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Diabetic wounds account for 25 to 50 percent of total diabetic health care costs annually, and present overall healing rates of less than 50 percent. Because delayed diabetic wound healing is associated with impaired fibroblast function, the authors hypothesize that tyrosine kinase Met (cMet) agonistic monoclonal antibody will promote diabetic wound healing by means of stable activation of hepatocyte growth factor/cMet signaling.Two 6-mm dorsal wounds were created in each mouse (6-week-old, male BKS.Cg-Dock7 m +/+Lepr db /J; n = 5). After subcutaneous injections of agonist (20 mg/kg) at 0 and 72 hours, the wound sizes were measured at days 0, 1, 3, 6, and 10. Histologic and immunohistochemical analyses were performed at day 10 (cMet, α-smooth muscle actin, CD68, and transforming growth factor-β). In vitro cytotoxicity and migration tests with diabetic fibroblasts were performed with or without agonist treatment (1 or 10 nM). cMet pathway activation of fibroblasts was confirmed through p-p44/42 mitogen-activated protein kinase, p-mTOR, p-cMet, and ROCK-1 expression.The cMet agonistic monoclonal antibody-treated group showed 1.60-fold lower wound area ( p = 0.027), 1.54-fold higher collagen synthesis ( p = 0.001), and 1.79-fold lower inflammatory cell infiltration ( p = 0.032) than the saline-treated control. The agonist increased cMet (1.86-fold; p = 0.029), α-smooth muscle actin (1.20-fold; p = 0.018), and vascular endothelial growth factor (1.68-fold, p = 0.029) expression but suppressed CD68 (1.25-fold; p = 0.043), transforming growth factor-β (1.25-fold; p = 0.022), and matrix metalloproteinase-2 (2.59-fold; p = 0.029) expression. In vitro agonist treatment (10 nM) of diabetic fibroblasts increased their migration by 8.98-fold ( p = 0.029) and activated the hepatocyte growth factor/cMet pathway.Tyrosine kinase Met agonistic monoclonal antibody treatment improved diabetic wound healing in mice and reduced wound-site inflammatory cell infiltration. These results need to be validated in large animals before piloting human trials.Although further clinical studies are necessary to evaluate its therapeutic efficacy, our study suggested that cMet agonistic monoclonal antibody can be the alternative modality in order to improve wound healing cascade in diabetic foot patients.

Two-dimensional (2D) histopathology based on the observation of thin tissue slides is the current paradigm in diagnosis and prognosis. However, labeling strategies in conventional histopathology are limited in compatibility with 3D imaging combined with tissue clearing techniques. Here, we present a rapid and efficient volumetric imaging technique of pathological tissues called 3D tissue imaging through de novo formation of fluorophores, or 3DNFC, which is the integration of citrate-based fluorogenic reaction DNFC and tissue clearing techniques. 3DNFC markedly increases the fluorescence intensity of tissues by generating fluorophores on nonfluorescent amino-terminal cysteine and visualizes the 3D structure of the tissues to provide their anatomical morphology and volumetric information. Furthermore, the application of 3DNFC to pathological tissue achieves the 3D reconstruction for the unbiased analysis of diverse features of the disorders in their natural context. We suggest that 3DNFC is a promising volumetric imaging method for the prognosis and diagnosis of pathological tissues.

Zinc oxide nanoparticles (ZnO) have attracted much attention as promising antibacterial agents due to their ability to generate reactive oxygen species (ROS) that effectively eliminate bacteria. However, when they are delivered inside the body, this distinct characteristic of ROS is restricted due to the limited penetration depth of external light, which is required for the photocatalysis of particles. To produce ROS without any light source when the particles are implanted, we introduced catechol-ZnO complexes to a hyaluronic acid (HA) hydrogel platform, which can self-generate sufficient ROS in the bacteria-infected tissue. Catechol-ZnO complexes enhanced ROS generation via electron transfer from the formation of complexes and o-semiquinone, and a hydrogel structure was created by coordinate bonds between functionalized catechol groups in HA and ZnO simultaneously. This hydrogel demonstrated different behaviors in terms of physical properties compared to chemically cross-linked HA hydrogels containing ZnO. This hydrogel showed a higher swelling ratio, enzymatic degradation resistance, and tissue adhesive strength. Enhanced ROS generation was confirmed using electron paramagnetic resonance (EPR), H2O2 concentration, glutathione depletion, and intracellular ROS detection. The improved antibacterial performance of hydrogels from ROS production was also confirmed through in vitro bacterial testing against two bacterial strains, E. coli and S. aureus. Furthermore, an in vivo experiment using an infected mouse model to analyze colony formation, histologic analysis, and hematological inflammatory markers revealed the effective antibacterial effects of catechol-ZnO complexes. Overall, the potential of the hydrogel created via catechol-ZnO complexes for antibacterial therapy was demonstrated through the capability to enhance ROS generation and eradicate bacteria.

Patients with diabetes experience delayed wound healing because of the uncontrolled glucose level in their bloodstream, which leads to impaired function of white blood cells, poor circulation, decreased production and repair of new blood vessels. Treatment using polydeoxyribonucleotide (PDRN), which is a DNA extracted from the sperm cells of salmon, has been introduced to accelerate the healing process of diabetic wounds. To accelerate the wound-healing process, sustained delivery of PDRN is critical. In this study, taking advantage of the non-invasive gelation property of alginate, PDRN was loaded inside the hydrogel (Alg-PDRN). The release behavior of PDRN was altered by controlling the crosslinking density of the Alg hydrogel. The amount of PDRN was the greatest inside the hydrogel with the highest crosslinking density because of the decreased diffusion. However, there was an optimal degree of crosslinking for the effective release of PDRN. In vitro studies using human dermal fibroblasts and diabetes mellitus fibroblasts and an in ovo chorioallantoic membrane assay confirmed that the Alg-PDRN hydrogel effectively induced cell proliferation and expression of angiogenic growth factors and promoted new blood vessel formation. Its effectiveness for accelerated diabetic wound healing was also confirmed in an in-vivo animal experiment using a diabetic mouse model.

The nucleotide-binding and oligomerization domain (NOD) is an innate pattern recognition receptor that recognizes pathogen- and damage-associated molecular patterns. The 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) is a matrix degradation product found in the synovial fluids of patients with osteoarthritis (OA). We investigated whether NOD2 was involved in 29-kDa FN-f-induced pro-catabolic gene expression in human chondrocytes. The expression of mRNA and protein was measured using quantitative real-time polymerase chain reaction (qrt-PCR) and Western blot analysis. Small interfering RNAs were used for knockdown of NOD2 and toll-like receptor 2 (TLR-2). An immunoprecipitation assay was performed to examine protein interactions. The NOD2 levels in human OA cartilage were much higher than in normal cartilage. NOD1 and NOD2 expression, as well as pro-inflammatory cytokines, including interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α), were upregulated by 29-kDa FN-f in human chondrocytes. NOD2 silencing showed that NOD2 was involved in the 29-kDa FN-f-induced expression of TLR-2. Expressions of IL-6, IL-8, matrix metalloproteinase (MMP)-1, -3, and -13 were also suppressed by TLR-2 knockdown. Furthermore, NOD2 and TLR-2 knockdown data demonstrated that both NOD2 and TLR-2 modulated the expressions of their adaptors, receptorinteracting protein 2 (RIP2) and myeloid differentiation 88, in 29-kDa FN-f-treated chondrocytes. 29-kDa FN-f enhanced the interaction of NOD2, RIP2 and transforming growth factor beta-activated kinase 1 (TAK1), an indispensable signaling intermediate in the TLR-2 signaling pathway, and activated nuclear factor-κB (NF-κB), subsequently leading to increased expressions of pro-inflammatory cytokines and cartilagedegrading enzymes. These results demonstrate that 29-kDa FN-f modulated pro-catabolic responses via cross-regulation of NOD2 and TLR-2 signaling pathways. [BMB Reports 2019; 52(6): 373-378].

The cGAS-STING pathway plays an important role in pathogen-induced activation of the innate immune response. The 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) found predominantly in the synovial fluid of osteoarthritis (OA) patients increases the expression of catabolic factors via the toll-like receptor-2 (TLR-2) signaling pathway. In this study, we investigated whether 29-kDa FN-f induces inflammatory responses via the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon gene (STING) pathway in human primary chondrocytes. The levels of cGAS and STING were elevated in OA cartilage compared with normal cartilage. Long-term treatment of chondrocytes with 29-kDa FN-f activated the cGAS/STING pathway together with the increased level of gamma-H2AX, a marker of DNA breaks. In addition, the expression of pro-inflammatory cytokines, including granulocytemacrophage colony-stimulating factor (GM-CSF/CSF-2), granulocyte colony-stimulating factor (G-CSF/CSF-3), and type I interferon (IFN-α), was increased more than 100-fold in 29-kDa FN-f-treated chondrocytes. However, knockdown of cGAS and STING suppressed 29-kDa FN-f-induced expression of GM-CSF, G-CSF, and IFN-α together with the decreased activation of TANK-binding kinase 1 (TBK1), interferon regulatory factor 3 (IRF3), and inhibitor protein κBα (IκBα). Furthermore, NOD2 or TLR-2 knockdown suppressed the expression of GM-CSF, G-CSF, and IFN-α as well as decreased the activation of the cGAS/STING pathway in 29-kDa FN-f-treated chondrocytes. These data demonstrate that the cGAS/STING/TBK1/IRF3 pathway plays a critical role in 29-kDa FN-f-induced expression of pro-inflammatory cytokines. [BMB Reports 2019; 52(5): 336-341].

Currently, dermal fillers are largely based on commercialized cross-linked hyaluronic acid (HA) injections, which require a large injection force. Additionally, HA can be easily decomposed by enzymes, and HA-treated tissues present a risk of developing granuloma. In this study, a chitosan-based dermal filler is presented that operates on a liquid-to-gel transition and allows the injection force to be kept ≈4.7 times lower than that required for HA injections. Evaluation of the physical properties of the chitosan filler indicates high viscoelasticity and recovery rate after gelation at 37 °C. Furthermore, in an in vivo evaluation, the liquid injection-type chitosan filler transitions to a gel state within 5 min after injection into the body, and exhibits a compressive strength that is ≈2.4 times higher than that of cross-linked HA. The filler also exhibits higher moldability and maintains a constant volume in the skin for a longer time than the commercial HA filler. Therefore, it is expected that the chitosan filler will be clinically applicable as a novel material for dermal tissue restoration and supplementation.

Keloids, tumor-like lesions that result from excessive scar formation, have no definitive treatment modality. Activation of c-mesenchymal-epithelial transition factor (c-Met) promotes cell proliferation and survival. Selective c-Met inhibitors, such as PHA-665752, may attenuate the activity of keloid fibroblasts and reduce keloid formation. Here, we aimed to evaluate the effect of PHA-665752, a second-generation selective small-molecule inhibitor of c-Met, on human keloid fibroblasts in vitro and in a mouse model. We performed in vitro cytotoxicity assays, scratch tests, western blotting, and immunofluorescence on human keloid fibroblasts. We also injected human fibroblasts into severe combined immunodeficient mice and measured the degree of nodule formation and skin histologic characteristics. We found that keloid fibroblast migration was inhibited by PHA-665752. Inhibitor treatment was also associated with lower expression of members of the hepatocyte growth factor/c-Met pathway, and lower fibroblast activity and collagen synthesis. In the in vivo experiments, PHA-665752—treated mice had lower nodule volumes and weights, accompanied by less inflammatory cell infiltration and collagen deposition, than those in control mice. These findings showed that although an in vivo model may not accurately represent the pathophysiology of human keloid development, PHA-665752 suppressed keloid fibroblast activity by inhibiting the c-Met—related tyrosine kinase pathway.

Introduction Proximal gastrectomy is an alternative treatment modality for gastric cancer in the upper third of the stomach. Though several reconstruction methods have been introduced, there is no standardization. We investigated the outcomes of laparoscopic proximal gastrectomy with double tract reconstruction (LPG-DTR). Aim To investigate the outcomes of LPG-DTR. Material and methods We evaluated 37 patients who underwent curative LPG with DTR between December 2013 and December 2018. Less than half of the proximal stomach was laparoscopically resected. We performed LPG-DTR after resection. Results A total of 37 patients were included in this study, 25 (70%) of whom were male and 12 (30%) of whom were female. Overall, 31 (83.7%) patients were diagnosed with gastric cancer, 5 (13.5%) with gastrointestinal stromal tumors, and 1 (2.8%) with leiomyoma. There were 3 (9.6%) complications. However, there were no complications of grade 3 or above. We did not observe postoperative mortality or recurrence after surgery. All patients underwent postoperative endoscopic surveillance successfully. None of the patients had postoperative reflux esophagitis or stenosis. The body weight and hemoglobin levels of the patients were lowest 12 months after surgery and gradually increased thereafter. Similarly, their vitamin B12 levels were lowest 6 months after surgery. However, iron been increased after surgery until 24 months after surgery. Conclusions LPG-DTR is a favorable treatment modality for gastric cancer in the upper third of the stomach.

The Land Surface Temperature (LST) is one of the significant factors to understand the water and energy cycles between the land surface and atmosphere. However, few previous studies for spatio-temporal variations of LST has been investigated. In this study, we conducted comparative analyses between the Communication, Ocean and Meteorological Satellite (COMS) and MOderate-Resolution Imaging Spectroradiometer (MODIS) LST data. We compared COMS data with observations to identify the accuracy and found relative underestimated patterns of the COMS data as compared to observations. We also found that COMS LST were underestimated in compare to MODIS LST. The Terra LST was verified to have more similar trends with the COMS LST rather than Aqua LST. While we identified the applicability of COMS based on the results of similar tendencies of two comparisons, more intensive validation research at a variety of field conditions should be conducted to gurantee current COMS LST.

Damage caused by natural disasters has been increasing throughout the world due to climate change. As part of adaptation strategies, concept of vulnerability to climate change is introduced abroad to identify flood hotspots and to establish improvement measures. However, there has been very few research on vulnerability domestically. In addition, vulnerability studies conducted at a country level was also reported to be unsuitable for sub-country level. In this study, we conduct vulnerability assessment using entropy method for the Nakdong River Basin as part of domestic application. Vulnerability for present condition are first determined, and then changes in vulnerability due to climate change and large-scale river restoration project to identify their impacts. To achieve this, proxy variables that make up the vulnerability index were selected based on literature survey and data availability. Using data collected for this area, weights for proxy variables were then determined by entropy method. Finally, vulnerability was calculated and mapped for the basin. Reviewing the vulnerability maps showed that vulnerability hotspots were consistent with historical flooded areas for present condition. This suggests that the methodology employed is reasonable. The results for future scenario also showed that there were areas where vulnerability is mitigated by the large-scale river restoration project but also increased by climate change. However, the effect of the project in the future is found to be minimal because the decrease in water level by the project is offset by the increase in runoff due to climate change. The methodology presented in this study is thought to be useful in establishing the disaster prevention plan and climate-change adaptation policy by identifying hotspots.

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