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Abstract Background Diabetes mellitus (DM) causes bone dysfunction due to poor bone quality, leading to severe deterioration in patient of quality of life. The mechanisms of bone metabolism in DM remain unclear, although chemical and/or mechanical factors are known to disrupt the homeostasis of osteoblasts and osteoclasts. The purpose of this study was to identify the changes of osteoblasts and osteoclasts under long-term hyperglycaemic conditions, using a mouse fracture model of long-term hyperglycemia (LT-HG). Methods C57BL/6J mice and green fluorescent protein (GFP) -positive bone marrow transplanted C57BL/6J mice with LT-HG, maintained under a state of hyperglycaemia for 2 months, were used in this study. After the experimental fracture, we examined the immunohistochemical expression of proinsulin and tumor necrosis factor (TNF) -α at the fracture site. C57BL/6J fracture model mice without hyperglycaemia were used as controls. Results In the LT-HG mice, chondrocyte resorption was delayed, and osteoblasts showed an irregular arrangement at the callus site. The osteoclasts were scattered with a decrement in the number of nuclei. The expression of proinsulin was confirmed in bone marrow derived cells (BMDCs) with neovascularization 2 and 3 weeks after fracture. Immunopositivity for TNF-α was also confirmed in immature chondrocytes and BMDCs with neovascularization at 2 weeks, and the number of positive cells was not decreased at 3 weeks. Examination of GFP-grafted hyperglycaemic mice showed that the majority of cells at the fracture site were GFP-positive. Immunohistochemistry showed that the rate of double positives was 15% for GFP and proinsulin and 47% for GFP and TNF-α. Conclusion LT-HG induces an increase in the number of proinsulin and TNF-α positive cells derived from BMDCs. We suggest that proinsulin and TNF-α positive cells are involved in both bone formation and bone resorption after fracture under hyperglycaemic conditions, resulting in the delay of bone healing.

Chronic expanding hematoma (CEH) is a rare anatomical condition that gradually expands due to trauma or surgery. We report the case of a 56-year-old woman who developed CEH 25 years after metal-on-polyethylene total hip arthroplasty. She presented with swelling and radiating pain in the right inguinal region. Tocilizumab was administered for treating rheumatoid arthritis and renal amyloid A amyloidosis. Diagnostic imaging and partial resection revealed a soft tissue mass and a CEH, respectively. The symptoms recurred 6 months later; dialysis was initiated, and the CEH was resected under general anesthesia, leading to improvement. This case report emphasizes the importance of prompt diagnosis and intervention in CEH management for preventing further complications and improving the patient's quality of life.

Abstract Uranyl acetate (UA) has been routinely used as a staining solution for ultrathin sections used in biological electron microscopy. As a radioactive nuclear material, UA is subject to strict international regulations. To develop an alternative and easy-to-use staining method for ultrathin sections, we examined various commercial light microscopic dyes. We found that Mayer’s hematoxylin followed by Reynold’s lead citrate solution showed staining results comparable to UA and Reynold’s lead citrate solution, and this method is therefore suggested as a reliable and promising alternative to UA staining.

ABSTRACT Gelsolin (GSN) is a structural actin-binding protein that is known to affect actin dynamics in the cell. Using mass spectrometry, we identified GSN as a novel Vpr-interacting protein. Endogenous GSN protein was expressed at detectable levels in monocyte-derived macrophages (MDM) and in THP-1 cells, but it was undetectable at the protein level in other cell lines tested. The HIV-1 infection of MDM was associated with a reduction in GSN steady-state levels, presumably due to the Vpr-induced degradation of GSN. Indeed, the coexpression of GSN and Viral protein R (Vpr) in transiently transfected HEK293T cells resulted in the Vpr-dependent proteasomal degradation of GSN. This effect was observed for Vprs from multiple virus isolates. The overexpression of GSN in HEK293T cells had no effect on Gag expression or particle release, but it reduced the expression and packaging of the HIV-1 envelope (Env) glycoprotein and reduced viral infectivity. An analysis of the HIV-1 splicing patterns did not reveal any GSN-dependent differences, suggesting that the effect of GSN on Env expression was regulated at a posttranscriptional level. Indeed, the treatment of transfected cells with lysosomal inhibitors reversed the effect of GSN on Env stability, suggesting that GSN reduced Env expression via enhanced lysosomal degradation. Our data identify GSN as a macrophage-specific host antiviral factor that reduces the expression of HIV-1 Env. IMPORTANCE Despite dramatic progress in drug therapies, HIV-1 infection remains an incurable disease that affects millions of people worldwide. The virus establishes long-lasting reservoirs that are resistant to currently available drug treatments and allow the virus to rebound whenever drug therapy is interrupted. Macrophages are long-lived cells that are relatively insensitive to HIV-1-induced cytopathicity and thus could contribute to the viral reservoir. Here, we identified a novel host factor, gelsolin, that is expressed at high levels in macrophages and inhibits viral infectivity by modulating the expression of the HIV-1 Env glycoprotein, which is critical in the spread of an HIV-1 infection. Importantly, the viral protein Vpr induces the degradation of gelsolin and thus counteracts its antiviral activity. Our study provides significant and novel insights into HIV-1 virus-host interactions and furthers our understanding of the importance of Vpr in HIV-1 infection and pathogenesis.

Human mannose receptor 1 (hMRC1) is a transmembrane glycoprotein that belongs to the C-type lectin family and is expressed on the surface of most tissue macrophages. hMRC1 contributes to the binding and transmission of HIV-1 and is involved in the endocytic uptake of HIV-1 for subsequent antigen presentation. We previously reported that hMRC1 functions as an antiviral factor by inhibiting virus release through a BST-2-like mechanism. The inhibition of virus release was not virus isolate-specific and, surprisingly, was not Env-dependent. We now report on another hMRC1 antiviral function that affects the infectivity of viral particles. Unlike its effect on virus release, the inhibition of viral infectivity by hMRC1 was virus isolate-specific. An analysis of chimeric Env revealed that the Env V3 region was a critical determinant for the inhibitory effect of hMRC1. Of note, exogenously expressed hMRC1 was packaged into viral particles in an Env-independent manner. Co-immunoprecipitation studies revealed a strong interaction of the hMRC1-sensitive NL43 Env with hMRC1, while the hMRC1-insensitive Envs of AD8 and 49.5 isolates interacted poorly if at all with hMRC1. An analysis of a panel of Transmitted/Founder (T/F) viruses revealed that all of them were R5-tropic, and more than half of them were inhibited by hMRC1. The detailed mechanism of how hMRC1 inhibits viral infectivity remains to be investigated. However, the high-affinity binding of hMRC1 to Env may cause a conformational change around the Env V3 region or obstruct the Env V3 region and may make it inaccessible for subsequent interaction with the coreceptor during virus entry.

Introduction: Posterior lumbar interbody fusion (PLIF) is a widely used effective, safe, and established treatment for degenerative spinal disorders. Adjacent segment disease (ASD) is one of the serious concerns governing the clinical results following spinal fusion surgery. Cortical bone trajectory (CBT) is an alternative and less-invasive technique for lumbar pedicle screw placement. Its unique medial and caudal entry point has the potential to prevent an iatrogenic facet joint violence leading to the ASD; however, the incidence of ASD following PLIF using the CBT technique (CBT-PLIF) remains unknown. Methods: Among patients surgically treated with CBT-PLIF in our institute, 52 consecutive patients (13 males, 39 females) with single-level degenerative lumbar spondylolisthesis (DLS) who were followed up for at least 24 months were exclusively enrolled. Their clinical and radiological features, including the incidence of radiographical and symptomatic ASD and significantly associated factor for the developing radiographical ASD, were retrospectively measured. Results: In the present study, we could confirm significant neurological improvement and reduction of the spondylolisthesis with mean follow-up period of 43 months. Radiographical and symptomatic ASD was observed in 14 (27%) and 2 (3.8%) cases, respectively. We compared these two groups and found that the latest lumbar lordosis was significantly different between the two groups, but not in age, body mass index, and Japan Orthopaedic Association score. Two patients with symptomatic ASD required additional surgical treatment around 1 year following the initial surgery. Conclusions: The present study, even though it is preliminary, revealed that CBT-PLIF can achieve a neurological improvement and an effective reduction of spondylolisthesis for the treatment of single-level DLS. The CBT technique is capable of reducing the incidence of ASD compared with the traditional technique; however, we must keep in mind that appropriate postoperative lumbar lordosis should be achieved. Larger, longer-term follow-up studies are required to elucidate the clinical output of CBT-PLIF.

The characterization of forest harvesting and subsequent vegetation recovery provides valuable insights for effective forest management. Although Landsat time series data offers spatially explicit information regarding large-area forest disturbance and recovery, the detailed characterization of country-wide harvest and post-harvest recovery is insufficient. Despite the importance of planting in harvest areas, the mapping of areas replanted by forest management after harvest are not usually considered. This study investigated an approach to detect harvest and other forest disturbance areas country-wide and to characterize post-harvest recovery using annual Landsat time series data from 1984 to 2020. To do so, a random forest algorithm was used to classify disturbance agents and stable land cover classes using predictor variables derived from the LandTrendr temporal segmentation of five spectral indices and topographic and climate data. Post-harvest recovery was characterized as forest species composition (i.e., coniferous/broadleaved forests) and then used to link replanted areas in harvest areas. The disturbance agents/stable land cover classification achieved producer’s and user’s accuracies of 80.1% (±4.8%) and 93.8% (±3.8%), respectively, for the forest harvest class. The overall accuracy of post-harvest recovery was high (83.9%) and a comparison with statistical data of replanted areas for entire Japan showed good agreement in trends and estimated area with a root mean square error of 2687.8 ha (15.2%). Overall, harvested forest area accounted for 4.6% of the total land area in Japan during the past 35 years. The results indicated that approximately 60.0% of coniferous plantation forests were recovered as coniferous forests. The spatial and temporal distribution of coniferous forests after harvest presumably represented the replanting activity of forest management in harvest areas. The approach presented in this study has the potential to provide valuable information on country-wide replantation activity in harvest areas using Landsat time series data.

Introduction Pacing-induced cardiomyopathy occasionally occurs in patients undergoing pacemaker implantation. Although compared with right ventricular (RV) apical pacing, RV septal pacing can attenuate left ventricular dyssynchrony; the success rate of lead placement on the RV septum using the stylet system is low. Additionally, no randomised controlled trial has addressed the issue regarding the accuracy of RV lead placement on the RV septum using the stylet and delivery catheter systems. This study hypothesises that a newly available delivery catheter system can improve the accuracy of RV lead placement on the RV septum.Methods and analysis In a multicentre, prospective, randomised, single-blind, controlled trial, 70 patients with pacemaker indication owing to atrioventricular block will be randomised to either the delivery catheter or stylet group before the pacemaker implantation procedure. The position of the RV lead tip will be assessed using ECG-gated cardiac CT in all patients within 4 weeks after pacemaker implantation. Lead tip positions are classified into three groups: (1) RV septum, (2) anterior/posterior edge of the RV septal wall and (3) RV free wall. The primary endpoint will be the success rate of RV lead tip placement on the RV septum, which will be evaluated using cardiac CT.Ethics and dissemination This study will be conducted according to the stipulations of the Helsinki Declaration and the institutional review board of Hamamatsu University School of Medicine. The results of the study will be disseminated at several research conferences and will be published in peer-reviewed journals.Trial registration number jRCTs042200014; Pre-results.

Developing accurate methods for estimating forest structures is essential for efficient forest management. The high spatial and temporal resolution data acquired by CubeSat satellites have desirable characteristics for mapping large-scale forest structural attributes. However, most studies have used a median composite or single image for analyses. The multi-temporal use of CubeSat data may improve prediction accuracy. This study evaluates the capabilities of PlanetScope CubeSat data to estimate canopy height derived from airborne Light Detection and Ranging (LiDAR) by comparing estimates using Sentinel-2 and Landsat 8 data. Random forest (RF) models using a single composite, multi-seasonal composites, and time-series data were investigated at different spatial resolutions of 3, 10, 20, and 30 m. The highest prediction accuracy was obtained by the PlanetScope multi-seasonal composites at 3 m (relative root mean squared error: 51.3%) and Sentinel-2 multi-seasonal composites at the other spatial resolutions (40.5%, 35.2%, and 34.2% for 10, 20, and 30 m, respectively). The results show that RF models using multi-seasonal composites are 1.4% more accurate than those using harmonic metrics from time-series data in the median. PlanetScope is recommended for canopy height mapping at finer spatial resolutions. However, the unique characteristics of PlanetScope data in a spatial and temporal context should be further investigated for operational forest monitoring.

Regulation of capsid disassembly is crucial for efficient HIV-1 cDNA synthesis after entry, yet host factors involved in this process remain largely unknown. Here, we employ genetic screening of human T-cells to identify maternal embryonic leucine zipper kinase (MELK) as a host factor required for optimal uncoating of the HIV-1 core to promote viral cDNA synthesis. Depletion of MELK inhibited HIV-1 cDNA synthesis with a concomitant delay of capsid disassembly. MELK phosphorylated Ser-149 of the capsid in the multimerized HIV-1 core, and a mutant virus carrying a phosphorylation-mimetic amino-acid substitution of Ser-149 underwent premature capsid disassembly and earlier HIV-1 cDNA synthesis, and eventually failed to enter the nucleus. Moreover, a small-molecule MELK inhibitor reduced the efficiency of HIV-1 replication in peripheral blood mononuclear cells in a dose-dependent manner. These results reveal a previously unrecognized mechanism of HIV-1 capsid disassembly and implicate MELK as a potential target for anti-HIV therapy.

Recent studies have identified host cell factors that regulate early stages of HIV-1 infection including viral cDNA synthesis and orientation of the HIV-1 capsid (CA) core toward the nuclear envelope, but it remains unclear how viral DNA is imported through the nuclear pore and guided to the host chromosomal DNA. Here, we demonstrate that N-terminally truncated POM121C, a component of the nuclear pore complex, blocks HIV-1 infection. This truncated protein is predominantly localized in the cytoplasm, does not bind to CA, does not affect viral cDNA synthesis, reduces the formation of 2-LTR and diminished the amount of integrated proviral DNA. Studies with an HIV-1-murine leukemia virus (MLV) chimeric virus carrying the MLV-derived Gag revealed that Gag is a determinant of this inhibition. Intriguingly, mutational studies have revealed that the blockade by N-terminally-truncated POM121C is closely linked to its binding to importin-β/karyopherin subunit beta 1 (KPNB1). These results indicate that N-terminally-truncated POM121C inhibits HIV-1 infection after completion of reverse transcription and before integration, and suggest an important role for KPNB1 in HIV-1 replication.

Developing countries that intend to implement the United Nations REDD-plus (Reducing Emissions from Deforestation and forest Degradation, and the role of forest conservation, sustainable management of forests, and enhancement of forest carbon stocks) framework and obtain economic incentives are required to estimate changes in forest carbon stocks based on the IPCC guidelines. In this study, we developed a method to support REDD-plus implementation by estimating tropical forest aboveground biomass (AGB) by combining airborne LiDAR with very-high-spatial-resolution satellite data. We acquired QuickBird satellite images of Kampong Thom, Cambodia in 2011 and airborne LiDAR measurements in some parts of the same area. After haze reduction and atmospheric correction of the satellite data, we calibrated reflectance values from the mean reflectance of the objects (obtained by segmentation from areas of overlap between dates) to reduce the effects of the observation angle and solar elevation. Then, we performed object-based classification using the satellite data (overall accuracy = 77.0%, versus 92.9% for distinguishing forest from non-forest land). We used a two-step method to estimate AGB and map it in a tropical environment in Cambodia. First, we created a multiple-regression model to estimate AGB from the LiDAR data and plotted field-surveyed AGB values against AGB values predicted by the LiDAR-based model (R2 = 0.90, RMSE = 38.7 Mg/ha), and calculated reflectance values in each band of the satellite data for the analyzed objects. Then, we created a multiple-regression model using AGB predicted by the LiDAR-based model as the dependent variable and the mean and standard deviation of the reflectance values in each band of the satellite data as the explanatory variables (R2 = 0.73, RMSE = 42.8 Mg/ha). We calculated AGB of all objects, divided the results into density classes, and mapped the resulting AGB distribution. Our results suggest that this approach can provide the forest carbon stock per unit area values required to support REDD-plus.

For serving green tea, there are two prominent methods: steeping the leaf or the powdered leaf (matcha style) in hot water. The purpose of the present study was to reveal chemical and functional differences before and after the powdering process of green tea leaf, since powdered green tea may contribute to expanding the functionality because of the different ingesting style. In this study, we revealed that the powdering process with a ceramic mill and stirring in hot water increased the average extracted concentration of epigallocatechin gallate (EGCG) by more than three times compared with that in leaf tea using high-performance liquid chromatography (HPLC) and liquid chromatography–tandem mass Spectrometry (LC-MS/MS) analyses. Moreover, powdered green tea has a higher inhibition effect of reactive oxygen species (ROS) production in vitro compared with the same amount of leaf tea. Our data suggest that powdered green tea might have a different function from leaf tea due to the higher catechin contents and particles.

Objective: Although the discrimination of crude drugs by metabolomics analysis using liquid chromatography--mass spectrometry (LC--MS) has been widely reported, there are few reports on natural products analysis using supercritical fluid chromatography (SFC)--MS. This study aimed to compare the results of metabolomic analysis of licorice obtained using SFC--MS and LC--MS. Methods: We analyzed 14 CHCl3 extracts of licorice roots (Glycyrrhiza glabra, G. uralensis, and G. inflata) using SFC-time-of-flight (TOF)/MS and LC-TOF/MS in conjunction with principal component analysis (PCA). The three species were assessed for discrimination based on the PCA results. The characteristic constituents of each variety were purified, and their chemical structures were elucidated using various spectroscopic analyses. Results: The PCA score plots of the results of both SFC and LC classified the licorice samples into three groups. Seven compounds (six known compounds and one new phenolic compound, glyglabone) were isolated from G. glabra, five known compounds were isolated from G. uralensis, and four known compounds were isolated from G. inflata. Conclusions: The 16 isolated compounds may be characteristic compounds in the three licorice species. The SFC and LC methods showed comparable results in distinguishing the three species. SFC was highly effective for directly analyzing CHCl3 extracts, reducing analysis time and organic solvent consumption by over 50% compared to the LC analysis. Thus, SFC-TOF/MS in combination with PCA can be used to differentiate between G. glabra, G. uralensis, and G. inflata. [ABSTRACT FROM AUTHOR]