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.     

Cytoskeletal Protein 4.1G Is Essential for the Primary Ciliogenesis and Osteoblast Differentiation in Bone Formation

The primary cilium is a hair-like immotile organelle with specific membrane receptors, including the receptor of Hedgehog signaling, smoothened. The cilium organized in preosteoblasts promotes differentiation of the cells into osteoblasts (osteoblast differentiation) by mediating Hedgehog signaling to achieve bone formation. Notably, 4.1G is a plasma membrane-associated cytoskeletal protein that plays essential roles in various tissues, including the peripheral nervous system, testis, and retina. However, its function in the bone remains unexplored. In this study, we identified 4.1G expression in the bone. We found that, in the 4.1G-knockout mice, calcium deposits and primary cilium formation were suppressed in the trabecular bone, which is preosteoblast-rich region of the newborn tibia, indicating that 4.1G is a prerequisite for osteoblast differentiation by organizing the primary cilia in preosteoblasts. Next, we found that the primary cilium was elongated in the differentiating mouse preosteoblast cell line MC3T3-E1, whereas the knockdown of 4.1G suppressed its elongation. Moreover, 4.1G-knockdown suppressed the induction of the cilia-mediated Hedgehog signaling and subsequent osteoblast differentiation. These results demonstrate a new regulatory mechanism of 4.1G in bone formation that promotes the primary ciliogenesis in the differentiating preosteoblasts and induction of cilia-mediated osteoblast differentiation, resulting in bone formation at the newborn stage.     

Application of Homology Modeling by Enhanced Profile–Profile Alignment and Flexible-Fitting Simulation to Cryo-EM Based Structure Determination

Application of cryo-electron microscopy (cryo-EM) is crucially important for ascertaining the atomic structure of large biomolecules such as ribosomes and protein complexes in membranes. Advances in cryo-EM technology and software have made it possible to obtain data with near-atomic resolution, but the method is still often capable of producing only a density map with up to medium resolution, either partially or entirely. Therefore, bridging the gap separating the density map and the atomic model is necessary. Herein, we propose a methodology for constructing atomic structure models based on cryo-EM maps with low-to-medium resolution. The method is a combination of sensitive and accurate homology modeling using our profile–profile alignment method with a flexible-fitting method using molecular dynamics simulation. As described herein, this study used benchmark applications to evaluate the model constructions of human two-pore channel 2 (one target protein in CASP13 with its structure determined using cryo-EM data) and the overall structure of Enterococcus hirae V-ATPase complex.     

Therapeutic Targets and Emerging Treatments in Advanced Chondrosarcoma

Simple SummaryChondrosarcomas develop chemoresistance to standard anticancer drugs, making it difficult to control unresectable or metastatic chondrosarcomas. To improve the clinical outcomes of chondrosarcoma, new treatment approaches, such as molecule-targeting agents and immunotherapy, are needed. Recent research has revealed promising biomarkers and therapeutic targets for chondrosarcoma. In addition, several molecule-targeting agents have shown favorable antitumor activities in several clinical studies in patients with advanced sarcomas, including chondrosarcoma. This review summarizes recent basic studies on biomarkers and therapeutic targets and recent clinical studies on treating chondrosarcomas.AbstractDue to resistance to standard anticancer agents, it is difficult to control the disease progression in patients with metastatic or unresectable chondrosarcoma. Novel therapeutic approaches, such as molecule-targeting drugs and immunotherapy, are required to improve clinical outcomes in patients with advanced chondrosarcoma. Recent studies have suggested several promising biomarkers and therapeutic targets for chondrosarcoma, including IDH1/2 and COL2A1. Several molecule-targeting agents and immunotherapies have shown favorable antitumor activity in clinical studies in patients with advanced chondrosarcomas. This review summarizes recent basic studies on biomarkers and molecular targets and recent clinical studies on the treatment of chondrosarcomas.     

A PSS‐Free PEDOT Conductive Film Supported by a Hierarchical Nanoporous Layer Glass

The importance of transparent conductive film is increasing due to its use in applications such as touch‐panel devices. Although indium tin oxide is widely used because of its high conductivity and transparency, conductive polymers are being studied as alternative materials that avoid the use of rare metals and the brittleness associated with existing systems. Polyethylene dioxythiophene (PEDOT)/polyethylene sulfonic acid (PSS) is drawing a lot of attention due to its well‐balanced conductivity, transparency, film formability, and chemical stability. The nonconductive PSS reportedly covers the conductive PEDOT. The PSS shell provides carrier and film‐formability to PEDOT but is also a barrier that hinders electrical conductivity. Therefore, the PEDOT film formability is explored supported by a substrate without the addition of PSS. The “hierarchical nanoporous layer glass” holds the PSS‐free PEDOT with its nanopores to form a homogeneous, transparent film. The PSS‐free PEDOT film thus achieves transparency of over 85% and resistivity of below 500 Ω sq^?1.     

Detail analysis of fusion neutronics benchmark experiment for iron

In order to specify the best nuclear data on iron, the fusion neutronics benchmark experiment on iron at Japan Atomic Energy Agency (JAEA)/Fusion Neutronics Source (FNS) was analyzed in detail with MCNP-4C and the latest nuclear data libraries, JENDL-3.3, FENDL-2.1, JEFF-3.1 and ENDF/B-VII.0. As a result, totally the calculation result with ENDF/B-VII.0 agreed with the measurement best, except that it underestimated the measured neutron flux above 10 MeV with the depth. It was noted that the calculation result with JENDL-3.3 overestimated the measured neutrons below a few keV. Through the DORT calculations based on the iron data in ENDF/B-VII.0, it was found out that the first inelastic scattering cross-section data of ⁵⁷Fe in JENDL-3.3 caused the overestimation.     

Effect of Deposition Rate on the Stress Evolution of Plasma-Sprayed Yttria-Stabilized Zirconia

The deposition rate plays an important role in determining the thickness, stress state, and physical properties of plasma-sprayed coatings. In this article, the effect of the deposition rate on the stress evolution during the deposition (named evolving stress) of yttria-stabilized zirconia coatings was systematically studied by varying the powder feed rate and the robot-scanning speed. The evolving stress during the deposition tends to increase with the increased deposition rate, and this tendency was less significant at a longer spray distance. In some cases, the powder feed rate had more significant influence on the evolving stress than the robot speed. This tendency can be associated with a deviation of a local deposition temperature at a place where sprayed particles are deposited from an average substrate temperature. At a further higher deposition rate, the evolving stress was relieved by introduction of macroscopic vertical cracks as well as horizontal branching cracks.     

Effects of fatty acid and emulsifier on the complex formation and in vitro digestibility of gelatinized potato starch

The effects of fatty acid, monoacylglycerol, and polyglycerol fatty acid ester with varying chain length in their acyl chains on the extent of complex formation (complex index) and in vitro enzymatic digestibility of gelatinized potato starch were investigated. The complex index increased with increase in the concentration of the ligands (fatty acid, monoacylglycerol, and polyglycerol fatty acid ester), with the plateau in the complex index value depending on the type of ligands. In comparison of complex index among fatty acid-samples, the complex index maximum increased as the chain length increased up to octanoic acid and then decreased. In comparison of complex index among fatty acid-, monoacylglycerol-, and polyglycerol fatty acid ester-samples at each acyl chain, the complex index maximum followed the order polyglycerol fatty acid ester > monoacylglycerol > fatty acid. Fatty acid, monoacylglycerol, and polyglycerol fatty acid ester with long acyl chains greatly reduced the enzymatic hydrolysis of starch. Polyglycerol fatty acid ester with palmitic acid chains was the strongest inhibitor of starch hydrolysis, suggesting that further complex formation may occur during the hydrolysis of gelatinized starch (enzyme-annealing).     

Occurrence and sources of perfluorinated surfactants in rivers in Japan

We analyzed perfluorinated surfactants (PFSs) in 20 river samples and 5 wastewater secondary effluent samples in Japan to reveal their occurrence and sources. Nine PFS species were determined: perfluorooctanesulfonate (PFOS), perfluorooctane sulfonamide (FOSA), perfluoroheptanoate (PFHpA), perfluorooctanoate (PFOA), perfluorononanoate (PFNA), perfluorodecanoate (PFDA), perfluoroundecanoate (PFUA), perfluorododecanoate (PFDDA), and perfluorotridecanoate (PFTDA). PFSs were detected in all rivers, revealing nationwide contamination of rivers. In particular, 11 out of 20 river samples exceeded New Jersey guidance for PFOA in drinking water (40 ng/L). PFOS, PFHpA, PFOA, and PFNA were major species in Japan. Concentrations of PFOS, PFHpA, and PFNA in rivers were strongly correlated with population density, suggesting that the chemicals were derived from urban activities. PFOA showed a significant but weak correlation. We used crotamiton, a marker of sewage effluent, for further source analysis. Concentrations of PFOS, PFHpA, and PFNAwere strongly correlated with those of crotamiton, and plots of secondary effluents fell near the regression lines of rivers, indicating that the PFOS, PFHpA, and PFNA in rivers were derived from sewage effluent. On the other hand, PFOA was found at remarkably high levels (54-192 ng/L) in seven river samples containing low levels of crotamiton, suggesting that it was derived from nonsewage point sources, as well as sewage effluent. The total fluxes of sewage-derived PFOS, PFHpA, PFOA, and PFNA from Japan were estimated to be 3.6, 2.6, 5.6, and 2.6 t/year, respectively. This is the first report to identify PFOA in several rivers, derived from nonsewage point sources, by using a marker of sewage effluent.