Elevated Expression of CCN3 in Articular Cartilage Induces Osteoarthritis in Hip Joints Irrespective of Age and Weight Bearing

Osteoarthritis (OA) occurs not only in the knee but also in peripheral joints throughout the whole body. Previously, we have shown that the expression of cellular communication network factor 3 (CCN3), a matricellular protein, increases with age in knee articular cartilage, and the misexpression of CCN3 in cartilage induces senescence-associated secretory phenotype (SASP) factors, indicating that CCN3 promotes cartilage senescence. Here, we investigated the correlation between CCN3 expression and OA degenerative changes, principally in human femoral head cartilage. Human femoral heads obtained from patients who received total hip arthroplasty were categorized into OA and femoral neck fracture (normal) groups without significant age differences. Gene expression analysis of RNA obtained from femoral head cartilage revealed that CCN3 and MMP-13 expression in the non-weight-bearing part was significantly higher in the OA group than in the normal group, whereas the weight-bearing OA parts and normal cartilage showed no significant differences in the expression of these genes. The expression of COL10A1, however, was significantly higher in weight-bearing OA parts compared with normal weight-bearing parts, and was also higher in weight-bearing parts compared with non-weight-bearing parts in the OA group. In contrast, OA primary chondrocytes from weight-bearing parts showed higher expression of CCN3, p16, ADAMTS4, and IL-1β than chondrocytes from the corresponding normal group, and higher ADAMTS4 and IL-1β in the non-weight-bearing part compared with the corresponding normal group. Acan expression was significantly lower in the non-weight-bearing group in OA primary chondrocytes than in the corresponding normal chondrocytes. The expression level of CCN3 did not show significant differences between the weight-bearing part and non-weight-bearing part in both OA and normal primary chondrocytes. Immunohistochemical analysis showed accumulated CCN3 and aggrecan neoepitope staining in both the weight-bearing part and non-weight-bearing part in the OA group compared with the normal group. The CCN3 expression level in cartilage had a positive correlation with the Mankin score. X-ray analysis of cartilage-specific CCN3 overexpression mice (Tg) revealed deformation of the femoral and humeral head in the early stage, and immunohistochemical analysis showed accumulated aggrecan neoepitope staining as well as CCN3 staining and the roughening of the joint surface in Tg femoral and humeral heads. Primary chondrocytes from the Tg femoral head showed enhanced expression of Ccn3, Adamts5, p16, Il-6, and Tnfα, and decreased expression of Col2a1 and -an. These findings indicate a correlation between OA degenerative changes and the expression of CCN3, irrespective of age and mechanical loading. Furthermore, the Mankin score indicates that the expression level of Ccn3 correlates with the progression of OA.     

State-of-the-Art Molecular Dynamics Simulation Studies of RNA-Dependent RNA Polymerase of SARS-CoV-2

Molecular dynamics (MD) simulations are powerful theoretical methods that can reveal biomolecular properties, such as structure, fluctuations, and ligand binding, at the level of atomic detail. In this review article, recent MD simulation studies on these biomolecular properties of the RNA-dependent RNA polymerase (RdRp), which is a multidomain protein, of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are presented. Although the tertiary structures of RdRps in SARS-CoV-2 and SARS-CoV are almost identical, the RNA synthesis activity of RdRp of SARS-CoV is higher than SARS-CoV-2. Recent MD simulations observed a difference in the dynamic properties of the two RdRps, which may cause activity differences. RdRp is also a drug target for Coronavirus disease 2019 (COVID-19). Nucleotide analogs, such as remdesivir and favipiravir, are considered to be taken up by RdRp and inhibit RNA replication. Recent MD simulations revealed the recognition mechanism of RdRp for these drug molecules and adenosine triphosphate (ATP). The ligand-recognition ability of RdRp decreases in the order of remdesivir, favipiravir, and ATP. As a typical recognition process, it was found that several lysine residues of RdRp transfer these ligand molecules to the binding site such as a “bucket brigade.” This finding will contribute to understanding the mechanism of the efficient ligand recognition by RdRp. In addition, various simulation studies on the complexes of SARS-CoV-2 RdRp with several nucleotide analogs are reviewed, and the molecular mechanisms by which these compounds inhibit the function of RdRp are discussed. The simulation studies presented in this review will provide useful insights into how nucleotide analogs are recognized by RdRp and inhibit the RNA replication.     

ARSlice: Head-Mounted Display Augmented with Dynamic Tracking and Projection

Computed tomography (CT) generates cross-sectional images of the body. Visualizing CT images has been a challenging problem. The emergence of the augmented and virtual reality technology has provided promising solutions. However, existing solutions suffer from tethered display or wireless transmission latency. In this paper, we present ARSlice, a proof-of-concept prototype that can visualize CT images in an untethered manner without wireless transmission latency. Our ARSlice prototype consists of two parts, the user end and the projector end. By employing dynamic tracking and projection, the projector end can track the user-end equipment and project CT images onto it in real time. The user-end equipment is responsible for displaying these CT images into the 3D space. Its main feature is that the user-end equipment is a pure optical device with light weight, low cost, and no energy consumption. Our experiments demonstrate that our ARSlice prototype provides part of six degrees of freedom for the user, and a high frame rate. By interactively visualizing CT images into the 3D space, our ARSlice prototype can help untrained users better understand that CT images are slices of a body.     

Genome Editing Using Cas9 Ribonucleoprotein Is Effective for Introducing PDGFRA Variant in Cultured Human Glioblastoma Cell Lines

Many variants of uncertain significance (VUS) have been detected in clinical cancer cases using next-generation sequencing-based cancer gene panel analysis. One strategy for the elucidation of VUS is the functional analysis of cultured cancer cell lines that harbor targeted gene variants using genome editing. Genome editing is a powerful tool for creating desired gene alterations in cultured cancer cell lines. However, the efficiency of genome editing varies substantially among cell lines of interest. We performed comparative studies to determine the optimal editing conditions for the introduction of platelet-derived growth factor receptor alpha (PDGFRA) variants in human glioblastoma multiforme (GBM) cell lines. After monitoring the copy numbers of PDGFRA and the expression level of the PDGFRα protein, four GBM cell lines (U-251 MG, KNS-42, SF126, and YKG-1 cells) were selected for the study. To compare the editing efficiency in these GBM cell lines, the modes of clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein 9 (Cas9) delivery (plasmid vs. ribonucleoprotein (RNP)), methods of transfection (lipofection vs. electroporation), and usefulness of cell sorting were then evaluated. Herein, we demonstrated that electroporation-mediated transfer of Cas9 with single-guide RNA (Cas9 RNP complex) could sufficiently edit a target nucleotide substitution, irrespective of cell sorting. As the Cas9 RNP complex method showed a higher editing efficiency than the Cas9 plasmid lipofection method, it was the optimal method for single-nucleotide editing in human GBM cell lines under our experimental conditions.     

Microorganisms in landfill bioreactors for accelerated stabilization of solid wastes

Landfill bioreactors (LBRs) with management of leachate and biogas have presented numerous advantages such as accelerated stabilization of solid wastes, reduced amount of leachate, and in situ leachate treatment. Such advantages have minimized environmental risks, have allowed extension of the useful life of the landfill site, and have fostered cost reduction. LBRs of three types have been developed using both anaerobic and aerobic modes: anaerobic, aerobic, and hybrid. Microorganisms in landfills cause various reactions related with organic fractions and heavy metals. Such functions have been stimulated in LBRs by recirculation of leachate with or without aeration. To date, most studies of microorganisms in LBRs have analyzed bacteria and archaea based on 16S rRNA genes and have analyzed fungi based on 18S rRNA genes from a taxonomical viewpoint. Indicator genes for specific functions in LBRs such as nitrification, denitrification, and methane production have also been monitored. The population dynamics of microorganisms in LBRs have been partially clarified, but the obtained data remain limited because of highly heterogeneous features of solid wastes inside LBRs. Systematic monitoring of microorganisms should be established to improve LBR performance.     

Compound-specific factors influencing sorption nonlinearity in natural organic matter

Nonlinear sorption by natural organic matter may have a significant impact on the behavior of organic contaminants in soils and sediments. This study presents a molecular probe approach based on linear solvation energy relationships (LSERs) to identify and quantify the molecular interactions causing concentration-dependent sorption and proposes estimation methods for sorption nonlinearities. Sorption isotherms ranging over concentrations of more than 4 orders of magnitude were determined in batch systems for 23 and 16 chemically diverse probe compounds in a lignite sample and a peat soil, respectively. Each sorbent showed characteristic nonlinear sorption with Freundlich exponents (1/n) being 0.7-1. The LSER-based analysis revealed that the strength of nonspecific interactions did not vary with concentration for both sorbents. In lignite, specific interactions did not affect sorption nonlinearity either, suggesting that compound-independent factors of lignite were responsible for the nonlinear sorption. In the peat soil, by contrast, the specific interactions related to the solute polarizability/dipolarity parameter (S) decreased with increasing concentration. Consequently, compounds of higher S values were more susceptible to nonlinear sorption in the peat soil. Phenol probes have shown that hydrogen bond donating properties of sorbate compounds have a substantial impact on the overall strength of sorption with organic matter, but no significant influence on sorption nonlinearity. Heterocyclic aromatic compounds appear to undergo additional interactions that are not accounted for by the LSER. These additional interactions considerably enhance both sorption capacity and nonlinearity.     

Elevated Alpha 1(I) to Alpha 2(I) Collagen Ratio in Dermal Fibroblasts Possibly Contributes to Fibrosis in Systemic Sclerosis

Systemic sclerosis (SSc) is characterized by excessive collagen deposition in the skin and internal organs. Activated fibroblasts are the key effector cells for the overproduction of type I collagen, which comprises the α1(I) and α2(I) chains encoded by COL1A1 and COL1A2, respectively. In this study, we examined the expression patterns of α1(I) and α2(I) collagen in SSc fibroblasts, as well as their co-regulation with each other. The relative expression ratio of COL1A1 to COL1A2 in SSc fibroblasts was significantly higher than that in control fibroblasts. The same result was observed for type I collagen protein levels, indicating that α2(I) collagen is more elevated than α2(I) collagen. Inhibition or overexpression of α1(I) collagen in control fibroblasts affected the α2(I) collagen levels, suggesting that α1(I) collagen might act as an upstream regulator of α2(I) collagen. The local injection of COL1A1 small interfering RNA in a bleomycin-induced SSc mouse model was found to attenuate skin fibrosis. Overall, our data indicate that α2(I) collagen is a potent regulator of type I collagen in SSc; further investigations of the overall regulatory mechanisms of type I collagen may help understand the aberrant collagen metabolism in SSc.     

Additive effects of methyl ammonium bromide or formamidinium bromide in methylammonium lead iodide perovskite solar cells using decaphenylcyclopentasilane

Journal of Materials Science: Materials in Electronics - Fabrication and characterization of methylammonium lead iodide perovskite solar cells incorporated with methylammonium bromide (MABr),...     

Dynamic Power Integrity Control of ATE for Eliminating Overkills and Underkills in Device Testing

This paper proposes a power integrity control technique for dynamically controlling power supply voltage fluctuations for a device under test (DUT), and demonstrates its effectiveness for eliminating the overkills/underkills due to the difference of power supply impedance between an automatic test equipment (ATE) and a practical operating environment of the DUT. The proposed method injects compensation currents into the power supply nodes on the ATE system in a feed-forward manner such that the ATE power supply waveform matches with the one on the customer’s operating environment of the DUT. A method for calculating the compensation current is also described. Experimental results show that the proposed method can emulate the power supply voltage waveform under a customer’s operating condition and eliminate 95 % of overkills/underkills in the maximum operating frequency testing with 105 real silicon devices. Limitations and applications of the proposed method are also discussed.     

Estimating the creep behavior of polycarbonate with changes in temperature and aging time

Thermoplastic resins are typically used without any kind of physical aging treatment. For such materials, creep behavior and physical aging, which depend on time and temperature, occur simultaneously. The effects of these processes are evident after quenching and are recorded in the material as a thermal history. This history strongly influences mechanical properties and creep behavior in particular. Thus, a more thorough understanding of the physical aging process is desirable. We examined the creep deformation of polycarbonate (PC) to reveal the effects of physical aging on creep behavior. The effects were dependent on both time and temperature. The relationship between physical aging and creep behavior exemplified superposition principles with regard to time and both pre-test aging time and pre-test aging temperature. The superposition principles allowed the calculation of creep deformations at a given temperature; the calculated results were corroborated by experimental data.