Pavel Kulich

Veterinary Research Institute

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Jaroslav Tuřánek

Charles University

Josef Mašek

Veterinary Research Institute

Milan Raška

Palacký University Olomouc

František Hubatka

International Clinical Research Center, St. Anne's University Hospital Brno

Eliška Bartheldyová

International Clinical Research Center, St. Anne's University Hospital Brno

Jan Kotouček

Veterinary Research Institute

Miroslav Machala

Veterinary Research Institute

Pavlína Turánek Knötigová

St. Anne's University Hospital Brno

Andrew D. Miller

Mendel University in Brno

ŠK

Štěpán Koudelka

Veterinary Research Institute

Cooperative Institutions

Veterinary Research Institute

135

Masaryk University

117

Czech Academy of Sciences

University of Veterinary Sciences Brno

Charles University

Central European Institute of Technology

Brno University of Technology

Mendel University in Brno

University Hospital Brno

St. Anne's University Hospital Brno

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A murine model of the effects of inhaled CuO nanoparticles on cells of innate and adaptive immunity – a kinetic study of a continuous three-month exposure

Nanotoxicology (2019)

Vladimı́r Holáň Eliška Javorková Kristýna Vrbová Zbyněk Večeřa Pavel Mikuška

The inhalation or application of nanoparticles (NPs) has serious impacts on immunological reactivity. However, the effects of NPs on the immune system are influenced by numerous factors, which cause a high variability in the results. Here, mice were exposed to a three month continuous inhalation of copper oxide (CuO) NPs, and at different time intervals (3, 14, 42 and 93 days), the composition of cell populations of innate and adaptive immunity was evaluated in the spleen by flow cytometry. The ability of spleen cells from exposed and control mice to respond to stimulation with T- or B-cell mitogens by proliferation and by production of cytokines IL-2, IL-6, IL-10, IL-17 and IFN-γ was characterized. The results showed that the inhalation of CuO NPs predominantly affects the cells of innate immunity (changes in the proportion of eosinophils, neutrophils, macrophages and antigen-presenting cells) with a minimal effect on the percentage of T and B lymphocytes. However, the proliferative and secretory activity of T cells was already significantly enhanced after 3 days from the start of inhalation, decreased on day 14 and normalized at the later time intervals. There was no correlation between the impacts of NPs on the cells of innate and adaptive immunity. The results have shown that the inhalation of CuO NPs significantly alters the composition of cell populations of innate immunity and modulates the proliferation and production of cytokines by cells of the adaptive immune system. However, the immunomodulatory effects of inhaled NPs strongly depend on the time of inhalation.

10.1080/17435390.2019.1602679

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Citations (18)

Pinching Off Syndrome in Two White-Tailed Sea Eagles (Haliaeetus albicilla) in the Czech Republic

Journal of Raptor Research (2008)

Lucia Fasungová Ivan Literák Oldřich Sychra Ladislav Novotný Veronika Grymová

White (mutation)

10.3356/jrr-07-03.1

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Citations (6)

Characterisation of Pasteurella multocida isolated from rabbits in the Czech Republic

Veterinární Medicína (2006)

Zoran Jaglic Z. Kučerová Kateřína Nedbalcová Pavel Kulich P. Alexa

Twenty seven Pasteurella multocida rabbit isolates were characterised by pulsed-field gel electrophoresis (PFGE) generated by restriction endonuclease ApaI, and examined for the presence of capsule and fimbriae, and tested of haemolytic activity and endotoxin release in presence of rabbit plasma. Among the isolates a high number (n = 20) of different PFGE types was observed. All isolates were found to be encapsulated and four of them also possessed fimbriae. Although the presence of the aphA gene, conferring a haemolytic phenotype for Escherichia coli under anaerobic conditions, was indicated in all of the isolates by PCR, clear haemolytic activity was observed in only one isolate when grown anaerobically. All of the isolates grew easily in the presence of rabbit plasma which showed a high capacity to bind the released endotoxin.

Pasteurella

Strain (injury)

10.17221/5547-vetmed

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Citations (8)

Optimizing Tubulin Yield from Porcine Brain Tissue

Journal of Visualized Experiments (2024)

Adam Norek Michaela Šťastná Jan Gečnuk Pavel Kulich Jan Gebauer

Neural tissue from any source is an excellent material for tubulin isolation, as the dendrites and axons of neurons are rich in microtubules. Here, we present a procedure for extracting tubulin that can be employed, with minor modifications, for neural tissue from multiple sources. In the presented protocol, a new clarification step of the crude lysate has been introduced, which led to a significant reduction in the initial amount of insoluble debris before the first polymerization step occurred. This additional step allowed the processing of additional tissue while using the same instrumentation and, thus, increasing the relative volume of the processed homogenate. The newly introduced step has no significant effect on the quality of purified tubulin, as confirmed by in vitro activity assays and transmission electron microscopy. The described procedure contains all crucial steps, including tissue collection, transport, tissue homogenization, tubulin isolation cycles, and final polishing by ion exchange chromatography using FPLC and subsequent activity measurement assays. The homogeneity of purified tubulin was more than 97%, as confirmed by MS/MS analysis utilizing electrospray ionization and MALDI-TOF.

10.3791/64925

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Rapid spread of mouse mammary tumor virus in cultured human breast cells

Retrovirology (2007)

Stanislav Indik Walter H. Günzburg Pavel Kulich Brian Salmons Francoise Rouault

The role of mouse mammary tumor virus (MMTV) as a causative agent in human breast carcinogenesis has recently been the subject of renewed interest. The proposed model is based on the detection of MMTV sequences in human breast cancer but not in healthy breast tissue. One of the main drawbacks to this model, however, was that until now human cells had not been demonstrated to sustain productive MMTV infection. Here, we show for the first time the rapid spread of mouse mammary tumor virus, MMTV(GR), in cultured human mammary cells (Hs578T), ultimately leading to the infection of every cell in culture. The replication of the virus was monitored by quantitative PCR, quantitative RT-PCR and immunofluorescence imaging. The infected human cells expressed, upon cultivation with dexamethasone, MMTV structural proteins and released spiked B-type virions, the infectivity of which could be neutralized by anti-MMTV antibody. Replication of the virus was efficiently blocked by an inhibitor of reverse transcription, 3'-azido-3'-deoxythymidine. The human origin of the infected cells was confirmed by determining a number of integration sites hosting the provirus, which were unequivocally identified as human sequences. Taken together, our results show that human cells can support replication of mouse mammary tumor virus.

Provirus

Mammary tumor

Infectivity

Immunofluorescence

10.1186/1742-4690-4-73

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Citations (65)

Immobilization of histidine-tagged proteins on monodisperse metallochelation liposomes: Preparation and study of their structure

Analytical Biochemistry (2010)

Josef Mašek Eliška Bartheldyová Zina Korvasová Michaela Škrabalová Štěpán Koudelka

Dispersity

Gel permeation chromatography

10.1016/j.ab.2010.08.023

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Citations (31)

Development of 3D Printed Multi-Layered Orodispersible Films with Porous Structure Applicable as a Substrate for Inkjet Printing

Pharmaceutics (2023)

Jan Elbl Martin Veselý Dagmar Blaháčková Jaroslav Ondruš Pavel Kulich

The direct tailoring of the size, composition, or number of layers belongs to the advantages of 3D printing employment in producing orodispersible films (ODFs) compared to the frequently utilized solvent casting method. This study aimed to produce porous ODFs as a substrate for medicated ink deposited by a 2D printer. The innovative semi-solid extrusion 3D printing method was employed to produce multilayered ODFs, where the bottom layer assures the mechanical properties. In contrast, the top layer provides a porous structure for ink entrapment. Hydroxypropyl methylcellulose and polyvinyl alcohol were utilized as film-forming polymers, glycerol as a plasticizer, and sodium starch glycolate as a disintegrant in the bottom matrix. Several porogen agents (Aeroperl® 300, Fujisil®, Syloid® 244 FP, Syloid® XDP 3050, Neusilin® S2, Neusilin® US2, and Neusilin® UFL2) acted as porosity enhancers in the two types of top layer. ODFs with satisfactory disintegration time were prepared. The correlation between the porogen content and the mechanical properties was proved. A porous ODF structure was detected in most samples and linked to the porogen content. SSE 3D printing represents a promising preparation method for the production of porous ODFs as substrates for subsequent drug deposition by 2D printing, avoiding the difficulties arising in casting or printing medicated ODFs directly.

Plasticizer

Polyvinyl Alcohol

10.3390/pharmaceutics15020714

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Citations (8)

Inhalation of ZnO Nanoparticles: Splice Junction Expression and Alternative Splicing in Mice

Toxicological Sciences (2018)

Pavel Rössner Kristýna Vrbová Simona Strapáčová Andrea Rössnerová Antonín Ambrož

Despite the wide application of nanomaterials, toxicity studies of nanoparticles (NP) are often limited to in vitro cell models, and the biological impact of NP exposure in mammals has not been thoroughly investigated. Zinc oxide (ZnO) NPs are commonly used in various consumer products. To evaluate the effects of the inhalation of ZnO NP in mice, we studied splice junction expression in the lungs as a proxy to gene expression changes analysis. Female ICR mice were treated with 6.46 × 104 and 1.93 × 106 NP/cm3 for 3 days and 3 months, respectively. An analysis of differential expression and alternative splicing events in 298 targets (splice junctions) of 68 genes involved in the processes relevant to the biological effects of ZnO NP was conducted using next-generation sequencing. Three days of exposure resulted in the upregulation of IL-6 and downregulation of BID, GSR, NF-kB2, PTGS2, SLC11A2, and TXNRD1 splice junction expression; 3 months of exposure increased the expression of splice junctions in ALDH3A1, APAF1, BID, CASP3, DHCR7, GCLC, GCLM, GSR, GSS, EHHADH, FAS, HMOX-1, IFNγ, NF-kB1, NQO-1, PTGS1, PTGS2, RAD51, RIPK2, SRXN1, TRAF6, and TXNRD1. Alternative splicing of TRAF6 and TXNRD1 was induced after 3 days of exposure to 1.93 × 106 NP/cm3. In summary, we observed changes of splice junction expression in genes involved in oxidative stress, apoptosis, immune response, inflammation, and DNA repair, as well as the induction of alternative splicing in genes associated with oxidative stress and inflammation. Our data indicate the potential negative biological effects of ZnO NP inhalation.

GCLM

10.1093/toxsci/kfy288

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Citations (29)

Rekonstrukce železniční tratě Vsetín - Velké Karlovice mezi km 17,000 a km 19,950 včetně návrhu technologie prací

Pavel Kulich

Source

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Preparation of MS2 Phage-Like Particles and Their Use As Potential Process Control Viruses for Detection and Quantification of Enteric RNA Viruses in Different Matrices

Frontiers in Microbiology (2016)

Pavel Mikel Petra Vašíčková Radek Tesařı́k Hana Malenovská Pavel Kulich

The detection and quantification of enteric RNA viruses is based on isolation of viral RNA from the sample followed by quantitative reverse transcription polymerase chain reaction (RT-qPCR). To control the whole process of analysis and in order to guarantee the validity and reliability of results, process control viruses (PCV) are used. The present article describes the process of preparation and use of such PCV- MS2 phage-like particles (MS2 PLP) - in RT-qPCR detection and quantification of enteric RNA viruses. The MS2 PLP were derived from bacteriophage MS2 carrying a unique and specific de novo-constructed RNA target sequence originating from the DNA of two extinct species. The amount of prepared MS2 particles was quantified using four independent methods - UV spectrophotometry, fluorimetry, transmission electron microscopy and a specifically developed duplex RT-qPCR. To evaluate the usefulness of MS2 PLP in routine diagnostics different matrices known to harbor enteric RNA viruses (swab samples, liver tissue, serum, feces, and vegetables) were artificially contaminated with specific amounts of MS2 PLP. The extraction efficiencies were calculated for each individual matrix. The prepared particles fulfill all requirements for PCV - they are very stable, non-infectious, and are genetically distinct from the target RNA viruses. Due to these properties they represent a good morphological and physiochemical model. The use of MS2 PLP as a PCV in detection and quantification of enteric RNA viruses was evaluated in different types of matrices.

Bacteriophage MS2

Enteric virus

10.3389/fmicb.2016.01911

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Citations (35)