Systematic Metabolomic Analysis of Eicosanoids after Omega‑3 Polyunsaturated Fatty Acid Supplementation by a Highly Specific Liquid Chromatography–Tandem Mass Spectrometry-Based Method

Omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) have beneficial effects in many pathological processes, especially cardiovascular disease, and their protective eicosanoid metabolites are thought to play important roles. However, how ω-3 PUFAs affect the eicosanoid profile has not been elucidated comprehensively. Here, we systematically analyzed the eicosanoid metabolites induced by ω-3 PUFA supplementation. We developed an LC–MS/MS-based method covering 32 arachidonic acid (ARA) metabolites and 37 ω-3 PUFA-derived products. The limits of detection for eicosanoids were between 0.0625 and 1 pg and the detection specificity was optimized. We then quantified eicosanoids in mouse and human plasma and mouse aorta samples after ω-3 PUFA supplementation. Levels of EPA hydroxyl products, 4-HDoHE, 17,18-EEQ, 17,18-DiHETE, TXB2, and LXA4 were significantly changed in both mouse samples, and those of 2-series PGs, EDPs and DHA hydroxyl products were changed in aorta samples. Correlation network analysis of mouse plasma data revealed that some eicosanoids had higher connection degree or betweenness centrality score than others after ω-3 PUFA supplementation. Eicosanoids in human plasma were profiled across five time points after ω-3 PUFA supplementation. Fuzzy c-mean clustering algorithm suggested that the time curves of eicosanoid activity could be described with three kinetic patterns: sustained upregulation, short-term upregulation, and downregulation. This is the first systematic profiling of eicosanoids with ω-3 PUFA supplementation. The highly specific eicosanoid metabolomic and related data analysis methods would be powerful tools for comprehensive eicosanoid study

Technology implementation: What does the future hold for construction?

The BRANZ supported research project, Technology adoption roadmap for the New Zealand construction sector, began in 2019 and involved extensive data collection. A number of insights were identified from the research that will prove helpful for industry planning to adopt new technology. It includes a survey of technology use internationally and in NZ

Anharmonic Densities of States for Vibrationally Excited I^–(H₂O), (H₂O)₂, and I^–(H₂O)₂

Monte Carlo sampling calculations were performed to determine the anharmonic sum of states, <i>N</i>_anh(<i>E</i>), for I^–(H₂O), (H₂O)₂, and I^–(H₂O)₂ versus internal energy up to their dissociation energies. The anharmonic density of states, ρ_anh(<i>E</i>), is found from the energy derivative of <i>N</i>_anh(<i>E</i>). Analytic potential energy functions are used for the calculations, consisting of TIP4P for H₂O···H₂O interactions and an accurate two-body potential for the I^–···H₂O fit to quantum chemical calculations. The extensive Monte Carlo samplings are computationally demanding, and the use of computationally efficient potentials was essential for the calculations. Particular emphasis is directed toward I^–(H₂O)₂, and distributions of its structures versus internal energy are consistent with experimental studies of the temperature-dependent vibrational spectra. At their dissociation thresholds, the anharmonic to harmonic density of states ratio, ρ_anh(<i>E</i>)/ρ_h(<i>E</i>), is ∼2, ∼ 3, and ∼260 for I^–(H₂O), (H₂O)₂, and I^–(H₂O)₂, respectively. The large ratio for I^–(H₂O)₂ results from the I^–(H₂O)₂ → I^–(H₂O) + H₂O dissociation energy being more than 2 times larger than the (H₂O)₂ → 2H₂O dissociation energy, giving rise to highly mobile H₂O molecules near the I^–(H₂O)₂ dissociation threshold. This work illustrates the importance of treating anharmonicity correctly in unimolecular rate constant calculations

Targeting interleukin 4 and interleukin 13: a novel therapeutic approach in bullous pemphigoid

Aim: Bullous pemphigoid (BP) is an organ-specific autoimmune bullous disease characterized by autoantibodies that target the cellular adhesion molecules BP180 and BP230. Both immunoglobulin (Ig)G and IgE are involved in the induction of subepidermal blisters. Specifically, IgE autoantibodies are presumed to be responsible for the pruritic and erythematous features of BP. Histologically, eosinophil infiltration is a prominent feature in BP. Eosinophils and IgE are mostly associated with the Th2 immune response. Th2 cytokines, particularly interleukin (IL)-4 and IL-13, are presumed to contribute to the pathology of BP. The aim of this review is to discuss the role of IL-4/13 in the pathogenesis of BP and the potential of using IL-4/13 antagonists for treatment.Methods: After searching in PubMed and Web of Science databases using ‘bullous pemphigoid’, ‘interleukin-4/13’, and ‘dupilumab’ as keywords, studies related was compiled and examined.Results: Overall, IgE, eosinophils, IL-4, and IL-13 may interact with each other in the pathogenesis of BP; these potential interactions provide clues concerning targets for molecular treatment.Conclusion: Anti-IL-4/13 treatment has been experimentally used in patients with BP, with satisfactory outcomes and few side effects. However, before this novel therapy can be approved for regular usage, further studies are needed concerning the long-term safety and systemic usage of IL-4/13 monoclonal antibody treatment in BP.KEY MESSAGESBP is an autoimmune skin disease with Th2-mediated autoimmune response involvement.As typical Th2 cytokines, IL-4 and IL-13 may contribute to the pathogenesis of BP in multiple ways, such as promoting Th2 cell polarization, driving the immunoglobulin class switching, recruiting eosinophils and basophils, and inducing pruritus.As a promising therapeutic approach for BP, IL-4/13 antagonists have shown satisfactory outcomes in preliminary clinical studies. BP is an autoimmune skin disease with Th2-mediated autoimmune response involvement. As typical Th2 cytokines, IL-4 and IL-13 may contribute to the pathogenesis of BP in multiple ways, such as promoting Th2 cell polarization, driving the immunoglobulin class switching, recruiting eosinophils and basophils, and inducing pruritus. As a promising therapeutic approach for BP, IL-4/13 antagonists have shown satisfactory outcomes in preliminary clinical studies.</p

Unmasking Rare, Large-Amplitude Motions in D₂‑Tagged I^–·(H₂O)₂ Isotopomers with Two-Color, Infrared–Infrared Vibrational Predissociation Spectroscopy

We describe a two-color, isotopomer-selective infrared–infrared population-labeling method that can monitor very slow spectral diffusion of OH oscillators in H‑bonded networks and apply it to the I^–·(HDO)·(D₂O) and I^–·(H₂O)·(D₂O) systems, which are cryogenically cooled and D₂-tagged at an ion trap temperature of 15 K. These measurements reveal very large (>400 cm^–1), spontaneous spectral shifts despite the fact that the predissociation spectra in the OH stretching region of both isotopologues are sharp and readily assigned to four fundamentals of largely decoupled OH oscillators held in a cyclic H-bonded network. This spectral diffusion is not observed in the untagged isotopologues of the dihydrate clusters that are generated under the same source conditions but does become apparent at about 75 K. These results are discussed in the context of the large-amplitude “jump” mechanism for H-bond relaxation dynamics advanced by Laage and Hynes in an experimental scenario where rare events can be captured by following the migration of OH groups among the four available positions in the quasi-rigid equilibrium structure

Presentation_1.PDF

<p>Phosphorus (P) is an important nutrient, whose plant-available form phosphate is often low in natural forest ecosystems. Mycorrhizal fungi mine the soil for P and supply their host with this resource. It is unknown how ectomycorrhizal communities respond to changes in P availability. Here, we used young beech (Fagus sylvatica L.) trees in natural forest soil from a P-rich and P-poor site to investigate the impact of P amendment on soil microbes, mycorrhizas, beech P nutrition, and photosynthesis. We hypothesized that addition of P to forest soil increased P availability, thereby, leading to enhanced microbial biomass and mycorrhizal diversity in P-poor but not in P-rich soil. We expected that P amendment resulted in increased plant P uptake and enhanced photosynthesis in both soil types. Young beech trees with intact soil cores from a P-rich and a P-poor forest were kept in a common garden experiment and supplied once in fall with triple superphosphate. In the following summer, labile P in the organic layer, but not in the mineral top soil, was significantly increased in response to fertilizer treatment. P-rich soil contained higher microbial biomass than P-poor soil. P treatment had no effect on microbial biomass but influenced the mycorrhizal communities in P-poor soil and shifted their composition toward higher similarities to those in P-rich soil. Plant uptake efficiency was negatively correlated with the diversity of mycorrhizal communities and highest for trees in P-poor soil and lowest for fertilized trees. In both soil types, radioactive P tracing (H₃³³PO₄) revealed preferential aboveground allocation of new P in fertilized trees, resulting in increased bound P in xylem tissue and enhanced soluble P in bark, indicating increased storage and transport. Fertilized beeches from P-poor soil showed a strong increase in leaf P concentrations from deficient to luxurious conditions along with increased photosynthesis. Based on the divergent behavior of beech in P-poor and P-rich forest soil, we conclude that acclimation of beech to low P stocks involves dedicated mycorrhizal community structures, low P reserves in storage tissues and photosynthetic inhibition, while storage and aboveground allocation of additional P occurs regardless of the P nutritional status.</p

Table_1_Genome-wide survey of Calcium-Dependent Protein Kinases (CPKs) in five Brassica species and identification of CPKs induced by Plasmodiophora brassicae in B. rapa, B. oleracea, and B. napus.xlsx

Calcium-dependent protein kinase (CPK) is a class of Ser/Thr protein kinase that exists in plants and some protozoa, possessing Ca2+ sensing functions and kinase activity. To better reveal the roles that Brassica CPKs played during plant response to stresses, five Brassica species, namely Brassica rapa (B. rapa), Brassica nigra (B. nigra), Brassica oleracea (B. oleracea), Brassica juncea (B. juncea), and Brassica napus (B. napus) were selected and analyzed. In total, 51 BraCPK, 56 BniCPK, 56 BolCPK, 88 BjuCPK, and 107 BnaCPK genes were identified genome wide and phylogenetics, chromosomal mapping, collinearity, promoter analysis, and biological stress analysis were conducted. The results showed that a typical CPK gene was constituted by a long exon and tandem short exons. They were unevenly distributed on most chromosomes except chromosome A08 in B. napus and B. rapa, and almost all CPK genes were located on regions of high gene density as non-tandem form. The promoter regions of BraCPKs, BolCPKs, and BnaCPKs possessed at least three types of cis-elements, among which the abscisic acid responsive-related accounted for the largest proportion. In the phylogenetic tree, CPKs were clustered into four primary groups, among which group I contained the most CPK genes while group IV contained the fewest. Some clades, like AT5G23580.1(CPK12) and AT2G31500.1 (CPK24) contained much more gene members than others, indicating a possibility that gene expansion occurred during evolution. Furthermore, 4 BraCPKs, 14 BolCPKs, and 31 BnaCPKs involved in the Plasmodiophora brassicae (P. brassicae) defense response in resistant (R) or susceptible (S) materials were derived from online databases, leading to the discovery that some R-specific induced CPKs, such as BnaC02g08720D, BnaA03g03800D, and BolC04g018270.2J.m1 might be ideal candidate genes for P. brassicae resistant research. Overall, these results provide valuable information for research on the function and evolution of CDK genes.</p

The issue of disability in the Czech Republic

The bachelor thesis "The issue of disability in the Czech Republic" deals with the system of disability assesment and assigning of disability pension and the next social help, which is designed for the handicapped people, with concentration on the changes, which happened in 2010 and 2012. The aim of this thesis is to assess the system, show how the system works in practice and present the advantages and disadvantages of the system. The first part shows the question in theory, explain the basic terms and relevant legislation and tries to assess on basis of existent articles or interviews with experts. The second part deals with analysis of interviews, which were made with disabled people, medical assessor and officer from OSSZ. Aim is to show how the system works in practice and how is assessed by mentioned participants and by the author

Table_2_Genome-wide survey of Calcium-Dependent Protein Kinases (CPKs) in five Brassica species and identification of CPKs induced by Plasmodiophora brassicae in B. rapa, B. oleracea, and B. napus.xls

Calcium-dependent protein kinase (CPK) is a class of Ser/Thr protein kinase that exists in plants and some protozoa, possessing Ca2+ sensing functions and kinase activity. To better reveal the roles that Brassica CPKs played during plant response to stresses, five Brassica species, namely Brassica rapa (B. rapa), Brassica nigra (B. nigra), Brassica oleracea (B. oleracea), Brassica juncea (B. juncea), and Brassica napus (B. napus) were selected and analyzed. In total, 51 BraCPK, 56 BniCPK, 56 BolCPK, 88 BjuCPK, and 107 BnaCPK genes were identified genome wide and phylogenetics, chromosomal mapping, collinearity, promoter analysis, and biological stress analysis were conducted. The results showed that a typical CPK gene was constituted by a long exon and tandem short exons. They were unevenly distributed on most chromosomes except chromosome A08 in B. napus and B. rapa, and almost all CPK genes were located on regions of high gene density as non-tandem form. The promoter regions of BraCPKs, BolCPKs, and BnaCPKs possessed at least three types of cis-elements, among which the abscisic acid responsive-related accounted for the largest proportion. In the phylogenetic tree, CPKs were clustered into four primary groups, among which group I contained the most CPK genes while group IV contained the fewest. Some clades, like AT5G23580.1(CPK12) and AT2G31500.1 (CPK24) contained much more gene members than others, indicating a possibility that gene expansion occurred during evolution. Furthermore, 4 BraCPKs, 14 BolCPKs, and 31 BnaCPKs involved in the Plasmodiophora brassicae (P. brassicae) defense response in resistant (R) or susceptible (S) materials were derived from online databases, leading to the discovery that some R-specific induced CPKs, such as BnaC02g08720D, BnaA03g03800D, and BolC04g018270.2J.m1 might be ideal candidate genes for P. brassicae resistant research. Overall, these results provide valuable information for research on the function and evolution of CDK genes.</p

Exploring the Influence of Biologically Relevant Ions on the Corrosion Behavior of Biodegradable Zinc in Physiological Fluids

This work presents a study on the influence of biologically relevant ions on the corrosion of zinc (Zn) in physiological fluids. Electrochemical techniques were used to investigate the degradation of pure Zn exposed to different physiological electrolytes containing chlorides, carbonates, sulfates, and phosphates. The corrosion behavior of Zn in the solutions over a 7-day period was also assessed. SEM, EDS, and FTIR were used to analyze corrosion products. With respect to corrosion, the most aggressive ions are chlorides, which induce localized corrosion, while carbonates and phosphates reduce the corrosive attack of the chloride on Zn while inducing uniform corrosion. Sulfates reduce the corrosion rate by disrupting Zn’s passive layer. The overall corrosion rate of Zn changed in each electrolyte depending on the nature of the solution and the corrosion product formed. These findings will be useful in predicting the in-service behavior of future biodegradable Zn medical implants