Antibacterial lanostane triterpenoids from Ganoderma tsugae.

A phytochemical investigation on the 80% EtOH extract of the fruiting bodies of Ganoderma tsugae resulted into the isolation of two previously undescribed lanostane triterpenoids, 7,11-dioxo-3ß-acetyloxy-26,27-dihydroxy-lanosta-8,24-diene (1) and 7,20-dioxo-3ß-acetyloxy-11ß,15α-dihydroxy-22,23,24,25,26,27-hexanorlanosta-8-ene (2), togeher with one known lanostane triterpenoid ganodermanontriol (3). Structural elucidation of all the compounds were performed by spectral methods such as 1D and 2D (1H-1H COSY, HMQC, and HMBC) NMR spectroscopy. All the triterpenoids were in vitro evaluated for their antibacterial activities against six pathogenic microorganisms. Compound 3 exhibited some activities against three Gram positive bacteria with MIC values less than 30 µg/ml.

Adhesion Properties of Recycled High-Viscosity Asphalt-Aggregate Interface under Dynamic Water Erosion.

The drainage of asphalt pavement requires the use of a large amount of high-viscosity-modified asphalt, which faces the service environment under dynamic water erosion. The feasibility of recycling high-viscosity-modified asphalt should be investigated to facilitate sustainable infrastructure construction. This study used ultrasonic equipment to simulate dynamic water erosion test conditions and tested the adhesion performance of different types of recycled high-viscosity asphalt at various environmental temperatures. The adhesion energy index and microstructure of recycled high-viscosity asphalt were analyzed using the contact angle test and AFM test. The results demonstrate that the higher the environmental temperature, the worse the anti-stripping performance of recycled high-viscosity asphalt. From the perspective of adhesion performance indicators, a 6% recycling agent dosage is more conducive to restoring the performance of aged high-viscosity -modified asphalt. The AFM test showed that the microstructure of high-viscosity -modified asphalt represented significant changes with an increase in the recycling agent content, and the change in the adhesion force of recycled high-viscosity -modified asphalt was consistent with the results of macroscopic adhesion performance tests. This study illustrates the applicability of implementing regeneration technology for the recycling of aged drainage asphalt pavement.

Experimental and Numerical Investigation of Joints for a Pultruded Fiber-Reinforced Polymer Truss.

Bolted connections usually govern the structural rigidity and load-carrying capacity of pultruded glass fiber-reinforced polymer (GFRP) truss structures. In this study, a novel bolted integrated gusset plate (IGP) connection is proposed to enhance the stiffness and capacity of GFRP truss structures. Nine double-lap shear tests of GFRP joints and numerical simulation were conducted to investigate the influence of variable design parameters of the bolted GFRP joints (number of bolts, width and thickness of GFRP, edge distance of bolts, and the employment of adhesive). Three full-scale GFRP truss joints were tested under static loading to study the response of a typical bolted connection, a bolted gusset plate connection, and the proposed IGP connection. The nine double-lap shear tests showed that the bolted-bonded mixed connection has 50% higher shear stiffness and 27% higher ductility compared with bolted joints, and bearing failure dominated the capacity of most specimens, which agreed well with numerical simulation results. Tests on the three full-scale GFRP truss joints showed that the bolted gusset plate can substantially reduce the number of cracks and improve the initial stiffness, but the maximum bearing capacity of the joints did not increase because the shear fracture of pultruded GFRP webs governs the capacity. The proposed IGP substantially increased the stiffness and capacity compared with the bolted connection and typical bolted gusset plate connection. The full-scale GFRP joint test is suggested to be used together with direct shear tests to study the performance of joints of the GFRP truss.

Influence of the Cross-Sectional Shape and Corner Radius on the Compressive Behaviour of Concrete Columns Confined by FRP and Stirrups.

Axial compression tests were carried out on 72 FRP (fiber reinforced polymer)-stirrup composite-confined concrete columns. Stirrups ensure the residual bearing capacity and ductility after the FRP fractures. To reduce the effect of stress concentration at the corners of the confined square-section concrete columns and improve the restraint effect, an FRP-stirrup composite-confined concrete structure with rounded corners is proposed. Different corner radii of the stirrup and outer FRP were designed, and the corner radius of the stirrup was adjusted accurately to meet the designed corner radius of the outer FRP. The cross-section of the specimens gradually changed from square to circular as the corner radius increased. The influence of the cross-sectional shape and corner radius on the compressive behaviour of FRP-stirrup composite-confined concrete was analysed. An increase in the corner radius can cause the strain distribution of the FRP to be more uniform and strengthen the restraint effect. The larger the corner radius of the specimen, the better the improvement of mechanical properties. The strength of the circular section specimen was greatly improved. In addition, the test parameters also included the FRP layers, FRP types and stirrup spacing. With the same corner radius, increasing the number of FRP layers or densifying the stirrup spacing effectively improved the mechanical properties of the specimens. Finally, a database of FRP-stirrup composite-confined concrete column test results with different corner radii was established. The general calculation models were proposed, respectively, for the peak points, ultimate points and stress-strain models that are applicable to FRP-, stirrup- and FRP-stirrup-confined concrete columns with different cross-sectional shapes under axial compression.

Compressive Behavior of Bamboo Sheet Twining Tube-Confined Concrete Columns.

This study experimentally investigated various axial compressive parameters of a new type of confined concrete, which is termed bamboo sheet twining tube-confined concrete (BSTCC). This new composite structure was composed of an outer bamboo composite tube (BCT) jacket and a concrete core. Under axial compression, the parameters of thirty-six specimens include concrete strength (i.e., C30 and C50) and BCT thickness (i.e., 6, 12, 18, 24, and 30 layers). The mechanical properties of the BSTCC specimens from the perspective of the failure mode, stress-strain relationship, effect of BCT thickness and dilation behavior were analyzed. The results showed that, in compression, with an increase in BCT thickness in the range of 18-layers of bamboo sheets, the strength increased remarkably. When the strength of the concrete core was high, the confinement effect of the BCT was reduced. In addition, the BCT thickness relieved the dilation of the BSTCC specimens. Finally, the experimental results were compared with predictions obtained from 7 existing FRP-confined concrete models. All the predictions had good agreement with the test results, which further confirmed that the models developed for FRP-confined concrete can provide an acceptable approximation of the ultimate strength of the BSTCC specimens.

What is the role of the nitrate reductase (euknr) gene in fungi that live in nitrate-free environments? A targeted gene knock-out study in Ampelomyces mycoparasites.

Mycoparasitic fungi can be utilized as biocontrol agents (BCAs) of many plant pathogens. Deciphering the molecular mechanisms of mycoparasitism may improve biocontrol efficiency. This work reports the first functional genetic studies in Ampelomyces, widespread mycoparasites and BCAs of powdery mildew fungi, and a molecular genetic toolbox for future works. The nitrate reductase (euknr) gene was targeted to reveal the biological function of nitrate assimilation in Ampelomyces. These mycoparasites live in an apparently nitrate-free environment, i.e. inside the hyphae of powdery mildew fungi that lack any nitrate uptake and assimilation system. Homologous recombination-based gene knock-out (KO) was applied to eliminate the euknr gene using Agrobacterium tumefaciens-mediated transformation. Efficient KO of euknr was confirmed by PCR, and visible phenotype caused by loss of euknr was detected on media with different nitrogen sources. Mycoparasitic ability was not affected by knocking out euknr as a tested transformant readily parasitized Blumeria graminis and Podosphaera xanthii colonies on barley and cucumber, respectively, and the rate of mycoparasitism did not differ from the wild type. These results indicate that euknr is not involved in mycoparasitism. Dissimilatory processes, involvement in nitric oxide metabolism, or other, yet undiscovered processes may explain why a functional euknr is maintained in Ampelomyces.

Trichoderma reesei Isolated From Austrian Soil With High Potential for Biotechnological Application.

Fungi of the genus Trichoderma are of high importance for biotechnological applications, in biocontrol and for production of homologous and heterologous proteins. However, sexual crossing under laboratory conditions has so far only been achieved with the species Trichoderma reesei, which was so far only isolated from tropical regions. Our isolation efforts aimed at the collection of Trichoderma strains from Austrian soils surprisingly also yielded 12 strains of the species T. reesei, which was previously not known to occur in Europe. Their identity was confirmed with tef1- and rpb2-sequencing and phylogenetic analysis. They could clearly be distinguished from tropical strains including the common laboratory wildtypes by UP-PCR and genetic variations adjacent to the mating type locus. The strains readily mated with reference strains derived from CBS999.97. Secreted cellulase and xylanase levels of these isolates were up to six-fold higher than those of QM6a indicating a high potential for strain improvement. The strains showed different responses to injury in terms of induction of sporulation, but a correlation to alterations in the nox1-gene sequence was not detected. Several synonymous SNPs were found in the sequence of the regulator gene noxR of the soil isolates compared to QM6a. Only in one strain, non-synonymous SNPs were found which impact a PEST sequence of NoxR, suggesting altered protein stability. The availability of sexually fertile strains from middle Europe naturally producing decent amounts of plant cell wall degrading enzymes opens up novel perspectives for non-GMO strain improvement and biological pretreatment of plant biomass for bioethanol production. Moreover, the varied response of these strains to injury in terms of sporulation, which is independent of Nox1 and NoxR suggests that additional regulators impact this phenomenon in T. reesei.

NO and N2 O transformations of diverse fungi in hypoxia: evidence for anaerobic respiration only in Fusarium strains.

Fungal denitrification is claimed to produce non-negligible amounts of N2 O in soils, but few tested species have shown significant activity. We hypothesized that denitrifying fungi would be found among those with assimilatory nitrate reductase, and tested 20 such batch cultures for their respiratory metabolism, including two positive controls, Fusarium oxysporum and Fusarium lichenicola, throughout the transition from oxic to anoxic conditions in media supplemented with NO 2 - . Enzymatic reduction of NO 2 - (NIR) and NO (NOR) was assessed by correcting measured NO- and N2 O-kinetics for abiotic NO- and N2 O-production (sterile controls). Significant anaerobic respiration was only confirmed for the positive controls and for two of three Fusarium solani cultures. The NO kinetics in six cultures showed NIR but not NOR activity, observed through the accumulation of NO. Others had NOR but not NIR activity, thus reducing abiotically produced NO to N2 O. The presence of candidate genes (nirK and p450nor) was confirmed in the positive controls, but not in some of the NO or N2 O accumulating cultures. Based on our results, we conclude that only the Fusarium cultures were able to sustain anaerobic respiration and produced low amounts of N2 O as a response to an abiotic NO production from the medium.

Inhibition of MiR-92a May Protect Endothelial Cells After Acute Myocardial Infarction in Rats: Role of KLF2/4.

BACKGROUND This study was designed to investigate the effects of microRNA-92 (miR-92), Kruppel-like factor 2 (KLF2), and Kruppel-like factor 4 (KLF4) on endothelial injury after acute myocardial infarction (AMI). MATERIAL AND METHODS Blood samples were collected from 50 AMI patients for detection of cardiac troponin I (cTnI), heart-type fatty acid-binding protein (H-FABP), and von Willebrand factor (vWF). The Sprague-Dawley rat models of AMI (n=30) were established by ligating their left anterior descending coronary artery. The cardiac markers of AMI patients and rat models were analyzed with enzyme-linked immunosorbent assay and immunohistochemistry. Human umbilical vein endothelial cells were processed into 5 groups: control, negative control, miR-92a inhibitors, miR-92a inhibitors + KLF2 small interfering RNA (siRNA), and miR-92a inhibitors + KLF4 siRNA. Cell proliferation and apoptosis were detected using MTT assay and flow cytometry. RT-PCR and Western blot were conducted to analyze KLF2 and KLF4 expressions. RESULTS AMI patients exhibited significantly higher expression of both endothelial injury markers (e.g., cTnI, H-FABP, vWF) and miR-92a in blood samples, when compared with controls (P<0.05). Model rats also had similar expressional tendencies, along with lower KLF2 and KLF4 expressions (P<0.05). Further, it could be observed in cellular experiments that treatment of miR-92a mimics can further upregulate endothelial injury markers, and miR-92a and both KLF2 and KLF4 were downregulated by miR-92a mimics (all, P<0.05). Also, the luciferase activity assay confirmed the direct binding of miR-92a to 3' UTR of KLF2/4. CONCLUSIONS MiR-92a was involved in the endothelial injury process after AMI and was able to suppress KLF2 and KLF4 expression.