BACKGROUND: 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD: dioxin) is a potent environmental toxicant that alters various cell function. Both reactive oxygen species (ROS) and mitogen-activated protein kinases (MAPK) mediate dioxin-induced cytotoxicity. Since dioxin was shown to increase renal cell fibronectin secretion in a dose-dependent manner and ROS and MAPK also play roles in fibronectin upregulation in renal cells, the present study examined whether ROS and/or MAPK activation play a role in dioxin-induced fibronectin upregulation in tubular epithelial cells. METHODS: Madin-Darby canine kidney (MDCK) cells were cultured with minimum essential medium (MEM) containing 10% fetal bovine serum. Growth arrested and synchronized MDCK cells by serum deprivation were stimulated with dioxin 1 nM in the presence or absence of extracellular signal-regulated protein kinase (ERK) inhibitor PD98059 50 microM, p38 MAPK inhibitor 100 nM, trolox 500 microM, or taurine 500 microM for up to 48 hours. Dichlorofulorescein (DCF)-sensitive cellular ROS was measured by FACScan and fibronectin in the media and cellular MAPK by a Western blot analysis. RESULTS: Dioxin 1 nM significantly increased cellular ROS and fibronectin in MDCK cells. Antioxidants, trolox and taurine, effectively inhibited dioxin-induced cellular ROS and fibronectin secretion. Dioxin increased phosphorylation of ERK at 5 minutes and P38 MAPK at 48 hours. Dioxin did not affect c-Jun NH2-terminal kinase (JNK) activation for up to 48 hours. Both PD98059 and p38 MAPK inhibitor suppressed dioxin-induced fibronectin secretion by MDCK cells. CONCLUSION: These data suggest that dioxin increases fibronectin secretion by renal distal tubular epithelial cells through ROS and MAPK (ERK and p38 MAPK) and this may lead to renal fibrosis.
Antioxidants ; Blotting, Western ; Epithelial Cells ; Fibronectins* ; Fibrosis ; Kidney ; Madin Darby Canine Kidney Cells* ; Mitogen-Activated Protein Kinases* ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; Phosphotransferases ; Protein Kinases ; Reactive Oxygen Species* ; Taurine ; Up-Regulation

BACKGROUND: High glucose upregulates MCP-1 expression in rat glomerular mesangial cells and in human peritoneal mesothelial cells. However, the role of high glucose-induced MCP-1 on the development and progression of diabetic renal injury and peritoneal injury during peritoneal dialysis(PD) using high glucose PD solutions are not clear. Since MCP-1 was shown to upregulate transforming growth factor-beta1(TGF-beta1) and collagen expression in lung fibroblasts, the present study investigated the effects of MCP-1 on fibronectin secretion by mouse mesangial cells(MMC), human peritoneal mesothelial cells (HPMC), and human peritoneal fibroblasts(HPFB). METHODS: Synchronized cells were stimulated by different concentrations of MCP-1(0.1-100 ng/mL) or TGF-beta1(0.1-10 ng/mL) for 48 hours. Fibronectin protein secreted into the media was analyzed by Western blot analysis. RESULTS: MCP-1 up to 100 ng/mL did not affect fibronectin secretion by MMC. TGF-beta1 10 ng/mL, however, increased fibronectin secretion by MMC 2.8 fold that of control. MCP-1 up to 100 ng/mL did not affect fibronectin secretion by HPMC. But, TGF-beta1 0.1 ng/mL increased fibronectin secretion by HPMC 1.8 fold compared to control. On the other hand, MCP-1 increased fibronectin secretion by HPFB in a dose-dependent manner. MCP-1 at 1-10 ng/mL significantly increased fibronectin when compared to M199 control. 100 ng/mL MCP-1 further increased fibronectin secretion by HPFB compared to 0.1-10 ng/mL MCP-1. CONCLUSION: These results suggest a possible role for MCP-1 in the development and progression of peritoneal fibrosis and support the view that in addition to recruiting inflammatory cells MCP-1 may play a role in tissue fibrosis in other organs.
Animals ; Blotting, Western ; Chemokine CCL2* ; Collagen ; Fibroblasts* ; Fibronectins* ; Fibrosis ; Glucose ; Hand ; Humans* ; Lung ; Mesangial Cells ; Mice ; Monocytes* ; Peritoneal Fibrosis ; Rats ; Transforming Growth Factor beta1

High glucose activates protein kinase C, induces reactive oxygen species generation, and upregulates expression of transforming growth factor-beta1(TGF-beta1) and fibronectin by human peritoneal mesothelial cells(HPMC). High glucose also induces premature senescence in mesothelial cells. Mesothelial cells shrink after exposure to hypertonic medium and intracellular uptake of amino acids increase to ensure subsequent volume increase. Based on these observations, new and more biocompatible peritoneal dialysis solutions that are glucose free and/or iso-osmolar have been developed. We investigated the effects of different osmolality and different osmotic agents including glucose, mannitol, and icodextrin on viability and proliferation of HPMC. HPMC were obtained from the omental tissues of consenting patients undergoing Cesarean section or elective abdominal surgery. All experiments were performed using cells in the 2nd or 3rd passage. Near-confluent HPMC grown in culture dishes were incubated with serum-free medium for 48 hours to arrest and synchronize cell growth. Lactate dehydrogenase(LDH) release was measured for cell viability and [3H]-thymidine incorporation for proliferation of cultured HPMC, after exposing HPMC to different concentrations of glucose, mannitol, and icodextrin for up to 96 hours. High glucose and mannitol at concentrations up to 100 mM(375 mOsm) did not increase LDH release up to 96 hours compared to control M199. When HPMC were exposed to 2, 4, 7.5, and 9% of icodextrin for 24-96 hours, LDH release did not increase. Glucose at 30, 50, and 100 mM significantly inhibited [3H]-thymidine incorporation by HPMC at 24 and 48 hours. Mannitol at 30, 50, and 100 mM for 24 hours and at only 100 mM for 48 hours also significantly inhibited cell proliferation. Icodextrin 9% (305 mOsm) inhibited cell proliferation compared with control M-199 at 24 hours. In conclusion, high osmolality per se dose not appear to increase HPMC death. However, high osmolality appears to inhibit HPMC proliferation at early stage. In addition, high glucose appears to inhibit HPMC proliferation independent of osmolality since high glucose continues to inhibit cell proliferation at 48 and 72 hours when mannitol at the same concentration did not. Icodextrin 9% of which osmolality is 305 mOsm inhibits HPMC proliferation at early stage but does not appear to increase HPMC death.
Humans

Purpose: To explore the scope and method of applying standardized nursing terminologies to nursing diagnosis and problems used in nursing practice. Methods: A descriptive study was done with a retrospective analysis of the nursing records of 141,420 patients that were hospitalized in 4 tertiary hospitals. The nursing diagnosis and problems collected from the records were standardized, and the standardized nursing diagnosis and problems cross mapped with NANDA-I, confirmed in a nursing focus group. Results: 65 (67.7%) of the 96 standardized nursing diagnosis and problems were equal with NANDA-I and included in the 10 domains of NANDA-I. Among 86 nursing diagnosis and problems excluded from the cross mapping with NANDA-I, the 63 terms (73.3%) related to surgery/procedure were the most common. Conclusion It is meaningful that multi-tertiary hospital nursing diagnosis and problems were standardized and cross mapping with standard nursing terminologies was performed. As for the method of applying standardized nursing terminologies in nursing practice, it is appropriate to use several standardized nursing terminologies complementarily.

Traumatic thyroid hematoma is a rare problem in cases of blunt trauma, but it can be a life-threatening condition. We report here on a case of thyroid hematoma that was complicated with unexpected airway obstruction due to blunt trauma on the anterior neck. A 23-year-old man who developed neck painful swelling and dyspnea was admitted to the emergency room with a blunt neck injury after motor-bicycle accident, and he hit his anterior neck against the handle-bar. He had no previous thyroid pathology and other medical problems. On the initial examination, his vital signs were stable and the oxygen saturation was 97.8%. A computed tomography (CT) scan of the neck showed fragmentation and extensive hematoma within the right thyroid gland and dislocation of the trachea. No definite bleeding focus was identified on the angiography that was done through the carotid artery. Two hours later, the patient experienced sudden respiratory distress and the oxygen saturation dropped rapidly. The patient needed emerg encyendotracheal intubation and surgical intervention. He successfully underwent right lobectomy of the thyroid gland, and was discharged on the 6th hospital day without any complications. The thyroid function was normal after 3 months.
Airway Obstruction* ; Angiography ; Carotid Arteries ; Dislocations ; Dyspnea ; Emergency Service, Hospital ; Hematoma* ; Hemorrhage ; Humans ; Intubation ; Neck ; Neck Injuries ; Neck Pain ; Oxygen ; Pathology ; Thyroid Gland* ; Trachea ; Vital Signs ; Young Adult

PURPOSE: Bone morphogenic protein (BMP)-7, a member of TGF-beta1 superfamily, is an endogenous antifibrotic protein highly expressed in normal kidney. It is not known, however, whether human peritoneal mesothelial cells (HPMC) express BMP-7 or if BMP-7 protects against peritoneal fibrosis and by what mechanism. We examined the effect of BMP-7 overexpression in TGF-beta1-induced epithelial-mesenchymal transition (EMT) of HPMC and in TGF-beta1 signaling in HPMC to elucidate the mechanisms of antifibrotic effect of BMP-7. METHODS: Growth arrested and synchronized HPMC were stimulated with 2 ng/mL of TGF-beta1 to induce EMT. HPMC were transiently transfected with adenovirus-mediated human BMP-7 (AdBMP-7) or with GFP (AdGFP). EMT was defined as downregulation of E-cadherin and upregulation of alpha-smooth muscle actin (SMA). RESULTS: HPMC constitutively expressed BMP-7 mRNA and protein. BMP-7 mRNA and protein expression were significantly inhibited by 50 mM D-glucose, 2x diluted commercial peritoneal dialysis solution, and 2 ng/ml of TGF-beta1. Transfection of AdBMP-7 resulted in 2.5-fold increase in BMP-7 mRNA expression in HPMC. TGF-beta1 significantly decreased E-cadherin and increased alpha-SMA expression in GFP transfected cells. BMP-7 overexpression effectively reversed TGF-beta1-induced E-cadherin and alpha-SMA expression and significantly suppressed TGF-beta1-induced phosphorylation of Smad2/3, ERK1/2, JNK, and p38 MAPK in HPMC as compared to GFP transfected cells. CONCLUSION: BMP-7 is an endogenous antifibrotic protein and downregulation of BMP-7 in HPMC by high glucose, PD solution, and TGF-beta1 may permit the development of peritoneal fibrosis during long-term PD. Our data demonstrate that BMP-7 overexpression reverses TGF-beta1-induced EMT of HPMC and consequent peritoneal fibrosis possibly through inhibition of Smad2/3 and MAPK phosphorylation.
Actins ; Bone Morphogenetic Protein 7* ; Cadherins ; Down-Regulation ; Epithelial-Mesenchymal Transition ; Glucose ; Humans ; Kidney ; p38 Mitogen-Activated Protein Kinases ; Peritoneal Dialysis ; Peritoneal Fibrosis ; Peritoneum* ; Phosphorylation ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; Up-Regulation

BACKGROUND: The transforming growth factor-beta1 (TGF-beta1) plays a key role in lung fibrosis. However, the mole?cular mechanisms involved in TGF-beta1-induced lung fibrosis are unclear. TGF-beta1 is the key inducer of myofibroblast transdifferentiation via de novo synthesis of alphasmooth muscle actin (alpha-SMA). Since TGF-beta1 signals through reactive oxygen species (ROS) and ROS have been shown to induce accumulation of extracellular matrix (ECM) in various tissues, this study examined if ROS play a role in TGF-beta1-induced fibronectin secretion and alpha-SMA expression in human lung fibroblasts, MRC-5 cells. METHODS: Growth arrested and synchronized MRC-5 cells were stimulated with TGF-beta1 (0.2-10 ng/ml) in the presence or absence of N-acetylcysteine (NAC) or diphenyleneiodonium (DPI) for up to 96 hours. Dichlorofluorescein (DCF)- sensitive cellular ROS were measured by FACScan and secreted fibronectin and cellular alpha-SMA by Western blot analysis. RESULTS: TGF-beta1 increased the level of fibronectin secretion and alpha-SMA expression in MRC-5 cells in a dose- dependent manner. Both NAC (20 and 30 mM) and DPI (1 and 5 microM significantly inhibited TGF-beta1-induced fibronectin and alpha-SMA upregulation. The TGF-beta1-induced cellular ROS level was also significantly reduced by NAC and DPI. CONCLUSIONS: The results suggest that NADPH oxidase-dependent ROS play an important role in TGF-beta1-induced fibronectin secretion and alpha-SMA expression in MRC-5 cells, which leads to myofibroblast transdifferentiation and progressive lung fibrosis.
Acetylcysteine ; Actins* ; Blotting, Western ; Extracellular Matrix ; Fibroblasts* ; Fibronectins* ; Fibrosis ; Humans* ; Lung* ; Muscle, Smooth* ; Myofibroblasts ; NADP ; Pulmonary Fibrosis ; Reactive Oxygen Species* ; Transforming Growth Factor beta1 ; Transforming Growth Factors* ; Up-Regulation

Staphylococcus aureus is able to utilize efficiently transferrin-bound iron as an iron source, whereas other staphylococci are not. The reason for this difference remains unclear. We compared the activity of siderophore-mediated iron-uptake systems among S. aureus, S. epidermidis, and S. saprophyticus. S. aureus was more susceptible to streptonigrin than the other two staphylococci. S. aureus was able to utilize efficiently transferrin-bound iron in proportion to the level of iron-saturation and produced siderophores in an inverse relation to iron-saturation. In contrast to S. aureus, S. epidermidis and S. saprophyticus were able to utilize only holotransferrin (HT; about 80% iron- saturated) and produced siderophores only in media containing HT. Moreover, they utilized HT less efficiently than S. aureus, though they produced greater amount of siderophores than S. aureus in media containing HT. The ability of the equivalent siderophores per se to capture iron from HT was not significantly different among the three species. Nevertheless, the siderophores from S. aureus stimulated the growth of the staphylococci to a greater degree than did the siderophores from S. epidermidis and S. saprophyticus. The siderophores from S. epidermidis and S. saprophyticus also stimulated the growth of S. aureus to a greater degree than those of the original bacteria which produced them. This indicates that S. aureus possesses a greater ability to produce more-efficient siderophores responding to very low iron-availability, as well as a greater ability to utilize iron-siderophore complexes, than the other two staphylococci. This explains in part the higher virulence of S. aureus compared to other staphylococci.
Bacteria ; Iron* ; Siderophores ; Staphylococcus aureus ; Streptonigrin ; Transferrin ; Virulence

PURPOSE: The purpose of this study were to identify the influential factors of bullying of intensive care nurses and to suggest a final structural model based on identified relationships between nursing organizational culture, authentic leadership, self-esteem, and bullying in the workplace. METHODS: Data were collected from 221 nurses at intensive care units in eight general hospitals using structured questionnaires and analyzed by structural equation modeling. RESULTS: In this study, the average of bullying in the workplace was 1.34±0.40, nursing organizational culture was 3.31±0.47, self-esteem was 2.79±0.44, and authentic leadership was 3.61±0.60. The factors affecting nursing organizational culture were authentic leadership (β=.54, p<.001) and self-esteem (β=.24, p=.002) that had direct positive effects on the nursing organizational culture. The nursing organizational culture had also a direct effect on reducing workplace bullying (β=−.45, p<.001). Authentic leadership (β=−.24, p=.004) and self-esteem (β=−.11, p=.004) had indirect effects on workplace bullying, which was mediated by the nursing organizational culture. CONCLUSION To understand and reduce workplace bullying, evaluating a nursing organizational culture should be preceded. Based on the finding of this study, an intervention for increasing authentic leadership and self-esteem of nurses can positively help to create the nursing organizational culture and then reduce workplace bullying.

Purpose: The aim of this study was to investigate the outcomes of endodontic treatment of mandibular second molar with C-shaped canal in elderly patients and related factors affecting the survival of the tooth. Materials and Methods: From 2010 to 2015, the survival rate following endodontic treatment was evaluated in elderly patients over 60 years who visited the Veterans Health Service Medical Center for endodontic treatment. The presence of C-shaped canals was confirmed using clinical records and radiographic features. Patientʼs age, sex, systemic diseases, tooth location, vitality, signs and symptoms, fractures, caries, apical radiolucency, canal shaping methods, sealer leakage, filling voids, and restoration of prosthesis were included in the analyses as confounding variables. The survival rate of teeth was analyzed using Kaplan–Meire analysis and the relationship between the survival rate and variables was analyzed using Simple and Multiple Cox regression analysis.Result: In total, 107 teeth in elderly patients had C-shaped canal. The survival rate of teeth that received endodontic treatment was 63.70%. None of the factors investigated significantly influenced the survival rate (P>0.05). Conclusion In elderly patients with C-shaped canal, the survival rate after root canal treatment was not significantly different from that of other mandibular molars.