Dual-Function Semaphorin 4D Released by Platelets: Suppression of Osteoblastogenesis and Promotion of Osteoclastogenesis

Effects of the antiosteoblastogenesis factor Semaphorin 4D (Sema4D), expressed by thrombin-activated platelets (TPs), on osteoblastogenesis, as well as osteoclastogenesis, were investigated in vitro. Intact platelets released both Sema4D and IGF-1. However, in response to stimulation with thrombin, platelets upregulated the release of Sema4D, but not IGF-1. Anti-Sema4D-neutralizing monoclonal antibody (mAb) upregulated TP-mediated osteoblastogenesis in MC3T3-E1 osteoblast precursors. MC3T3-E1 cells exposed to TPs induced phosphorylation of Akt and ERK further upregulated by the addition of anti-sema4D-mAb, suggesting the suppressive effects of TP-expressing Sema4D on osteoblastogenesis. On the other hand, TPs promoted RANKL-mediated osteoclastogenesis in the primary culture of bone-marrow-derived mononuclear cells (BMMCs). Among the known three receptors of Sema4D, including Plexin B1, Plexin B2 and CD72, little Plexin B2 was detected, and no Plexin B1 was detected, but a high level of CD72 mRNA was detected in RANKL-stimulated BMMCs by qPCR. Both anti-Sema4D-mAb and anti-CD72-mAb suppressed RANKL-induced osteoclast formation and bone resorptive activity, suggesting that Sema4D released by TPs promotes osteoclastogenesis via ligation to a CD72 receptor. This study demonstrated that Sema4D released by TPs suppresses osteogenic activity and promotes osteoclastogenesis, suggesting the novel property of platelets in bone-remodeling processes.     

STUDIES ON THE MYCOFLORA OF MALT

The mycoflora of malt which caused gushing beer has been investigated and compared with the mycoflora of malt from nine malting plants in Sweden, Denmark, Finland and England. Malt which caused gushing beer was found to contain a high proportion of grains contaminated by Aspergillus, Penicillium and Rhizopus. The commonest species isolated were found to be Aspergillus fumigatus and Aspergillus amstelodami. These fungi occurred on all malt samples investigated independent of their origin. The mycoflora on malt differs greatly from that found on barley. Evidence is presented showing that barley becomes contaminated during the malting process.     

Role of acidic sphingomyelinase in thymol‐mediated dendritic cell death

Scope : Thymol is a component of several plants with antimicrobial activity. Little is known about the effects of thymol on immune cells of the host. This study addressed the effects of thymol on dendritic cells (DCs), regulators of innate and adaptive immunity. Methods and results : Immunohistochemistry, Western blotting and fluorescence‐activated cell sorting analysis were performed in bone marrow‐derived DCs either from wild‐type mice or from mice lacking acid sphingomyelinase (ASM^?/?) treated and untreated for 24 h with thymol (2–100 μg/mL). Thymol treatment resulted in activation of ASM, stimulation of ceramide formation, downregulation of anti‐apoptotic Bcl‐2 and Bcl‐xL proteins, activation of caspase 3 and caspase 8, DNA fragmentation as well as cell membrane scrambling. The effects were dependent on the presence of ASM and were lacking in ASM^?/? mice or in wild‐type DCs treated with sphingomyelinase inhibitor amitriptyline. Conclusion : Thymol triggers suicidal DC death, an effect mediated by and requiring activation of ASM.     

Wettability and bonding quality of exterior coatings on jabon and sengon wood surfaces

Wettability and bonding quality of exterior coatings on fast-growing wood surfaces were studied. Samples of air-dried flat-grained (tangential surface) and edge-grained (radial surface) pattern of jabon (Anthocephalus cadamba) and sengon (Paraserianthes falcataria) woods were used. Before application of exterior coatings, the surfaces of the lumber samples were sanded. To provide wood surfaces with various degrees of roughness, abrasive papers of 120, 240, and 360 grits were used for the surface preparation. The wettability of two exterior coatings (water-based acrylic and oil-based alkyd varnishes) on the sanded wood surfaces was measured using a sessile drop contact angle method. The Shi and Gardner (S/G) model was used to evaluate and compare the wettability of the surface coatings on the wood. The sanded wood samples were coated with the two coatings (two layers each). Bonding quality of the coating layers was measured using a crosscut tape test method. Experimental results show that constant contact angle change rate (K value) of the S/G model decreased as the grit number of abrasive paper increased. This indicates that the wettability decreased as the roughness of the surface decreased (surface becomes smoother). There was no evidence of differences in wettability between tangential and radial wood surfaces. The oil-based alkyd coating generated better wettability compared to the water-based acrylic. The crosscut tests showed that the bonding quality of the coating films on both jabon and sengon wood decreased as the surface became smoother. The sengon wood compared to jabon wood provided better coating wettability and bonding quality. Wettability in terms of the K values was a good indication for determining the bonding quality of the two varnish layers.     

How We Make DNA Origami

DNA origami has attracted substantial attention since its invention ten years ago, due to the seemingly infinite possibilities that it affords for creating customized nanoscale objects. Although the basic concept of DNA origami is easy to understand, using custom DNA origami in practical applications requires detailed know‐how for designing and producing the particles with sufficient quality and for preparing them at appropriate concentrations with the necessary degree of purity in custom environments. Such know‐how is not readily available for newcomers to the field, thus slowing down the rate at which new applications outside the field of DNA nanotechnology may emerge. To foster faster progress, we share in this article the experience in making and preparing DNA origami that we have accumulated over recent years. We discuss design solutions for creating advanced structural motifs including corners and various types of hinges that expand the design space for the more rigid multilayer DNA origami and provide guidelines for preventing undesired aggregation and on how to induce specific oligomerization of multiple DNA origami building blocks. In addition, we provide detailed protocols and discuss the expected results for five key methods that allow efficient and damage‐free preparation of DNA origami. These methods are agarose‐gel purification, filtration through molecular cut‐off membranes, PEG precipitation, size‐exclusion chromatography, and ultracentrifugation‐based sedimentation. The guide for creating advanced design motifs and the detailed protocols with their experimental characterization that we describe here should lower the barrier for researchers to accomplish the full DNA origami production workflow.     

PLA/metal oxide biocomposites for antimicrobial packaging application

ABSTRACT

The effects of metal oxide addition in polylactic acid (PLA) film were investigated in this study. PLA/Metal Oxide biocomposites were prepared by solvent casting method. The incorporation of metal oxide resulted in no chemical structure change of neat PLA. PLA/ZnO and PLA/MgO exhibited the widest inhibition while PLA/TiO₂ resulted in the smallest inhibition zone. The addition of metal oxides caused fracture and void on the surface of PLA. The addition of metal oxide into PLA films decreased the tensile strength at the concentration higher than 0.1 per hundred resin (phr). Incorporation of metal oxides accelerated the thermal degradation of PLA and tends to increase the biodegradability of PLA.     

Hydroxylation of phenol with hydrogen peroxide catalyzed by Fe- and AIFe-Bentonite

The paper described pillarisations of natural bentonite from Pacitan East Java of Indonesia by using AI and Fe. Intercalation process by using surfactant molecule has also been carried out. Natural bentonite was intercalated with HDTMA-Br （hexadecyltrimethylammonium-bromide） 1, 5% solution before pillared with AI and Fe metal to give HDTMA-bentonite forms. The ratio of bentonite and intercalating agent or pillaring agent was 1 gr/50 mL. The mixture was agitated, and then the solid phase was washed with distilled water. Then it was dried and calcined at 450℃ for 4 hours. Their catalytic activity and selectivity were studied for phenol hydroxylation using tlzOz （30%）. The reaction condition of this reaction was as follows： ratio of phenol/ H202 = 1：1 （molar ratio）, concentration of phenol = 1 M, reaction temperature was 60℃, and ratio of catalyst/phenol was 1：10. The products were hydroquinone and cathecol.     

Potential Activity of Fevicordin-A from Phaleria macrocarpa (Scheff) Boerl. Seeds as Estrogen Receptor Antagonist Based on Cytotoxicity and Molecular Modelling Studies

Fevicordin-A (FevA) isolated from Phaleria macrocarpa (Scheff) Boerl. seeds was evaluated for its potential anticancer activity by in vitro and in silico approaches. Cytotoxicity studies indicated that FevA was selective against cell lines of human breast adenocarcinoma (MCF-7) with an IC₅₀ value of 6.4 μM. At 11.2 μM, FevA resulted in 76.8% cell death of T-47D human breast cancer cell lines. Critical pharmacophore features amongst human Estrogen Receptor-α (hERα) antagonists were conserved in FevA with regard to a hypothesis that they could make notable contributions to its pharmacological activity. The binding stability as well as the dynamic behavior of FevA towards the hERα receptor in agonist and antagonist binding sites were probed using molecular dynamics (MD) simulation approach. Analysis of MD simulation suggested that the tail of FevA was accountable for the repulsion of the C-terminal of Helix-11 (H11) in both agonist and antagonist receptor forms. The flexibility of loop-534 indicated the ability to disrupt the hydrogen bond zipper network between H3 and H11 in hERα. In addition, MM/GBSA calculation from the molecular dynamic simulations also revealed a stronger binding affinity of FevA in antagonistic action as compared to that of agonistic action. Collectively, both the experimental and computational results indicated that FevA has potential as a candidate for an anticancer agent, which is worth promoting for further preclinical evaluation.     

Cover Feature: Exploring the Flexibility of the Glycopeptide Antibiotic Crosslinking Cascade for Extended Peptide Backbones (ChemBioChem 6/2023)

The glycopeptide antibiotics (GPAs) are a clinically approved class of antimicrobial agents that classically function through the inhibition of bacterial cell-wall biosynthesis by sequestration of the precursor lipid II. The oxidative crosslinking of the core peptide by cytochrome P450 (Oxy) enzymes during GPA biosynthesis is both essential to their function and the source of their synthetic challenge. Thus, understanding the activity and selectivity of these Oxy enzymes is of key importance for the future engineering of this important compound class. Recent reports of GPAs that display an alternative mode of action and a wider range of core peptide structures compared to classic lipid II-binding GPAs raises the question of the tolerance of Oxy enzymes for larger changes in their peptide substrates. In this work, we explore the ability of Oxy enzymes from the biosynthesis pathways of lipid II-binding GPAs to accept altered peptide substrates based on a vancomycin template. Our results show that Oxy enzymes are more tolerant of changes at the N terminus of their substrates, whilst C-terminal extension of the peptide substrates is deleterious to the activity of all Oxy enzymes. Thus, future studies should prioritise the study of Oxy enzymes from atypical GPA biosynthesis pathways bearing C-terminal peptide extension to increase the substrate scope of these important cyclisation enzymes.     

Differences between vehicle lateral displacement on the road and in a fixed-base simulator

This work investigates differences in lateral displacement when driving on curved and straight road sections in real-road and simulator conditions. We observed 100 licensed drivers on a rural road and 100 in a fixed-base simulator. Speed and lateral position on the real road were measured using videocameras. The analysis indicates that the mean vehicle lateral displacement is in general higher on the real road than in the simulator. However, these differences decrease for higher speeds at curved sections and for lower speeds at straight sections. It was also found that the standard deviation of the vehicle lateral displacement is significantly lower on the real road than the corresponding values in the simulator, at either curved or straight sections. Actual or potential applications of this research are related to a more realistic assessment of driving behavior scenarios derived on the basis of simulation experiment results.