Adventitial Tertiary Lymphoid Organs as Potential Source of MicroRNA Biomarkers for Abdominal Aortic Aneurysm

Abdominal aortic aneurysm (AAA) is an inflammatory disease associated with marked changes in the cellular composition of the aortic wall. This study aims to identify microRNA (miRNA) expression in aneurysmal inflammatory cells isolated by laser microdissection from human tissue samples. The distribution of inflammatory cells (neutrophils, B and T lymphocytes, mast cells) was evaluated in human AAA biopsies. We observed in half of the samples that adventitial tertiary lymphoid organs (ATLOs) with a thickness from 0.5 to 2 mm were located exclusively in the adventitia. Out of the 850 miRNA that were screened by microarray in isolated ATLOs (n = 2), 164 miRNAs were detected in ATLOs. The three miRNAs (miR-15a-3p, miR-30a-5p and miR-489-3p) with the highest expression levels were chosen and their expression quantified by RT-PCR in isolated ATLOs (n = 4), M1 (n = 2) and M2 macrophages (n = 2) and entire aneurysmal biopsies (n = 3). Except for the miR-30a-5p, a similar modulation was found in ATLOs and the two subtypes of macrophages. The modulated miRNAs were then evaluated in the plasma of AAA patients for their potential as AAA biomarkers. Our data emphasize the potential of miR-15a-3p and miR-30a-5p as biomarkers of AAA but also as triggers of ATLO evolution. Further investigations will be required to evaluate their targets in order to better understand AAA pathophysiology.     

Structure and analgesic properties of layered double hydroxides intercalated with low amounts of ibuprofen

Ibuprofen‐intercalated layered double hydroxides (LDH‐IBU) have been successfully synthesized via a coprecipitation method with a nominal [Al³⁺]/[Mg²⁺] ratio of 0.5 and a variable molar IBU/([Al³⁺]+[Mg²⁺]) ratio of 0, 0.15, 0.18, 0.24, 0.36, and 0.72. After an accurate determination of the composition, the nature of the intercalated species and the effective intercalation yield from to IBU, it is shown that the synthesis route used allows a good control of the quantity of intercalated IBU within the LDH framework. This results in different samples with full or partial IBU intercalation in the interlayer space in exchange of nitrate anions. The analysis of the X‐ray diffraction basal reflections reveals that the intercalation of IBU in the framework only increases the basal distances with no alteration of the brucite‐type layers. Also, a computational study used to model the positions and shapes of the basal reflections showed that the structure of the nonfully intercalated compounds follows a random interstratification scheme. Finally, three samples ranging from slightly to fully IBU‐intercalated galleries were selected for preliminary in vivo assays. These tests showed a strong tendency that after 24 hours the low yield of IBU‐intercalated compounds are almost as efficient as the fully intercalated sample.     

Simulation des Mikrorisswachstums unter Schwingbeanspruchung. Teil 1: Modell und Simulationsergebnisse,Mikrorisswachstum, Streuung,Einfluss von Beanspruchungsart und ‐höhe,Risskeimdichte und Korngröße

Simulation of fatigue microcrack growth. Part 1: Modelling and results of simulation, microcrack growth, scatter, effect of load condition, density of microcrack seeds and grain size Presented is a simulation of microcrack growth under alternating stresses. Microstructural barriers and the state of stress play a dominant role in the early stages of crack growth of metals. The polycristalline metal was modeled as an aggregat of hexagonal grains with a different crystallographic orientation of each grain. The effect of grain bounderies on stage I crack growth is considered in the model. The mode of shear crack growth is analyzed on the basis of microstructural crack growth within the first few grains, where the crack growth decelerates as the crack tip gets closer to the grain boundery. Normal stress crack growth has been considered for those cracks which are longer than microstructurelly short cracks, so‐called physically small cracks. Furthermore the transition from stage I to stage II growth is considered. The model is applied for thinwalled tubular specimens of the ferritic steel AlSI 1015 and the aluminium alloy Al Mg Si 1 subjected to tension and torsion as well as combined tension‐torsion loading. Also different load sequences are investigated. The microstructural crack pattern and crack distribution can sucessfully simulated with the model. The crack growth behaviour and the effect of lifetime until a crack length of 500 μm ist presented for numerous parameters.     

Stability and growth characteristics of GFPuv-labeled Cronobacter sakazakii isolated from foods

Stability of ultraviolet green fluorescent protein (GFPuv)-labeled Cronobacter sakazakii and C. muytjensii isolated from foods and the effects of the plasmid GFPuv (pGFPuv) on growth were analyzed. PCR analysis and microscopic observation showed that C. sakazakii and C. muytjensii isolates took up the plasmid into cells and expressed the GFPuv gene. All Cronobacter transformants maintained this plasmid after frozen storage and 2 consecutive subcultures. The C. sakazakii FWHd16 transformant was the most stable, while the C. muytjensii FWHd11 transformant was the least stable. All transformants showed nearly identical growth curves during lag, log and stationary phases, compared to wild type parental isolates. The maximum bacterial growth rates (μ_max) of the transformants and parents were similar, indicating that the presence of pGFPuv in transformants did not affect cell growth. Stable GFPuv-labeled Cronobacter has potential for use in tracking bacterial behavior during food handling and drying.     

Multiwalled Carbon Nanotubes for Heterogeneous Nanocatalytic Ozonation

Multiwalled carbon nanotubes functionalized by plasma oxygen (CNTs) have been used as heterogeneous catalysts for the ozonation of methyl orange (MO) dye (CI 13025) in aqueous solutions. It was found that the addition of CNTs significantly enhanced the dye decolorization as compared to ozone alone or when activated carbon was used at the same dose as CNTs. Both the initial ozone concentration and catalyst dosage enhanced the removal of MO. However, ozone gas concentrations higher than 6 g/m³ NTP did not further improve the decolorization rates. The removal efficiency of MO increased with pH in the range 2 to 3, while a reverse trend was observed when the pH increased from 3 to 9. The addition of a radical scavenger resulted in only a limited change in the decolorization rates suggesting that molecular ozone was the main pathway by which MO decolorization occurred in solution. However, under favorable conditions for MO attraction to CNT surface (pH = 3), the decolorization rate has significantly increased. At higher pH than the pKa value of MO (3.47) and the point of zero charge of CNT (3.87), a condition that favors the electrostatic repulsion of MO from CNT, the rates were reduced in the presence of CNT as compared to ozone alone possibly due to loss of part of the supplied ozone in un-useful parallel reactions.     

Structures of HIV TAR RNA–Ligand Complexes Reveal Higher Binding Stoichiometries

Target TAR by NMR : Tripeptides containing arginines as terminal residues and non‐natural amino acids as central residues are good leads for drug design to target the HIV trans‐activation response element (TAR). The structural characterization of the RNA–ligand complex by NMR spectroscopy reveals two specific binding sites that are located at bulge residue U23 and around the pyrimidine‐stretch U40‐C41‐U42 directly adjacent to the bulge.

     

Impact of incomplete DNase I treatment on human macrophage proteome analysis

The aim of our study was to analyze the proteomic pattern of human macrophages obtained over a 4 year period from blood donors. The purpose was to simulate a long-term clinical study to assess the application of 2-D DIGE technique for differential proteomic analysis of these scarce samples. Bioinformatic analysis of 2-D DIGE gels of 19 different cultures of macrophages assessed whether they did or did not contain at least specific five spots identified by MS as being or containing bovine deoxyribonuclease I (DNase I). Bovine DNase I was used during sample treatment to remove nucleic acids from protein extracts. Macrophages were classified in two groups, which appeared to be differentiated by the completeness of DNase I treatment. Further detailed analysis revealed a different proteomic pattern of macrophage protein samples according to the completeness of this treatment. The major group of proteins affected, accounting for one third of the differentially expressed proteins, included proteins involved in cell motion and actin cytoskeleton reorganization. The use of DNase I for the removal of nucleic acids from protein samples must be avoided in proteomic studies since it can generate bias in the analysis of protein expression patterns.     

Effects of the counter-cation nature and preparation method on the structure of La2Zr2O7

Lanthanum zirconate with a pyrochlore structure exhibits interesting thermal and structural properties for thermal barrier coating applications operating at high temperature. This study focused on the synthesis of La₂Zr₂O₇ powders by soft chemistry (citrate, coprecipitation, and sol–gel methods), and the effects of the solvent and the nature of the precursors used on the powder properties were investigated. The nature of the counter-cation influenced the structural phase of the compound. A low polarizable counter-cation (chloride) permitted the elaboration of a pyrochlore structure within a larger range of parameters compared with highly polarizable counter-cations (nitrate and acetate). Moreover, the nature of the solvent, and specifically the strength of the interactions within it, had an impact on the particle size. Specifically, the use of water, which is a solvent with strong interactions, led to a smaller particle size than the use of ethanol, which has weaker interactions. Based on these results, the crystalline phase of the powders can be predicted as a function of the precursor and the solvent used for the different synthesis routes, and the particle size can be used to evaluate the reactivity of the powders.     

Regrowth and survival of indicator microorganisms on the surfaces of household containers used for the storage of drinking water in rural communities of South Africa

The present study covered two rural communities of South Africa: Ncera and Ntselamanzi villages. Raw water from Ncera river is used by the community of Ncera village for drinking, while the community of Ntselamanzi receives their drinking water from Alice purification system. Treated water is supplied to the community by a public standpipe system. In rural communities of South Africa, many households use polyethylene (PE) and galvanized steel (GS) containers for the storage of their drinking water. To investigate the regrowth and survival of indicator microorganisms on the surface of household containers during the storage of drinking water, PE and GS slides were suspended in the appropriate household containers for a period of 48 h. This period of 48 h was chosen as the study period because results from the questionnaire indicated that the largest percentage (62%) of households store their water for that length of time. The experiment was performed to test drinking water as it is collected and stored by rural communities. No disinfection of household containers or slides was done during the study period. Attached coliphages (F-RNA (FP) and somatic phage (SP), coliform bacteria (total coliform (TC), presumptive Escherichia coli (EC), Salmonella (Sal) and Clostridium perfringens (CP) were measured during the study period. With the exception of CP, attached indicator microorganisms consisted of TC, presumptive E. coli and Salmonella, somatic and F-RNA coliphages, although the yield (average count) for the last four groups (EC: < 1-3 cfu cm(-2), Sal: < 1-15 cfu cm(-2), FP: < 1-7 pfu cm(-2), SP: < 1-7pfu cm(-2)) was lower than that of TC (3-183 cfu cm(-2)). However, the lowest yield of indicator microorganisms was noted for presumptive E. coli. Whereas the occurrence and survival of TC was noted on the surface of household containers during the entire period of the experimental study, other indicator microorganisms occurred from time to time. The regrowth of indicator microorganisms occurred 48 h after the exposure of slides to both types of test waters. This length of time mostly resulted in the regrowth of TC (with an increase in bacterial counts) while the persistence of other indicator organism groups on the surface of the slides was apparent. A comparison between PE and GS containers showed that more TC (average count) regrew on PE than on GS containers (for river water, PE: from 36 to 55 cfu cm(-2), GS: from 25 to 26 cfu cm(-2); for standpipe water, PE: from 147 to 183 cfu cm(-2), GS from 3 to 4 cfu cm(-2)). This study revealed that both types of household containers supported the growth and survival of indicator microorganisms due to the bad quality of the intake water before storage. The storage of drinking water for 48 h mainly resulted in the regrowth of TC. Nevertheless, the persistence of other indicator microorganisms was observed on the surface of household containers.