Recently, we reported violapyrones B, C, H and I, unusual 3, 4, 6-trisubstituted alpha-pyrone derivatives, from the culture broth of the marine Streptomyces sp. 112CH148. In previous studies, violapyrones have been shown to have antibacterial and antitumor activities. However, the anti-inflammatory effect of violapyrones has not been reported yet. As part of our ongoing study for the discovery of bioactive metabolites from marine microorganisms, we found that violapyrones also have anti-inflammatory activity. In this study, we investigated the effect of violapyrones on LPS-induced inflammatory responses in vitro. Violapyrones B and C did not affect the viability of RAW 264.7 cells at concentrations up to 25 microM. However, violapyrones B and C inhibited the production of NO compared to the LPS-induced control. In addition, violapyrones B and C down-regulated the expression of iNOS protein in LPS-stimulated RAW 264.7 cells. To the best of our knowledge, this is the first report on the anti-inflammatory activity of violapyrones B and C.
Streptomyces*

Proper seat belt use saves lives; however, the use rate decreased in Korea. This study aimed to measure the magnitude of the preventive effect of seat belt on case-fatality across drivers and passengers. We used the Emergency Department based Injury In-depth Surveillance (EDIIS) database from 17 EDs between 2011 and 2012. All of adult injured patients from road traffic injuries (RTI) in-vehicle of less than 10-seat van were eligible, excluding cases with unknown seat belt use and outcomes. Primary and secondary endpoints were in-hospital mortality and intracranial injury. We calculated adjusted odds ratios (AORs) of seat belt use and driving status for study outcomes adjusting for potential confounders. Among 23,698 eligible patients, 15,304 (64.6%) wore seat belts. Driver, middle aged (30-44 yr), male, daytime injured patients were more likely to use seat belts (all P < 0.001). In terms of clinical outcome, no seat belt group had higher proportions of case-fatality and intracranial injury compared to seat belt group (both P < 0.001). Compared to seat belt group, AORs (95% CIs) of no seat belt group were 10.43 (7.75-14.04) for case-fatality and 2.68 (2.25-3.19) for intracranial injury respectively. In the interaction model, AORs (95% CIs) of no seat belt use for case-fatality were 11.71 (8.45-16.22) in drivers and 5.52 (2.83-14.76) in non-driving passengers, respectively. Wearing seat belt has significantly preventive effects on case-fatality and intracranial injury. Public health efforts to increase seat belt use are needed to reduce health burden from RTIs.
Accidents, Traffic/mortality/*prevention & control ; Adult ; Aged ; Craniocerebral Trauma/prevention & control ; Databases, Factual ; Emergency Service, Hospital ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Motor Vehicles ; Odds Ratio ; Republic of Korea/epidemiology ; Seat Belts/*utilization ; Young Adult

We investigated the synergistic apoptotic effects of co-treatments with Chios gum mastic (CGM) and eugenol on G361 human melanoma cells. An MTT assay was conducted to investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and analyses of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate expression and translocation of apoptosis-related proteins following CGM and eugenol co-treatment. Proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed.The results indicated that the co-treatment of CGM and eugenol induces multiple pathways and processes associated with an apoptotic response in G361 cells. These include nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, an increase of Bax and decrease of Bcl-2, a decreased DNA content, cytochrome c release into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of 40 microg/ml CGM or 300 microM eugenol for 24 hours did not induce apoptosis. Our present data thus suggest that a combination therapy of CGM and eugenol is a potential treatment strategy for human melanoma.
Apoptosis ; Blotting, Western ; Caspase 3 ; Caspase 7 ; Caspase 9 ; Cytochromes c ; Cytosol ; DNA ; DNA Fragmentation ; Electrophoresis ; Eugenol ; Gingiva ; Humans ; Melanoma ; Membrane Potential, Mitochondrial ; Proteasome Endopeptidase Complex ; Proteins ; Resins, Plant

The influence of spinal cord injury (SCI) on protein expression in the rat urinary bladder was assessed by proteomic analysis at different time intervals post-injury. After contusion SCI between T9 and T10, bladder tissues were processed by 2-DE and MALDI-TOF/MS at 6 hr to 28 days after SCI to identify proteins involved in the healing process of SCI-induced neurogenic bladder. Approximately 1,000 spots from the bladder of SCI and sham groups were visualized and identified. At one day after SCI, the expression levels of three protein were increased, and seven spots were down-regulated, including heat shock protein 27 (Hsp27) and heat shock protein 20 (Hsp20). Fifteen spots such as S100-A11 were differentially expressed seven days post-injury, and seven proteins including transgelin had altered expression patterns 28 days after injury. Of the proteins with altered expression levels, transgelin, S100-A11, Hsp27 and Hsp20 were continuously and variably expressed throughout the entire post-SCI recovery of the bladder. The identified proteins at each time point belong to eight functional categories. The altered expression patterns identified by 2-DE of transgelin and S100-A11 were verified by Western blot. Transgelin and protein S100-A11 may be candidates for protein biomarkers in the bladder healing process after SCI.
Animals ; Biological Markers/metabolism ; Electrophoresis, Gel, Two-Dimensional ; Female ; HSP20 Heat-Shock Proteins/metabolism ; HSP27 Heat-Shock Proteins/metabolism ; Microfilament Proteins/metabolism ; Muscle Proteins/metabolism ; Proteome/*biosynthesis ; Proteomics ; Rats ; Rats, Sprague-Dawley ; S100 Proteins/metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Spinal Cord Injuries/*metabolism/pathology ; Urinary Bladder/*metabolism ; *Wound Healing

Among the several rotenoids, amorphigenin is isolated from the leaves of Amopha Fruticosa and it is known that has anti-proliferative effects and anti-cnacer effects in many cell types. The main aim of this study was to investigate the effects of amorphigenin on osteoclast differentiation in vitro and on LPS treated inflammatory bone loss model in vivo. We show here that amorphigenin inhibited RANKL-induced osteoclast differentiation from bone marrow macrophages in a dose dependent manner without cellular toxicity. Anti-osteoclastogenic properties of amorphigenin were based on a down-regulation of c-fos and NFATc1. Amorphigenin markedly inhibited RANKL-induced p38 and NF-kappaB pathways, but other pathways were not affected. Micro-CT analysis of the femurs showed that amorphigenin protected the LPS-induced bone loss. We concluded that amorphigenin can prevent inflammation-induced bone loss. Thus we expect that amorphigenin could be a treatment option for bone erosion caused by inflammation.
Bone Marrow ; Down-Regulation ; Femur ; Inflammation ; Macrophages ; NF-kappa B ; Osteoclasts ; Osteoporosis ; Rotenone ; T-Lymphocytes

Chios gum mastic (CGM) is obtained from the stem and leaves of Pistacia lentiscus trees and has been extensively used for centuries in Mediterranean and Middle Eastern countries, both as a dietary supplement and herbal remedy. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying modulation of cell cycle and induction of apoptosis in YD9 human oral squamous carcinoma cell line treated with CGM. The viability of YD9 cells and human normal keratinocyes (HaCaT cells), and the growth inhibition of YD9 cells were assessed by the MTT assay and clonogenic assay respectively. The hoechst staining and DNA electrophoresis were conducted to observe the YD9 cells undergoing apoptosis. YD9 cells were treated with CGM, and Western blotting, immunocytochemistry, confocal microscopy and FACScan flow cytometry were conducted. Mitochondrial membrane potential change and proteasome activity were measured. CGM treatment on YD9 cells resulted in a does-dependent inhibition of cell growth and induced apoptotic cell death. And tested YD9 cells showed several lines of apoptotic manifestation. Flow cytometric analysis revealed that CGM resulted in G1 arrest in cell cycle progression which was associated with decrease in the protein expression of cyclin D1, cyclin D3, Cdk2 and Cdk4, and increase in the protein expression of p21(WAF1/CIP1) and p53. These results demonstrate that CGM induces G1 the cell cycle arrest via the modulation of cell cycle-related proteins, and apoptosis via mitochondria and caspase pathway in YD9 cells, suggesting that CGM can be considered as a novel therapeutic strategy for human oral squamous cell carcinoma from its strong cell cycle arrest and apoptosis-inducing activity.
Apoptosis ; Blotting, Western ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Death ; Cell Line ; Cyclin D1 ; Cyclin D3 ; Dietary Supplements ; DNA ; Electrophoresis ; Flow Cytometry ; Gingiva ; Humans ; Immunohistochemistry ; Membrane Potential, Mitochondrial ; Microscopy, Confocal ; Mitochondria ; Pistacia ; Proteasome Endopeptidase Complex ; Proteins ; Resins, Plant ; Trees

Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. Recently it reported that CGM induced apoptosis in a few cancer cells in vitro. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with a natural product, CGM and a CDCA derivative, HS-1200 on human osteosarcoma (HOS) cells. To investigate whether the co-treatment of CGM and HS-1200 compared with each single treatment efficiently reduced the viability of HOS cells, MTT assay was conducted. Induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and DNA hypoploidy, Westen blot analysis and immunofluorescent staining were performed to study the alterations of the expression level and translocation of apoptosis-related proteins in co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) change were also assayed. In this study, HOS cells co-treated with CGM and HS-1200 showed several lines of apoptotic manifestation whereas each single treated HOS cells did not. Although the single treatment of 40 microgram/mL CGM or 25 micrometer HS-1200 for 24 h did not induce apoptosis, the cotreatment of them induced prominently apoptosis. Therefore our data provide the possibility that combination therapy of CGM and HS-1200 could be considered as a novel therapeutic strategy for human osteosarcoma.
Apoptosis ; Chenodeoxycholic Acid ; DNA ; Electrophoresis ; Exudates and Transudates ; Gingiva ; Humans ; Membrane Potential, Mitochondrial ; Osteosarcoma ; Pistacia ; Proteasome Endopeptidase Complex ; Proteins ; Resins, Plant ; Trees

BACKGROUND AND OBJECTIVES: Myocardial ruptures, including: ventricular free wall rupture (VFWR) and ventricular septal rupture (VSR), after acute myocardial infarction (AMI), are fatal complications. Recently, ubiquitous use of echocardiographs, and other imaging techniques, allows us to diagnose these complications in the antemortem period. Thus, this study retrospectively evaluated the clinical characteristics of patients with myocardial ruptures following AMI. SUBJECTS AND METHODS: 620 patients that had had AMIs, between January 1999 and June 2001, were analysed for the purpose of this study. Myocardial ruptures were diagnosed from their clinical symptoms, echocardiographs, and pericardiocenteses or cardiac catheterizations. The clinical characteristics of the patients with myocardial ruptures (n=15) were compared to those patients with myocardial infarction, without rupture (n=397), from their Q waves. RESULTS: The patients with myocardial ruptures were older than those without (67+/-9.7 years vs 60+/-11.7 years, p<0.05), and ruptures were more frequent in women (66.7% vs 25.2%, p<0.001). The frequency of systemic hypertension, DM, and the distribution of infarction sites were similar in both groups. Also, clinical characteristics between patients with VFWR, and those with VSR, were similar. Of the patients with VFWR (n=8), 7 suddenly died, and 1 was alive directly following surgery. Of the patients with VSR (n=7), 4 died. CONCLUSION: Myocardial rupture is a fatal complication of AMI, which is more frequent in women, and the patients with ruptures, in our study, were older than those without.
Cardiac Catheterization ; Cardiac Catheters ; Female ; Heart Rupture ; Humans ; Hypertension ; Infarction ; Myocardial Infarction* ; Pericardiocentesis ; Retrospective Studies ; Rupture* ; Ventricular Septal Rupture

BACKGROUND: Tei index is a Doppler-derived index of combined systolic and diastolic myocardial performance proposed to be a promising noninvasive measurement of overall cardiac function, calculated as the sum of isovolumic contraction (ICT) and isovolumic relaxation times (IRT) divided by ejection time (ET). This study aimed to investigate the effect of changes in preload on Tei index. SUBJECTS AND METHOD: The study population was 10 patients with chronic renal failure who are on regular hemodialysis (HD). They were 3 men and 7 women with a mean age of 45.3+/-12.4 years old. The complete 2-dimensional and Doppler echocardiography including recordings of mitral inflow and left ventricular outflow were performed using HP SONOS 1500 equipment before and after HD. Body weight was measured using electric scale before and after HD. Blood pressure and pulse rate were measured by an automated cuff measurement every 1 hour throughout HD. Hematocrit was monitored noninvasively and continuously using the Crit-Line instrument (In-Line Diagnostics, Riverdale, UT, USA) during HD. RESULTS: There were no significant changes in systolic and diastolic blood pressure, RR interval, left ventricular end-diastolic and end-systolic dimension, left atrial dimension, and left ventricular ejection fraction during HD. The mean body weight loss during HD was 2.1+/-0.65 kg. The percentage of reduction of body weight was 4.1+/-1.56%. Changes in total blood volume at the end of HD were compared with that of baseline -11.79+/-5.51%. The percent change in total blood volume during the HD correlated significantly with the amount of change in body weight (r=0.678, p<0.05) and percent change in body weight (r=0.835, p<0.01), respectively. ICT before HD and at the end of HD were 39+/-13.0, 42+/-24.1 msec (p=NS). IRT before HD and at the end of HD were 76+/-29.1, 95+/-28.5 msec (p<0.01) and , ET before HD and at the end of HD were 317+/-18.1, 289+/-9.0 msec (p<0.01), respectively. Tei index before HD was 0.36+/-0.10, and the index at the end of HD were 0.47+/-0.13 (p<0.001). CONCLUSION: When there is a change in preload during a short period, it seems that the effect of change in preload should be considered when using Tei index to assess ventricular function.
Blood Pressure ; Blood Volume ; Body Weight ; Echocardiography, Doppler ; Female ; Heart Rate ; Hematocrit ; Humans ; Kidney Failure, Chronic ; Male ; Relaxation ; Renal Dialysis ; Stroke Volume ; Ventricular Function

OBJECTIVES: In spite of the improvement in therapeutic strategy, the mortality rate from sepsis is still high. The purpose of this study was to examine the clinical characteristics and prognostic factors of sepsis to get help in treatment and estimation of prognosis of sepsis. METHODS: We analyzed the clinical and bacteriologic data of 313 admitted patients with bacteremia at Pusan National University Hospital from Jan., 1996 to Dec., 1997 retrospectively and all patients were categorized into 4 groups (bacteremia, sepsis, severe sepsis, septic shock) by the definition from American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference, 1992. RESULTS: 1) Male to female ratio was 1.22: 1 and mean age was 52 years. 2) The overall mortality rate was 32.3% and the mortality rates of bacteremia, sepsis, severe sepsis and septic shock were 14.9%, 22.8%, 52.0%, and 95.2%, respectively. 3) Underlying diseases predisposing to bacteremia were diabetes mellitus(14.4%), solid cancer (13.1%), hematologic malignancy(10.2%) and liver cirrhosis(9.9%), but the most common was no underlying disease. 4) Among the total 80 species of isolated microorganisms, gram-positive organisms were responsible in 169 cases, gram-negative organisms in 218 cases. E. coli was isolated most frequently, followed by S. aureus, S. epidermidis, K. pneumoniae, Enterococcus spp., Enterobacter spp., Str. viridans group, CNS, P. aeruginosa and S. typhi. 5) Although the source of bacteremia could not be identified in 28.8% of the patients, the others had the primary site of infections ; skin(17.9%), gastrointestinal tract(16.9%), respiratory tract(12.5%), urinary tract(9.9%) and biliary tract(7.3%). 6) The mortality rate in patients with gram-positive bacteremia was 39.4%, with gram-negative bacteremia was 20.9% and with polymicrobial bacteremia was 33.3%. MRSA sepsis showed the highest mortality rate(58.8%), followed by Enterococcus spp.(50.0%), K. pneumoniae (35.0%), P. aeruginosa(27.3%) and E. coli(18.8%). 7) There was significant relation between etiologic organisms of bacteremia and the primary site of infections. 8) Using logistic regression analysis, mortality was predicted by disseminated intravascular coagulation, severity of sepsis and severity of underlying diseases. CONCLUSION: Sepsis occurred in 16 patients among 1,000 adult admitted patients and overall mortality rate was 32.3%, still high. The mortality rate had positive correlation with the severity of sepsis. Among 16 evaluated risk factors of mortality, mortality was predicted by disseminated intravascular coagulation, severity of sepsis and severity of underlying disease.
Adult ; Bacteremia* ; Busan ; Consensus ; Critical Care ; Disseminated Intravascular Coagulation ; Enterobacter ; Enterococcus ; Female ; Humans ; Liver ; Logistic Models ; Male ; Methicillin-Resistant Staphylococcus aureus ; Mortality ; Pneumonia ; Prognosis ; Retrospective Studies ; Risk Factors ; Sepsis* ; Shock, Septic ; Thorax