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Mycophenolate mofetil (MMF) is an immunosuppressant drug approved for prophylaxis of transplant rejection in patients undergoing solid organ transplantation and is further employed in management of various autoimmune disorders. MMF exhibits notable pharmacokinetic inter- and intraindividual variability necessitating tailored therapeutic approaches to achieve optimal therapeutic outcomes while mitigating risks of adverse effects. The objective of this review was to summarize factors that influence the pharmacokinetics of MMF and its active metabolite mycophenolic acid in order to deduce recommendations for personalized treatment strategies. Presumed predictors were analysed in relation to each of the four pharmacokinetic phases, providing tools and targets for MMF dosing optimization amenable to clinical implementation.

The experiments were carried out on 5 different bioproducts for control of Verticillium longisporum in oilseed rape. For in vitro trials, greenhouse trials and field trials, the bioproducts used were based on bacteria (Bacillus amyloliquefaciens, Pseudomonas veronii) and fungi (Pythium oligandrum, Trichoderma asperellum, Coniothyrium minitans). In in vitro trials, some products (based on Trichoderma asperellum) showed a 100% inhibition effect on the pathogen for a whole trial period of 45 days. The greenhouse trial showed significant differences in bioproducts effectiveness (P < 0.00723) compared to control. Three bioproducts (based on Coniothyrium minitans, Trichoderma asperellum and Pseudomonas veronii) showed a 100% inhibition effect against the pathogen. In field trials conducted in 3 locations, there were some differences in yield, which can be important for growers and practice use. The highest yield (19.1% higher than the control) was achieved with a bioproduct based on Trichoderma asperellum. In trials, it is possible to see that there are promising results that can be used for further testing.

The increasing concentration of atmospheric carbon dioxide (CO2) demands innovative solutions for its capture and sequestration. Composite and nanocomposite materials have gained attention for their high surface area, adjustable properties, and improved adsorption capabilities. This manuscript explores the preparation, characterization, and performance assessment of a novel composite material consisting of polyamide 6 nanofibers combined with two types of zeolites, designed for CO2 capture. We detail the fabrication process, as well as the structural, chemical, and morphological characterization of the composites. The results confirmed the successful modification of polyamide 6 nanofibers with both types of zeolites. CO2 sorption measurements revealed that the composite with zeolite Y exhibited a notably high sorption capacity, with a maximum of 35.8 cm³/g, significantly surpassing the performance of amine-modified nanofibers, which demonstrated sorption capacities approximately three times lower. These findings highlight the potential of this composite for efficient CO2 capture, providing valuable insights into its practical application for reducing greenhouse gas emissions. While the CO2 sorption capacities of the composite are lower than those of pure zeolites, the composite offers practical advantages, such as easier handling and simplified application.

The Erysimum latent virus (ELV), a tymovirus, was first isolated from several wild and cultivated brassicas in Germany. Its virions were shown to be serologically distinct from those of the turnip yellow mosaic virus (TYMV), which is also found in wild and cultivated plants in several European countries but also in other parts of the world. TYMV and ELV were among the first plant viruses to have had their genomes sequenced, and when other tymovirus genomes were sequenced, it was found that, in phylogenies, ELV is probably the basal outlier to all other tymoviruses. Here, we report the near-complete genomic sequence of another isolate of ELV from Czechia. This isolate was found in 1990 in Sisymbrium altissimum plants showing mosaic symptoms. It was detected using ELISA tests and electron microscopy. We have now sequenced the full coding sequence of this isolate using contemporary high throughput methods and found that the German and Czech isolates of ELV are closely related and are of the same virus species.

Background: Ivacaftor is a modern drug used in the treatment of cystic fibrosis. It is highly lipophilic and exhibits a strong positive food effect. These characteristics can be potentially connected to a pronounced lymphatic transport after oral administration.Methods: A series of studies was conducted to describe the basic pharmacokinetic parameters of ivacaftor in jugular vein cannulated rats when dosed in two distinct formulations: an aqueous suspension and an oil solution. Additionally, an anesthetized mesenteric lymph duct cannulated rat model was studied to precisely assess the extent of lymphatic transport.Results: Mean ± SD ivacaftor oral bioavailability was 18.4 ± 3.2% and 16.2 ± 7.8%, respectively, when administered as an aqueous suspension and an oil solution. The relative contribution of the lymphatic transport to the overall bioavailability was 5.91 ± 1.61% and 4.35 ± 1.84%, respectively.Conclusion: Lymphatic transport plays only a minor role in the process of ivacaftor intestinal absorption, and other factors are, therefore, responsible for its pronounced positive food effect.

Layered crystal structures tend to form flat platelet-like crystallites, and nanofibers having such a structure exhibit strip-like morphology. Crystallographic plane forming the dominant flat surface of the nanofibers can be used for surface modification with catalytically active nanoparticles capable of anchoring to the dominant flat surface. In this study, polyvinylidene fluoride (PVDF) nanofibers exhibiting strip-like morphology and longitudinal folding were prepared using wire electrospinning, and surface modified with CeO2 nanoparticles. Experimental characterization of the CeO2/PVDF membrane using (high-resolution) scanning electron microscopy and X-ray photoelectron spectroscopy was supplemented by a force field-based molecular modeling. The modeling has shown that the dominant PVDF(100) plane is suitable for anchoring the CeO2 nanoparticles. In this respect, the PVDF(100) plane is comparable to the less exposed fluorine-oriented PVDF(010) plane, and both planes show stronger interaction with CeO2 compared to hydrogen-oriented PVDF(010) plane. Molecular modeling also revealed preferred crystallographic orientations of anchored CeO2 nanoparticles: these are the catalytically active planes (100), (110), and (111). The successful surface modification and the finding that CeO2 nanoparticles on the dominant PVDF(100) surface can preferentially exhibit these crystallographic orientations thus provides the possibility of various practical applications of the CeO2/PVDF membrane.

Allexivirus (Alphaflexiviridae) was first described in 1970 by Razvjazkina. Since then, Allexivirus species have been detected in many countries. Although this genus primarily infects plants in the Amaryllidaceae, other hosts include plants in the Fabaceae, Rosaceae and Orchidaceae. Thirteen Allexivirus species have been assigned. Eight of these infect Allium hosts, and these include: shallot virus X (ShVX), garlic virus A (GarV-A), garlic virus B (GarV-B), garlic virus C (GarV-C), garlic virus D (GarV-D), garlic virus E (GarV-E), garlic virus X (GarV-X), and garlic-mite filamentous virus (GarMbFV). Five have been described from non-Allium hosts, including blackberry virus E (BVE), vanilla latent virus (VLV), alfalfa virus S (AVS), Arachis pintoi virus (ApV), and Senna severe yellow mosaic virus (SSYMV). This review analyzes the taxonomic positions of the thirteen recognized species and four unassigned species (Allexivirus DS-2013/CZE isolate, shallot mite-borne latent virus (SMbLV), cassia mild mosaic virus (CaMMV), and papaya virus A (PaVA)). Based on the inspection of data, we have concluded that PaVA is an Allexivirus, DS-2013/CZE is an isolate of GarV-D, and SMbLV is an isolate of ShVX. Current knowledge of the host ranges, symptoms, genome structure and modes of transmission of these viruses is also summarized, and control measures employed against them are outlined.

Data on gastric pH in rats to be used in preclinical models for pH-dependent drug absorption are still limited or contradictory. The aim of this study was to describe gastric pH in rats at fasted state and to evaluate its changes induced by pentagastrin or omeprazole in order to mimic gastric pH at fasted and fed human subjects. Twenty Wistar rats, fasting for 12 h, were randomly assigned into four treatment groups (n=5): control, pre-treated with omeprazole 2 h before pH measurement, pre-treated with omeprazole 12 h before pH measurement, and pre-treated with pentagastrin 20 min before pH measurement. An incision on the stomach wall was made in anesthetized animals, and pH of gastric juice was measured. The observed pH values were significantly different among groups (p=0.0341), with the median (IQR) values of gastric pH of 3.5 (2.7–4.2), 6.7 (4.7–7.0), 5.6 (3.5–6.4) and 2.2 (1.6–3.1) in control, omeprazole 2 h, omeprazole 12 h and pentagastin group, respectively. We recommend using short interval pentagastrin and 2 h omeprazole pre-treatment in fasting animals to model similar gastric pH as is expected in human fasted and fed state pharmacokinetic studies, respectively.

The objective of this prospective study was to examine the exposure to the main active metabolites of ciprofloxacin in critically ill patients and to examine the factors (demographic, laboratory and genetic) that could potentially affect the drug metabolic conversion of ciprofloxacin. The secondary aim was to develop a population pharmacokinetic model for the metabolite showing the most associations with the abovementioned factors. A total of 29 patients were treated with intravenous infusion of ciprofloxacin and enrolled on this trial. Blood samples for pharmacokinetic analysis were taken at 1, 4, and 11.5 h following the completion of the infusion. Sex, age, body weight, height, serum creatinine and bilirubin levels, and creatinine clearance (CLCR) were recorded, and polymorphisms rs2032582 and rs1045642 in the ABCB1 gene, rs4148977 in the SLCO1A2 gene and rs762551 in the CYP1A2 gene were analyzed. A three-stage parent drug–metabolite population pharmacokinetic model was developed. Median (IQR) metabolite/parent ratios of the desethylene ciprofloxacin, formyl ciprofloxacin and oxociprofloxacin were 5.86 (4.09–9.87)%, 4.08 (3.38–6.92)% and 5.91 (3.42–13.65)%, respectively. The desethylene ciprofloxacin metabolic ratio was positively associated with height (r2 = 0.2277, p = 0.0089) and CLCR (r2 = 0.2023, p = 0.0144) and negatively associated with age (r2 = 0.2227, p = 0.0112). Males had a significantly higher oxociprofloxacin metabolic ratio than females (9.14 vs 3.42%, p = 0.0043). In the desethylene ciprofloxacin population PK model, the volume of distribution decreased with age, the parent drug-metabolite transfer rate constant increased with CLCR, and the metabolite elimination rate constant decreased with age and is increased in CYP1A2 rs762551 variant allele carriers. We therefore hypothesized that the CYP1A2 inhibition by ciprofloxacin is mediated by its metabolite desethylene ciprofloxacin.

One of the major concerns for all in vivo experiments is intra- and inter-subject variability, which can be a great source of inaccuracy. The aim of this study is, therefore, to estimate the ability of parallel vs. cross-over design studies in order to describe the relative pharmacokinetic performance of the studied drug formulations. We analyzed the data from a drug development program that examined the performance of innovative abiraterone acetate formulations against the identical reference product in three stages. In stages 1–3, groups A–F were dosed with the reference product once in a parallel manner. Stage 4 was performed to evaluate the intra-individual variability (IIV) by repeated administration of the reference product to the same animals. Although the geometric mean (90% CI) values of abiraterone AUClast in groups A–F were similar to the IIV group (24.36 (23.79–41.00) vs. 26.29 (20.56–47.00) mg/mL·min·g), the results generated in the isolated parallel groups provided imprecise estimates of the true AUClast values ranging from 9.62 to 44.62 mg/mL·min·g due to chance. Notably, in 4 out of 15 possible pair comparisons between the parallel groups, the confidence intervals did not include 100%, which is the true ratio for all comparisons tested after identical formulation administration to all groups. A cross-over design can significantly improve the methodology in short-term comparative pre-clinical pharmacokinetic studies, and can provide more precise and accurate results in comparison to more traditional pre-clinical study designs.

In the Czech Republic, demethylation inhibitors (DMIs) are used both as fungicides in controlling phoma stem canker and as growth regulators. This heavy use can result in the development of resistant isolates. A total of 45 and 286 Leptosphaeria maculans isolates were tested in vitro, using the mycelial growth and microtiter plate assays, respectively. The objective was to determine the sensitivity of L. maculans isolates collected in the Czech Republic to the fungicides tetraconazole, metconazole, and prochloraz. The mean EC50 values with the mycelial growth plate method were 1.33, 0.78, and 0.40 µg mL−1 for tetraconazole, metconazole, and prochloraz, respectively. The mean EC50 values for the microtiter plate assay were 3.01, 0.44, and 0.19 µg mL−1 for tetraconazole, metconazole, and prochloraz, respectively. All three fungicides also had high variation factors that may be due to inserts in the ERG11 promoter region. In addition, cross sensitivity among the three fungicides was observed. Overall, the high variation factors and the PCR (polymerase chain reaction) results observed in this study could signify the presence of resistant isolates in L. maculans Czech populations, especially in isolates tested for sensitivity to tetraconazole.

A total of 187 naturally late blight‑diseased potato leaves were sampled from 31 sites and five regions of the Czech Republic during the growing season in 2012–2014 and 2016 and examined microscopically for the ability of Phytophthora infestans to produce oospores in infected leaves under field conditions. Although the occurrence of the A1 and A2 mating types required for sexual reproduction of P. infestans was previously confirmed in the Czech Republic, no oospores were detected in this study. To study the effect of temperature on the survival of oospores produced from crosses of three pairs of P. infestans isolates, oospores in leaf discs and agar were exposed to temperatures ranging from –24 to 10 °C and then evaluated for viability with the plasmolysis test. Oospore viability ranged from 13.02 to 63.90 % and from 7.77 to 63.37 % for oospores produced in agar and leaf discs, respectively, with the highest frequencies of viable oospores occurring at 4 and 10 °C. To determine whether oospores may survive under field conditions, agar plates with oospores were buried in soil in Prague‑Suchdol and Svitavy for seven months during the overwintering period (October–May) in 2011–2015. There were statistically significant differences in oospore viability examined by the plasmolysis test between the localities, and oospore viability ranged from 29 to 43 % and from 15 to 44 % in Prague‑Suchdol and Svitavy, respectively.

The aim of this prospective PK study was to evaluate the pharmacokinetics of ciprofloxacin dosed within the first 36 h (early phase) and after 3 days of treatment (delayed phase) using individual and population PK analysis. The secondary aim of the study was to evaluate possible dosing implications of the observed PK differences between early and delayed phases to achieve a PK/PD target for ciprofloxacin of AUC24/MIC ≥ 125. Blood concentrations of ciprofloxacin (1 and 4 h after dose and trough) were monitored in critically ill adults in the early and delayed phases of the treatment. Individual and population PK analyses were performed. Complete concentration-time profiles in the early phase, delayed phase, and both phases were obtained from 29, 15, and 14 patients, respectively. No systematic changes in ciprofloxacin PK parameters between the early and delayed phases were observed, although variability was higher at the early phase. Both individual and population analyses provided similar results. Simulations showed that after standard dosing, it is practically impossible to reach the recommended ciprofloxacin PK/PD target (AUC/MIC ≥ 125) for pathogens with MIC ≥ 0.5 mg/L. A dosing nomogram utilizing patients’ creatinine clearance and MIC values was constructed. Both individual and population analyses provided similar results. Therapeutic drug monitoring should be implemented to safeguard the optimal ciprofloxacin exposure.

Plant viruses are important pathogens that cause significant crop losses. A plant protein extraction protocol that combines crushing the tissue by a pestle in liquid nitrogen with subsequent crushing by a roller-ball crusher in urea solution, followed by RuBisCO depletion, reduction, alkylation, protein digestion, and ZipTip purification allowed us to substantially simplify the sample preparation by removing any other precipitation steps and to detect viral proteins from samples, even with less than 0.2 g of leaf tissue, by a medium resolution nanoLC-ESI-Q-TOF. The presence of capsid proteins or polyproteins of fourteen important viruses from seven different families (Geminiviridae, Luteoviridae, Bromoviridae, Caulimoviridae, Virgaviridae, Potyviridae, and Secoviridae) isolated from ten different economically important plant hosts was confirmed through many identified pathogen-specific peptides from a protein database of host proteins and potential pathogen proteins assembled separately for each host and based on existing online plant virus pathogen databases. The presented extraction protocol, combined with a medium resolution LC-MS/MS, represents a cost-efficient virus protein confirmation method that proved to be effective at identifying virus strains (as demonstrated for PPV, WDV) and distinct disease species of BYDV, as well as putative new viral protein sequences from single-plant-leaf tissue samples. Data are available via ProteomeXchange with identifier PXD022456.

Apple mosaic, rose mosaic, plum and birch line pattern, horse chestnut yellow mosaic, and hazel mosaic are some of many diseases caused by one of the longest-known pome fruit viruses: Apple mosaic virus. The worldwide distribution of Apple mosaic virus and its negative impacts on fruit, nut and hop production are sufficient to cause international interest. This review endeavors to piece together all known information about the virus. We describe taxonomic position, virion structure, the host range and symptoms, modes of transmission and diagnostics for the virus, as well as distribution and control of the diseases it causes.

The aim of the study was to evaluate the effect of temperature on apoptosis of bovine blood eosinophil granulocytes in vitro. Heparinised bovine blood was incubated for 1, 4 and 24 h under following temperatures: 4, 23 and 37 °C. UV irradiation was used as positive control of apoptosis. Eosinophil granu­locytes apoptosis was detected by flow cytometry after simultaneous staining with Annexin-V and propidium iodide. From selected temperatures, 4 °C induced the eosinophil granulocytes apoptosis least. The proportion of apoptotic eosinophil granulocytes amounted to (mean ± SD) 1.65 ± 0.46%; 1.76 ± 0.36%; 4.78 ± 1.70% after 1, 4 and 24 h incubation, respectively.

The study was an in vitro analysis of bovine blood neutrophil apoptosis and necrosis based on the detection of their morphological and biochemical features. The experiment was carried out in six clinically healthy Holstein x Bohemian Red Pied crossbred virgin heifers aged 16 to 18 months. The fresh and in vitro cultivated blood were analysed by light and electron microscopy and flow cytometry. In fresh blood, non apoptotic or necrotic neutrophils were found by light microscopy. On the other hand, 3.34% apoptotic and 0.32% necrotic neutrophils were detected by flow cytometry. After four hours of incubation, 3.53% apoptotic and 1.37% necrotic neutrophils were found by light microscopy. In the same conditions, there were assessed 17.54% apoptotic and 0.58% necrotic neutrophils by flow cytometry. Correlation coefficient between light microscopy and flow cytometry in portion of apoptotic neutrophils was 0.625 (P

The aim of the student thesis was to review the effect of the significant bacterial pathogens, Staphylococcus aureus and Steptococcus uberis on programmed cell death – apoptosis in vitro, and to determine the dynamic of morphological changes during aging neutrophil granulocytes of heifers mammary gland in vitro. Using light and electron microscopy we have noted characteristic alterations of apoptotic neutrophils. These are running in three consequential phases. We found out, that the interaction of bacterial pathogens with neutrophils during the incubation have lead in expressive quantitative and qualitative changes in apoptotic cells proportion. Concretely, the influence of both pathogens on mammary gland neutrophils caused the defer of apoptosis expression. Here, S. aureus caused lower number of apoptotic neutrophils in comprasion with S. uberis. The outcomes testified, that S. aureus and S. uberis interaction with heifers mammary gland neutrophils in vitro causes alterations relating to apoptosis of these cells. Looking at the results of the study, we can conclude, that the pathogens S. aureus and S. uberis are not only significant heifers mammary gland – affection causers, but they significantly influence the cells of defensive system in their functions too. They significantly decrease the appereance of morphological apoptosis manifestations on neotrophils of tissue pool of the heifers mammary gland. The numbers of apoptotic cells in neutrophil population confirm, that during the interaction with mentioned pathogens the defer of morphological apoptosis manifestations happens. Then, higher number of apoptotic neutrophils in stages of apoptotic corpuscles implies increasing dynamic of this process. Beside that, the dynamic of apoptotic process is influenced by the specifity of certain bacterious actor too.

Recently, it has been shown that the stem nematode, Ditylenchus dipsaci (Nematoda: Anguinidae), is genetically more related to the gall-forming nematodes from genera Anguina, Heteroanguina, and Mesoanguina than to other members of the genus Ditylenchus. This finding was provided by molecular data written in the evolutionary variable, non-coding internal transcribed spacers (ITS1 and ITS2) of the ribosomal DNA (rDNA). In the current paper, we analyze the nucleotide sequences and predict the secondary structures of two expansion segments (D2, D3) of the 28S ribosomal RNA (rRNA)-coding gene for the plant-parasitic nematodes from the genus Ditylenchus and their related anguinids. In general, the expansion segment D2 appeared to be more variable than the segment D3 illustrating their different evolutionary constraints. Comparative analysis of the aligned sequences and predicted secondary structures revealed similar trend showing the tight relationships between the stem nematodes (D. dipsaci, D. gigas, D. weischeri) and gall-forming nematodes from the subfamily Anguininae. Phylogeny reconstructions disjoined the family Anguinidae into two monophyletic clusters (Clade 1 and 2). Clade 1 constitutes the stem nematodes (D. dipsaci, D. gigas, etc) and gall-forming nematodes from the genera Anguina, Heteroanguina, Subanguina and Mesoanguina, while clade 2 includes other Ditylenchus species like D. destructor and D. halictus. Collectively, deciphering the exact phylogenetic relationships within the family Anguinidae (Nematoda: Tylenchida) with respect to our results should provide a framework for a taxonomic revision in order to reflect biological history of these nematodes. In addition, we provide novel molecular data, which may be exploited in diagnostic tools for phytosanitary control of these economically important plant parasites.