning of biological systems. The body’s need for Zn increases during the weaning period, when sufficient intake of Zn serves to ensure proper bone development, antioxidant protection and immunity of the young organism. The absorption of Zn from the GIT is influenced by the composition of the feed ration. The form of Zn and fibre in the feed may influence not only the absorption of Zn but also the bioavailability of other trace elements in the body. Although the results of our in vitro and in vivo experiments did not show differences in the bioavailability of Zn from its different sources, its form in the feed influenced the solubility of Mn and Fe in the small intestine of pigs. The addition of potato fibre to the diet increased the absorption of Zn and Cu from the GIT of pigs, but had a significantly negative effect on Fe absorption. Supplementation of growing pig diets with Zn glycinate, increased the strength, mineralization and femur bone density of pigs compared to ZnSO4. The combination of an organic source of Zn and dietary fibre positively affected the nutritional and mineral status of young pigs, which may be particularly crucial during the weaning period characterised by reduced feed intake and insufficient absorption capacity of the digestive tract.

TOKARČÍKOVÁ, KatarínaČOBANOVÁ, KlaudiaTAKÁCSOVÁ, Margaréta – BARSZCZ, Marcin – TACIAK, Marcin – TUŚNIO, Anna – GREŠÁKOVÁ, Ľubomíra**. Trace mineral solubility and digestibility in the small intestine of piglets are affected by zinc and fibre sources. In Agriculture – Basel, 2022, vol. 12 no. 4, art. no. 517. (2021: 3.408 – IF, Q1 – JCR, 0.525 – SJR, Q2 – SJR, karentované – CCC). (2022 – Current Contents). ISSN 2077-0472. Dostupné na:

SKIBA, Grzegorz – RAJ, Stanislawa – SOBOL, Monika – KOWALCZYK, Pawel – BARSZCZ, Marcin – TACIAK, Marcin – TUŚNIO, Anna – ČOBANOVÁ, KlaudiaGREŠÁKOVÁ, Ľubomíra – GRELA, Eugeniusz Ryszard. Influence of the zinc and fibre addition in the diet on biomechanical bone properties in weaned piglets. In Animals, 2022, vol. 12, no. 2, art. no. 181. (2021: 3.231 – IF, Q1 – JCR, 0.610 – SJR, Q1 – SJR, karentované – CCC). (2022 – Current Contents). ISSN 2076-2615.