Bioceramic-mediated chondrocyte hypertrophy promotes calcified cartilage formation for rabbit osteochondral defect repair.

Osteochondral tissue is a highly specialized and complex tissue composed of articular cartilage and subchondral bone that are separated by a calcified cartilage interface. Multilayered or gradient scaffolds, often in conjunction with stem cells and growth factors, have been developed to mimic the respective layers for osteochondral defect repair. In this study, we designed a hyaline cartilage-hypertrophic cartilage bilayer graft (RGD/RGDW) with chondrocytes. Previously, we demonstrated that RGD peptide-modified chondroitin sulfate cryogel (RGD group) is chondro-conductive and capable of hyaline cartilage formation. Here, we incorporated whitlockite (WH), a Mg2+-containing calcium phosphate, into RGD cryogel (RGDW group) to induce chondrocyte hypertrophy and form collagen X-rich hypertrophic cartilage. This is the first study to use WH to produce hypertrophic cartilage. Chondrocytes-laden RGDW cryogel exhibited significantly upregulated expression of hypertrophy markers in vitro and formed ectopic hypertrophic cartilage in vivo, which mineralized into calcified cartilage in bone microenvironment. Subsequently, RGD cryogel and RGDW cryogel were combined into bilayer (RGD/RGDW group) and implanted into rabbit osteochondral defect, where RGD layer supports hyaline cartilage regeneration and bioceramic-containing RGDW layer promotes calcified cartilage formation. While the RGD group (monolayer) formed hyaline-like neotissue that extends into the subchondral bone, the RGD/RGDW group (bilayer) regenerated hyaline cartilage tissue confined to its respective layer and promoted osseointegration for integrative defect repair.

SARS-CoV-2 infection engenders heterogeneous ribonucleoprotein interactions to impede translation elongation in the lungs.

Translational regulation in tissue environments during in vivo viral pathogenesis has rarely been studied due to the lack of translatomes from virus-infected tissues, although a series of translatome studies using in vitro cultured cells with viral infection have been reported. In this study, we exploited tissue-optimized ribosome profiling (Ribo-seq) and severe-COVID-19 model mice to establish the first temporal translation profiles of virus and host genes in the lungs during SARS-CoV-2 pathogenesis. Our datasets revealed not only previously unknown targets of translation regulation in infected tissues but also hitherto unreported molecular signatures that contribute to tissue pathology after SARS-CoV-2 infection. Specifically, we observed gradual increases in pseudoribosomal ribonucleoprotein (RNP) interactions that partially overlapped the trails of ribosomes, being likely involved in impeding translation elongation. Contemporaneously developed ribosome heterogeneity with predominantly dysregulated 5 S rRNP association supported the malfunction of elongating ribosomes. Analyses of canonical Ribo-seq reads (ribosome footprints) highlighted two obstructive characteristics to host gene expression: ribosome stalling on codons within transmembrane domain-coding regions and compromised translation of immunity- and metabolism-related genes with upregulated transcription. Our findings collectively demonstrate that the abrogation of translation integrity may be one of the most critical factors contributing to pathogenesis after SARS-CoV-2 infection of tissues.

Efficacy of Minimally Invasive Oblique Lumbar Interbody Fusion Using Polyetheretherketone Cages for Lumbar Pyogenic Spondylodiscitis Treatment.

(1) Background: This study evaluated the efficacy and safety of a minimally invasive oblique lumbar interbody fusion (OLIF) using polyetheretherketone (PEEK) cages for the treatment of lumbar pyogenic spondylodiscitis. (2) Methods: Fifty-one patients with single-level lumbar pyogenic spondylodiscitis were included in the study. Patients were divided into two groups: anterior lumbar interbody fusion with a tri-cortical iliac bone graft (ALIF+ tri-cortical iliac bone graft) (n = 28) and OLIF using PEEK cages with an autologous bone graft (OLIF+ PEEK cages) (n = 23). Perioperative radiographic parameters, complications, and clinical outcomes in both groups were analyzed and compared. (3) Results: The postoperative and final follow-up LL (lumbar lordosis) and RL (regional lordosis) were improved in both groups (p < 0.001). But, compared with the ALIF group, the OLIF group had more improvement of the RL. The operation time was 79 min for the OLIF group and 101 min for the ALIF group (p < 0.05). The intraoperative blood loss was 92 mL for the OLIF group and 114 mL for the ALIF group (p < 0.05). Significant clinical improvement was observed in visual analogue scale scores for the back and Oswestry Disability Index in both groups (p < 0.001). There was no recurrence of infection. (4) Conclusions: Compared with the ALIF group, the OLIF group had more improvement in radiographic and clinical outcomes. Thus, OLIF using PEEK cages with an autologous bone graft could be proposed for the surgical treatment of lumbar pyogenic spondylodiscitis.

Effect of the High-Pressure Hydrogen Gas Exposure in the Silica-Filled EPDM Sealing Composites with Different Silica Content.

With the increasing interest in hydrogen energy, the stability of hydrogen storage facilities and components is emphasized. In this study, we analyzed the effect of high-pressure hydrogen gas treatment in silica-filled EPDM composites with different silica contents. In detail, cure characteristics, crosslink density, mechanical properties, and hydrogen permeation properties were investigated. Results showed that material volume, remaining hydrogen content, and mechanical properties were changed after 96.3 MPa hydrogen gas exposure. With an increase in the silica content, the crosslink density and mechanical properties increased, but hydrogen permeability was decreased. After treatment, high-silica-content composites showed lower volume change than low-silica-content composites. The crack damage due to the decompression caused a decrease in mechanical properties, but high silica content can inhibit the reduction in mechanical properties. In particular, EPDM/silica composites with a silica content of above 60 phr exhibited excellent resistance to hydrogen gas, as no change in their physical and mechanical properties was observed.

Filler Effects on H2 Diffusion Behavior in Nitrile Butadiene Rubber Blended with Carbon Black and Silica Fillers of Different Concentrations.

Filler effects on H2 diffusion in nitrile butadiene rubbers (NBRs) blended with carbon black and silica fillers of different concentrations are first investigated by employing a volumetric analysis. Total uptake, solubility, and diffusivity of hydrogen for ten filled-NBR, including neat NBR, are determined in an exposed pressure range of 1.3 MPa~92.6 MPa. Filler dependence on hydrogen uptake and diffusion is distinctly observed in the NBRs blended with high abrasion furnace (HAF) carbon black (CB) fillers compared to NBRs blended with medium thermal furnace (MT) CB and silica filler, which is related to the specific surface area of carbon black and interface structure. The HAF CB filled-NBR follows dual sorption behavior combined with Henry's law and the Langmuir model, responsible for two contributions of solubility from polymer and filler. However, a single gas sorption behavior coming from the polymer is observed satisfying Henry's law up to 92.6 MPa for NBR blended with MT CB filled-NBR and silica filled-NBR. Diffusion demonstrates Knudsen and bulk diffusion behavior below and above, respectively, at certain pressures. With increasing pressure, the filler effect on diffusion is reduced, and diffusivity converges to a value. The correlation observed between diffusivity and filler content (or crosslink density) is discussed.

Filler Influence on H2 Permeation Properties in Sulfur-CrossLinked Ethylene Propylene Diene Monomer Polymers Blended with Different Concentrations of Carbon Black and Silica Fillers.

Filler effects on H2 permeation properties in sulfur-crosslinked ethylene propylene diene monomer (EPDM) polymers blended with two kinds of carbon black (CB) and silica fillers at different contents of 20 phr-60 phr are investigated by employing volumetric analysis in the pressure exposure range of 1.2 MPa~9.0 MPa. A linear relationship is observed between the sorbed amount and pressure for H2 gas, which is indicative of Henry's law behavior. The hydrogen solubility of EPDM composites increases linearly with increasing filler content. The magnitude of hydrogen solubility for the filled EPDM composites is dependent on the filler type. The hydrogen solubility is divided into two contributions: hydrogen absorption in the EPDM polymer and hydrogen adsorption at the filler surface. Neat EPDM reveals pressure-dependent bulk diffusion behavior. However, with increasing filler content, the diffusivity for the filled EPDM composites is found to be independent of pressure. The magnitude of filler effects on the hydrogen permeation parameter is measured in the order of high abrasion furnace CB~semireinforcing furnace CB ˃ silica, whose effect is related to the specific surface area of CB particles and interfacial structure. The correlation between the permeation parameters and filler content (or crosslink density) is discussed.

Photoactivatable ribonucleosides mark base-specific RNA-binding sites.

RNA-protein interaction can be captured by crosslinking and enrichment followed by tandem mass spectrometry, but it remains challenging to pinpoint RNA-binding sites (RBSs) or provide direct evidence for RNA-binding. To overcome these limitations, we here developed pRBS-ID, by incorporating the benefits of UVA-based photoactivatable ribonucleoside (PAR; 4-thiouridine and 6-thioguanosine) crosslinking and chemical RNA cleavage. pRBS-ID robustly detects peptides crosslinked to PAR adducts, offering direct RNA-binding evidence and identifying RBSs at single amino acid-resolution with base-specificity (U or G). Using pRBS-ID, we could profile uridine-contacting RBSs globally and discover guanosine-contacting RBSs, which allowed us to characterize the base-specific interactions. We also applied the search pipeline to analyze the datasets from UVC-based RBS-ID experiments, altogether offering a comprehensive list of human RBSs with high coverage (3,077 RBSs in 532 proteins in total). pRBS-ID is a widely applicable platform to investigate the molecular basis of posttranscriptional regulation.

Influence of Carbon Black and Silica Fillers with Different Concentrations on Dielectric Relaxation in Nitrile Butadiene Rubber Investigated by Impedance Spectroscopy.

In neat nitrile butadiene rubber (NBR), three relaxation processes were identified by impedance spectroscopy: α and α' processes and the conduction contribution. We investigated the effects of different carbon black (CB) and silica fillers with varying filler content on the dielectric relaxations in NBR by employing a modified dispersion analysis program that deconvolutes the corresponding processes. The central frequency for the α' process with increasing high abrasion furnace (HAF) CB filler was gradually upshifted at room temperature, while the addition of silica led to a gradual downshift of the center frequency. The activation energy behavior for the α' process was different from that for the central frequency. The use of HAF CB led to a rapid increase in DC conductivity, resulting from percolation. The activation energy for the DC conductivity of NBRs with HAF CB decreased with increasing filler, which is consistent with that reported in different groups.

Chemical RNA digestion enables robust RNA-binding site mapping at single amino acid resolution.

RNA-binding sites (RBSs) can be identified by liquid chromatography and tandem mass spectrometry analyses of the protein-RNA conjugates created by crosslinking, but RBS mapping remains highly challenging due to the complexity of the formed RNA adducts. Here, we introduce RBS-ID, a method that uses hydrofluoride to fully cleave RNA into mono-nucleosides, thereby minimizing the search space to drastically enhance coverage and to reach single amino acid resolution. Moreover, the simple mono-nucleoside adducts offer a confident and quantitative measure of direct RNA-protein interaction. Using RBS-ID, we profiled ~2,000 human RBSs and probed Streptococcus pyogenes Cas9 to discover residues important for genome editing.

SERPINA3 is a key modulator of HNRNP-K transcriptional activity against oxidative stress in HCC.

Most studies about serpin peptidase inhibitor, clade A member 3 (SERPINA3) has been limited to its inhibitory functions and mechanisms. Herein, we report a novel role of SERPINA3 in transcriptional regulation of HCC progression-related genes. Among 19 selected genes through HCC cell isolation system based on telomere length, microarray analyses, and cell-based studies, SERPINA3 was the strongest determinant of increases in telomere length, HCC cell proliferation, survival, migration, and invasion. We also found that SERPINA3 strongly interacted with heterogeneous nuclear ribonucleoprotein K (HNRNP-K) under H2O2 exposure, and the oxidation-elicited SERPINA3-HNRNP-K complex enhanced the promoter activities and transcript levels of a telomere-relating gene (POT1) and HCC-promoting genes (UHRF1 and HIST2H2BE). Intriguingly, the inhibition of SERPINA3 oxidation rendered the transcriptional activity of the SERPINA3-HNRNP-K complex suppressed. Moreover, the co-immunoprecipitated HNRNP-K with SERPINA3 quantitatively correlated with not only the level of SERPINA3 oxidation but also the level of POT1, UHRF1, and HIST2H2BE transcripts and telomere length in HCC tissues. Therefore, the upregulated transcriptional activity of HNRNP-K mediated by SERPINA3 promotes HCC cell survival and proliferation and could be an indicator of poor prognosis for HCC patients.