Dasatinib Inhibits Lyn and Fyn Src-Family Kinases in Mast Cells to Suppress Type I Hypersensitivity in Mice

Mast cells (MCs) are systemically distributed and secrete several allergic mediators such as histamine and leukotrienes to cause type I hypersensitivity. Dasatinib is a type of anti-cancer agent and it has also been reported to inhibit human basophils. However, dasatinib has not been reported for its inhibitory effects on MCs or type I hypersensitivity in mice. In this study, we examined the inhibitory effect of dasatinib on MCs and MC-mediated allergic response in vitro and in vivo. In vitro, dasatinib inhibited the degranulation of MCs by antigen stimulation in a dose-dependent manner (IC 50 , ~34 nM for RBL-2H3 cells; ~52 nM for BMMCs) without any cytotoxicity. It also suppressed the secretion of inflammatory cytokines IL-4 and TNF-α by antigen stimulation. Furthermore, dasatinib inhibited MC-mediated passive cutaneous anaphylaxis (PCA) in mice (ED 50 , ~29 mg/kg). Notably, dasatinib significantly suppressed the degranulation of MCs in the ear tissue. As the mechanism of its effect, dasatinib inhibited the activation of Syk and Syk-mediated downstream signaling proteins, LAT, PLCγ1, and three typical MAP kinases (Erk1/2, JNK, and p38), which are essential for the activation of MCs. Interestingly, in vitro tyrosine kinase assay, dasatinib directly inhibited the activities of Lyn and Fyn, the upstream tyrosine kinases of Syk in MCs. Taken together, dasatinib suppresses MCs and PCA in vitro and in vivo through the inhibition of Lyn and Fyn Src-family kinases. Therefore, we suggest the possibility of repositioning the anti-cancer drug dasatinib as a treatment for various MC-mediated type I hypersensitive diseases.

A New Histone Deacetylase Inhibitor, MHY4381, Induces Apoptosis via Generation of Reactive Oxygen Species in Human Prostate Cancer Cells

Histone deacetylase (HDAC) inhibitors represent a novel class of anticancer agents, which can be used to inhibit cell proliferation and induce apoptosis in several types of cancer cells. In this study, we investigated the anticancer activity of MHY4381, a newly synthesized HDAC inhibitor, against human prostate cancer cell lines and compared its efficacy with that of suberoylanilide hydroxamic acid (SAHA), a well-known HDAC inhibitor. We assessed cell viability, apoptosis, cell cycle regulation, and other biological effects in the prostate cancer cells. We also evaluated a possible mechanism of MHY4381 on the apoptotic cell death pathway. The IC50 value of MHY4381 was lower in DU145 cells (IC50=0.31 μM) than in LNCaP (IC50=0.85 μM) and PC-3 cells (IC50=5.23 μM). In addition, the IC50 values of MHY4381 measured in this assay were significantly lower than those of SAHA against prostate cancer cell lines. MHY4381 increased the levels of acetylated histones H3 and H4 and reduced the expression of HDAC proteins in the prostate cancer cell lines. MHY4381 increased G2/M phase arrest in DU145 cells, and G1 arrest in LNCaP cells. It also activated reactive oxygen species (ROS) generation, which induced apoptosis in the DU145 and LNCaP cells by increasing the ratio of Bax/Bcl-2 and releasing cytochrome c into the cytoplasm. Our results indicated that MHY4381 preferentially results in antitumor effects in DU145 and LNCaP cells via mitochondria-mediated apoptosis and ROS-facilitated cell death pathway, and therefore can be used as a promising prostate cancer therapeutic.

Association Between High Neutrophil to Lymphocyte Ratio and Delayed Recovery From Bell's Palsy

OBJECTIVES: Factors predictive of the severity of and recovery from Bell's palsy remain unclear. This study evaluated the association between neutrophil to lymphocyte ratio (NLR) and the severity of and recovery from Bell's palsy. METHODS: This retrospective study included 51 patients who were hospitalized with Bell's palsy from 2015 to 2017. Degree of paralysis was assessed by House-Brackmann (H-B) grade. Patients with H-B grades 2–4 were classified as having mild to moderate palsy and patients with H-B grade 5 or 6 were classified as having severe palsy. Patients were evaluated for obesity, hypertension and diabetes mellitus, and blood tests were performed to determine NLR and platelet to lymphocyte ratio. Patients were treated with steroids and antiviral agents. H-B grade was assessed 1 week, 1 month, and 3 months after treatment. RESULTS: NLR was significantly higher in patients with severe than with mild to moderate palsy (P=0.048). Recovery time was significantly longer in patients with high NLR than low NLR (P=0.045). CONCLUSION: Higher NLR in patients with Bell's palsy was associated with longer recovery time. NLR may be prognostic of recovery time in patients with Bell's palsy.

An Anti-Cancer Drug Candidate CYC116 Suppresses Type I Hypersensitive Immune Responses through the Inhibition of Fyn Kinase in Mast Cells

Mast cells are the most prominent effector cells of Type 1 hypersensitivity immune responses. CYC116 [4-(2-amino-4-methyl-1,3-thiazol-5-yl)-N-[4-(morpholin-4-yl)phenyl] pyrimidin-2-amine] is under development to be used as an anti-cancer drug, but the inhibitory effects of CYC116 on the activation of mast cells and related allergy diseases have not reported as of yet. In this study, we demonstrated, for the first time, that CYC116 inhibited the degranulation of mast cells by antigen stimulation (IC₅₀, ∼1.42 µM). CYC116 also inhibited the secretion of pro-inflammatory cytokines including TNF-α (IC₅₀, ∼1.10 µM), and IL-6 (IC₅₀, ∼1.24 µM). CYC116 inhibited the mast cell-mediated allergic responses, passive cutaneous anaphylaxis (ED50, ∼22.5 mg/kg), and passive systemic anaphylaxis in a dose-dependent manner in laboratory experiments performed on mice. Specifically, CYC116 inhibited the activity of Fyn in mast cells and inhibited the activation of Syk and Syk-dependent signaling proteins including LAT, PLCγ, Akt, and MAP kinases. Our results suggest that CYC116 could be used as an alternative therapeutic medication for mast cell-mediated allergic disorders, such as atopic dermatitis and allergic rhinitis.

The Association of Patient-Reported Local Allergic Symptoms with Postoperative Outcomes of Chronic Rhinosinusitis

Implication of Porphyromonas gingivalis in colitis and homeostasis of intestinal epithelium / 한국실험동물학회지

Emerging evidences have reported that periodontitis can be a risk factor for the pathogenesis of various systemic diseases. Porphyromonas gingivalis (Pg), one of the crucial pathogens in chronic periodontitis, has been spotlighted as a potential cause for the promotion and acceleration of periodontitis-associated systemic disorders. To investigate the association between Pg and intestinal disease or homeostasis, we treated Pg-derived lipopolysaccharide (LPS) in murine colitis model or intestinal organoid, respectively. Pg-derived LPS (Pg LPS) was administrated into chemically induced murine colitis model and disease symptoms were monitored compared with the infusion of LPS derived from E. coli (Ec LPS). Organoids isolated and cultured from mouse small intestine were treated with Pg or Ec LPS and further analyzed for the generation and composition of organoids. In vivo observations demonstrated that both Pg and Ec LPS exerted slight protective effects against murine colitis. Pg LPS did not affect the generation and growth of intestinal epithelial organoids. Among subtypes of epithelial cells, markers for stem cells, goblet cells or Paneth cells were changed in response to Pg LPS. Taken together, these results indicate that Pg LPS leads to partial improvement in colitis and that its treatment does not significantly affect the self-organization of intestinal organoids but may regulate the epithelial composition.

Intramural Gastric Hematoma after Acute Necrotizing Pancreatitis: A Case Report and Review of Imaging Findings

Intramural hematoma of the gastrointestinal tract is a rare disease entity. Pancreatitis-induced intramural gastric hematoma (IGH) is far more seldom reported. Here, we report a rare case of a giant IGH occurring as a delayed complication of pancreatitis in a 51-year-old man. The diagnosis was made using computed tomography (CT) and endoscopic ultrasonography. The patient was conservatively managed, and follow-up abdominal CT showed marked decreases in the size of the IGH.

Microglial involvement in the development of olfactory dysfunction

Olfactory impairment is the most common clinical manifestation among the elderly, and its prevalence increases sharply with age. Notably, growing evidence has shown that olfactory dysfunction is the first sign of neurodegeneration, indicating the importance of olfactory assessment as an early marker in the diagnosis of neurological disorders. In this review, we describe the nature of olfactory dysfunction and the advantage of using animal models in olfaction study, and we include a brief introduction to olfactory behavior tests widely used in this field. The contribution of microglia in the neurodegenerative processes including olfactory impairment is then discussed to provide a comprehensive description of the physiopathological role of interactions between neurons and microglia within the olfactory system.

Lysophosphatidic acid protects against acetaminophen-induced acute liver injury

We investigated the effect of lysophosphatidic acid (LPA) in experimental acetaminophen (APAP)-induced acute liver injury. LPA administration significantly reduced APAP-challenged acute liver injury, showing attenuated liver damage, liver cell death and aspartate aminotransferase and alanine aminotransferase levels. APAP overdose-induced mortality was also significantly decreased by LPA administration. Regarding the mechanism involved in LPA-induced protection against acute liver injury, LPA administration significantly increased the glutathione level, which was markedly decreased in APAP challenge-induced acute liver injury. LPA administration also strongly blocked the APAP challenge-elicited phosphorylation of JNK, ERK and GSK3β, which are involved in the pathogenesis of acute liver injury. Furthermore, LPA administration decreased the production of TNF-α and IL-1β in an experimental drug-induced liver injury animal model. Mouse primary hepatocytes express LPA₁(,)₃–₆, and injection of the LPA receptor antagonist KI16425 (an LPA₁(,)₃-selective inhibitor) or H2L 5765834 (an LPA₁(,)₃(,)₅-selective inhibitor) did not reverse the LPA-induced protective effects against acute liver injury. The therapeutic administration of LPA also blocked APAP-induced liver damage, leading to an increased survival rate. Collectively, these results indicate that the well-known bioactive lipid LPA can block the pathogenesis of APAP-induced acute liver injury by increasing the glutathione level but decreasing inflammatory cytokines in an LPA₁(,)₃(,)₅-independent manner. Our results suggest that LPA might be an important therapeutic agent for drug-induced liver injury.

Inhibition by miR-410 facilitates direct retinal pigment epithelium differentiation of umbilical cord blood-derived mesenchymal stem cells

Retinal pigment epithelium (RPE) is a major component of the eye. This highly specialized cell type facilitates maintenance of the visual system. Because RPE loss induces an irreversible visual impairment, RPE generation techniques have recently been investigated as a potential therapeutic approach to RPE degeneration. The microRNA-based technique is a new strategy for producing RPE cells from adult stem cell sources. Previously, we identified that antisense microRNA-410 (anti-miR-410) induces RPE differentiation from amniotic epithelial stem cells. In this study, we investigated RPE differentiation from umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) via anti-miR-410 treatment. We identified miR-410 as a RPE-relevant microRNA in UCB-MSCs from among 21 putative human RPE-depleted microRNAs. Inhibition of miR-410 induces overexpression of immature and mature RPE-specific factors, including MITF, LRAT, RPE65, Bestrophin, and EMMPRIN. The RPE-induced cells were able to phagocytize microbeads. Results of our microRNA-based strategy demonstrated proof-of-principle for RPE differentiation in UCB-MSCs by using anti-miR-410 treatment without the use of additional factors or exogenous transduction.

Direct Conversion of Human Umbilical Cord Blood into Induced Neural Stem Cells with SOX2 and HMGA2

Recent advances have shown the direct reprogramming of mouse and human fibroblasts into induced neural stem cells (iNSCs) without passing through an intermediate pluripotent state. Thus, direct reprogramming strategy possibly provides a safe and homogeneous cellular platform. However, the applications of iNSCs for regenerative medicine are limited by the restricted availability of cell sources. Human umbilical cord blood (hUCB) cells hold great potential in that immunotyped hUCB units can be immediately obtained from public banks. Moreover, hUCB samples do not require invasive procedures during collection or an extensive culture period prior to reprogramming. We recently reported that somatic cells can be directly converted into iNSCs with high efficiency and a short turnaround time. Here, we describe the detailed method for the generation of iNSCs derived from hUCB (hUCB iNSCs) using the lineage-specific transcription factors SOX2 and HMGA2. The protocol for deriving iNSC-like colonies takes 1~2 weeks and establishment of homogenous hUCB iNSCs takes additional 2 weeks. Established hUCB iNSCs are clonally expandable and multipotent producing neurons and glia. Our study provides an accessible method for generating hUCB iNSCs, contributing development of in vitro neuropathological model systems.

Immunologic properties of differentiated and undifferentiated mesenchymal stem cells derived from umbilical cord blood

The expression of immunogenic markers after differentiation of umbilical cord blood (UCB)-derived mesenchymal stem cells (MSC) has been poorly investigated and requires extensive in vitro and in vivo testing for clinical application. The expression of human leukocyte antigen (HLA) classes on UCB-derived MSC was tested by Fluorescence-activated cell sorting analysis and immunocytochemical staining. The undifferentiated MSC were moderately positive for HLA-ABC, but almost completely negative for HLA-DR. The MSC differentiated to chondrocytes expressed neither HLA-ABC nor HLA-DR. The proliferation of MSC was not significantly affected by the allogeneic lymphocytes stimulated with concanavalin A. The responder lymphocytes showed no significant decrease in proliferation in the presence of the MSC, but the apoptosis rate of the lymphocytes was increased in the presence of MSC. Taken together, these findings indicate that UCB-derived MSC differentiated to chondrocytes expressed less HLA class I and no class II antigens. The MSC showed an immunomodulatory effect on the proliferation and apoptosis of allogeneic lymphocytes. These data suggest that the differentiated and undifferentiated allogeneic MSC derived from umbilical cord blood can be a useful candidate for allogeneic cell therapy and transplantation without a major risk of rejection.

A New Histone Deacetylase Inhibitor, MHY219, Inhibits the Migration of Human Prostate Cancer Cells via HDAC1

Histone deacetylase (HDAC) inhibitors are considered novel agents for cancer chemotherapy. We previously investigated MHY219, a new HDAC inhibitor, and its potent anticancer activity in human prostate cancer cells. In the present study, we evaluated MHY219 molecular mechanisms involved in the regulation of prostate cancer cell migration. Similar to suberanilohydroxamic acid (SAHA), MHY219 inhibited HDAC1 enzyme activity in a dose-dependent manner. MHY219 cytotoxicity was higher in LNCaP (IC50=0.67 muM) than in DU145 cells (IC50=1.10 muM) and PC3 cells (IC50=5.60 muM) after 48 h of treatment. MHY219 significantly inhibited the HDAC1 protein levels in LNCaP and DU145 cells at high concentrations. However, inhibitory effects of MHY219 on HDAC proteins levels varied based on the cell type. MHY219 significantly inhibited LNCaP and DU145 cells migration by down-regulation of matrix metalloprotease-1 (MMP-1) and MMP-2 and induction of tissue inhibitor of metalloproteinases-1 (TIMP-1). These results suggest that MHY219 may potentially be used as an anticancer agent to block cancer cell migration through the repression of MMP-1 and MMP-2, which is related to the reduction of HDAC1.

Characterization and clinical application of mesenchymal stem cells from equine umbilical cord blood

Tendinitis of the superficial digital flexor tendon (SDFT) is a significant cause of lameness in horses; however, recent studies have shown that stem cells could be useful in veterinary regenerative medicine. Therefore, we isolated and characterized equine umbilical cord blood mesenchymal stem cells (eUCB-MSCs) from equine umbilical cord blood obtained from thoroughbred mares during the foaling period. Horses that had tendinitis of the SDFT were treated with eUCB-MSCs to confirm the therapeutic effect. After eUCB-MSCs transplantation, the core lesion in the SDFT was found to decrease. These results suggest that transplantation using eUCB-MSCs could be another source of cell treatment.

Isolation and characterization of equine amniotic membrane-derived mesenchymal stem cells

Recent studies have shown that mesenchymal stem cells (MSCs) are able to differentiate into multi-lineage cells such as adipocytes, chondroblasts, and osteoblasts. Amniotic membrane from whole placenta is a good source of stem cells in humans. This membrane can potentially be used for wound healing and corneal surface reconstruction. Moreover, it can be easily obtained after delivery and is usually discarded as classified waste. In the present study, we successfully isolated and characterized equine amniotic membrane-derived mesenchymal stem cells (eAM-MSCs) that were cultured and maintained in low glucose Dulbecco's modified Eagle's medium. The proliferation of eAM-MSCs was measured based on the cumulative population doubling level (CPDL). Immunophenotyping of eAM-MSCs by flow cytometry showed that the major population was of mesenchymal origin. To confirm differentiation potential, a multi-lineage differentiation assay was conducted. We found that under appropriate conditions, eAM-MSCs are capable of multi-lineage differentiation. Our results indicated that eAM-MSCs may be a good source of stem cells, making them potentially useful for veterinary regenerative medicine and cell-based therapy.

Trends of CT Use in the Adult Emergency Department in a Tertiary Academic Hospital of Korea during 2001-2010

OBJECTIVE: We wanted to assess the trends of CT examinations that were conducted in an adult emergency department (ED). MATERIALS AND METHODS: We searched the medical database to identify adult patients (> or = 18 years) who had visited the ED and the number of CT examinations of the patients during the period from January 2001 to December 2010. We also analyzed the types of CT scans performed in terms of body parts, they were as follows; head CTs, facial bone CTs, neckl CTs, chest CTs, abdominal CTs, and miscellaneous CTs. Further, miscellaneous CTs were subdivided as CT angiography and others. RESULTS: A total of 113656 CT scans were examined for 409439 adult ED patients during a 10-year period, and the number of CT scans increased by 255% (from 4743 CTs in 2001 to 16856 CTs in 2010), while the adult ED patient volume increased by 34% during the same period. Although the head CTs proportionally occupied the most, the facial bone CTs had the largest rate of increase (3118%), followed by cervical CTs (1173%), chest CTs (455%), miscellaneous CTs (388%; 862% and 84% for CT angiography and others, respectively), abdominal CTs (315%) and head CTs (95%) per 1000 patients during the decade. CONCLUSION: CT use in adult ED has increased at a rate that far exceeds the growth of ED patient volume, with facial bone CTs and cervical CTs having the largest increasing rate, followed by chest CTs, miscellaneous CTs, abdominal CTs and head CTs.

Treatment of Non-union Distal Humerus Fractures after Operation

PURPOSE: This study is a retrospective analysis of patients who had undergone surgical treatment for non-union of distal humerus fracture. We evaluated them in terms of causes of injury, radiologic findings, and clinical outcomes such as prognosis. MATERIALS AND METHODS: Seven consecutive radiologic patients who were confirmed to have nonunion of a distal humerus fracture underwent reoperations. These patients had already undergone operations for distal humerus fractures. This survey was held from 2005 to 2010. The average period up to diagnosis of non-union after the first operation was 7.4 months (4 to 16 months). The mean follow-up period was 24.6 months (12 to 65 months). Each patient was graded functionally according to the Mayo Elbow Performance Score and the Disabilities of the Arm, Shoulder and Hand Score. RESULTS: Osteosynthesis was performed by internal fixation with plates and screws and then a bone graft for non-union of the distal humerus fracture. The average range of motion within the elbow joints was found to be a flexion contracture of 18.8 degrees (0~30 degrees) and further flexion of 120.2 degrees (102~140 degrees). Among postoperative complications, three cases of medium-degree stiffness, two cases of medial column nonunion, and one case of dissociation of the internal fixator were reported. CONCLUSION: Stable internal fixation for maintenance reduction status is essential after accurate initial anatomical reduction. We concluded that nonunion could be prevented by additional surgical treatment such as autogenous bone graft, if it is necessary.

Granular Cell Tumor Originating from Gallbladder / 대한내과학회지

Granular cell tumors are rare benign tumors that arise from Schwann cells. They are especially rare in the gallbladder; indeed, only four cases have been reported in the English-language literature. This paper reports a granular cell tumor in the gallbladder and is the first such report in a Korean woman. She was admitted to the hospital with jaundice and fever and was diagnosed with acute hepatitis A. While hospitalized, a well-demarcated round mass was incidentally found in her gallbladder. The acute hepatitis A improved with conservative care, but the mass did not change in size after 4 months. The tumor was resected with the gallbladder during laparoscopic surgery, and was found to be a granular cell tumor. The tumor was composed of sheets or irregular fascicles of large polygonal or spindle cells with plump eosinophilic granular cytoplasm that was immunohistochemically positive for S-100 and neuron-specific enolase and negative for neurofilament.

Single Portal Endoscopic Carpal Tunnel Release in Patients Older than 65 Years

PURPOSE: There have been few reports about the endoscopic carpal tunnel release (ECTR) in elderly patients and its efficacy and safety are not well-known. We evaluated the clinical outcomes of ECTR using Agee technique in patients older than 65 years. MATERIALS AND METHODS: From October 2000 to January 2007, thirty-five patients (42 hands) who underwent ECTR using Agee technique were enrolled. The average age of the patients was 67.2 years (range, 65-71 years). The duration of symptoms averaged 10 months (range, 6-33 months). For evaluation of the clinical outcomes, physical examination and subjective assessment of the hand function using the Boston carpal tunnel questionnaire were performed at postoperative 1-year follow-up and compared with those obtained at preoperative evaluation. The mean follow-up period was 18 months (range, 12-24 months). RESULTS: There were no neurovascular injury and scar tenderness. At postoperative 1-year follow-up, paresthesia, numbness, Phalen's sign, tinel sign, two point discrimation, and grip power were significantly improved compared with those obtained at preoperation. According to the Boston questionnaire, symptom severity score improved from 3.43 preoperatively to 1.89 postoperatively, and functional status score improved from 3.18 preoperatively to 2.21 postoperatively (p<0.05). Thenar atrophy still remained in 32 hands (76.2%). CONCLUSION: Although thenar atrophy did not improve in many cases, symptom severity and functional status scores improved in most patients treated with ECTR. The single portal ECTR is a safe and efficacious treatment option in elderly patients with carpal tunnel syndrome.

A Case of Silicosis and Pneumothorax in a Workplace Producing Dental Porcerlain / 대한산업의학회지

BACKGROUND: Silicosis is more likely to occur in people working in the mining industry. However, workers suffering from silicosis have recently been reported frequently in other areas. We present a case of silicosis occuring in a 43-year-old man who had worked for 20 years in a workplace producing dental porcelain. CASE: The man was admitted to the emergency room with acute chest pain caused by pneumothorax. Chest X-ray indicated numerous small opacities spread over the whole lung field and a large opacity in the right middle lung field. According to ILO classification, the shape of the small opacities was t/s, the profusion rate was 2/3 and the large opacity was classified into the B category. Following this diagnosis of silicosis, the patient's medical history and work exposure history were examined. According to his medical history, he had undergone closed thoracostomy in 2006 because he had suffered pneumothorax twice (in 2005 and 2006) and his smoking history was 7 pack years. In particular, he had been exposed to silica dust for 20 years in his workplace. CONCLUSION: Despite the absence of any specific risk factor that caused pneumothorax, the patient suffered this condition three times. All clinical results and the progress of his physical symptoms, including radiologic findings from chest X-ray and computed tomography, clearly supported the diagnosis of silicosis. Except for exposure to silica dust in the workplace, no other risk factors causing silicosis were found. Therefore, he was finally diagnosed as having silicosis caused by exposure to silica dust in the workplace and followed by pneumothorax.