PURPOSE: To investigate the effects of dipyridamole (DPD) on the production of reactive oxygen species (ROS) and oxidative stress in cultured human trabecular meshwork cells (HTMC). METHODS: Antioxidant activity of DPD was determined by DPPH assay. Primarily cultured HTMC were exposed to 0, 20, and 50 microm DPD using serum-deprived media. The effect of DPD on the production of ROS was assessed with the DCHFDA assay. The effect of DPD on the t-butyl hydroperoxide (tBHP)-induced oxidative stress was assessed with resazurin assay. RESULTS: DPD showed significant antioxidant activity. DPD significantly decreased the production of ROS (p < 0.05) and improved cellular activity significantly after treatment with t-BHP (p < 0.05). DPD did not affect the generation of nitric oxides. CONCLUSIONS: DPD suppressed the formation of ROS and possessed cytoprotective activity against the oxidative stress in HTMC.
Dipyridamole ; Humans ; Oxazines ; Oxidative Stress ; Reactive Oxygen Species ; tert-Butylhydroperoxide ; Trabecular Meshwork ; Xanthenes

PURPOSE: To evaluate the protective effect of creatine on the survival of retinal ganglion cells after serum deprivation. METHODS: RGC-5 cells were exposed to 5 mM creatine with serum-free media for 4 days. Cellular survival and mitochondrial respiratory activity were measured with MTT assay and resazurin assay, respectively. Degree of apoptosis was evaluated with vital staining using acridine orange/Hoechest 33342 and flow cytometric analysis using annexin/PI, respectively. RESULTS: Creatine increased cellular survival of RGC-5 cells significantly after serum deprivation. Additionally, creatine increased mitochondrial respiratory activity and inhibited apoptosis of RGC-5 cells. CONCLUSIONS: The energy precursor creatine increased survival of retinal ganglion cells after serum deprivation. Creatine could be relevant for the cytoprotection of retinal ganglion cells.
Apoptosis ; Creatine ; Culture Media, Serum-Free ; Cytoprotection ; Oxazines ; Retinal Ganglion Cells ; Xanthenes

This paper specifically discusses the risk assessment on the pesticide residues in vegetables collected from traditional markets, big marts and departments in the southern part of Seoul. Vegetable samples were 6,583 cases from January to December in 2009. Monte-Carlo simulation was used to calculate the uncertainty for the risk index using pesticide residues, average dietary intake for vegetables and acceptable daily intake. Deterministic risk indexes were 7.33% of diethofencarb, 5.13% of indoxacarb, 3.96% of EPN, 3.92% of diniconazole and 3.09% of chlorothalonil, respectively. And other pesticides were below 3%. Distributions of risk indexes obtained by the Monte-Carlo simulations were similar to the deterministic values, even though the confidence intervals for 95% were very wide. We confirmed that health risks caused by eating vegetables exceeded maximum residue limits of pesticide are very low and the population is generally safe, judging from the risk indexes located between 0.07 to 9.49%.
Eating ; Korea ; Nitriles ; Oxazines ; Pesticide Residues ; Pesticides ; Phenylcarbamates ; Risk Assessment ; Triazoles ; Uncertainty ; Vegetables

AIMTo assess that metabolic status of spermatozoa could provide a useful tool for evaluation of semen quality.

METHODSThe accuracy of the spectrophotometric application of the resazurin reduction assay was assessed using receiver operating characteristic (ROC) analysis.

RESULTSAreas under ROC curves (AUC) for motile sperm concentration and sperm index (SI) (sperm concentration multiplied by the square root of percentage sperm motility multiplied by the percentage normal sperm morphology) were 0.922. The best discrimination between poor and good semen samples according to the SI was achieved at a cut-off point of A610 = 0.209, where high sensitivity (94.1%) and specificity (91.7%) were calculated. The assay was less accurate when motile sperm concentration was used as the criterion value, yielding sensitivity of 88.2% and specificity of 87.5%, respectively. Likelihood ratios (LR) indicate that absorbances lower than 0.209 were at least 11.3 times as likely to be found in good semen samples than those in poor according to the SI, whereas in the case of motile sperm concentration, the LR was calculated to be 7.06.

CONCLUSIONThese results show that the resazurin reduction assay combined with spectrophotometry is an accurate method of assessing the quality of boar semen.

Animal Husbandry ; Animals ; Area Under Curve ; Male ; Oxazines ; pharmacokinetics ; pharmacology ; Semen ; physiology ; Sperm Motility ; Swine ; Xanthenes ; pharmacokinetics ; pharmacology

Animals ; Humans ; Oxazines ; metabolism ; Phenylpropionates ; metabolism ; Receptors, Cytoplasmic and Nuclear ; agonists ; antagonists & inhibitors ; Thiazolidinediones ; metabolism ; Transcription Factors ; agonists ; antagonists & inhibitors

PURPOSE: The aim of this in vitro study was to investigate the adhesion of initial colonizer, Streptococcus sanguis, on resin, titanium and zirconia under the same surface polishing condition. MATERIALS AND METHODS: Specimens were prepared from Z-250, cp-Ti and 3Y-TZP and polished with 1 microm diamond paste. After coating with saliva, each specimen was incubated with Streptococcus sanguis. Scanning electron microscope, crystal violet staining and measurement of fluorescence intensity resulting from resazurin reduction were performed for quantifying the bacterial adhesion. RESULTS: Surface of resin composite was significantly rougher than that of titanium and zirconia, although all tested specimens are classified as smooth. The resin specimens showed lower value of contact angle compared with titanium and zirconia specimens, and had hydrophilic surfaces. The result of scanning electron microscopy demonstrated that bound bacteria were more abundant on resin in comparison with titanium and zirconia. When total biofilm mass determined by crystal violet, absorbance value of resin was significantly higher than that of titanium or zirconia. The result of relative fluorescence intensities also demonstrated that the highest fluorescence intensity was found on the surface of resin. Absorbance value and fluorescence intensity on titanium was not significantly different from those on zirconia. CONCLUSION: Resin specimens showed the roughest surface and have a significantly higher susceptibility to adhere Streptococcus sanguis than titanium and zirconia when surfaces of each specimen were polished under same condition. There was no significant difference in bacteria adhesion between titanium and zirconia in vitro.
Bacteria ; Bacterial Adhesion ; Biofilms ; Colon ; Diamond ; Electrons ; Fluorescence ; Gentian Violet ; Microscopy, Electron, Scanning ; Oxazines ; Saliva ; Streptococcus sanguis ; Titanium ; Xanthenes ; Zirconium

OBJECTIVETo establish a rapid and sensitive high-performance liquid chromatography (HPLC) method for detecting pazufloxacin concentrations in the saliva, gingival crevicular fluid and serum of healthy adults.

METHODSSamples of saliva, gingival crevicular fluid and serum were obtained from healthy adults receiving intravenous infusion of pazufloxacin. The concentrations of pazufloxacin in the samples were quantified by HPLC equipped with a reversed-phase column (Agilent Zorbax SB-C18 5 µm, 250 mm×4.6 mm). The mobile phase for pazufloxacin was a mixture of acetonitrile and 0.5% phosphoric acid containing 1% triethylamine (155:850), and 20 µl of the resulting solution was injected into the HPLC system at a flow rate of 1.0 ml/min. The detection wavelength was set at 245 nm. The samples were first deproteinized by precipitation with methanol followed by supernatant drying; the residue was reconstituted with the mobile phase and centrifuged, and the supernatants were directly injected into the HPLC system.

RESULTSPazufloxacin in the samples were totally separated without interference by any endogenous substances. The calibration curves showed a good linear regression (r>0.999). The detection limit was 10 ng/ml with within-day and between-day coefficients of variation performance all below 5% and recovery rates all above 91%.

CONCLUSIONHPLC is both sensitive and selective for quantification of pazufloxacin in saliva, gingival crevicular fluid and serum.

Adult ; Chromatography, High Pressure Liquid ; methods ; Female ; Fluoroquinolones ; analysis ; blood ; Gingival Crevicular Fluid ; chemistry ; Humans ; Male ; Oxazines ; analysis ; blood ; Saliva ; chemistry ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo illustrate the effects of the site and method of liposuction on differentiation of human preadipocytes.

METHODSForty-two fatty samples were obtained with liposuction, which were then divided into four groups according to operation sites (abdomen, hip or extremity) and the methods (conventional negative-pressure or syringe method). Each sample was treated with collagenase I to release preadipocytes for in vitro culture. Affected by the differentiation-induced agents for 9 days or 28 days, the cultured adipocytes were stained with Nile red and observed under a fluorescent microscope. The differentiation rates were examined with flow cytometric analysis and the quantity of intracytoplasmic lipids was determined with oil red O staining. The results were analyzed with independent samples t-test (Mann-Whitney) using SPSS 10.0.

RESULTSThere was no statistical difference in differentiation at the 9th day or 28th day among the preadipocytes obtained from the abdomen, hip or extremity with the negative-pressure method. The preadipocytes from the abdomen differentiated more at the 28th day than the 9th day (P < 0.05), which was not observed in the hip or the extremity. The preadipocytes obtained from the abdomen with the negative-pressure method differentiated more than those with the syringe method (P < 0.05).

CONCLUSIONNo essential difference was found in preadipocyte differentiation among the liposuction sites, while the abdomen might have some superiority. The negative-pressure method of liposuction is the first option in future research of tissue engineering. The flow cytometric analysis is a convenient way to study preadipocyte differentiation.

Abdomen ; surgery ; Adipocytes ; cytology ; Adult ; Cell Differentiation ; Cells, Cultured ; Female ; Flow Cytometry ; Hip ; surgery ; Humans ; Lipectomy ; methods ; Male ; Microscopy, Fluorescence ; Middle Aged ; Oxazines

To evaluate prevalence of HBsAg in the liver tissue of various liver diseases among Koreans, a total of 614 cases of liver biopsies; hepatitis (104), cirrhosis (99), hepatocellular carcinoma (78), cholangiocarcinoma (3), metastatic carcinoma (35), HBsAg carrier (15) and miscellaneous (280) ; were studied by histochemical methods of Shikata's modified orcein, Gomori's aldehyde-fuchsin, aldehyde thionine, Weigert's resorcin-fuchsin, alcian blue, and Verhoeff's elastic staining. It was found that the modified orcein and Gomori's aldehyde-fuchsin methods gave the best and most reliable results. The prevalence of orcein positive materials in various liver diseases showed constantly negative results in acute hepatitis, portal and biliary cirrhosis, cholangiocarcinoma and metastatic tumors, and positive results in 27.5% of chronic persistent hepatitis, 56.8% in chronic active hepatitis, 56% in chronic active hepatitis leading to cirrhosis, 59.5% in adjacent liver tissue of hepatocellular carcinoma, 11.5% in tumor cells of hepatocellular carcinoma, 60% in asymptomatic carriers, and 0.7% in the miscellaneous disease group. The ground glass appearing hepatocytes which were present in various numbers in various liver diseases gave intense positive reaction with orcein staining, especially in asymptomatic carriers. These results strongly indicate that chronic hepatitis, macronodular cirrhosis and hepatocellular carcinoma among Koreans are closely related with hepatitis B virus infection, and the percentage reported here might well be lower than actual prevalence since the specimens are mostly needle biopsy materials and the distribution of orcein positive materials are often spotty.
Hepatitis B Surface Antigens/analysis* ; Human ; Korea ; Liver/immunology ; Liver/pathology ; Liver Diseases/immunology* ; Liver Diseases/pathology ; Oxazines/diagnostic use ; Staining and Labeling

PURPOSE: Indoxacarb insecticide poisoning causes methemoglobinemia, which is occasionally life-threatening. However, there is limited data on indoxacarb effects after human ingestion. The purpose of this study was to examine the clinical features, complications, management, and medical outcome of patients with indoxacarb insecticide poisoning. METHODS: We retrospectively reviewed the medical records of 10 patients with indoxacarb insecticide poisoning who had visited our emergency centers from January 2008 to December 2011. We collected data on the general characteristics of the patients, their clinical symptoms and signs, laboratory data, management of their condition, and clinical results. RESULTS: Among the 10 patients, 8 were diagnosed with methemoglobinemia. The clinical manifestations of indoxacarb insecticide poisoning were hypotension (3 patients), altered mentality (5 patients), cyanosis (5 patients), dyspnea (2 patients), seizure (3 patients), and cardiac arrest (2 patients). Four patients had a poisoning severity score of 3 and 2 patients had a poisoning severity score of 2. Four patients were treated with methylene blue for methemoglobinemia and one patient was treated with a high dose (150 mg/kg) of ascorbic acid. The serum methemoglobin saturation of five patients who were treated with methylene blue or a high dose of ascorbic acid was nearly normalized. Four patients experienced rhabdomyolysis, pneumonia, hemolytic anemia, acute pancreatitis, and heart failure as a complication of indoxacarb insecticide poisoning. CONCLUSION: We observed a variety of clinical features, complications, management, medical outcome, and clinical course of patients with indoxacarb insecticide poisoning. We could also ascertain the efficacy of methylene blue and high dose ascorbic acid for indoxacarb-induced metheglobinemia.
Anemia, Hemolytic ; Ascorbic Acid ; Cyanosis ; Dyspnea ; Eating ; Emergencies ; Heart Arrest ; Heart Failure ; Humans ; Hypotension ; Medical Records ; Methemoglobin ; Methemoglobinemia ; Methylene Blue ; Oxazines ; Pancreatitis ; Pneumonia ; Retrospective Studies ; Rhabdomyolysis ; Seizures