Porcine MHC classical class I genes are coordinately expressed in superantigen-activated mononuclear cells

The expression of the major histocompatibility complex (MHC) classical class I genes is important for the adaptive immune response to target virus-infected cells and cancer cells. The up-regulation of the MHC is achieved by hormonal/cytokine signals including IFN-γ-inducible elements. The swine leukocyte antigen (SLA), the MHC class I region of pigs, consists of the duplicated classical class I genes, SLA-1, SLA-2 and SLA-3, but the molecular mechanisms involved in their up-regulation after T cell stimulation have not been fully elucidated. In order to better understand some of the putative regulatory mechanisms of SLA class I gene expression in activated T cells, we examined the coordinated expression of the SLA classical class I, IFN-γ and interferon regulatory factor-1 (IRF-1) genes in the peripheral blood mononuclear cells (PBMCs) of SLA homozygous Clawn miniature swine stimulated for 72h with either IFN-γ or an enterotoxin produced by Staphylococcus aureus. This enterotoxin, toxic shock syndrome-1 (TSST-1), is known to act as a superantigen (sAG) to activate the T cells in various vertebrate species. We showed by using mAbs and flow cytometry that the CD4(+)CD25(+) cell number of swine PBMCs was also increased by TSST-1 and to a lesser degree by IFN-γ. Time course analyses of the expression of the IFN-γ, IRF-1 and the three classical class I genes, SLA-1, SLA-2, and SLA-3, in PBMCs by quantitative real-time PCR revealed a transitory response to TSST-1 or IFN-γ stimulation. The IFN-γ mRNA levels in the PBMCs were continuously up-regulated over the first 48h by TSST-1 or IFN-γ. In contrast, SLA class I expression moderately increased at 24h and then decreased to a baseline level or less at 72h of IFN-γ or TSST-1 stimulation. The three classical SLA class I genes showed similar expression kinetics, although SLA-3 mRNA level was consistently lower than those of SLA-1 and -2. The expression of IRF-1, a modulator of SLA expression, showed similar kinetics to those of the three classical SLA class I genes. The expression profiles detected by flow cytometry of the SLA molecules on the cell surface of PBMCs were maintained at a consistently high level during cell stimulation with either TSST-1 or IFN-γ, which was distinct from the kinetics of mRNA expression. These results showed that miniature swine SLA class I mRNA expression was effectively and equally up-regulated among the three loci and coordinately with IRF-1 gene expression after stimulation of T cell activation by sAG or IFN-γ.     

The effect of copper nanoparticles and copper (II) salt on redox reactions and epigenetic changes in a rat model

The aim of the study was to evaluate the effects of a diet containing different levels of Cu in two different chemical forms (carbonate and nanoparticles) on redox reactions and epigenetic changes in a rat model. For 4 weeks, five experimental groups (eight rats in each) were fed diets with two dosages of added Cu (standard—6.5 mg/kg or half of the standard dosage—3.25 mg/kg, and as a negative control no additional Cu in the mineral mixture) in two forms (standard—CuCO₃ and copper nanoparticles). Addition of Cu nanoparticles resulted in higher Cp (ceruloplasmin) activity and LOOH (lipid peroxides) and MDA (malondialdehyde) content, as well as decrease the CAT (catalase) activity and level of PC (protein carbonyl), 3‐NT (3‐nitrotyrosine), 8‐OHdG (8‐hydroxydeoxyguanosine), GSH + GSSG (total glutathione) and DNA methylation. Reducing the dose of copper resulted in a decrease in the level of LOOH and GSH + GSSG as well as CAT activity, but increased the level of PC and methylated DNA. Based on these evidence, we concluded that addition of copper nanoparticles in the diet reduces protein oxidation and nitration as well as DNA oxidation and methylation. Lowering the level of Cu in the diet increases the oxidation of proteins and DNA methylation.     

Role of tumor necrosis factor-alpha and nitric oxide in luteolysis in cattle

Although prostaglandin (PG) F2alpha is known to be a principal luteolytic factor, its action on the bovine corpus luteum (CL) is mediated by other intra-ovarian factors. Tumor necrosis factor-alpha (TNFalpha) and its specific receptors are present in the bovine CL with the highest expressions at luteolysis. TNFalpha in combination with interferon-gamma reduced progesterone (P4) secretion, increased PGF2alpha and leukotriene C4 (LTC4) production, and induced apoptosis of the luteal cells in vitro. Low concentrations of TNFalpha caused luteolysis, which resulted in a decreased level of P4, and increased levels of PGF2alpha, LTC4 and nitrite/nitrate (stable metabolites of nitric oxide-NO) in the blood. Inhibition of local NO production counteracts spontaneous and PGF2alpha-induced luteolysis. Therefore, NO is a likely candidate for the molecule that mediates PGF2alpha and TNFalpha actions during luteolysis. Both PGF2alpha and TNFalpha increase NO concentrations in blood, and stimulate NO synthase expression on protein level in the bovine CL cells. NO stimulates PGF2alpha and LTC4 secretion, inhibits P4 production and reduces the number of viable luteal cells. TNFalpha and NO induce apoptotic death of the CL by modulating expression of bcl-2 family genes and by stimulating expression and activity of caspase-3. The above findings indicate that TNFalpha and NO play crucial roles in functional and structural luteolysis in cattle.     

Influence of oral co-administration of a preparation containing calcium and magnesium and food on enrofloxacin pharmacokinetics

The objective of this study has been to determine the influence of food and ions on the pharmacokinetics of enrofloxacin (ENRO) in turkeys, administered per os at a dose of 10 mg/kg of body weight (b.w.). Co-administration of ENRO with ions or with food significantly retarded its absorption, and the interaction was more pronounced when the drug was given together with food. The bioavailability of ENRO was 65.78 ± 7.81% and 47.99 ± 9.48% with ions and food, respectively. The maximum concentration (C_max) in plasma of animals exposed to ions reached 0.87 ± 0.26 μg/ml in a t_max of 2.07 ± 0.76 h; in animals which were fed while medicated, the analogous parameters were 0.36 ± 0.13 μg/ml and 8.06 ± 3.08 h. The PK/PD analysis demonstrated that a decrease in the concentration of ENRO in turkeys’ blood due to the interaction with ions or food might impair the drug's clinical efficacy toward some pathogenic microorganisms in turkeys if a routine dose of 10 mg ENRO/kg b.w. is administered.     

Loss in macronutrient pools in bilberry and lingonberry in mesic Scots pine forests after Northern red oak introduction

Forest transformation from coniferous monocultures to mixed stands is being promoted worldwide, including the introduction of fast-growing broadleaved tree species within native stands. Here, we studied how enrichment of temperate European Scots pine (Pinus sylvestris) forest by North-American northern red oak Quercus rubra impacted macronutrient concentrations in two long-lived and dominant components of the forest understory: bilberry Vaccinium myrtillus and lingonberry V. vitis-idaea. Study sites were located in forest complexes (central Poland) which occupy continuously reforested lands (hereafter ancient forests) as well as post-agricultural lands (recent forests), all suitable for mesic pine forests. Samples of bilberry and lingonberry leaves, stems, and fruits were collected in pine stands and in adjacent Scots pine-red oak stands, in both ancient and recent forests. Concentrations of macronutrients (C, N, P, K, Ca, S, and Mg) in aboveground biomass components were analysed using standardized chemical procedures. The study revealed intra- and interspecific (bilberry vs. lingonberry) differences in concentrations of all nutrients in leaves, stems, and fruits, except for invariable C concentrations. Macronutrient accumulations in plants were decreased by land-use discontinuity and favoured by enrichment of tree stands by Q. rubra. The estimated macronutrient pools were much higher for V. myrtillus than V. vitis-idaea in all forest types studied. They were lower in forests enriched with Q. rubra, both ancient (up to 25.5% for bilberry and 99.9% for lingonberry) and recent (46.9% and 99.9%, respectively), as well as in recent pine forest (46.6% and 81.1%, respectively) than in ancient pine forest. Higher K and S pools (39.3% and 6.5%, respectively) noted for bilberry in an ancient forest with Q. rubra were exceptions. Despite more effective accumulations of elements at the species level, macronutrient pools of Vaccinium myrtillus and V. vitis-idaea decreased significantly in the presence of introduced Q. rubra due to negative impacts of this broadleaved tree on bilberry and lingonberry cover and biomass. Therefore, the limitation of alien Q. rubra planting in sites of mesic pine forest with the abundant occurrence of V. myrtillus and/or V. vitis-idaea is recommended.

Graphic abstract

     

Species proportions by area in mixtures of Scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) and European beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.)

Scots pine (Pinus sylvestris L.) and European beech (Fagus sylvatica L.) dominate many of the European forest stands. Also, mixtures of European beech and Scots pine more or less occur over all European countries, but have been scarcely investigated. The area occupied by each species is of high relevance, especially for growth evaluation and comparison of different species in mixed and monospecific stands. Thus, we studied different methods to describe species proportions and their definition as proportion by area. 25 triplets consisting of mixed and monospecific stands were established across Europe ranging from Lithuania to Spain in northern to southern direction and from Bulgaria to Belgium in eastern to western direction. On stand level, the conclusive method for estimating the species proportion as a fraction of the stand area relates the observed density (tree number or basal area) to its potential. This stand-level estimation makes use of the potential from comparable neighboring monospecific stands or from maximum density lines derived from other data, e.g. forest inventories or permanent observations plots. At tree level, the fraction of the stand area occupied by a species can be derived from the proportions of their crown projection area or of their leaf area. The estimates of the potentials obtained from neighboring monospecific stands, especially in older stands, were poorer than those from the maximum density line depending on the Martonne aridity index. Therefore, the stand-level method in combination with the Martonne aridity index for potential densities can be highly recommended. The species’ proportions estimated with this method are best approximated by the proportions of the species’ leaf areas. In forest practice, the most commonly applied method is an ocular estimation of the proportions by crown projection area. Even though the proportions of pine were calculated here by measuring crown projection areas in the field, we found this method to underestimate the proportion by 25% compared to the stand-level approach.     

Forest recovery in set-aside windthrow is facilitated by fast growth of advance regeneration

Key message

The disturbance of a research plot by a windstorm allowed us to study the role of the seedling bank in the regeneration processes. The released advance regeneration dominated among the saplings; taller individuals retained their position until the end of the study. Pioneer species occurred sporadically. Seven years after the disturbance, the windthrow was covered by a dense thicket of young trees.

Context

The dominant role played by advance regeneration in natural regeneration processes after intense wind disturbances is still a matter of dispute.

Aims

We took advantage of a windstorm in one of our research plots to study the role of the seedling bank released by the disturbance in the regeneration processes.

Methods

We collected data in 70 plots, recording the survivorship of seedlings, annual height growth, and signs of browsing. The height ranking was analyzed with Kendall’s concordance coefficient, and the height growth rates were compared using Dunn’s test.

Results

The density of seedlings increased from 6.7/m² in 2008 to 8.1/m² in 2010 and then decreased to 1.2/m² in 2015. The density of saplings increased continuously from 0.14 to 1.9/m². The highest size differentiation occurred in sycamore maple; the individuals which were taller before the windstorm retained their position until the year 2015. The only species that was recruited mainly from germinants was European hornbeam.

Conclusion

The advance regeneration released by the windstorm played a major role in the regeneration process, while pioneer species occurred only sporadically. Seven years after the disturbance, the windthrow was already covered by a dense thicket of young trees.

     

Studies of enzymatic oxidation of TMP-fibers and lignin model compounds by a Laccase–Mediator-System using different 14C and 13C techniques

In this work, the results of the enzymatic oxidation of TMP-fibers (thermomechanical pulp) and a well-structured lignin model compound, the dehydropolymer (DHP), were investigated by different ¹⁴C and ¹³C methods, caused by a Laccase–Mediator-System (LMS). These methods are the nuclear magnetic resonance spectroscopy (¹³C-NMR) with DHP (unmarked) and the determination of the ¹⁴CO₂ release of ¹⁴C-marked DHP and TMP-fibers. The ¹³C-NMR measurements were chosen to analyze the structural changes of the LMS-treated DHP model compounds and TMP-fibers qualitatively and quantitatively. The data of ¹⁴CO₂ release give an explanation of the demethylation of DHP and TMP-fibers. The effect of the LMS is shown by comparing the results in respect of DHP and TMP-fibers, which were only treated with laccase and of an inactivated LMS as the control. Comparing the results of the ¹³C-NMR method, in particular the use of the Mediator during the enzymatical treatment, showed significant changes in the structure of the DHP. Also, the TMP-fibers were materially influenced by the LMS. The analysis of the ¹⁴CO₂ release data of the ¹⁴C-marked DHP and TMP-fibers revealed that the rate of ¹⁴CO₂ increases in the ¹⁴C-2 atom as well as in the O¹⁴CH₃ group within the first hour of Laccase–Mediator incubation. Therefore, the ¹⁴CO₂ release from the DHP was higher than from the TMP-fibers.     

Mechanisms of cuticular uptake of xenobiotics into living plants: evaluation of a logistic-kinetic penetration model

The objective of this study was to determine whether a logistic-kinetic penetration model could be applied to whole plant uptake. Uptake over 24 h was determined for three model compounds, applied in the presence and absence of surfactants, into the leaves of two plant species. Data for two time intervals were used in the model to predict uptake at intermediate intervals and compared with experimental results. Overall, the model fit the whole plant uptake data well. The study confirmed that an increase (or decrease) in active ingredient (ai) concentration or an increase in contact area will have no effect on the penetration rate factor, q, within the normal working concentration range. This enabled uptake to be predicted at different times for concentrations of ai not already studied, having first derived q for one concentration of the formulation of interest and having 24 h (maximum) uptake results for all formulations and concentrations of interest. The advantages of the models and equations described are that few variables are required, and they are simple to measure.     

Volatile compounds in Spanish, French, and American oak woods after natural seasoning and toasting

The volatile composition (volatile phenols, phenolic aldehydes, furanic compounds, lactones, phenyl ketones, and other related compounds) of Spanish oak heartwood of Quercus robur, Quercus petraea,Quercus pyrenaica, and Quercus faginea was studied by gas chromatography/mass spectrometry, in relation to the processing in barrels cooperage and in relation to the French oak of Quercus robur (Limousin) and Quercus petraea (Allier) and American oak of Quercus alba (Missouri), which are habitually used in cooperage. The volatile composition of seasoned oak woods varied according to individual trees, species, and origins, and the differences were more significant in Spanish species with respect to American species than with respect to French species. The toasting process influenced the volatile composition of wood. It led to high increases in the concentration of volatile phenols, furanic aldehydes, phenyl ketones, and other related structures, but the effect on w-lactones levels depended on species and origin. The volatile composition in Spanish oak species evolved during toasting like in French and American oak, but quantitative differences were found, especially important in American species with respect to European species.