The cell culture became an invaluable tool for studying cell behaviour, development, function, gene expression, toxicity of compounds and efficacy of novel drugs. Although most results were obtained from cell cultivation in two-dimensional (2D) systems, in which cells are grown in a monolayer, three-dimensional (3D) cultures are more promising as they correspond closely to the native arrangement of cells in living tissues. In our study, we focused on three types of 3D in vitro systems used for cultivation of one cell type. Cell morphology, their spatial distribution inside of resulting multicellular structures and changes in time were analysed with histological examination of samples harvested at different time periods. In multilayered cultures of WRL 68 hepatocytes grown on semipermeable membranes and non-passaged neurospheres generated by proliferation of neural progenitor cells, the cells were tightly apposed, showed features of cell differentiation but also cell death that was observable in short-term cultures. Biogenic scaffolds composed of extracellular matrix of the murine tibial anterior muscle were colonized with C2C12 myoblasts in vitro. The recellularized scaffolds did not reach high cell densities comparable with the former systems but supported well cell anchorage and migration without any signs of cell regression. and H. Hrebíková, D. Čížková, J. Chvátalová, R. Pisal, R. Adamčik, P. Beznoska, D. Díaz-Garcia, J. Mokrý
Karcinom prostaty je nejčastějším solidním zhoubným nádorem u mužů v řadě vyspělých zemí světa. V naší studii jsme hodnotili schopnost detekovat pravděpodobnost přítomnosti karcinomu prostaty (KP) ve vzorcích lidské moči čichem speciálně trénovaného psa. Definitivní potvrzení KP je vždy nutné ověřit histologicky. Jedním z alternativních biomarkerů, které pes v moči cítí, jsou pak VOC (volatile organic compounds). Předmětem našeho dalšího výzkumu bude zjišťování, na jakou látku v moči pes vlastně reaguje a díky tomu je schopen označit pozitivní vzorek., Prostate cancer is the most common solid malign tumour in men in many developed countries of the world. Our study evaluates the ability to detect the presence of prostate cancer in human urine samples using olfactory cells of specifically trained dog. The definitive confirmation of prostate cancer should be verified histologically. One of the alternative biomarkers that can be detected by the dog are so called volatile organic compounds. In our research we are trying to identify the specific agent in urine, which is detected by the dog and thereby a positive sample is marked, and Vyhnánková V., Pacík D., Urbanová L., Nečas A., Tučková M.
Huntingtonova nemoc (HD) je neurodegenerativní porucha způsobená elongací CAG repetic v genu kódující protein huntingtin (Htt). U pacientů jsou v postižených tkáních přítomny vedle monomerní formy hlavně N‑koncové fragmenty, oligomery a polymery mutovaného huntingtinu (mtHtt), oproti tomu samotná monomerní forma mtHtt je exprimována v podstatě ve všech buňkách. Nejvíce postižené tkáně jsou bazální ganglia a mozková kůra. V této studii jsme analyzovali přítomnost N‑koncových fragmentů a oligomerů Htt v různých tkáních 24- a 36měsíčních transgenních (TgHD) miniprasat exprimujících N‑koncovou část lidského mutovaného huntingtinu a jejich zdravých sourozenců. Zjistili jsme, že mozková kůra a varlata na rozdíl od svalu a srdce TgHD miniprasat obsahují kromě monomerní formy i N‑koncové fragmenty a oligomerní smíry. Ve svalech z 36 měsíčních TgHD miniprasat však již začíná mírná fragmentace. Tato zjištění napodobují časnou progresi onemocnění u lidí, a proto miniprase poskytuje slibný model pro terapeutické testování HD. Klíčová slova: Huntingtonova nemoc – transgenní miniprasečí model – mutovaný huntingtin –proteinové fragmenty – oligomerní struktury Autoři deklarují, že v souvislosti s předmětem studie nemají žádné komerční zájmy. Redakční rada potvrzuje, že rukopis práce splnil ICMJE kritéria pro publikace zasílané do biomedicínských časopisů., Huntington’s disease (HD) is a neurodegenerative disorder caused by the the elongation of CAG triplet repeat in the gene encoding the huntingtin protein (Htt). In patients, in addition to the monomeric form of huntingtin, N‑terminal fragments, oligomers, and polymers are present mostly in the affected tissues, even though the mutated huntingtin (mtHtt) is expressed basically in all cells. The most affected tissues are basal ganglia and cerebral cortex. In this study we analyzed the presence of N‑terminal fragments and oligomers of Htt in different tissues of 24 and 36 months old experimental animals. This was done in our large animal model of HD, which uses transgenic (TgHD) minipigs expressing N‑terminal part of human mtHtt. Among all the tissues tested, we found cortex and testes to contain N‑terminal fragments as well as oligomeric smears in TgHD minipigs compared to wild type siblings. On the other hand, we did not detect any fragments or oligomers in muscle and heart of TgHD minipigs, only starting fragmentation in muscles of 36 months old animals. These findings mimic the early progression of the disease in humans, hence presents minipig as a promising model for therapeutic testing of HD., and D. Vidinska, J. Motlik, Z. Ellederova
Huntingtonova nemoc (HN) je autozomálně dominantní neurodegenerativní onemocnění charakterizované poškozením volních a mimovolních pohybů, poruchami chování a zhoršením kognitivních funkcí. Společně s hlavními motorickými příznaky byly poruchy hlasu a řeči pozorovány u většiny pacientů s HN. Zvířecí prasečí model je často používán pro výzkum v preklinických studiích. I přes zjevné rozdíly v anatomii artikulačních orgánů mezi prasaty a lidmi lze očekávat stejné trendy u patofyziologických mechanizmů s ohledem na chrochtání i lidskou fonaci. Hlavním cílem této studie bylo proto navržení vhodného experimentu, který umožní získání dostatečně dlouhého záznamu chrochtání od co největšího počtu prasátek. Dalším cílem studie bylo zrealizovaní výsledné verze experimentu na celé databázi a vyhodnocení množství a kvality získaných nahrávek. Databáze použitá pro studii zahrnovala 17 HN transgenních miniprasátek a 16 zdravých sourozenců ze stejných vrhů. Testované varianty experimentu, provedené na části databáze zahrnující čtyři prasnice, byly rozděleni do čtyř podskupin: (a) pozitivní – krmení, (b) pozitivní – zvuková stimulace, (c) negativní – bránění v pohybu, (d) negativní – nepříjemné doteky. Hodnocení kvality získaných nahrávek bylo provedeno s pomocí audio softwaru, ve kterém bylo izolováno čisté prasečí chrochtání a všechny akustické artefakty vymazány. Nejlepších výsledků bylo dosaženo s použitím experimentu při kterém: (i) je záznamové zařízení upevněno na tělo prasátka, (ii) prasátko je ponecháno několik minut o samotě v místnosti, aby se uklidnilo a (iii) osoba vstoupí do místnosti a snaží se nabízet prasátku krmivo zatímco před ním couvá. V důsledku tohoto jednání prasátko následuje osobu s krmením, což je doprovázeno chrochtáním. Dostatečně dlouhé (20 chrochtnutí a více) a čisté nahrávky byly získány od 24 z 33 prasátek (73 %). Závěrem lze tedy říci, že experiment je proveditelný. Klíčová slova: Huntingtonova nemoc – chrochtání – transgenní model miniprasat – animální modely – poruchy hlasu a řeči Autoři deklarují, že v souvislosti s předmětem studie nemají žádné komerční zájmy. Redakční rada potvrzuje, že rukopis práce splnil ICMJE kritéria pro publikace zasílané do biomedicínských časopisů., Huntington’s disease (HD) is an autosomal-dominant neurodegenerative disorder characterized by the impairment of voluntary and involuntary movements, behavioral disorders and cognitive decline. Besides the main motor symptoms, voice and speech disorders have been documented in a large majority of patients with HD. The animal model of pigs is often used in preclinical studies. Although there are obvious differences in the anatomy of the articulation organs between pigs and humans, the same trends in pathophysiological mechanisms can be expected in both grunting and human phonation. The main aim of the study was therefore to design a suitable experiment that would allow for acquisition of a sufficiently long recording of grunting from as many pigs as possible. The second goal was to perform the final version of the experiment in all available pigs and to evaluate the amount and quality of the acquired recordings. The database consists of 17 HD transgenic minipigs and 16 healthy siblings. Tested variants of the experiment, performed on subgroup of four sows, were divided into four subgroups: (a) positive – feeding, (b) positive – sound stimulation, (c) negative – hindering in movement, (d) negative – unpleasant touch. The evaluation of the quality of the elicited recording was performed using audio software where pure pig grunting was selected and all acoustic artefacts deleted. The best results were reached using the experiment in which: (i) a recording device is put on the pig’s body, (ii) the pig is left alone for few minutes in the pen in order to calm down, and (iii) a person enters the room and tries to offer the pig food while walking backwards. As a result, the pig follows the person and grunts. Sufficiently long (20 single grunts or more) and clear recordings were received from 24 out of 33 pigs (73%). The realisation of the experiment is therefore possible. Key words: Huntington’s disease – grunting – transgenic pigs – animal models – voice and speech disorders The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE “uniform requirements” for biomedical papers., and T. Tykalova, J. Hlavnicka, M. Macakova, M. Baxa, R. Cmejla, J. Motlik, J. Klempir, J. Rusz
Melatonin is a hormone with strong antioxidant properties. In this experiment, Freund's complete adjuvant was used as a stressogenic substance given to laboratory outbred mice, whereas melatonin was investigated as a protectant against the stressogenic effect. Levels of low molecular weight antioxidants, thiobarbituric acid reactive substances, and tumor necrosis factor α and activity of glutathione reductase were determined in blood from the animals. Surprisingly, melatonin was not involved in direct regulation of antioxidants, thiobarbituric acid reactive substances and tumor necrosis factor α. On the other hand, melatonin regulated glutathione reductase activity. We can conclude on regulation of metabolism caused by melatonin in the model. The effect was more important than the expected regulation of immunity and basal oxidative homeostasis. and M. Pohanka, B. Ruttkay-Nedecký, J. Fusek, V. Adam, R. Kizek
Pohybová inaktivita je průkazně spojena s manifestací chronických, kvalitu a prognózu života člověka významně negativně modifikujících onemocnění. Benefity pohybové aktivity zprostředkovává jistě mnoho více či méně provázaných patofyziologických mechanizmů, které dosud nebyly ve své složitosti zcela prozkoumány. Koncem 20. století se podařilo prokázat, že pracující příčně pruhovaný sval skutečně reguluje metabolickou a fyziologickou odezvu v ostatních orgánech. Jedná o několik stovek substancí, které mají autokrinní, parakrinní a endokrinní účinek. Tyto proteiny a peptidy, jsou-li vylučovány do krevního oběhu, ovlivňují významným způsobem metabolizmus vzdálených orgánů. Byly klasifikovány jako „myokiny“ (cytokiny produkované myocyty). Mezi identifikované myokiny lze zařadit např.: interleukiny – IL4, IL6, IL7, IL15, myostatin, LIF (leucemia inhibitory factor), BDNF (brain-derived neurotropic factor), IGF1 (insulin-like growth factor), FGF2 (fibroblastový růstový faktor 2), FGF21, FSTL1 (folistatin-related protein 1), irisin, EPO (erytropoetin) a BAIBA (β-aminoisobutyric acid). Myokiny mají v lidském organizmu v prvé řadě imunoregulační roli. Další významnou úlohou myokinů je, shodou okolností rovněž v interakci s tukovou tkání, regulace energetické homeostázy. Ovlivňují také růst svalových vláken a jejich regeneraci, stimulují angiogenezi, uplatňují se v regulaci metabolizmu glukózy a mají prokázaný efekt na lipidy. S ohledem na svoji rozmanitou funkci představují myokiny do budoucna terapeutický cíl v léčbě poruch svalového růstu regenerace a také obezity. Další recentní výzkum směřuje k odhalení tzv. myokinové rezistence jako rezultátu dlouhodobé svalové inaktivity a její asociaci s chronickým subklinickým zánětem., Physical inactivity is demonstrably related to the manifestation of chronic diseases which significantly modify the quality and prognosis of life in a negative way. The benefits of exercise are surely mediated by many pathophysiological mechanisms interrelated in varying degrees, which have not yet been fully examined in their complexity. In the late 20th century it was positively proven that a working striated muscle really regulates the metabolic and physiological response in the other organs. These involve several hundred substances with autocrine, paracrine and endocrine effects. These proteins and peptides, if released into the blood stream, substantially affect the metabolism of distant organs. They were classified as “myokines“ (cytokines produced by myocytes). The identified myokines include e.g. IL4, IL6, IL7, IL15, myostatin, LIF (leukemia inhibitory factor), BDNF (brain-derived neurotrophic factor), IGF1 (insulin-like growth factor), FGF2 (fibroblast growth factor 2), FGF21, FSTL1 (follistatin-related protein 1), irisin, EPO (erythropoetin) and BAIBA (β-aminoisobutyric acid). Myokines have first of all an immunoregulatory role in the human body. Another important effect of myokines is, coincidentally also in the interaction with adipose tissue, the regulation of energy homeostasis. They also affect the growth of muscle fibres and their regeneration, stimulate angiogenesis, they are involved in the regulation of glucose metabolism and have a proven effect on lipids. Considering their diverse function, myokines present a prospective therapeutic goal in the treatment of disorders of muscle growth and regeneration as well as obesity. Another recent research moves toward uncovering of the “myokine resistance” as a result of long-term muscle inactivity and its association with chronic subclinical inflammation., and Zuzana Stránská, Štěpán Svačina
BACKGROUND: The method of continual determination of the rat blood cholinesterase activity was developed to study the changes of the blood cholinesterases following different intervetions. AIMS: The aim of this study is registration of cholinesterase activity in the rat blood and its changes to demonstrate detoxification capacity of rats to inactivate sarin or VX in vivo. METHODS: The groups of female rats were premedicated (ketamine and xylazine) and cannulated to a. femoralis. Continual blood sampling (0.02 ml/min) and monitoring of the circulating blood cholinesterase activity were performed. Normal activity was monitored 1-2 min and then the nerve agent was administered i.m. (2×LD50). Using different time intervals of the leg compression and relaxation following the agent injection, cholinesterase activity was monitored and according to the inhibition obtained, detoxification capacity was assessed. RESULTS: Administration of sarin to the leg, then 1 and 5 min compression and 20 min later relaxation showed that further inhibition in the blood was not observed. On the other hand, VX was able to inhibit blood cholinesterases after this intervention. CONCLUSIONS: The results demonstrated that sarin can be naturally detoxified on the contrary to VX. Described method can be used as model for other studies dealing with changes of cholinesterases in the blood following different factors. and J. Bajgar, J. Cabal, J. Kassa, M. Pavlík
The aim of this study project was to prepare our own method of porcine oesophageal manometry. Ten mature experimental pigs entered the study. Conventional water-perfused system was decided for manometry. Porcine resting and relaxed pressures of the lower oesophageal sphincter are fully comparable with healthy human subjects. Evocable swallowing is doable and oesophageal peristalsis is quantifiable. Basic manometric parameters were different in male and female animals. Oesophageal manometry in experimental pigs is feasible. Porcine oesophageal manometry will be usable for preclinical studies in future. and I. Tachecí, V. Radochová, J. Květina, S. Rejchrt, M. Kopáčová, J. Bureš
AIM/BACKGROUND: Pistacia terebinthus is used as a coffee substitute in the East and Southern Anatolia regions of Turkey. It contains unsaturated fatty acids, tocopherols, polyphenols and carotenoids. P. terebinthus has anti-inflammatory and potential antioxidant activity. In this study we evaluated the protective effects of P. terebinthus coffee (PTC) on thioacetamide (TAA)-induced liver injury in rats. MATERIALS AND METHODS: Twenty-eight male Sprague-Dawley rats were equally randomized into four groups. Chronic liver injury was induced with TAA (100 mg/kg i.p. three times weekly). The first group of rats served as control and received only tap water (G1), and the remaining groups of rats received PTC, p.o (G2); TAA (G3); TAA plus PTC, p.o (G4), respectively. RESULTS: After 8 weeks, PTC intake significantly reduced fibrosis/inflammation scores (p PTC intake reduced transforming growth factor beta (TGF-β) concentrations in the liver (p PTC intake. DISCUSSION AND CONCLUSION: PTC intake provided beneficial effects against TAA-induced liver injury in rats. PTC probably suppresses the proinflammatory cytokines through NF-κB signaling pathway. and I. H. Bahcecioglu, M. Ispiroglu, M. Tuzcu, C. Orhan, M. Ulas, U. Demirel, M. Yalniz, I. H. Özercan, N. Ilhan, K. Sahin