We established an in vitro experimental system using the following procedure. We first introduced tritium-labeled norepinephrine ([3H]-NE) into PC12 cells. The [3H]-NE incorporated-PC12 cells were stimulated by a high concentration (60 mM) of K+ buffer during 12 minutes. Then, we collected 100 microliter supernatant and counted the amount of [3H]-NE release from PC12 cells with a scintillation counter. After screening fungal, Streptomyces spp. or bacterial product using this experimental sytem, we obtained FS390 from Streptomyces spp. which inhibited [3H]-NE release from PC12 cells. FS390 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons. The inhibitory effect was seen even when the PC12 cells were treated with low K+ buffer containing ionomycin (1 micrometer) as an ionopore. This result suggests that the inhibitory action of FS390 on neurotransmitter release appeared after the influx of Ca2+.
Adenosine Triphosphate ; Animals ; Exocytosis ; Ionomycin ; Mass Screening* ; Neurons ; Neurotransmitter Agents* ; Norepinephrine ; PC12 Cells* ; Rats ; Scintillation Counting ; Streptomyces

BACKGROUND: The establishment of safe and functional intravenous lines is of fundamental importance in premature sick neonates. Due to technical difficulties and various potential complications, central venous catheterization(CVC) has become an important part of long term care for prematurity. In terms of safety, it is believed by some that the central vein catheter tip should not be in the right atrium to avoid fatal complications. However, catheter performance and durability can be improved by positioning the catheter tip within the right atrium. Here, we evaluated the effectiveness of intraatrial catheter tip positioning for prematurities, to resolve this controversy between patient safety and catheter performance. METHODS: Premature neonates in whom CVC had been attempted at a nursery and pediatric intensive care unit were enrolled in this study. We successfully performed eighteen CVCs in twenty patients from September 2003 to December 2004. CVC tips were in positioned within the right atrium, and central venous route, central venous catheter depth, duration of catheterization, and any complications during or after catheterization were evaluated. RESULTS: Eighteen CVC were successful among twenty attempted in premature neonates. Two attempts failed due to guidewire insertion failure. A left supraclavicular venous approach was selected for 16 patients and a right approach for two. For intraatrial tip positioning, distances from skin to catheter tip were adjusted to an average depth of 5 cm in 11 patients and 6 cm in three patients. Two arterial punctures were the only minor complications. Mean duration of catheterization was 19 days without any long term complicaions. CONCLUSIONS: To prematue long term CVC performance, left supraclavicular subclavian vein catheterization and intraatrial catheter tip positioning should be considered a first option in premature neonates.
Catheterization* ; Catheterization, Central Venous ; Catheters* ; Central Venous Catheters ; Heart Atria ; Humans ; Infant, Newborn ; Intensive Care Units ; Long-Term Care ; Nurseries ; Patient Safety ; Punctures ; Skin ; Subclavian Vein ; Veins*

Crohn 's disease is characterized by a chronic relapsing inflammation of the bowel in which pro-inflammatory cytokines play an important role. Rebamipide is an anti-gastric ulcer drug with anti-inflammatory properties in vivo and in vitro. The effects of rebamipide on Crohn 's disease have not been carefully evaluated. This study investigated the potential of rebamipide to protect Crohn 's disease using a murine model of colitis induced by trinitrobenzene sulfonic acid (TNBS). Rebamipide dramatically improved histopathological symptom involving myeloperoxidase (MPO)activation and increase of microscopic damage score in TNBS induced colitis. Rebamipide suppressed IL-8 secretion, ICAM-1 induction and nuclear factor-kappaB (NF-kappaB) activation by TNF-alpha and induced heme oxygenase-1(HO-1)in HT-29 cells. HO-1 inducer cobalt protoporphyrin IX (CoPPIX)suppressed NF-kappaB activation by TNF-alpha in HT-29 cells like rebamipide, and mimicked the protective effects of rebamipide on TNBS induced colitis. This suggests that rebamipide exerts anti-inflammatory effects by down-regulating NF-kappaB activity via inducting HO-1 expression. In conclusion, this study suggests that rebamipide represents a potential therapeutic agent and HO-1 is an important therapeutic target for the treatment of Crohn's disease.
Cobalt ; Colitis ; Colon* ; Crohn Disease ; Cytokines ; Down-Regulation* ; Heme Oxygenase (Decyclizing)* ; Heme* ; HT29 Cells ; Humans ; Inflammation ; Intercellular Adhesion Molecule-1 ; Interleukin-8 ; NF-kappa B* ; Peroxidase ; Tumor Necrosis Factor-alpha ; Ulcer

BACKGROUND: Patients with ischemic heart disease are frequently maintained on a regimen of aspirin because of its ability to reduce variable thrombotic complications. However, aspirin has been recognized as a causative factor of increased perioperative bleeding. This study was aimed to determine whether aprotinin maintain its efficacy at reducing blood loss in patients on aspirin undergoing off-pump coronary artery bypass grafting (OPCAB). METHODS: In the prospective, double blind, randomized study, we investigated 30 patients on preoperative aspirin medication undergoing OPCAB surgery. Patients received aprotinin (1 x 106 KIU bolus for loading plus 0.5 x 106 KIU/hr, n = 15, the aprotinin group) or saline solution (n = 15, the control group). Operation time, total transfusion amount during operation, and plasma D-dimer levels immediately after the induction of anesthesia and immediately after operation were investigated and compared between and within groups where possible. RESULTS: Both group showed no comparable demographic and operation data such as operation time and number of grafts. The aprotinin group showed significantly less transfusion amount than the control group (packed RBC 352.7 +/- 89.2 ml vs 478.0 +/- 132.1 ml). Compared with that immediately after anesthetic induction value, postoperative D-dimer significantly increased in the control group but not in the aprotinin group. CONCLUSIONS: The above results suggest that aprotinin significantly reduces transfusion amount and probably inhibits fibrinolysis in patients with aspirin undergoing OPCAB.
Anesthesia ; Aprotinin* ; Aspirin* ; Coronary Artery Bypass, Off-Pump* ; Fibrinolysis ; Hemorrhage ; Hemostasis* ; Humans ; Myocardial Ischemia ; Plasma ; Prospective Studies ; Sodium Chloride ; Transplants*

Heme oxygenase-1 (HO-1) is a stress-inducible enzyme with anti-inflammatory activity, but the mechanisms underlying this activity are incompletely understood. Nuclear transcription factor kappa B (NF-kappa B) activation is an important factor in the pathogenesis of inflammatory bowel disease (IBD). We investigated the suppressive effects of HO-1 on the activation of NF-kappa B by pro-inflammatory cytokines in cultured colonic epithelial cells and by trinitrobenzene sulfonic acid (TNBS) in the colon of mice. The expression level of HO-1 in the colonic epithelium of a patient with inflammatory bowel disease and pseudo-membranous colitis was lower than that in a healthy control subject. In cultured human colonic epithelial HT-29 cells, pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha ) and IL-1 beta down-regulate HO-1 expression. The HO-1 inducer, cobalt protoporphyrin IX (CoPPIX), dramatically down-regulated NF-kappa B activation in HT-29 cells by TNF-alpha. In addition, bilirubin-a product of heme catabolism by HO-1-and the carbon monoxide donor tricarbonyldichlororuthenium (II) dimer also suppressed NF-kappa B activation by TNF-alpha. However, iron, another heme metabolite, did not suppress NF-kappa B activation by TNF-alpha. Furthermore, CoPPIX diminished the macroscopic and histopathological symptoms of TNBS-induced colitis and down-regulated NF-kappa B activation in mice. In conclusion, this study suggests that HO-1 plays an important role in the down-regulation of NF-kappa B activation, which is a key factor in the pathogenesis of IBD and is thus an excellent therapeutic target for the treatment of IBD.
Animals ; Carbon Monoxide ; Cobalt ; Colitis ; Colon* ; Cytokines ; Down-Regulation* ; Epithelial Cells ; Epithelium* ; Heme Oxygenase-1* ; Heme* ; HT29 Cells ; Humans ; Inflammation* ; Inflammatory Bowel Diseases ; Interleukin-1beta ; Iron ; Metabolism ; Mice ; NF-kappa B ; Tissue Donors ; Transcription Factors* ; Tumor Necrosis Factor-alpha

Discovery of Nod2 has brought to light the significance of mononuclear cells as well as epithelial cells in inflammatory bowel disease (IBD) pathogenesis. Similarly, CCL20 is expressed in both mononuclear cells and epithelial cells and is likely to link innate and acquired immunity. We therefore asked whether CCL20 expression is altered in the peripheral blood mononuclear cells (PBMCs) from patients with ulcerative colitis (UC), a major type of IBD in Korea, and is correlated with the disease activity. The expression levels of CCL20 mRNA were significantly high in the PBMCs from the patients with UC. CCL20 protein expression was also up-regulated in the mucosal epithelium in UC but not in normal controls. Interestingly, however, disease activity index (DAI) revealed that untreated UC groups express higher expression levels of CCL20 mRNA than treated UC groups, implying that CCL20 may be a potential target for the anti-inflammatory treatments. In an agreement with this, three months follow up study revealed that the UC patients who were treated with 5-amino salicylic acid (5-ASA) and glucocorticoid showed dramatic decrease in their CCL20 mRNA levels as compared to untreated ones. Moreover, TNF-alpha-or IL-1beta-induced CCL20 secretion in human epithelial HT-29 cells was significantly diminished by the treatment with 5-ASA and/or dexamethasone, suggesting that CCL20 may be one of the central targets of the anti-inflammatory drugs. Collectively, these results suggest that CCL20 expression in UC may be associated with altered immune and inflammatory responses in the blood as well as the intestinal mucosa and further implied a potential for CCL20 as an important diagnostic marker for UC.
Adaptive Immunity ; Blood Cells* ; Colitis, Ulcerative* ; Crohn Disease ; Dexamethasone ; Epithelial Cells ; Epithelium ; Follow-Up Studies ; Gene Expression* ; HT29 Cells ; Humans ; Inflammatory Bowel Diseases ; Intestinal Mucosa ; Korea ; RNA, Messenger ; Salicylic Acid ; Sulfasalazine* ; Ulcer*

Aspergillus funigatus and other pathogenic fungi synthesize a toxic epidithi-odiopiperzine (ETP) metabolite, namely gliotoxin. Gliotoxin commonly react with sulfhydryl groups, and then, forms hydrogen peroxide. These fungal toxins induce apoptotic cell death in various cells. Apoptosis induced by gliotoxin need calcium. Effect of calcium preconditioning was not reported in gliotoxin-induced apoptosis. To examine the effect of protein kinase C (PKC) and calcium which was regulate caspase-3, PKC and calcium preconditioning before gliotoxin treatment, apoptotic agents such as bcl-2 family, caspase-3 and DNA fragmentation in A7r5 cell line from rat smooth muscle cell were studied. These results showed that gliotoxin induces the expression of bad of bcl-2 family, caspase-3 activation and DNA fragmentation in A7r5 cells. Gliotoxin treatment followed by calcium and PKC preconditioning suppress the Bad of bcl-2 family, and inhibited caspase-3 activation, respectively. These results suggest that PKC and calcium preconditioning protect the gliotoxin-induced apoptosis, through the protection of pro-apoptotic bcl-2 family in A7r5 cells.
Animals ; Apoptosis ; Aspergillus ; Calcium ; Caspase 3 ; Cell Death ; Cell Line ; DNA Fragmentation ; Fungi ; Gliotoxin ; Humans ; Hydrogen Peroxide ; Muscle, Smooth* ; Mycotoxins ; Myocytes, Smooth Muscle* ; Protein Kinase C ; Rats*

Paclitaxel (Taxol) is known as effective drug for inhibition of cell cycle encouraging in human cancer cells. This drug named an antimicrotubule agent which simulate the mitotic arrest towards an apoptosis. The influence of phorbol 12 myristate 13 acetate (PMA) activated protein kinase C (PKC) and nitric oxide (NO) on taxol-induced apoptosis, is poorly understood. To investigate the effects of PMA and NO on the signal transduction in taxol-induced apoptosis in C6-glial cells, the viability and caspase-3 activity of C6-glial cells were analyzed. Pretreatement with PKC activatior (PMA) protected taxol-induced cell death in C6-glial cells, by inhibited caspases-3 activity. On the other hand, the taxol-induced apoptosis was highly enhanced by sodium nitroprusside (SNP) and lipopolysaccharide (LPS), as NO activator. These results suggest that PMA strongly blocks the apoptotic effect of taxol, while nitric oxide has no protective effects in the process of toxol-induced apoptosis in C6-glial cells.
Apoptosis ; Caspase 3 ; Cell Cycle ; Cell Death ; Hand ; Humans ; Myristic Acid ; Nitric Oxide* ; Nitroprusside ; Paclitaxel ; Protein Kinase C ; Signal Transduction

BACKGROUND: Falls in the elderly can lead to disability, hospitalizations, and premature death. Even if the fall does not cause significant injury, it may lead to fear of falling, loss of self confidence and restriction of ambulation. Thus, we conducted this study to examine the risk factors of falls in the elderly. METHODS: The study population consisted of 70 persons, older than 65 years. Subjocts were subdivided into 3 groups according to their experience of falls, during the past 2 year period. Among them, 30 persons had no previous experience, 20 had one fall, and 20 more than one fall. Age, past history, situations surrounding falls, MMSE K, GDS short form, ADL, IADL, Mini Nutritional assessment (MNA) were reviewed, and the Get up and go test, evaluation of orthostic hypotension and hearing were done. T-test, chi square, ANOVA, logistic regression test using the SAS program was performed. RESULTS: The mean age of the participants was 75.4 years with 62.9% malas. The factors associated with falling were age (P=0.01), scores of MNA (P=0.04), Get up and go test (P=0.004), past history (P=0.01), MMSE K (P=0.02), GDS short form (P=0.001), ADL (P=0.003) and IADL (P=0.002). Those in the group who fell once occured mostly while doing a riskful task, while those who fell more than once happened mostly during positional change (p<0.0001). Independent predictors of falls were get up and go test results and GDS short form scores. CONCLUSION: The independent predictors of falls were Get up and go test results and Geriatric Depression Scale scores.
Activities of Daily Living ; Aged* ; Depression ; Hearing ; Hospitalization ; Humans ; Hypotension ; Logistic Models ; Mortality, Premature ; Nutrition Assessment ; Risk Factors* ; Walking

Aspergillus funigatus and other pathogenic fungi synthesize a toxic epidithi- odiopiperzine (ETP) metabolite called gliotoxin. Gliotoxin is an epidithiodiopiperzine compound which can both react with sulfhydryl groups and form hydrogen peroxide. The fungal toxin gliotoxin induces apoptotic cell death in a variety of cells. Apoptosis induced by gliotoxin need calcium but effect of calcium preconditioning is unknown by gliotoxin. We studied the effect of protein kinase C and calcium preconditioning on gliotoxin-induced apoptosis in H9c2 cell. PKC and calcium preconditiong inhibited DNA fragmentation by gliotoxin. From this above results suggest that gliotoxin induce apoptosis via caspase-3 activation, because caspase-3 inhibitor (DEVD-CHO) didn't induce apoptosis in gliotoxin treated H9c2 clls. Calcium and PKC preconditioning inhibit caspase-3 activation by gliotoxin. These data means that PKC preconditioning is related with caspase-3 regulate in gliotoxin-induced apoptosis.
Apoptosis* ; Aspergillus ; Calcium ; Caspase 3 ; Cell Death ; DNA Fragmentation ; Fungi ; Gliotoxin* ; Hydrogen Peroxide ; Protein Kinase C* ; Protein Kinases*