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

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

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

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

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

In the present study, we determined the protective mechanism of HSP90 against neuronal cell death induced by Abeta. For the evaluation of protective role of HSP90, we used human neuroblastoma SK-N-SH cell lines, examined AlamarBlue assay, Western blot analysis and immunofluorescence assay. Incubation of SK-N-SH cells with Abeta significantly induced neuronal cell death. However, HSP90 induced by mild heat shock could attenuate neuronal apoptosis in Abeta treated condition. To identify the role of HSP90, we determined localization of HSP90 in SK-N-SH cells. Interestingly, HSP90 was increased and localized in mitochondria as treatment of mild heat shock. Also, treatment or increase of HSP90 largely elevated level of Bcl-2 expression, whereas inhibition of HSP90 with HSP90 antisense oligonucleotide significantly decreased Bcl-2 expression. In contrast to Bcl-2, Bax expression was regulated independently by HSP90. Moreover, increase of HSP90 could attenuate collapse of mitochondrial membrane potential induced by Abeta. However, HSP90 antisense oligonucleotide largely increase breakdown of mitochondrial membrane potential induced by Abeta. These data suggest that HSP90 as chaperone protein significantly attenuates neuronal damage and protects neuroanl cells from neurotoxin such as Abeta.
Apoptosis ; Blotting, Western ; Cell Death ; Cell Line ; Fluorescent Antibody Technique ; Hot Temperature ; Humans ; Membrane Potential, Mitochondrial ; Mitochondria ; Neuroblastoma ; Neurons ; Oxazines ; Shock ; Xanthenes

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

Indoxacarb is an oxadiazine insecticide with selective lethality through blockade of neuronal voltage-dependent sodium channels. It has a low mammalian toxicity, and few cases of human toxicity after indoxacarb ingestion can be found in the literature. A 36 year-old male patient visited our ED after a generalized tonic clonic seizure, which was witnessed by his mother. His past medical history was nonspecific. On initial presentation, he showed a decreased level of consciousness with a Glasgow coma score of 5/15 (E1V1M3), unprotected airway, hypoxia, and cyanosis. The saturation gap and cyanosis after intubation and mechanical ventilation was strongly suggestive of methemoglobinemia due to poisoning. Finally, the methemogobin (metHb) level was 27.4%. Therefore, the patient received 100 mg of methylene blue (2 mg/kg, 1% solution) and 50 g of charcoal. The insecticide was found to be ingested xenobiotic (Steward Gold(R); 5% indoxacarb; 95% inert ingredients and other components). On the second hospital day, the patient became alert. The patient's metHb level was 0.1%. The endotracheal tube was removed. On the fifth hospital day, he was discharged in good condition. Herein we present a case of indoxacarb poisoning with methemoglobiemia and seizure, which are unusual presentations.
Anoxia ; Charcoal ; Coma ; Consciousness ; Cyanosis ; Eating ; Humans ; Intubation ; Male ; Methemoglobinemia ; Methylene Blue ; Mothers ; Neurons ; Oxazines ; Respiration, Artificial ; Seizures ; Sodium Channels ; Wit and Humor as Topic

Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease predominantly affecting diarthroidal joints. Following the successful application of biologic agents, several small molecule inhibitors are currently under clinical trials. Small molecule inhibitors have several strengths compared with biologics. First, they can target several inflammatory cytokines together by blocking common signal transduction pathways. Second, they can be taken orally. Third, the price can be made flexible. Among the several small molecule inhibitors in the development process, fostamatinib and tofacitinib are the closest to the clinics at the moment. Fostamatinib, which is a Syk inhibitor, showed superior efficacy over placebo with tolerable safety signals. Diarrhea, hypertension and infection are representative adverse events. Tofacitinib, which is JAK inhibitor, is now finishing phase 3 clinical trials. It showed clinical efficacy comparable to Adalimumab and similar adverse effect profiles to the biologics, which include opportunistic infections. For laboratory abnormalities, leukopenia, anemia, increase of LDL and serum Cr were reported, which, however, were stabilized with prolonged use. Other classes of small molecule inhibitors did not show impressive efficacy as these small molecule inhibitors. In conclusion, small molecule inhibitors are promising novel therapeutic agents for the treatment of RA. They will be able to change the treatment paradigm of RA if they can show long-term safety.
Anemia ; Antibodies, Monoclonal, Humanized ; Arthritis, Rheumatoid ; Biological Agents ; Cytokines ; Diarrhea ; Hypertension ; Joints ; Leukopenia ; Opportunistic Infections ; Oxazines ; Piperidines ; Pyridines ; Pyrimidines ; Pyrroles ; Signal Transduction