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Abstract Mature osteoclasts degrade bone matrix by exocytosis of active proteases from secretory lysosomes through a ruffled border. However, the molecular mechanisms underlying lysosomal trafficking and secretion in osteoclasts remain largely unknown. Here, we show with GeneChip analysis that RUN and FYVE domain-containing protein 4 (RUFY4) is strongly upregulated during osteoclastogenesis. Mice lacking Rufy4 exhibited a high trabecular bone mass phenotype with abnormalities in osteoclast function in vivo. Furthermore, deleting Rufy4 did not affect osteoclast differentiation, but inhibited bone-resorbing activity due to disruption in the acidic maturation of secondary lysosomes, their trafficking to the membrane, and their secretion of cathepsin K into the extracellular space. Mechanistically, RUFY4 promotes late endosome-lysosome fusion by acting as an adaptor protein between Rab7 on late endosomes and LAMP2 on primary lysosomes. Consequently, Rufy4-deficient mice were highly protected from lipopolysaccharide- and ovariectomy-induced bone loss. Thus, RUFY4 plays as a new regulator in osteoclast activity by mediating endo-lysosomal trafficking and have a potential to be specific target for therapies against bone-loss diseases such as osteoporosis.

Abstract Osteoarthritis (OA) is a progressive and irreversible degenerative joint disease that is characterized by cartilage destruction, osteophyte formation, subchondral bone remodeling, and synovitis. Despite affecting millions of patients, effective and safe disease-modifying osteoarthritis drugs are lacking. Here we reveal an unexpected role for the small molecule 5-aminosalicylic acid (5-ASA), which is used as an anti-inflammatory drug in ulcerative colitis. We show that 5-ASA competes with extracellular-matrix collagen-II to bind to osteoclast-associated receptor (OSCAR) on chondrocytes. Intra-articular 5-ASA injections ameliorate OA generated by surgery-induced medial-meniscus destabilization in male mice. Significantly, this effect is also observed when 5-ASA was administered well after OA onset. Moreover, mice with DMM-induced OA that are treated with 5-ASA at weeks 8–11 and sacrificed at week 12 have thicker cartilage than untreated mice that were sacrificed at week 8. Mechanistically, 5-ASA reverses OSCAR-mediated transcriptional repression of PPARγ in articular chondrocytes, thereby suppressing COX-2-related inflammation. It also improves chondrogenesis, strongly downregulates ECM catabolism, and promotes ECM anabolism. Our results suggest that 5-ASA could serve as a DMOAD.

Electron paramagnetic resonance (EPR) dosimetry for a tooth from an individual exposed is well known as retrospective dosimetry in radiological accidents. A major constraint of the conventional X-band tooth-EPR dosimetry is the necessity to extract the tooth of the exposed patient for dose assessment. In this study, to conduct the dose assessments of exposed patients through part-extraction of tooth enamel, the minimum detectable dose (MDD) of the tooth enamel was evaluated based on the amount of mass. Further, a field test was conducted via intercomparison using various dose assessment methods to verify the feasibility of X-band tooth-EPR dosimetry using the minimum mass of tooth enamel. The intercomparison results demonstrated that effective dose determination via X-band tooth-EPR dosimetry is reliable. Consequently, it was determined that the minimum mass of tooth enamel required to evaluate an absorbed dose above 0.5 Gy is 15 mg. Thus, EPR dosimetry using 15 mg of tooth enamel can be applied in the triage and initial medical response stages for patients exposed during radiological accidents. This approach represents an advancement in managing radiological accidents by offering a more efficient and less invasive method of dose assessment.

The Korea Retrospective Dosimetry network (KREDOS) performed an inter-laboratory comparison to confirm the harmonization and reliability of the results of retrospective dosimetry using mobile phone. The mobile phones were exposed to 192Ir while attached to the human phantoms in the field experiment, and the exposure doses read by each laboratory were compared. This paper describes the reference dosimetry performed to present the reference values for inter-comparison and to obtain additional information about the dose distribution. Reference dosimetry included both measurement using LiF:Mg,Cu,Si and calculation via MCNP simulation to allow a comparison of doses obtained with the two different methodologies. When irradiating the phones, LiF elements were attached to the phones and phantoms and irradiated at the same time. The comparison results for the front of the phantoms were in good agreement, with an average relative difference of about 10%, while an average of about 16% relative difference occurred for the back and side of the phantom. The differences were attributed to the different characteristics of the physical and simulated phantoms, such as anatomical structure and constituent materials. Nevertheless, there was about 4% of under-estimation compared to measurements in the overall linear fitting, indicating the calculations were well matched to the measurements.

The counting efficiencies obtained using anthropomorphic physical phantoms are generally used in whole-body counting measurements to determine the level of internal contamination in the body. Geometrical discrepancies between phantoms and measured individuals affect the counting efficiency, and thus, considering individual physical characteristics is crucial to improve the accuracy of activity estimates. In the present study, the counting efficiencies of whole-body counting measurements were calculated considering individual physical characteristics by employing Monte Carlo simulation for calibration. The NaI(Tl)-based stand-up and HPGe-based bed type commercial whole-body counters were used for calculating the counting efficiencies. The counting efficiencies were obtained from 19 computational phantoms representing various shapes and sizes of the measured individuals. The discrepancies in the counting efficiencies obtained using the computational and physical phantoms range from 2% to 33%, and the results indicate that the counting efficiency depends on the size of the measured individual. Taking into account the body size, the equations for estimating the counting efficiencies were derived from the relationship between the counting efficiencies and the body-build index of the subject. These equations can aid in minimizing the size dependency of the counting efficiency and provide more accurate measurements of internal contamination in whole-body counting measurements.

Abstract Vacuum bell therapy has been acceptable substitute for pectus excavatum patients who want to improve their appearance but avoid surgical correction. The aim of this study was to assess the pre-treatment characteristics of patients with pectus excavatum and to establish characteristics that can potentially help identify ideal candidates for vacuum bell therapy. Expected improvements in thoracic indices were evaluated using pre-treatment chest computed tomography, which was performed before and after applying a vacuum bell device. Treatment results after 1-year of application were evaluated using changes in the Haller index before and after treatment. The patients were categorized into two groups according the post- treatment changes in Haller index calculated using chest radiographs: those with changes in Haller index less than 0.5 (Group 1) and those with greater than or equal to 0.5 (Group 2). Pre-treatment Haller index was significantly lower in Group 1 than in Group 2 (3.1 ± 0.46 vs. 4.2 ± 1.14, respectively, p

Obesity is a disease in which fat is abnormally or excessively accumulated in the body, and many studies have been conducted to overcome it with various techniques. In this study, we evaluated whether micro-current stimulation (MCS) can be applied to prevent obesity by regulating the adipogenesis through 3T3-L1 cells and ob/ob mice. To specify the intensity of MCS, Oil Red O staining was conducted with various intensities of MCS. Based on these, subsequent experiments used 200 and 400 μA for the intensity of MCS. The expressions of insulin signaling pathway-related proteins, including phosphorylation of IGF-1 and IR, were decreased in all MCS groups, and in turn, downstream signals such as Akt and ERK were decreased. In addition, MCS reduced the nucleus translocation of PPAR-γ and decreased the protein expression of C/EBP-α. In the ob/ob mouse model, MCS reduced body weight gain and abdominal adipose tissue volume. In particular, the concentration of triglycerides in serum was also decreased. Taken together, our findings showed that MCS inhibited lipid accumulation by regulating insulin signaling in 3T3-L1, and it was effective at reducing body weight and adipose tissue volume in ob/ob mice. These suggest that MCS may be a useful treatment approach for obesity.

Autophagy appears to be involved in maintaining normal intracellular insulin content by accelerating the insulin degradation rate in β-cells (Marsh et al., 2007) [1]. 2-deoxy-d-glucose (2-DG) is metabolized by hexokinase, and acts as an inhibitor of glycolysis. 2-DG triggers glucose deprivation without altering other nutrients or metabolic pathways (Aghaee et al., 2012) [2], and appears to be an ideal tool for studying autophagy. Rapamycin induced upregulation of autophagy in both cultured isolated islets and pancreatic β-cells (Tanemura et al., 2012) [3]. Here, we examined that 2-DG or rapamycin-induced autophagy may decrease the production of intracellular insulin in INS-1E insulinoma cells. Data showed that autophagy was increased by 2-DG or rapamycin by Western blotting and Immunofluorescence staining analyses. Also, intracellular insulin decreased by 2-DG or rapamycin. Furthermore, the autophagy inhibitors, bafilomycin A1 and/or 3-methyladenine, in the presence or absence of 2-DG or rapamycin increased intracellular insulin in INS-1E insulinoma cells. Keywords: Insulin, Autophagy, 2-Deoxy-D-glucose, Rapamycin, INS-1E Insulinoma cells

Background: Magnolia officinalis cortex has been traditionally used to treat stomach and intestine diseases in traditional Korean medicine. In this study, we investigated the effect of water extract of M. officinalis cortex (WEMC) on osteoclast differentiation and function. Methods: Phytochemical characterization of WEMC was performed by high-performance liquid chromatography analysis. Osteoclast differentiation of bone marrow-derived macrophages was determined by tartrate-resistant acid phosphatase activity assay. Receptor activator of nuclear factor-κB ligand (RANKL) signaling factors and transcription factors regulating osteoclast differentiation were analyzed by Western blot and real-time polymerase chain reaction. Bone resorption function of mature osteoclasts was examined by using culture plate coated with inorganic crystalline calcium phosphate. Furthermore, the in vivo effect of WEMC on osteoporosis was examined using RANKL-induced bone loss model, characterized by micro-computed tomography and bone metabolism marker analysis. Results: WEMC inhibited RANKL-induced osteoclast differentiation and the bone resorbing activity of mature osteoclasts. WEMC contains gallic acid and honokiol as active constituents contributing to the inhibitory effect of WEMC on osteoclast differentiation. Further, WEMC suppressed RANKL-induced activation of p38 and nuclear factor-κB pathways and expression of osteoclastogenic transcription factors such as c-Fos for AP-1 and nuclear factor of activated T cells cytoplasmic 1. Ectopic overexpression of a constitutive active form of nuclear factor of activated T cells cytoplasmic 1 rescued the antiosteoclastogenic effect of WEMC. Consistent with the in vitro results, WEMC suppressed RANKL-induced trabecular bone loss in mice. Conclusion: WEMC might have a therapeutic potential to treat pathological bone diseases due to increased osteoclast differentiation and function.

A biomechanical test is a good evaluation method that describes the structural, functional, and pathological differences in the bones, such as osteoporosis and fracture. The tensile test, compression test, and bending test are generally performed to evaluate the elastic modulus of the bone using mice. In particular, the femoral head compression test is mainly used for verifying the osteoporosis change of the femoral neck. This study conducted bone mineral density analysis using in vivo microcomputed tomography (micro-CT) to observe changes in osteoporosis over time. It proposed a method of identifying the elastic modulus of the femur in the normal group (CON group) and the osteoporotic group (OVX group) through finite element analysis based on the femoral head compression test and also conducted a comparative analysis of the results. Through the femoral head compression test, it was verified that the CON group’s ultimate and yield loads were significantly higher than those of the OVX group. It was considered that this result was caused by the fact that the bone mineral density change by osteoporosis occurred in the proximal end more often than in the femur diaphysis. However, the elastic modulus derived from the finite element analysis showed no significant difference between the two groups.

Seat comfort is one of the main factors that consumers consider when purchasing a car. In this study, we develop a dummy with a shear-force sensor to evaluate seat comfort. The sensor has dimensions of 25 mm × 25 mm × 26 mm and is made of S45C. Electroless nickel plating is employed to coat its surface in order to prevent corrosion and oxidation. The proposed sensor is validated using a qualified load cell and shows high accuracy and precision (measurement range: -30-30 N; sensitivity: 0.1 N; linear relationship: R = 0.999; transverse sensitivity:

Ultrasound Contrast Agents (UCAs) were developed to maximize reflection contrast so that organs can be seen clearly in ultrasound imaging. UCAs increase the signal to noise ratio (SNR) by linear and non-linear mechanisms and thus help more accurately visualize the internal organs and blood vessels. However, the UCAs on the market are not only expensive, but are also not optimized for use in various therapeutic research applications such as ultrasound-aided drug delivery. The UCAs fabricated in this study utilize conventional lipid and albumin for shell formation and perfluorobutane as the internal gas. The shape and density of the UCA bubbles were verified by optical microscopy and Cryo SEM, and compared to those of the commercially available UCAs, Definity® and Sonovue®. The size distribution and characteristics of the reflected signal were also analyzed using a particle size analyzer and ultrasound imaging equipment. Our experiments indicate that UCAs composed of spherical microbubbles, the majority of which were smaller than 1 um, were successfully synthesized. Microbubbles 10 um or larger were also identified when different shell characteristics and filters were used. These laboratory UCAs can be used for research in both diagnoses and therapies.

This study aimed to predict bone fracture risk during skeletal unloading through a subject-specific finite element (FE) analysis considering heterogeneous tissue modulus. Twelve male and 14 female Institute of Cancer Research (ICR) mice (6 weeks old) were allocated into skeletal unloading and normal groups (each gender in both groups). The right hind tibia of each mouse in the skeletal unloading groups was subjected to sciatic neurectomy (denervation) and was scanned before and at 2 week after denervation using µCT. Bone volume (BV) and the distribution of Young's moduli (E) were measured from µCT images. The apparent modulus (Eapp) was calculated using the subject-specific (FE) analysis considering the heterogeneous tissue modulus derived by Hounsfield unit. At 2 weeks after denervation, the Eapp was significantly decreased in both genders (p < 0.05), and the distributions of E differed between the skeletal unloading and normal groups. However, the BV in females was significantly decreased (p < 0.05), while that in male was unchanged (p &gt 0.05). These results indicated that skeletal unloading reduced bone strength, leading to increased bone fracture risk.

This article builds a model of cumulative growth in order to analyze the relationship between intellectual property rights (IPRs) and economic growth in 38 countries from 1980 to 2005. The analysis focuses on the impacts of IPRs on the growth gap between countries using a catch-up model and USPTO database. The empirical results show that the strengthening of IPRs has a positive impact on innovation in developed and developing countries in Asia, while we fail to find evidence in Latin America. Secondly, similar to assertions made by De Long and Summers (1991) and Dowrick and Nguyen (1989), investment in fixed capital plays a critical role in growth gap dynamics. What needs to be emphasized here is the cumulative causal relationship between investment and growth: investment in fixed capital improves productivity and encourages economic growth thereby triggering even more investment. This paper confirms that the strengthening of IPRs and investment in fixed capital contribute to the widening of the economic development gap for the 1980–2005 period.

This study investigated B16F10 melanoma cell death induced by melatonin combined with endoplasmic reticulum (ER) stress through the PI3K/Akt/mTOR pathway. Cell viability was significantly decreased after treatment with melatonin combined with ER stress from thapsigargin or tunicamycin compared to no treatment or treatment with melatonin only. Combined melatonin and ER stress also significantly reduced expression of p85β, p-Akt (Ser473, Thr308), and p-mTOR (Ser2448, Ser2481) compared to treatment with melatonin only. The ER stress protein p-PERK and p-eIF2α were significantly increased under combined melatonin and ER stress treatment compared to no treatment or treatment with melatonin only. Combined melatonin and ER stress significantly reduced Bcl-2 protein and augmented Bax protein compared to melatonin-only treatment. Also, the combined treatment significantly lowered expression of catalase, Cu/Zn-SOD, and Mn-SOD proteins compared to melatonin only. Expression of p85β was significantly more decreased under treatment with melatonin and thapsigargin or tunicamycin plus the PI3K inhibitors LY294002 or wortmannin than under treatment with only melatonin or a PI3K inhibitor. The PI3K downstream target p-Akt (Ser473, Thr308) showed significantly decreased expression under treatment with melatonin and thapsigargin or tunicamycin plus PI3K inhibitors than under treatment with melatonin or PI3K inhibitors only. These results indicate that survival of B16F10 melanoma cells after combined treatment with melatonin and ER stress inducers is suppressed through regulation of the PI3K/Akt/mTOR pathway. Melatonin combined with thapsigargin or tunicamycin appears to be a promising strategy for effective melanoma treatment.

Cyclosporine A (CsA) is a powerful immunosuppressive drug with side effects including the development of chronic nephrotoxicity. In this study, we investigated CsA treatment induced apoptotic and autophagic cell death in pituitary GH3 cells. CsA treatment (0.1 to 10 µM) decreased survival of GH3 cells in a dose-dependent manner. Cell viability decreased significantly with increasing CsA concentrations largely due to an increase in apoptosis, while cell death rates due to autophagy altered only slightly. Several molecular and morphological features correlated with cell death through these distinct pathways. At concentrations ranging from 1.0 to 10 µM, CsA induced a dose-dependent increase in expression of the autophagy markers LC3-I and LC3-II. Immunofluorescence staining revealed markedly increased levels of both LC3 and lysosomal-associated membrane protein 2 (Lamp2), indicating increases in autophagosomes. At the same CsA doses, apoptotic cell death was apparent as indicated by nuclear and DNA fragmentation and increased p53 expression. In apoptotic or autophagic cells, p-ERK levels were highest at 1.0 µM CsA compared to control or other doses. In contrast, Bax levels in both types of cell death were increased in a dose-dependent manner, while Bcl-2 levels showed dose-dependent augmentation in autophagy and were decreased in apoptosis. Manganese superoxide dismutase (Mn-SOD) showed a similar dose-dependent reduction in cells undergoing apoptosis, while levels of the intracellular calcium ion exchange maker calbindin-D9k were decreased in apoptosis (1.0 to 5 µM CsA), but unchanged in autophagy. In conclusion, these results suggest that CsA induction of apoptotic or autophagic cell death in rat pituitary GH3 cells depends on the relative expression of factors and correlates with Bcl-2 and Mn-SOD levels.

In this paper, a kinematically redundant parallel haptic device with large workspace is presented. The haptic device has a similar kinematic structure to the well-known Delta manipulator. However, it has a special arrangement of actuators and one redundant actuator added to the third leg. The proposed haptic device has essentially 4-DOF, however, only three translational DOF are used for 3-DOF positioning and force reflection, and one rotational DOF by the redundant actuator is used to increase well-conditioned workspace. If the redundant actuator's angle is controlled to follow x and y position, the haptic device has a large cylindrical workspace and can maintain good kinematic and statics performance over the whole workspace. The kinematics and workspace are analysed, and the optimal design method of finding minimum link lengths to satisfy prescribed workspace is presented. Finally, the prototype haptic device and control experiment result are presented.

Pasteurella spp. can cause fatal zoonotic infections in humans. We performed a multicenter study to investigate the prevalence and clinical features of Pasteurella infections in South Korea during 2018‒2022. We also conducted a collaborative systematic review and metaanalysis of the global burden of Pasteurella bacteremia. The study included 283 cases and found an increasing trend in Pasteurella infections. Blood cultures were positive in 8/35 (22.9%) cases sampled, for an overall bacteremia-associated rate of 2.8% (8/283). Aging was a significant risk factor for bacteremia (odds ratio 1.05 [95% CI 1.01–1.10]), according to multivariate analyses. For the meta-analysis, we included a total of 2,012 cases from 10 studies. The pooled prevalence of bacteremia was 12.4% (95% CI 7.3%–18.6%) and of mortality 8.4% (95% CI 2.7%–16.5%). Our findings reflect the need for greater understanding of the increase in Pasteurella infections and the global burden of Pasteurella bacteremia to determine appropriate case management. [ABSTRACT FROM AUTHOR]

Background: Clinical management of patients infected with hepatitis B virus (HBV) or hepatitis C virus (HCV) relies on the viral load (VL). The Cobas 5800 system (Roche Diagnostics) can determine VLs in 200 and 500 μL samples, but the performance of each protocol has not been compared. We evaluated the performance of both protocols for the HBV and HCV tests. Methods: Precision and linearity were verified using commercial panels. Probit analyses were used to determine limits of detection (LoDs). The results obtained with 336 samples were compared using the 200 and 500 μL protocols. Data from 6,737 retrospective HBV and 768 HCV samples were compared to estimate the effects of the different LoDs on the diagnostic results of the protocols. Correlations between protocols were tested with Spearman's rank correlation coefficients (rho). Results: The precision and linearity of both protocols were verified. The LoDs for the 200 and 500 μL protocols were 6.5 and 2.7 IU/mL for HBV and 29.7 and 8.2 IU/mL for HCV, respectively. The agreement between the protocols ranged from 0.8 to 1.0. The results obtained with the HBV and HCV tests showed a strong correlation (rho=0.994). Only 0.4% of HBV and 0.4% of HCV test results were affected by the LoDs of the 200 μL protocol. Conclusions: The Cobas 5800 200 and 500 μL protocols for the HBV DNA and HCV RNA tests demonstrated excellent performance. These findings establish the 200 μL protocol as a new option for low-volume samples, especially for pediatric and difficult-to-bleed patients. [ABSTRACT FROM AUTHOR]

Background: Staphylococcal cassette chromosome mec type V (SCCmec V) methicillin-resistant Staphylococcus aureus (MRSA) has been recovered from patients and livestock. Using comparative genomic analyses, we evaluated the phylogenetic emergence of SCCmec V after transmission from overseas donor strains to Korean recipient strains. Methods: Sixty-three complete MRSA SCCmec V genomes (including six Korean clinical isolates) were used to construct a phylogenetic tree. Single-nucleotide polymorphisms were identified using Snippy, and a maximum-likelihood-based phylogenetic tree was constructed using RAxML. The possible emergence of the most common ancestor was estimated using BactDating. To estimate mecA horizontal gene transfer (HGT) events, Rangerdtl was applied to 818 SCCmec V strains using publicly available whole-genome data. Results: The phylogenetic tree showed five major clades. German strains formed a major clade; their possible origin was traced to the 1980s. The emergence of Korean SCCmec V clinical isolates was traced to 2000-2010. mecA HGT events in Staphylococcus spp. were identified in seven strains. P7 (Hong Kong outbreak strain) served as the donor strain for two Korean sequence type (ST) 59 strains, whereas the other five recipient strains emerged from different SCCmec V donors. Conclusions: Most Korean SCCmec V strains may have emerged during 2000-2010. A unique MRSA SCCmec V strain, ST72 (a Korean common type of community-associated MRSA), was also identified. The genomic dynamics of this clone with a zoonotic background should be monitored to accurately understand MRSA evolution. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the efficacy of light-emitting diode (LED) and their dual-wavelengths as a treatment strategy for osteoarthritis. Methods: We induced osteoarthritis in male Sprague-Dawley rats by intra-articular injection of sodium iodoacetate into the right rear knee joint. The animals with lesions were divided into an untreated group and an LED-treated group (n=7 each). In the LED-treated group, the lesioned knee was irradiated with lasers (850 and 940 nm) and dose (3.15 J/cm2) for 20 minutes per session, twice a week for 4 weeks. Knee joint tissues were stained and scanned using an in vivo micro-computed tomography (CT) scanner. Serum interleukin (IL)-6 and IL-18 levels were determined using enzyme-linked immuno-sorbent assay. Several functional tests (lines crossed, rotational movement, rearing, and latency to remain rotating rod) were performed 24 hours before LED treatment and at 7, 14, 21, and 28 days after treatment. Results: LED-treated rats showed improved locomotor function and suppressed matrix-degrading cytokines. Micro-CT images indicated that LED therapy had a preserving effect on cartilage and cortical bone. Conclusion: LED treatment using wavelengths of 850 and 940 nm resulted in significant functional, anatomical, and histologic improvements without adverse events in a rat model. Further research is required to determine the optimal wavelength, duration, and combination method, which will maximize treatment effectiveness. [ABSTRACT FROM AUTHOR]