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As the process of bone regeneration is preceded by an inflammatory response, the immune system has long been considered important for fracture healing. Despite many studies on the contribution of immune cells to bone-related diseases, the role of immune cells in the regeneration therapy of lost bone is not well understood. In addition, various types of cells are involved in the clinical bone regeneration environment, but most of the osteo-biology studies are conducted in an osteoblast-only environment. Here, we investigated the effects of macrophages and dendritic cells on osteogenic differentiation in a co-culture environment involving human periosteal cell-derived osteoblasts, human monocyte-derived osteoclasts, and myeloid-derived cells. In addition, the cluster of myeloid immune cells involved in the clinical bone regeneration process was analyzed through bone defect rat modeling. We found that specific types of myeloid cells and related cytokines increased osteogenic differentiation. These results were confirmed in experiments using myeloid cells originating from human primitive peripheral blood mononuclear cells and by measuring the colonization of macrophages and dendritic cells in an in vivo bone defect environment. In addition, Next generation sequencing (NGS) analysis was performed through RNA sequencing for osteogenesis caused by macrophages and dendritic cells in vitro , which implemented a clinical bone regeneration environment. The results of these experiments suggest that the role of M2 macrophages or dendritic cells is markedly increased during osteogenic differentiation. Therefore, we propose that the exchange of bioactive factors between macrophages and dendritic cells during the bone formation metabolic process is a crucial step of tissue regeneration rather than limited to the initial inflammatory response. This study indicates that M2 macrophages, among myeloid cells, can be mediators that play a vital role in the effective bone regeneration process and shows the potential as a useful next-generation advanced cell therapy for bone regeneration treatment. [ABSTRACT FROM AUTHOR]

Background: It has been suggested that the anterolateral ligament (ALL) is an important anterolateral stabilizer of the knee joint which functions to prevent anterolateral subluxation and anterior subluxation at certain flexion angles in the knee. Purpose: To analyze and systematically interpret the biomechanical function of the ALL. Methods: An online search was conducted for human cadaveric biomechanical studies that tested function of the ALL in resisting anterolateral subluxation and anterior subluxation of the knee. Two reviewers independently searched Medline, Embase, and the Cochrane Database of Systematic Reviews for studies up to 25 September 2018. Biomechanical studies not reporting the magnitude of anterior tibial translation or tibial internal rotation in relation to the function of the ALL were excluded. Results: Twelve biomechanical studies using human cadavers evaluating parameters including anterior tibial translation and/or internal tibial rotation in anterior cruciate ligament (ACL)-sectioned and ALL-sectioned knees were included in the review. Five studies reported a minor increase or no significant increase in anterior tibial translation and internal tibial rotation with further sectioning of the ALL in ACL-deficient knees. Five studies reported a significant increase in knee laxity in tibial internal rotation or pivot shift with addition of sectioning the ALL in ACL-deficient knees. Two studies reported a significant increase in both anterior tibial translation and internal tibial rotation during application of the anterior-drawer and pivot-shift tests after ALL sectioning. Conclusion: There was inconsistency in the biomechanical characteristics of the ALL of the knee in resisting anterolateral and anterior subluxation of the tibia. [ABSTRACT FROM AUTHOR]

Background: The collagen ultrastructure of torn medial meniscus posterior roots (MMPRs) has not been precisely defined. Purpose: To investigate the ultrastructure of torn MMPRs, focusing on their collagen fibers, and to compare the collagen net architecture between intact and torn MMPRs using the Collagen Meniscal Architecture (CMA) scoring system. Study Design: Descriptive laboratory study. Methods: Forty-three human meniscal specimens were obtained from 32 patients with osteoarthritis during total knee arthroplasty between January 2018 and November 2018. There were 23 specimens taken from patients with an MMPR tear and 20 taken from patients without an MMPR tear served as a control group. The presence of an MMPR tear was defined as a complete radial tear within 9 mm of the posterior root attachment. The collagen ultrastructure of the meniscal specimens was assessed with transmission electron microscopy using the CMA scoring system. Patient demographics included sex, age, and body mass index, and radiographic assessments included the Kellgren-Lawrence (K-L) grading system and the mechanical axis angle. Results: The median CMA score was significantly higher in torn MMPRs (5.5 [interquartile range, 3.5-6.0]) than in intact MMPRs (2.0 [interquartile range, 1.5-3.8]) (P <.001). When the CMA scores were converted to CMA grading, 23 torn MMPRs had 1 grade I, 9 grade II, and 13 grade III menisci. In 20 intact MMPRs, there were 12 grade I, 7 grade II, and 1 grade III menisci. No significant differences in sex, age, body mass index, K-L grade, or mechanical axis angle were found between groups. Conclusion: This study showed that torn MMPRs had decreased numbers and disorganized courses of collagen fibers. The structural problem of torn MMPRs can negatively affect meniscal healing, function, and long-term survival after root repair. Clinical Relevance: These results might provide a histopathological reason for the low healing rate after MMPR repair. [ABSTRACT FROM AUTHOR]

• Based on the Kano model, we classified delighters, satisfiers, and dissatisfiers. • Personal and administrative quality lead to satisfaction and reduce dissatisfaction. • Procedural fairness is negatively related to both satisfaction and dissatisfaction. • Distributive and interactional fairness are positively related to satisfaction. • Satisfaction and dissatisfaction affect continuance intention in an opposite way. While college interdisciplinary programs are widespread in South Korea, little is known about what factors can increase college student satisfaction and/or dissatisfaction or whether satisfaction and dissatisfaction are opposite or different constructs. Thus, applying the Stimulus-Organization-Response theory and Kano model, this study seeks to employ service fairness and service quality as diagnostic tools to predict satisfaction and dissatisfaction, examine the asymmetric effect of those attributes on both emotions, and investigate how those emotions are related to the intention of continuing the program. The following results were produced: (1) personal and administrative quality are positively related to satisfaction and negatively related to dissatisfaction, working as satisfiers; (2) procedural fairness is negatively related to both satisfaction and dissatisfaction, working as a dissatisfier; (3) distributive and interactional fairness are positively related to satisfaction, working as delighters; and (4) satisfaction and dissatisfaction are significantly related to continuance intention in an opposite direction. [ABSTRACT FROM AUTHOR]

Abstract Influenza continues to pose a threat to public health by causing illness and mortality in humans. Discovering host factors that regulate influenza virus propagation is vital for the development of novel drugs. We have previously reported that sphingosine kinase (SphK) 1 promotes influenza A virus (IAV) replication in vitro. Here we demonstrate that the other isoform of SphK, SphK2 promotes the replication of influenza A virus (IAV) in cultured cells, and temporary inhibition of SphK1 or SphK2 enhances the host defense against influenza in mice. IAV infection led to an increased expression and phosphorylation of SphK2 in host cells. Furthermore, pharmacologic inhibition or siRNA-based knockdown of SphK2 attenuated IAV replication in vitro. Notably, oral administration of an SphK2-specific inhibitor substantially improved the viability of mice following IAV infection. In addition, the local instillation of an SphK1-specific inhibitor or an inhibitor that globally blocks SphK1 and SphK2 provided protection to IAV-infected mice. Collectively, our results indicate that both SphK1 and SphK2 function as proviral factors during IAV infection in vivo. Therefore, SphK1 and SphK2 represent potential host targets for therapeutics against influenza. Highlights • Sphingosine kinase (SK) 2 accelerates IAV replication in cultured cells. • IAV infection increases the expression and activation of SK2. • Oral administration of SK2 inhibitor improves the viability of mice following IAV infection. • Transient inhibition of SK1 provides protection to IAV-infected mice. [ABSTRACT FROM AUTHOR]

Bone morphogenetic proteins (BMPs) have tremendous therapeutic potential regarding the treatment of bone and musculoskeletal disorders due to their osteo-inductive ability. More than twenty BMPs have been identified in the human body with various functions, such as embryonic development, skeleton genesis, hematopoiesis, and neurogenesis. BMPs can induce the differentiation of MSCs into the osteoblast lineage and promote the proliferation of osteoblasts and chondrocytes. BMP signaling is also involved in tissue remodeling and regeneration processes to maintain homeostasis in adults. In particular, growth factors, such as BMP-2 and BMP-7, have already been approved and are being used as treatments, but it is unclear as to whether they are the most potent BMPs that induce bone formation. According to recent studies, BMP-9 is known to be the most potent inducer of the osteogenic differentiation of mesenchymal stem cells, both in vitro and in vivo. However, its exact role in the skeletal system is still unclear. In addition, research results suggest that the molecular mechanism of BMP-9-mediated bone formation is also different from the previously known BMP family, suggesting that research on signaling pathways related to BMP-9-mediated bone formation is actively being conducted. In this study, we performed a phosphorylation array to investigate the signaling mechanism of BMP-9 compared with BMP-2, another influential bone-forming growth factor, and we compared the downstream signaling system. We present a mechanism for the signal transduction of BMP-9, focusing on the previously known pathway and the p53 factor, which is relatively upregulated compared with BMP-2. [ABSTRACT FROM AUTHOR]

Background: The collagen ultrastructure of the discoid lateral meniscus (DLM) has not been precisely defined. Purpose: To investigate the ultrastructure of the DLM, focusing on its collagen fibers, and to compare the collagen net architecture between intact and torn DLMs using the Collagen Meniscal Architecture (CMA) scoring system. Study Design: Descriptive laboratory study. Methods: Thirty specimens were taken from 30 patients with a diagnosis of a complete DLM using a 1-piece technique. The collagen ultrastructure of the DLMs was assessed with transmission electron microscopy. To evaluate the meniscal ultrastructure, the degree of collagen disruption, intrafibrillar edema, loss of banding, degree of collagen packing, and fibril size variability were assessed and graded from 1 (normal) to 3 (severe disarray) according to the CMA scoring system. The DLM specimens were divided into 3 groups according to the intrasubstance tear: the intact group (group I) had no tear; the simple tear group (group S) had a radial, longitudinal, or horizontal tear; and the complicated tear group (group C) had a complicated horizontal tear. Intact normal meniscus specimens (group N) were used as the control group. Results: There were 10 specimens in group I, 8 in group S, 12 in group C, and 13 in group N. In group I, there were 5 grade 1 and 5 grade 2 menisci; group S had 2 grade 1, 3 grade 2, and 3 grade 3 menisci; group C had 1 grade 1, 4 grade 2, and 7 grade 3 menisci; and group N had 4 grade 1, 7 grade 2, and 2 grade 3 menisci. A significant difference in the CMA score was observed between the 4 groups (P = .009). The median CMA score was significantly lower in group I (2; range, 1-4) than in group S (4; range, 2-7) (P = .041) and group C (4.25; range, 1.5-7) (P = .018). No significant difference was found between groups S, C, and N. Conclusion: Variability existed in the collagen ultrastructure of the DLM, and some DLMs showed a nearly normal ultrastructural pattern. The degree of density and disorganization of the collagen architecture in the DLM was related to the tear. Clinical Relevance: The study results might provide a histological background for partial meniscectomy in the treatment of a symptomatic DLM. [ABSTRACT FROM AUTHOR]

Summary T cells rapidly undergo contraction upon viral clearance, but how T cell function and fate are determined during this phase is unclear. During the contraction phase of an acute infection with lymphocytic choriomeningitis virus, we found that virus-specific CD8 + T cells within the splenic red pulp (RP) had higher two-dimensional (2D) effective affinity than those within the white pulp (WP). This increased antigen recognition of RP-derived CD8 + T cells correlated with more efficient target cell killing and improved control of viremia. FoxP3 + regulatory T cells and cytokine TGF-β limited the 2D-affinity in the WP during the contraction phase. Anatomical location drove gene expression patterns in CD8 + T cells that led to preferential differentiation of memory precursor WP T cells into long-term memory cells. These results highlight that intricate regulation of T cell function and fate is determined by anatomic compartmentalization during the early immune contraction phase. [ABSTRACT FROM AUTHOR]

Purpose: The aim of the study was to analyse the change in length and tension of the reconstructed single-bundle posterior cruciate ligament (PCL) with three different femoral tunnels at different knee flexion angles by use of three-dimensional finite element method. Methods: The right knees of 12 male subjects were scanned with a high-resolution computed tomography scanner at four different knee flexion angles (0°, 45°, 90° and 135°). Three types of single-bundle PCL reconstruction were then conducted in a 90° flexion model: femoral tunnels were created in anterolateral (AL), central and posteromedial (PM) regions of the footprint. Length versus flexion curves and tension versus flexion curves were generated. Results: Between 0° and 90° of knee flexion, changes in length and tension in the PM grafts were not significant. Whereas the lengths and tension of the AL and central grafts significantly increased in the same flexion range. The length and tension of the PM grafts at 135° of knee flexion were significantly higher than those at 90° of knee flexion, whereas the AL and the central grafts showed only slight length changes beyond 90° of flexion. However, the tension of the AL graft increased significantly beyond 90° of flexion. Conclusions: Changes in the graft length, and tension were generally affected by different femoral tunnels and knee flexion angles. In groups with the AL and PM single-bundle reconstruction, the graft tension increased beyond 90° of knee flexion when the graft is tensioned at 90° of flexion. These data suggest that final fixation angle at 90° for the AL or PM graft would induce graft overtension in high knee flexion of 135°. Whereas central graft which is fixed in 90° of flexion is desirable in terms of prevention of graft overtension. Because the graft tension within it was relatively constant beyond 90° of flexion. [ABSTRACT FROM AUTHOR]

Zanthoxylum piperitum has been used as a traditional Asian medicine to treat hypertension, stroke, bruise and muscle pain. It has been known to induce detoxification; affect anti-bacterial, anti-oxidant, and tyrosinase activity; inhibit osteosarcoma proliferation; anti-osteoarthritis inflammation. In this study, we aim to identify the therapeutic effect of Z. piperitum 90% EtOH extract (ZPE-LR) on rheumatoid arthritis. We investigated the anti-rheumatoid arthritis and -immunomodulatory activities of the ZPE-LR in collagen-induced arthritic (CIA) mice, a rheumatoid arthritis animal model. In order to assess the analgesic effects of ZPE-LR in vivo , acetic acid injection, formaldehyde injection, hot plate model was used. The mechanism for anti-inflammatory activity of ZPE-LR was identified with LPS-stimulated Raw 264.7 cells. Pharmacologically, oral administration of ZPE-LR into CIA mice resulted in a significant and dose-dependent decrease in clinical arthritis score and paw swelling compared to untreated negative control. Pathologic examination showed that ZPE-LR prevented morphological change in cartilage and destruction of phalanges in CIA mice. This protective effect was associated with reduced pain, inflammatory mediators such as NO, TNF-α, IL-1β, and IL-6, as well as COX-2 and iNOS expression. Furthermore, reduction of phosphor-ERK and BDNF indicates a novel rheumatoid arthritis-regulating mechanism by ZPE-LR treatment. These data suggest that the administration of ZPE-LR remarkably inhibited CIA progression and might be helpful in suppressing inflammation and pain in rheumatoid arthritis. [Display omitted] • ZPE-LR have been effectively inhibited on rheumatoid arthritis-induced pain and inflammation. • ZPE-LR prevented morphological change in cartilage and destruction of phalanges in CIA mice on pathologic examination. [ABSTRACT FROM AUTHOR]

Sphingosine analogs display diverse immunoregulatory activities with curative potential in autoimmune diseases and viral infections. Recently, the sphingosine analog AAL-R was shown to increase DC activation upon TLR7 stimulation. Here, we investigated the effect of AAL-R on activation of dendritic cells (DCs) infected by lymphocytic choriomeningitis virus (LCMV). Concomitant treatment of LCMV-infected DCs with AAL-R enhanced DC maturation and DC ability to stimulate and expand antiviral CD8+ T cells. Importantly, AAL-R's stimulatory activity was abrogated in type I interferon (IFN) receptor-deficient DCs following LCMV infection. In support of this observation, AAL-R increased type I IFN production from DCs infected with LCMV. Taken together, the sphingosine analog could directly act on DCs to promote defensive host DC responses to the viral invasion via type I IFN signaling. [ABSTRACT FROM AUTHOR]

Abstract: Measles virus (MV) manipulates host factors to facilitate virus replication. Sphingosine kinase (SK) is an enzyme catalyzing the formation of sphingosine 1-phosphate and modulates multiple cellular processes including the host defense system. Here, we determined the role of SK1 in MV replication. Overexpression of SK1 enhanced MV replication. In contrast, inhibition of SK impaired viral protein expression and infectious virus production from cells expressing MV receptor, SLAM or Nectin-4. The inhibition of virus replication was observed when the cells were infected by vaccine strain or wild type MV or V/C gene-deficient MV. Importantly, SK inhibition suppressed MV-induced activation of NF-κB. The inhibitors specific to NF-κB signal pathway repressed the synthesis of MV proteins, revealing the importance of NF-κB activation for efficient MV replication. Therefore, SK inhibition restricts MV replication and modulates the NF-κB signal pathway, demonstrating that SK is a cellular factor critical for MV replication. [Copyright &y& Elsevier]

Tiron is a potent antioxidant that counters the pathological effects of reactive oxygen species (ROS) production due to oxidative stress in various cell types. We examined the effects of tiron on mitochondrial function and osteoblastic differentiation in human periosteum-derived cells (hPDCs). Tiron increased mitochondrial activity and decreased senescence-associated β-galactosidase activity in hPDCs; however, it had a detrimental effect on osteoblastic differentiation by reducing alkaline phosphatase (ALP) activity and alizarin red-positive mineralization, regardless of H2O2 treatment. Osteoblast-differentiating hPDCs displayed increased ROS production compared with non-differentiating hPDCs, and treatment with tiron reduced ROS production in the differentiating cells. Antioxidants decreased the rates of oxygen consumption and ATP production, which are increased in hPDCs during osteoblastic differentiation. In addition, treatment with tiron reduced the levels of most mitochondrial proteins, which are increased in hPDCs during culture in osteogenic induction medium. These results suggest that tiron exerts negative effects on the osteoblastic differentiation of hPDCs by causing mitochondrial dysfunction. [ABSTRACT FROM AUTHOR]

Purpose: To (1) evaluate the optimal drill orientation of the anterolateral ligament (ALL) femoral tunnel to minimize collision with the anterior cruciate ligament (ACL) femoral tunnel during anatomical ACL reconstruction according to the need for far-cortex drilling and (2) investigate the geometric factors that affect tunnel collision secondary to drill orientation of the ALL femoral tunnel. Methods: A three-dimensional femoral model of patients who underwent anatomical single-bundle ACL reconstruction between 2015 and 2016 was constructed, and the geometric factors were evaluated. Virtual ALL femoral tunnels were created to simulate 45 drilling conditions. For each condition, whether the virtual ALL femoral tunnel and its trajectory violated the femoral cortex and the minimum distance between tunnels was investigated. Results: Thirty-nine subjects were included. Overall violation rates of the femoral cortex by the ALL tunnels and its trajectories were 11.1% (195 of 1755 conditions) and 40.7% (714 of 1755 conditions), respectively. A drilling angle of axial 0° and coronal − 40° showed the longest minimum distance between tunnels without femoral cortex violation by the ALL tunnel (6.3 ± 4.0 mm; collision rate 2.6% [1 of 39 subjects]). With simultaneous consideration of the ALL tunnel's trajectory representing far-cortex drilling, a drill angle of axial 40° and coronal 10° showed the longest minimum distance between tunnels without femoral cortex violation (0.6 ± 3.9 mm; collision rate 38.5% [15 of 39 subjects]). For surgical techniques requiring far-cortex drilling, regression analyses were performed on geometric factors that could affect tunnel collision, which revealed that the sagittal inclination angle of the ACL and the distance between the ACL femoral tunnel's outlet and ALL's femoral attachment were associated with tunnel collision. Conclusion: The optimal drill orientations of the ALL femoral tunnel to minimize collision with the ACL femoral tunnel were axial 0° and coronal − 40° for surgical techniques not requiring far-cortex drilling and axial 40° and coronal 10° for techniques requiring far-cortex drilling. For techniques requiring far-cortex drilling, additional adjustment for orientation of the ACL femoral tunnel is required to reduce the risk of tunnel collision. Therefore, an individualized surgical strategy should be applied according to the graft fixation method of the ALL femoral tunnel. [ABSTRACT FROM AUTHOR]

Purpose: The purpose of this study was to describe a modified trans-tendon method of repairing partial articular surface lesions, which restored the footprint of the rotator cuff anatomically. Methods: The 24 consecutive patients with modified trans-tendon method which allowed a wider pressurized contact area by use of additional knotless anchor were included in this study. All patients were evaluated with ASES score and visual analog scale (VAS) preoperatively, postoperative 3 and 12 months. The strength was measured using Isobex digital strength analyzer preoperatively and postoperative 12 months. Results: The ASES scores significantly improved from preoperative 38 ± 13 to 63 ± 5 at 3 months, and 89 ± 5 at 12 months postoperatively. The VAS scores also significantly improved from preoperative 6.6 ± 1.1 to 2 ± 0.7 at 3 months, 0.6 ± 0.7 at 12 months. The strengths significantly increased postoperatively, and there were no significant differences between affected and unaffected shoulders at 12 months postoperatively ( P > 0.05). The 22 of 24 patients were either satisfied or very satisfied with postoperative result at 12 months postoperatively. Conclusion: This arthroscopic-modified trans-tendon suture bridge technique for partial-thickness articular side tears of the rotator cuff has shown excellent functional results and very high satisfaction rate of patient during the 12-month follow-up period. Level of evidence: Evidence therapeutic study, Level IV. [ABSTRACT FROM AUTHOR]

We describe a method of arthroscopically assisted, mini-open, anatomical reconstruction of the coracoclavicular ligament. This method restores both components of the native ligament with the aim of achieving maximum stability with minimal disruption of the normal anatomy. Using the same principles of ligament reconstruction that are employed in other joints, transosseous tunnels are created following the native footprints of the conoid and trapezoid ligaments and an autologous graft is fixed using a PEEK screw. Adequate healing of the ligament occurs within the bone, to prevent stress risers with an appropriate working length. This procedure is unique, as it replaces the torn ligament with a natural substitute, in the appropriate location, through a minimally invasive procedure. This technique would be suitable for treatment of patients with either grade III or V acute acromioclavicular dislocations. Clinical outcomes for the first 13 consecutive patients treated with this procedure are reported, revealing excellent satisfaction rates with a Constant score of 96.6 at final follow-up. [ABSTRACT FROM AUTHOR]

Abstract: Aeroelastic analyses are performed for a 2-D typical section model with multiple nonlinearities. The differences between a system with multiple nonlinearities in its pitch and plunge spring and a system with a single nonlinearity in its pitch are thoroughly investigated. The unsteady supersonic aerodynamic forces are calculated by the doublet point method (DPM). The iterative V-g method is used for a multiple-nonlinear aeroelastic analysis in the frequency domain and the freeplay nonlinearity is linearized using a describing function method. In the time domain, the DPM unsteady aerodynamic forces, which are based on a function of the reduced frequency, are approximated by the minimum state approximation method. Consequently, multiple structural nonlinearities in the 2-D typical wing section model are influenced by the pitch to plunge frequency ratio. This result is important in that it demonstrates that the flutter speed is closely connected with the frequency ratio, considering that both pitch and plunge nonlinearities result in a higher flutter speed boundary than a conventional aeroelastic system with only one pitch nonlinearity. Furthermore, the gap size of the freeplay affects the amplitude of the limit cycle oscillation (LCO) to gap size ratio. [Copyright &y& Elsevier]

Abstract: Sphingosine 1-phosphate (S1P)-metabolizing enzymes regulate the level of bioactive sphingolipids that have curative potential. Recently, S1P-metabolizing enzymes such as sphingosine kinase 1 and S1P lyase were shown to regulate influenza virus replication and the virus-induced cytopathogenicity. The mechanism appeared to employ a JAK/STAT type I interferon signaling pathway that induces anti-viral status. Further, sphingosine analogs altered cytokine responses upon influenza virus infection. This article focuses on recent discoveries about the sphingolipid system that influences on host protection from viral virulence and the involvement of cytokine signaling in its underlying mechanisms. Deciphering the steps of this pathway could help us envision how the modulation of sphingolipid metabolism can be applied as a therapeutic approach to overcome infectious diseases. [Copyright &y& Elsevier]

Abstract: Hypoxia is known to regulate the expression of genes involved in the migration of various cell types. Although many studies have shown that hypoxia increases cell migration, it still remains unclear whether hypoxia could modulate the stromal cell derived factor-1 (SDF-1)-dependent migration of leukemic cell. Herein, we demonstrated that the SDF-1-dependent migration of HL-60, was reduced under hypoxia with no comparable decrease of CXC-type chemokine receptor CXCR4, a cognate receptor for SDF-1. Furthermore, we showed that migration toward SDF-1 was reduced by inactivation of either serine/threonine kinase Akt or extracellular signal regulated kinase Erk, which was confirmed by selective pathway inhibitor LY294002 and PD98059. In our results, phosphorylation of Erk was increased under hypoxia, but phosphorylation of Akt was attenuated on the contrary. These results led us to conclusion that hypoxia could inhibit the SDF-1-dependent migration of HL-60 via blocking of Akt activation. [Copyright &y& Elsevier]

Purpose: The purpose of this study was to prospectively investigate osteotomy gap filling rates on serial plain radiographs, and to evaluate whether alignment correction is maintained after medial opening wedge high tibial osteotomy (MOWHTO) using a locking plate without bone graft. Methods: Between March 2014 and June 2017, MOWHTO was performed without bone graft regardless of gap size. Radiographs were taken preoperatively, postoperatively, at 1, 3, 6, 12, 18, and 24 months after surgery. Radiographic examinations included a weight bearing long-standing anteroposterior (AP) view of the whole lower extremity, as well as, the AP, lateral, and both oblique views of the knee. Bone healing was measured on the medial oblique view of the knee. The postoperative alignment correction and its maintenance were assessed using the three radiologic parameters of the weight-bearing line (WBL) ratio, the hip–knee–ankle angle (HKAA), and the medial proximal tibial angle (MPTA) on the weight-bearing long-standing AP view of the lower extremity. Results: Fifty-two consecutive patients underwent MOWHTO, but three patients failed to follow-up for more than 24 months. A total of 49 patients were assessed in this study. The median opening gap height was 10.0 mm (IQR, 8.0–12.0; range, 7–20). On immediate post-operative radiographs, the mean gap filling was 31.4 ± 3.6%. After 1, 3, 6, 12, 18, and 24 months, the mean gap filling rates increased to 38.7 ± 4.4%, 51.4 ± 6.6%, 66.5 ± 5.1%, 84.8 ± 7.0%, 92.4 ± 5.6%, and 97.8 ± 2.3%, respectively. Statistical differences were observed between all the follow-up evaluations (P < 0.001). Statistical differences in the WBL ratio, HKAA, and MPTA were observed between preoperatively and 1 month after surgery (P < 0.001). The mean PTSA increased significantly from preoperatively to postoperatively (P < 0.001). However, no statistical differences were found between the post-operative follow-up radiographs performed for these four values. Conclusion: MOWHTO using a locking plate without bone graft achieved at least 90% bone healing and had no loss in correction at 2 years postoperatively. Level of Evidence: III. [ABSTRACT FROM AUTHOR]

Abstract: A discoid medial meniscus is a relatively rare pathology of the knee joint. And bilateral cases are even more rare. In the review of the literature by the authors, Only 13 cases of bilateral medial discoid menisci have been reported. We present one case of bilateral discoid medial menisci. One knee had an incomplete type verified by both magnetic resonance imaging (MRI) and arthroscopy. The other knee showed a complete type of medial discoid meniscus in MRI. Therefore, we suggest that similar pathology in the contralateral knee should be suspected, even if the patient is asymptomatic, if a complete or incomplete discoid medial meniscus is identified. [Copyright &y& Elsevier]

With the emergence of COVID‐19 worldwide, interest in blended learning in higher education is rapidly increasing. Despite the fact that the unipolar view that satisfaction and dissatisfaction are measured separately has been emphasized in higher education, fewer efforts have been made in a blended learning context. Therefore, this study attempts to document the unipolar view of satisfaction and dissatisfaction in a blended learning context by adopting the Kano model. More specifically, the objectives of this study is to (1) conceptualize online and offline service dimensions in blended learning, (2) examine the asymmetric effect between satisfaction and dissatisfaction, and (3) apply those results to the Kano model. The following results are reported: (1) system quality and assurance are symmetric, functioning as a satisfier; and (2) information quality, responsiveness, and tangibility are asymmetric, functioning as a delighter. Theoretical and practical implications are addressed. [ABSTRACT FROM AUTHOR]

DNA methylation is an epigenetic modification that regulates gene expression and plays an essential role in hematopoiesis. UHRF1 and DNMT1 are both crucial for regulating genome-wide maintenance of DNA methylation. Specifically, it is well known that hypermethylation is crucial characteristic of acute myeloid leukemia (AML). However, the mechanism underlying how DNA methylation regulates the differentiation of AML cells, including THP-1 is not fully elucidated. In this study, we report that UHRF1 or DNMT1 depletion enhances the phorbol-12-myristate-13-acetate (PMA)-induced differentiation of THP-1 cells. Transcriptome analysis and genome-wide methylation array results showed that depleting UHRF1 or DNMT1 induced changes that made THP-1 cells highly sensitive to PMA. Furthermore, knockdown of UHRF1 or DNMT1 impeded solid tumor formation in xenograft mouse model. These findings suggest that UHRF1 and DNMT1 play a pivotal role in regulating differentiation and proliferation of THP-1 cells and targeting these proteins may improve the efficiency of differentiation therapy in AML patients. [ABSTRACT FROM AUTHOR]

Methylparaben is most frequently used as an antimicrobial preservative in pharmaceuticals and foods. Methylparaben has been subjected to toxicological studies owing to the increasing concern regarding its possible impact on the environment and human health. However, the cytotoxicity and underlying mechanisms of methylparaben exposure in human lung cells have not been explored. Here, we investigated the effect of methylparaben on cell cycle, apoptotic pathways, and changes in the transcriptome profiles in human lung cells. Our results demonstrate that treatment with methylparaben causes inhibition of cell growth. In addition, methylparaben induced S- and G2/M-phase arrest as a result of enhanced apoptosis. Transcriptome analysis using RNA-seq revealed that mRNA expression of ER stress- and protein misfolding-related gene sets was upregulated in methylparaben-treated group. RNA splicing- and maturation-related gene sets were significantly down-regulated by methylparaben treatment. Interestingly, RNA-seq analysis at the transcript level revealed that alternative splicing events, especially retained intron, were markedly changed by a low dose of methylparaben treatment. Altogether, these data show that methylparaben induces an early phase of apoptosis through cell cycle arrest and downregulation of mRNA maturation. [ABSTRACT FROM AUTHOR]

DNA methylation is an epigenetic modification that regulates gene expression and plays an essential role in hematopoiesis. UHRF1 and DNMT1 are both crucial for regulating genome-wide maintenance of DNA methylation. Specifically, it is well known that hypermethylation is crucial characteristic of acute myeloid leukemia (AML). However, the mechanism underlying how DNA methylation regulates the differentiation of AML cells, including THP-1 is not fully elucidated. In this study, we report that UHRF1 or DNMT1 depletion enhances the phorbol-12-myristate-13-acetate (PMA)-induced differentiation of THP-1 cells. Transcriptome analysis and genome-wide methylation array results showed that depleting UHRF1 or DNMT1 induced changes that made THP-1 cells highly sensitive to PMA. Furthermore, knockdown of UHRF1 or DNMT1 impeded solid tumor formation in xenograft mouse model. These findings suggest that UHRF1 and DNMT1 play a pivotal role in regulating differentiation and proliferation of THP-1 cells and targeting these proteins may improve the efficiency of differentiation therapy in AML patients. [ABSTRACT FROM AUTHOR]

This study aimed to assess the safety and effectiveness of the highly cross-linked hyaluronic acid—LBSA0103—in patients with knee osteoarthritis (OA) as per the prescribing information (PI) in South Korea. A total of 3,140 subjects aged ≥19 years were enrolled in this post-marketing surveillance (PMS) study from 2013 to 2019. The subjects received one or two injections of LBSA0103. The median duration of follow-up was 308 days. Adverse events (AEs), adverse drug reactions (ADRs), and serious AEs (SAEs) were monitored. Effectiveness was evaluated based on an index of effectiveness in accordance with the guidelines established by the Ministry of Food and Drug Safety and using a 100-mm visual analog scale (VAS) for weight-bearing pain. Overall, 250 subjects (7.96%) experienced 292 AEs and of these, unexpected AEs occurred in 114 subjects (3.63% [95% CI: 3.00–4.35]). Injection site pain was the most frequent AE reported by 81 subjects (2.58% [95% confidence intervals (CI): 2.05–3.20]). One hundred subjects experienced 108 ADRs (3.18% [95% CI: 2.60, 3.86]) and 15 unexpected ADRs were experienced by 13 subjects (0.41% [95% CI: 0.22–0.71]). Seventeen subjects experienced 22 SAEs (0.54% [95% CI: 0.32–0.87]) during the entire PMS period, and all were considered "unlikely" related to the study drug. Most AEs were mild in terms of severity and resolved during the study period. LBSA0103 was also effective in relieving symptomatic pain in knee OA patients. The condition in more than 80% of the subjects was considered to be improved when assessed by the investigators. LBSA0103 resulted in a significant reduction in the mean VAS score at 12 weeks after the first and second injections (24.79 (± 20.55) mm and 17.63 (±12.31) mm, respectively; p<0.0001). In conclusion, LBSA0103, used for the treatment of knee OA in a real-world setting, was well tolerated, with an acceptable safety profile and consistent therapeutic effect. [ABSTRACT FROM AUTHOR]

Influenza continues to pose a threat to humans by causing significant morbidity and mortality. Thus, it is imperative to investigate mechanisms by which influenza virus manipulates the function of host factors and cellular signal pathways. In this study, we demonstrate that influenza virus increases the expression and activation of sphingosine kinase (SK) 1, which in turn regulates diverse cellular signaling pathways. Inhibition of SK suppressed virus-induced NF-κB activation and markedly reduced the synthesis of viral RNAs and proteins. Further, SK blockade interfered with activation of Ran-binding protein 3 (RanBP3), a cofactor of chromosome region maintenance 1 (CRM1), to inhibit CRM1-mediated nuclear export of the influenza viral ribonucleoprotein complex. In support of this observation, SK inhibition altered the phosphorylation of ERK, p90RSK, and AKT, which is the upstream signal of RanBP3/CRM1 activation. Collectively, these results indicate that SK is a key pro-viral factor regulating multiple cellular signal pathways triggered by influenza virus infection. [ABSTRACT FROM AUTHOR]

Circular RNA (circRNA) is a non-coding RNA with a covalently closed loop structure and usually more stable than messenger RNA (mRNA). However, coding sequences (CDSs) following an internal ribosome entry site (IRES) in circRNAs can be translated, and this property has been recently utilized to produce proteins as novel therapeutic tools. However, it is difficult to produce large proteins from circRNAs because of the low circularization efficiency of lengthy RNAs. In this study, we report that we successfully synthesized circRNAs with the splint DNA ligation method using RNA ligase 1 and the splint DNAs, which contain complementary sequences to both ends of precursor linear RNAs. This method results in more efficient circularization than the conventional enzymatic method that does not use the splint DNAs, easily generating circRNAs that express relatively large proteins, including IgG heavy and light chains. Longer splint DNA (42 nucleotide) is more effective in circularization. Also, the use of splint DNAs with an adenine analog, 2,6-diaminopurine (DAP), increase the circularization efficiency presumably by strengthening the interaction between the splint DNAs and the precursor RNAs. The splint DNA ligation method requires 5 times more splint DNA than the precursor RNA to efficiently produce circRNAs, but our modified splint DNA ligation method can produce circRNAs using the amount of splint DNA which is equal to that of the precursor RNA. Our modified splint DNA ligation method will help develop novel therapeutic tools using circRNAs, to treat various diseases and to develop human and veterinary vaccines. [ABSTRACT FROM AUTHOR]

Prostate-specific antigen (PSA) is a protein highly expressed in cancer cells of the prostate gland. In this study, the PSA recombinant protein was fused to the human immunoglobulin G crystallizable fragment (Fc) with the endoplasmic reticulum (ER) Lys-Asp-Glu-Leu (KDEL) retention signal tag for the plant expression system. Agrobacterium-mediated plant transformation was applied to generate transgenic tobacco plants expressing PSA tagged to KDEL (PSAK), PSA-Fc, and PSA-Fc fused to KDEL (PSA-FcK). Western blot analysis showed that the PSA-FcK protein was the most highly expressed among the three different proteins in the transgenic plant leaf. The PSAK, PSA-Fc, and PSA- FcK proteins were successfully purified from plant leaves. Reduced and non-reduced sodium dodecyl sulfate–polyacrylamide gel electrophoresis revealed that the PSA proteins fused to Fc and FcK were dimerized, whereas the PSA itself was not dimerized. N-glycan analysis demonstrated that the N-glycan profile of the PSA-Fc was mainly the structure with α(1,3)-fucose (Fuc) or β(1,2)-xylose (Xyl) glycan residues, whereas the PSA-Fc tagged with the KDEL ER retention signal had mainly an oligomannose-type structure. ELISA and SPR showed that PSA-FcK had a higher binding activity to Fc gamma receptor I than PSA-Fc and human Fc. In addition, PSA-FcK induced anti-PSA, and thus has the potential to be utilized as a prostate cancer vaccine. [ABSTRACT FROM AUTHOR]

Background: It is unclear whether different anterior cruciate ligament (ACL) graft trajectories in the distal femur would have different effects on stress generated within the distal femur around the femoral tunnel during knee motion. Thus, the purpose of this study was to determine differences in stress patterns around the femoral tunnel created by trans-portal (TP) vs. modified trans-tibial (TT) technique in anatomical ACL reconstruction at different knee flexion angles.Methods: Twelve male subjects' right knees were scanned with a high-resolution computed tomography (CT) scanner (slice thickness: 1 mm) at four different knee flexion angles (0°, 45°, 90°, and 135°). Three-dimensional (3D) models of these four different flexion angles were created and manipulated with several modelling programs. For the TP group, the virtual femoral tunnelling procedure was performed in a 135° flexion model from the low far anteromedial (AM) portal. For the modified TT group, the same knee models were drilled through the modified TT technique at 90° of flexion separately. Virtual grafts under tension of 40 N were put into corresponding bone tunnel and fixed at the outer aperture of femoral tunnels to simulate the suspensory fixation, followed by fixation of the grafts at the middle of tibial tunnels in the 0° knee flexion models. Finally, the models were exported to a finite element analysis package and analysed using ABAQUS/Explicit code (ABAQUS, USA) to monitor the stress occurring at the node where stress distribution occurred most significantly in the femoral bone around the bone tunnel.Results: In general, both groups showed a high stress distribution in bony structures around inner and outer orifices of the femoral tunnel. Mean maximal stresses occurring at the lateral femoral condyle around the inner orifice of the femoral tunnel in the TP group were found to be significantly greater than those in the modified TT group at all flexion angles except 90° of flexion. Mean maximal stresses monitored around the outer orifice of the femoral tunnel in the TP group were also significantly greater than those in the modified TT group at all flexion angles.Conclusions: Different tunnelling technologies could yield different stress patterns in the lateral femoral condyle around the femoral tunnel. During knee motion, higher stresses were noticed in the TP group than in the modified TT group, especially around inner and outer orifices of the tunnel. Position of the tunnel after reconstruction with the TP technique can have a greater effect on the stress increase in the femur compared to that with the modified TT technique. [ABSTRACT FROM AUTHOR]

The article reports a study which was conducted to determine at which flexion angles the grafts should be fixed to best restore patellar stability for current medial patellofemoral ligament (MPFL) reconstruction during knee flexion in vivo. To obtain more patellar stability throughout range of knee motion, a femoral drill hole located at isometric points inside the anatomic MPFL attachment sites is suggested. The best angle for graft fixation would be near 30 degrees of knee flexion.

Abstract: Severe patellar infera accompanied by pathologic shortening and fibrosis of the patellar tendon is relatively an uncommmon condition but can cause recalcitrant joint stiffness and limited range of motion after knee surgery or injury. This report presents a case of a 49-year-old male with severe patellar infera and limping due to joint stiffness after three-time knee surgeries. We report a successful outcome along with a finding of the well-healed allograft after the reconstruction of a patellar tendon using Achilles tendon-bone allograft. [Copyright &y& Elsevier]

Abstract: The adsorption mechanisms of metal cations on the surfaces of clay minerals and other minerals are important for understanding their fate in contaminated soils and in waste water treatment. The reactions of Cu2+ and Pb2+ heavy metal cations with layered double hydroxide (LDH), has been investigated in aqueous solutions to elucidate their removal behaviors by LDH. The reaction of aqueous CuCl2 solution with Mg/Al-LDH resulted in precipitation of Cu hydroxide chloride (Cu7Cl4(OH)10·H2O) along with slight decomposition of Mg/AI-LDH. On the other hand, the reaction of aqueous PbCl2 solution led to steady decomposition of Mg/Al-LDH structure along with the formation of various Pb precipitates such as carbonate chloride (PbCO3PbCl2) and hydroxide chloride (Pb(OH)Cl). Experimental techniques such as X-ray diffraction, X-ray photoelectron spectroscopy and cation removal isotherms were used to decipher the mechanism of uptake. This study indicates that LDH removes Cu2+ and Pb2+ transition metal cations from aqueous solution mainly by surface adsorption and precipitation. [Copyright &y& Elsevier]

Mechanical forces are pervasive in the inflammatory site where dendritic cells (DCs) are activated to migrate into draining lymph nodes. For example, fluid shear stress modulates the movement patterns of DCs, including directness and forward migration indices (FMIs), without chemokine effects. However, little is known about the effects of biomechanical forces on the activation of DCs. Accordingly, here we fabricated a microfluidics system to assess how biomechanical forces affect the migration and activity of DCs during inflammation. Based on the structure of edema, we proposed and experimentally analyzed a novel concept for a microchip model that mimicked such vascular architecture. The intensity of shear stress generated in our engineered chip was found as 0.2–0.6 dyne/cm2 by computational simulation; this value corresponded to inflammation in tissues. In this platform, the directness and FMIs of DCs were significantly increased, whereas the migration velocity of DCs was not altered by shear stress, indicating that mechanical stimuli influenced DC migration. Moreover, DCs with shear stress showed increased expression of the DC activation markers MHC class I and CD86 compared with DCs under static conditions. Taken together, these data suggest that the biomechanical forces are important to regulate the migration and activity of DCs. [ABSTRACT FROM AUTHOR]

Bacteria utilize endoribonuclease-mediated RNA processing and decay to rapidly adapt to environmental changes. Here, we report that the modulation of hns mRNA stability by the endoribonuclease RNase G plays a key role in Salmonella Typhimurium pathogenicity. We found that RNase G determines the half-life of hns mRNA by cleaving its 5′ untranslated region and that altering its cleavage sites by genome editing stabilizes hns mRNA, thus decreasing S. Typhimurium virulence in mice. Under anaerobic conditions, the FNR-mediated transcriptional repression of rnc encoding RNase III, which degrades rng mRNA, and simultaneous induction of rng transcription resulted in rapid hns mRNA degradation, leading to the derepression of genes involved in the Salmonella pathogenicity island 1 (SPI-1) type III secretion system (T3SS). Together, our findings show that RNase III and RNase G levels-mediated control of hns mRNA abundance acts as a regulatory pathway upstream of a complex feed-forward loop for SPI-1 expression. Author summary: Recent studies have shown that pathogenic bacteria with ribonuclease mutations display attenuated virulence, impaired mobility, and reduced proliferation in host cells. However, the molecular mechanisms underlying ribonuclease-associated pathogenesis have not yet been characterised. Here, we provide strong experimental evidence that the coordinated modulation of endoribonuclease activity constitutes an additional regulatory layer upstream of a complex feed-forward loop controlling global regulatory systems in the Salmonella pathogenicity island 1 (SPI-1) type III secretion system (T3SS). In addition, we showed that this regulatory pathway plays a key role in the virulence of S. Typhimurium in the host. Thus, our study improves the understanding of the mechanisms through which bacterial pathogens sense the host environment and respond precisely by expressing gene products required for adaptation to that particular niche. [ABSTRACT FROM AUTHOR]

Chronic viral infections are often established by the exploitation of immune-regulatory mechanisms that result in nonfunctional T cell responses. Viruses that establish persistent infections remain a serious threat to human health. Sphingosine kinase 2 (SphK2) generates sphingosine 1-phosphate, which is a molecule known to regulate multiple cellular processes. However, little is known about SphK2's role during the host immune responses to viral infection. Here, we demonstrate that SphK2 functions during lymphocytic choriomeningitis virus Cl 13 (LCMV Cl 13) infection to limit T cell immune pathology, which subsequently aids in the establishment of virus-induced immunosuppression and the resultant viral persistence. The infection of Sphk2-deficient (Sphk2-/-) mice with LCMV Cl 13 led to the development of nephropathy and mortality via T cell-mediated immunopathology. Following LCMV infection, Sphk2-/- CD4+ T cells displayed increased activity and proliferation, and these cells promoted overactive LCMV Cl 13-specific CD8+ T cell responses. Notably, oral instillation of an SphK2-selective inhibitor promoted protective T cell responses and accelerated the termination of LCMV Cl 13 persistence in mice. Thus, SphK2 is indicated as an immunotherapeutic target for the control of persistent viral infections. [ABSTRACT FROM AUTHOR]

Background: There has been no gold standard of the initial treatment strategy for acute patellar dislocation (APD) with osteochondral fracture (OCF). Hence the study aim is firstly, to review and compare clinical outcomes of patients who underwent conservative treatment for APD with or without OCF. Secondly, to characterize the location and size of fracture fragment.Methods: Sixty-nine consecutive patients who were retrospectively evaluated after first-time APD over a 2- year period were divided into two groups (group 1 (n = 24): APD with OCF and group 2 (n = 45): APD only). Magnetic resonance imaging (MRI) was used to assess patients with APD and OCF from the medial patella. All patients were treated with a supervised course of immobilization followed by progressive range of motion and strength exercise protocol. History of a recurrent dislocation, radiologic and functional scores were analyzed.Results: Redislocation rate was 31.2% in group 1 and 26.6% in group 2, showing no significant difference between the two groups (p = 0.690). Intergroup differences in terms of final Kujala and IKDC scores were not significant (p = 0.117 and p = 0.283, respectively). Fracture sites of the patella in group 1 were classified as follows: patellar medial margin (12), inferomedial facet (7), and inferomedial facet involving central ridge (5). In the subgroup of patient with OCF of the inferomedial facet of the patella, the fragments were found in the lateral gutter and did not cause pain or mechanical symptoms. Thus, loose body removal was not performed. However, all five patients with large OCF involving the central ridge of the patella failed non-operative treatment with recurrent dislocations, ultimately requiring fragment refixation and medial retinacular imbrication.Conclusions: First, APD patients with OCFs of medial margin or inferomedial facet showed similar redislocation rates and functional knee scores with those without OCFs after conservative treatment. Second, initial conservative treatment failed in some APD patients with large OCF, especially when OCFs were fractured from inferomedial facet involving central ridge. Surgery should be considered with this type. [ABSTRACT FROM AUTHOR]

Myofibroblasts are the major cell type that is responsible for increase in the mechanical stiffness in fibrotic tissues. It has well documented that the TGF-β/Smad axis is required for myofibroblast differentiation under the rigid substrate condition. However, the mechanism driving myofibroblast differentiation in soft substrates remains unknown. In this research, we demonstrated that interaction of yes-associated protein (YAP) and acetylated microtubule via dynein, a microtubule motor protein drives nuclear localization of YAP in the soft matrix, which in turn increased TGF-β1-induced transcriptional activity of Smad for myofibroblast differentiation. Pharmacological and genetical disruption of dynein impaired the nuclear translocation of YAP and decreased the TGF-β1-induced Smad activity even though phosphorylation and nuclear localization of Smad occurred normally in α-tubulin acetyltransferase 1 (α-TAT1) knockout cell. Moreover, microtubule acetylation prominently appeared in the fibroblast-like cells nearby the blood vessel in the fibrotic liver induced by CCl4 administration, which was conversely decreased by TGF-β receptor inhibitor. As a result, quantitative inhibition of microtubule acetylation may be suggested as a new target for overcoming fibrotic diseases. [ABSTRACT FROM AUTHOR]

The blue-green alga Spirulina maxima is a microscopic filamentous cyanobacterium. Spirulina was recently reported to elicit beneficial effects such as reducing cholesterol and inducing weight loss; however, its effects on inflammation are unknown. To determine the effect of S. maxima extract (SME) on innate immunity, we investigated the NLRP3 inflammasome activation, which is a multiprotein scaffolding complex that plays important roles in innate immune responses to many pathogenic infections in macrophages. SME suppressed lipopolysaccharide (LPS)-induced upregulation of the pro-inflammatory cytokines tumor necrosis factor-α, interleukin (IL)-12, IL-1β, and IL-18 in RAW264.7 cells. In addition, SME attenuated LPS-induced NLRP3 inflammasome activation, and thus pro-IL-1β could not be cleaved to IL-1β by activated caspase-1, which is activated by the NLRP3 inflammasome in RAW264.7 cells. Moreover, SME inhibited LPS-induced phosphorylation of extracellular signal-regulated kinase (ERK) in RAW264.7 cells, and attenuated the generation of ERK1 induced-reactive oxygen species (ROS), resulting in decreased expression of NF-κB. These findings suggest that SME suppresses the effects of the NLRP3 inflammasome via regulation of extracellular signal-regulated kinase (ERK). In summary, we demonstrated that SME prevents activation of the NLRP3 inflammasome by inhibiting ERK signaling. [ABSTRACT FROM AUTHOR]

To investigate whether earthworm cellulases contribute to the innate immune system, the responsiveness of cellulase activity and mRNA expression to bacterial challenge was examined by zymography and RNA sequencing. A zymographic analysis revealed that the activity levels of earthworm cellulases were upregulated in response to either a bacterial (Bacillus subtilis or Escherichia coli) or LPS challenge. After the challenge, significant increases in cellulase 1 and cellulase 2 activity levels were observed within 8–16 and 16–24 h, respectively. In the coelomic fluid, both activities were significantly upregulated at 8 h post-injection with B. subtilis. Based on RNA sequencing, cellulase-related mRNAs encoding beta-1,4-endoglucanases were upregulated by 3-fold within 6 h after B. subtilis injection. Our results clearly demonstrated that earthworm cellulases are upregulated by bacterial challenge at the mRNA and protein levels. These results support the view that earthworm cellulases act as inducible humoral effectors of innate immunity against bacterial infection. • Cellulase activity levels were upregulated by a bacterial or LPS challenge. • RNA-seq discovered that cellulase-related mRNAs increased by B. subtilis injection. • Earthworm cellulases could act as immune effectors against bacterial infection. [ABSTRACT FROM AUTHOR]

• Elderly patients in urology receive potentially inappropriate medications (PIMs). • 73.7% of patients with lower urinary tract symptoms received PIMs at least once. • The average number of PIMs used per patient was 1.31 ± 1.25 (0–7). • Number of chronic diseases and polypharmacy were predictive factors for PIMs. To investigate polypharmacy and potentially inappropriate medications (PIMs) in elderly patients visiting the urology department for lower urinary tract symptoms (LUTS). We retrospectively analyzed digital medical records of individuals over the age of 65 who visited the urology department for LUTS. This cross-sectional study was conducted in 10 hospitals located in South Korea, between September 2017 and December 2017. All prescribed medications were analyzed using electronic medical records. The updated 2015 Beers criteria were used to identify and assess the appropriateness of the prescribed drugs in elderly patients. We analyzed a total of 2143 patients aged over 65 years from 10 institutions. The mean age was 74.2 ± 6.26 years (65–97), 1634 (76.2%) were men. Patients took a mean of 6.48 ± 2.46 medications (range 0–18), and polypharmacy was found in 1762 patients (82.2%). The number of patients who received PIMs at least once was 1579 (73.7%). The average number of PIMs used per patient was 1.31 ± 1.25 (0–7). PIM use ratio was 18.9 ± 0.15% (0–67%). The number of chronic diseases, and concurrent medication and polypharmacy were predictive factors associated with PIM use. Our multi-institutional results show that a substantial proportion of elderly patients took PIMs when visiting the urology department. Factors associated with PIMs were the number of chronic diseases and polypharmacy. Medication use in elderly patients, especially in urology, should be monitored carefully. [ABSTRACT FROM AUTHOR]

Influenza A virus (IAV) continues to threaten human health. To date, two classes of antiviral drugs have been approved to treat IAV infection, but the continuous emergence of the drug-resistant IAV mutant reinforces the need to develop new antiviral drugs. In this study, we aimed to investigate the anti-IAV activity of an aqueous mixture of Agrimonia pilosa and Galla rhois extracts (APRG64). We demonstrated that APRG64 significantly reduced the IAV-induced cytopathic effect, the transcription/expression of viral proteins, and the production of infectious viral particles. Among nine major components of APRG64, apigenin was identified as the main ingredient responsible for the anti-IAV activity. Interestingly, APRG64 and apigenin inhibited the cell attachment and entry of virus and polymerase activity. Importantly, intranasal administration of APRG64 or apigenin strongly reduced viral loads in the lungs of IAV-infected mice. Furthermore, oral administration of APRG64 significantly reduced the level of viral RNAs and the expression level of pro-inflammatory cytokines in the lungs, which protected mice from IAV-induced mortality. In conclusion, APRG64 could be an attractive antiviral drug to treat IAV infection. [Display omitted] • APRG64 and its compound apigenin inhibit IAV replication. • APRG64 reduces pulmonary viral burden and the level of inflammatory cytokines. • Oral administration of APRG64 protects mice from IAV-induced mortality. [ABSTRACT FROM AUTHOR]

Fermented soybean products are gaining attention in the food industry owing to their nutritive value and health benefits. In this study, we performed genomic analysis and physiological characterization of two Debaryomyces spp. yeast isolates obtained from a Korean traditional fermented soy sauce "ganjang". Both Debaryomyces hansenii ganjang isolates KD2 and C11 showed halotolerance to concentrations of up to 15% NaCl and improved growth in the presence of salt. Ploidy and whole-genome sequencing analyses indicated that the KD2 genome is haploid, whereas the C11 genome is heterozygous diploid with two distinctive subgenomes. Interestingly, phylogenetic analysis using intron sequences indicated that the C11 strain was generated via hybridization between D. hansenii and D. tyrocola ancestor strains. The D. hansenii KD2 and D. hansenii- hybrid C11 produced various volatile flavor compounds associated with butter, caramel, cheese, and fruits, and showed high bioconversion activity from ferulic acid to 4-vinylguaiacol, a characteristic flavor compound of soybean products. Both KD2 and C11 exhibited viability in the presence of bile salts and at low pH and showed immunomodulatory activity to induce high levels of the anti-inflammatory cytokine IL-10. The safety of the yeast isolates was confirmed by analyzing virulence and acute oral toxicity. Together, the D. hansenii ganjang isolates possess physiological properties beneficial for improving the flavor and nutritional value of fermented products. • Halotolerant Debaryomyces spp. strains were isolated from Korean soy sauce. • The D. hansenii isolates show distinctive genome features and flavour profiles. • Immunomodulatory activity and safety of the D. hansenii isolates were evaluated. • The yeast isolates are beneficial for quality and function of fermented products. [ABSTRACT FROM AUTHOR]

Background: In vivo length change behavior of native medial patellofemoral ligament (MPFL) fibers throughout the range of knee motion has not been reported in vivo. Purpose: To measure the length changes of various fibers of the MPFL and to determine their length change patterns during in vivo passive knee flexion. Study Design: Descriptive laboratory study. Methods: The right knees of 11 living subjects were scanned with a high-resolution computed tomography scanner at 0, 30, 60, 90, and 120 of knee flexion, and 3-dimensional (3D) models were constructed using customized software. Five patellar points were determined: 20% (point 20), 30% (point 30), 40% (point 40), 50% (point 50), and 60% (point 60) from the superior pole of the patella. The Schöttle femoral point (point F) was marked on a translucent 3D model of a true lateral view. Five virtual fibers connecting these points on the 3D knee model were created, and the lengths of various fibers were digitally measured. Results: The average length changes were 9.1 6 2.5 mm in F20, 9.1 6 2.5 mm in F30, 8.1 6 2.6 mm in F40, 6.9 6 2.4 mm in F50, and 6.9 6 1.7 mm in F60. There were significant differences in length changes of these 5 fibers (P\.001). The lengths of 2 superior fibers (F20 and F30) increased as the knee flexed from 0 to 30 and decreased as the knee flexed over 30. The lengths of a middle fiber (F40) and an inferior fiber (F50) increased from 0 to 30, reached a plateau from 30 to 60, and then decreased from 60 to 120. F60 showed an increase from 0 to 30, and then a plateau pattern from 30 to 90, followed by a decrease during further flexion. Conclusion: Superior fibers exhibited their maximum lengths at low flexion angles, and inferior fibers exhibited their maximum lengths at midflexion angles. The MPFL is a complex of functionally various fibers with some taut and others slack over the whole range of knee motion. Clinical Relevance: The results for lengths and length change patterns of various MPFL fibers are expected to serve as a theoretical background for anatomic double-bundle MPFL reconstruction. [ABSTRACT FROM AUTHOR]

Purpose: The purpose of this study was to prospectively evaluate clinical and radiographic outcomes following anatomic medial patellofemoral ligament (MPFL) reconstruction using patellar suture anchor fixation for recurrent patellar instability. Methods: Twenty patients (20 knees) were enrolled in this study. The median age was 21 years, and the median follow-up was 34.5 months. Reconstruction was performed using a hamstring autograft fixed with two suture anchors at native patellar site of the MPFL. No patient had undergone additional medial tibial tuberosity transfer. Clinical scores (Kujala, Lysholm, and Tegner score) and apprehension test were completed preoperatively and at the follow-up. Preoperative and follow-up radiographic assessments included modified Insall-Salvati ratio, congruence angle, and lateral patellofemoral angle. Results: The preoperative Kujala and Lysholm scores were 52.6 ± 12.4 and 49.2 ± 10.7, and at follow-up visits, corresponding values were 90.9 ± 4.5 ( p < 0.001) and 90.9 ± 5.2 ( p < 0.001). Tegner score increased from 3.0 (range 1-4) to 5.0 (range 4-7) ( p < 0.001). The apprehension test was positive in all patients preoperatively, but only positive in one patient at follow-up. All radiographic assessments were significantly improved; modified Insall-Salvati index (from 1.75 to 1.65) ( p = 0.002), congruence angle (from 6.3° to −7.0°) ( p < 0.001), and lateral patellofemoral angle (from 2.1° to 4.9°) ( p = 0.008). No patient experienced a patellar fracture or redislocation. Conclusion: These results are comparable to those of bone tunnel techniques reported in the literature. This study shows that anatomic MPFL reconstruction using two suture anchors is a reliable treatment option. Level of evidence: Case series with no comparison group, Level IV. [ABSTRACT FROM AUTHOR]

Elaeocarpus sylvestris var. ellipticus (ES), a plant that grows in Taiwan, Japan, and Jeju Island in Korea. ES root bark, known as "sanduyoung," has long been used in traditional oriental medicine. ES is also traditionally used to treat anxiety, asthma, arthritis, stress, depression, palpitation, nerve pain, epilepsy, migraine, hypertension, liver diseases, diabetes, and malaria. However, lack of efficacy and mechanism studies on ES. In the present study, we aim to investigate the VZV-antiviral efficacy, pain suppression, and the anti-inflammatory and antipyretic effects of ES. and methods: Inhibition of VZV was evaluated by hollow fiber assays. Analgesic and antipyretic experiments were conducted using ICR mice and SD Rats, and anti-inflammatory experiments were conducted using Raw264.7 cells. To evaluate the efficacy of ESE against VZV, we conducted antiviral tests. ESE inhibited cell death by disrupting virus and gene expression related to invasion and replication. In addition, ESE suppressed the pain response as measured by writhing and formalin tests and suppressed LPS-induced inflammatory fever. Further, ESE inhibited the phosphorylation of IκB and NF-κB in LPS-induced Raw264.7 cells and expression of COX-2, iNOS, IL-1β, IL-6, and TNF-α. E. sylvestris shows potential as a source of medicine. ESE had a direct effect on VZV and an inhibitory effect on the pain and inflammation caused by VZV infection. [Display omitted] • The activity of ESE was confirmed in nude mice using an HF model of VZV infection. • ESE exerted analgesic, antipyretic, and anti-inflammatory effects in vivo models. • ESE could be a natural drug for the treatment of VZV infection-related diseases. [ABSTRACT FROM AUTHOR]

Purpose: Our objective was to evaluate the effectiveness of arthroscopic distal clavicle resection in cases presenting with pain and subtle instability after neglected grade II acromioclavicular (AC) joint dislocation. Methods: From February 1998 to May 2006, 17 patients with symptomatic AC instability following chronic grade II AC joint injury were studied. Sixteen patients were male and one female, with a mean age of 48 years. An all-arthroscopic procedure comprising disk removal and distal clavicle resection was performed in all cases. All patients were reviewed clinically, preoperatively and at final follow-up (mean 38 months) using the pain score on a visual analog scale and the Constant score. Strength was measured using an Isobex digital strength analyzer. Patient's personal satisfaction after the procedure was documented as excellent, good or poor. Postoperative AC joint radiographs were routinely obtained in all patients to measure the amount of clavicle resection. Results: Two patients underwent additional reconstructive surgery for disabling pain and dysfunction even after the arthroscopic resection procedure. The remaining 15 patients were analyzed. The pain scores improved significantly ( p = 0.03). The mean pain score was 5.8 (range 5-9) before treatment and 1.6 (range 0-3) at follow-up. The Constant scores had improved significantly at the final follow-up ( p = 0.001). The median Constant score increased from 46 (range 36-69) preoperatively to 71 (range 48-84) postoperatively. Strength had improved at the last follow-up. Eleven patients were satisfied and six were not satisfied with this procedure at the final follow-up. Conclusions: Arthroscopic distal clavicle resection statistically improved the pain score, Constant score and strength for grade II AC injury with subtle distal clavicle instability. However, six patients (33.5 %) were not satisfied subjectively. This procedure seemed to be a reasonable initial treatment option with lower morbidity. [ABSTRACT FROM AUTHOR]

[Display omitted] Agrimonia pilosa (AP), Galla rhois (RG), and their mixture (APRG64) strongly inhibited SARS-CoV-2 by interfering with multiple steps of the viral life cycle including viral entry and replication. Furthermore, among 12 components identified in APRG64, three displayed strong antiviral activity, ursolic acid (1), quercetin (7), and 1,2,3,4,6-penta- O -galloyl- β - d -glucose (12). Molecular docking analysis showed these components to bind potently to the spike receptor-binding-domain (RBD) of the SARS-CoV-2 and its variant B.1.1.7. Taken together, these findings indicate APRG64 as a potent drug candidate to treat SARS-CoV-2 and its variants. [ABSTRACT FROM AUTHOR]

Abstract: Genetic polymorphism among patients with acute lymphoblastic leukemia (ALL) is an important factor in the effectiveness and toxicity of anti-leukemic drugs. Genotyping of various polymorphisms that impact the outcome of anti-leukemic drug therapy (pharmacogenetics) presents an attractive approach for developing individualized therapy. We developed an easy and accurate method of analyzing multiple genes using a small amount of DNA, which we termed TotalPlex amplification. We used 16 pairs of specific bulging specific primers (SBS primers) for simultaneous amplification of 16 loci in a single PCR tube. Sixteen single nucleotide polymorphisms (SNPs) (CYP3A4*1B A>G, CYP3A5*3 G>A, GSTP1 313 A>G, GSTM1 deletion, GSTT1 deletion, MDR1 exon 21 G>T/A, MDR1 exon 26 C>T, MTHFR 677 C>T, MTHFR 1298 A>C, NR3C1 1088 A>G, RFC 80 G>A, TPMT 238 G>C, TPMT 460 G>A, TPMT 719 A>G, VDR intron 8 G>A, VDR FokI T>C) that have been implicated in the pharmacogenetics of ALL therapy were analyzed by TotalPlex amplification and SNP genotyping. We successfully amplified specific gene fragments using 16 pairs of primers in one PCR reaction tube with minimal spurious amplification products using TotalPlex amplification coupled to a multiplexed bead array detection system. The genotypes of 16 loci from 34 different genomic DNA (gDNA) samples derived using the TotalPlex system were consistent with the results of several standard genotyping methods, including automatic sequencing, PCR restriction fragment length polymorphism (RFLP) analysis, PCR, and allele-specific PCR (AS-PCR). Thus, the TotalPlex system represents a useful method of amplification that can improve the time, cost, and sample size required for high-throughput pharmacogenetic analysis of SNPs. [Copyright &y& Elsevier]