PURPOSE: Cyanate, known as one of the uremic toxins and derived spontaneously from urea, has several effects on the biologic substances including erythropoietin, antioxidant and ceruloplasmin. To find out the protective materials from the hazardous effect of cyanate in osteoblast, we added twenty amino acids, albumin globulin and hemoglobin in the culture media containing osteoblastic cells with cyanate. METHODS: Osteoblastic ROS 17/2.8 cells, exposed to various concentrations of sodium cyanate, were used to analyze for the cytotoxicity. The cyanate-induced cytotoxicity was assessed by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay by measuring the absorbance of the reaction solution at 570 nm. Viability of the treated cells was expressed as A570 of sample/A570 of control. The degree of the carbamylation was measured using trinitrobenzenesulphonic acid. The degree of the carbamylation in amino acid was about 50% in average. RESULTS: The degree of the carbamylation in albumin was increased depending on the incubation time with cyanate and the concentration of the cyanate. The degree of the carbamylation in globulin and hemoglobin was nearly zero. Asp, Glu, Leu, Trp and Tyr among the twenty amino acids revealed the protective effect against the damage induced by cyanate. And only albumin among the three proteins revealed the protective effect. CONCLUSION: On the basis of these results, Asp, Glu, Leu, Trp, Tyr and albumin are useful tools for the protection against damages by cyanate carbamylation.
Albumins ; Amino Acids* ; Ceruloplasmin ; Culture Media ; Cyanates ; Erythropoietin ; Osteoblasts* ; Sodium ; Urea ; Viperidae

Three diisocyanates can cause occupational asthma (OA): toluene diisocyanate (TDI), 4,4 diphenylmethane diisocyanate (MDI), and 1,6-hexamethylene diisocyanate (HDI). We analyzed potential biomarkers of isocyanate-induced OA, based on investigated immunologic, genetic, neurogenic, and protein markers, because there is no serological testing method. The prevalence of serum IgG to cytokeratin (CK)18 and CK19 in TDI-OA was significantly higher than in controls, although the prevalence of these antibodies was too low for them to be used as biomarkers. Another candidate biomarker was serum IgG to tissue transglutaminase (tTG), because the prevalence of serum specific IgG to tTG was significantly higher in patients with TDI-OA than in controls. The human leukocyte antigen (HLA) DRB1*1501-DQB1*0602-DPB1*0501 haplotype may be used as a genetic marker for TDI-OA in Koreans via enhanced specific IgE sensitization in exposed subjects. The genetic polymorphisms of catenin alpha 3, alpha-T catenin (CTNNA3) were significantly associated with TDI-OA. Additionally, examining the neurokinin 2 receptor (NK2R) 7853G>A and 11424 G>A polymorphisms, the NK2R 7853GG genotype had higher serum vascular endothelial growth factor (VEGF) levels than the GA or AA genotypes among Korean workers exposed to TDI. To identify new serologic markers using a proteomic approach, differentially expressed proteins between subjects with MDI-OA and asymptomatic exposed controls in a Korean population showed that the optimal serum cutoff levels were 69.8 ng/mL for ferritin and 2.5 microg/mL for transferrin. When these two parameters were combined, the sensitivity was 71.4% and the specificity was 85.7%. The serum cytokine matrix metalloproteinase-9 (MMP-9) level is a useful biomarker for identifying cases of TDI-OA among exposed workers. Despite these possible biomarkers, more effort should be focused on developing early diagnostic biomarkers using a comprehensive approach based on the pathogenic mechanisms of isocyanate-induced OA.
Antibodies ; Asthma ; Asthma, Occupational ; Biomarkers ; Cyanates ; Ferritins ; Genetic Markers ; Genotype ; GTP-Binding Proteins ; Haplotypes ; Humans ; Immunoglobulin E ; Immunoglobulin G ; Isocyanates ; Keratins ; Leukocytes ; Matrix Metalloproteinase 9 ; Polymorphism, Genetic ; Prevalence ; Proteins ; Receptors, Neurokinin-2 ; Serologic Tests ; Toluene 2,4-Diisocyanate ; Transferrin ; Transglutaminases ; Vascular Endothelial Growth Factor A

Humans ; Pyridines ; Spectrophotometry ; methods ; Thiocyanates ; urine

OBJECTIVE: Accumulating evidence suggests that oxidative stress plays a role in the pathophysiology of schizophrenia and that the potent antioxidants may be potential therapeutic drugs for schizophrenia. This study was undertaken to examine the effects of the potent antioxidant sulforaphane (SFN), found in cruciferous vegetables, on behavioral abnormalities (e.g., hyperlocomotion and prepulse inhibition [PPI] deficits) in mice after a single administration of the N-methyl-D-aspartate (NMDA)-receptor antagonist phencyclidine (PCP). METHODS: Effects of SFN (3, 10, and 30 mg/kg, intraperitoneally [i.p.]) on hyperlocomotion and PPI deficits in the adult male ddY mice after administration of PCP (3.0 mg/kg, subcutaneously [s.c.]) were examined. RESULTS: Administration of SFN (30 mg/kg, intraperitoneally [i.p.]), but not low doses (3 and 10 mg/kg, i.p.), significantly attenuated hyperlocomotion in mice after PCP administration (3.0 mg/kg, subcutaneously [s.c.]). Furthermore, administration of SFN (3, 10, and 30 mg/kg, i.p.) attenuated the PPI deficits in mice after PCP administration (3.0 mg/kg, s.c.) in a dose-dependent manner. CONCLUSION: These results suggest that SFN has antipsychotic activity in an animal model of schizophrenia. Therefore, it is likely that SFN may be a potential therapeutic drug for schizophrenia.
Adult ; Animals ; Antioxidants ; Humans ; Male ; Mice ; Models, Animal ; N-Methylaspartate ; Oxidative Stress ; Phencyclidine ; Schizophrenia ; Thiocyanates ; Vegetables

Adult ; Female ; Food Industry ; manpower ; Humans ; Male ; Middle Aged ; Smoking ; epidemiology ; Thiocyanates ; urine

This article dealed with the effects of processing method and duration on the major bioactive components (sinigrin and sinapine thiocyanate) in Brassica juncea. The contents of sinigrin and sinapine thiocyanate in decoctions of raw and processed B. juncea were determined and compared by high performance liquid chromatography on a Alltima C18 column (4.6 mm x 250 mm, 5 microm) at 35 degrees C with the acetonitrile-0.1% phosphoric acid as the mobile phrase in gradient elution. The detection wavelength of sinigrin and sinapine thiocyanate was set at 227 nm and 326 nm, and the flow rate was 1.0 mL x min(-1). It was found that with the extended processing duration, the contents of sinigrin and sinapine thiocyanate first increased and then decreased: i.e., 0-2 minutes they increased gradually (for sinigrin, by 9.65% in processed products and 356. 10% in powder; for sinapine thiocyanate, by 12.82% in processed products and 3.41% in powder), and achieved their highest content at 2 min; then, decreased during the next 5 minutes (for sinigrin, by 80.35% in processed products and 82.09% in powder; for sinapine thiocyanate, by 14.29% in processed products and 17.54% in powder), suggesting that processing duration could significantly affect the contents of bioactive components in B. juncea, enzymatic hydrolysis of sinigrin when the seed is crushed in the present of moisture may be responsible for the content change. It is recommended that the slow fire should be the best processing method and the raw seed could be used directly in the water extracts related industrial production.
Brassica ; chemistry ; Choline ; analogs & derivatives ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Glucosinolates ; chemistry ; Powders ; chemistry ; Thiocyanates ; chemistry

OBJECTIVEThis study aimed to investigate the effect of a lactoperoxidase-peroxidase-thiocyanate (LPO-H2O-SCN-) system with different concentrations of iodine (I-) on Streptococcus mutans (S. mutans), particularly on various parameters, including growth, adhesion, glucosyltransferase (GTF) enzyme activity, and insoluble exopolysaccharide synthesis.

METHODSS. mutans ATCC 25175 was used as experimental species. Clonal formation unit (CFU) were counted to investigate the inhibitory effect on bacterial growth. The inhibition rate of bacterial adherence was calculated to analyze the effect on adhesion. Anthrone method was used to determine the content of insoluble exopolysaccharides and the amount of reducing saccharides. GTF activity and enzyme activity were then determined.

RESULTSThe inhibitory ability of the LPO-H2O2-SCN- system with I- on the cariogenicinity of S. mutans was strengthened as I- concentration was increased. At I- concentration > or = 100 micromol x L(-1) the antibacterial effects were significantly increased compared with those of the control group (P < 0.05). At I- concentration > or = 1,000 micromol x L(-1), the antibacterial effects were significantly improved compared with those of the group with SCN-only (P < 0.05). At I- concentration > or = 100 micromol x L(-1), the inhibition rate of bacterial adherence was > 50%; insoluble exopolysaccharide synthesis and GTF enzyme activity were reduced (P < 0.05).

CONCLUSIONThe antibacterial effects of the LPO-H2O2-I- system were enhanced by adding I- to overcome the antagonistic effect of physiological SCN- concentration. LPO-H2O2-SCN- system with different concentrations of I- showed statistically significant inhibitory effects on growth, adhesion, insoluble exopolysaccharide synthesis, and GTF enzyme activity.

Anti-Bacterial Agents ; Bacterial Adhesion ; Hydrogen Peroxide ; In Vitro Techniques ; Iodine ; Lactoperoxidase ; Oxidation-Reduction ; Streptococcus mutans ; Thiocyanates

According to the polarity of different components in Sanpian Decoction, two fingerprints were established. Then the substance benchmark freeze-dried powder of 15 batches of Sanpian Decoction was prepared, followed by the determination of the fingerprints, index component content, and dry extract rates, the identification of attribution of characteristic peaks, and the calculation of similarities between these fingerprints and the reference(R), the content and transfer rate ranges of ferulic acid, sinapine thiocyanate, liquiritin, and glycyrrhizic acid, and the dry extract rate range. The results showed that the similarities of 15 batches of the substance benchmark fingerprints with R were all greater than 0.900.Further summarization of the characteristic peaks revealed that there were a total of 20 characteristic peaks in fingerprint 1, among which, eight were from Sinapis Semen, four from Paeoniae Radix Alba, six from Chuanxiong Rhizoma, and two from Glycyrrhizae Radix et Rhizoma. A total of 16 characteristic peaks were observed in fingerprint 2, including one from Sinapis Semen, three from Paeoniae Radix Alba, eight from Chuanxiong Rhizoma, and four from Glycyrrhizae Radix et Rhizoma. The average dry extract rate of 15 batches of substance benchmarks was 18.25%, with a dry extract rate range of 16.28%-20.76%. The index component content and transfer rate ranges were listed as follows: 0.15%-0.18% and 38.81%-58.05% for ferulic acid; 0.26%-0.42% and 36.51%-51.02% for sinapine thiocyanate; 0.09%-0.15% and 48.80%-76.61% for liquiritin; 0.13%-0.24% and 23.45%-35.61% for glycyrrhizic acid. The fingerprint, dry extract rate, and index component content determination was combined for analyzing the quality value transfer of substance benchmarks in the classic prescription Sanpian Decoction.The established quality evaluation method for the substance benchmarks was stable and feasible, which has provided a basis for the quality control of Sanpian Decoction and the follow-up development of related preparations.
Benchmarking ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Glycyrrhizic Acid/analysis* ; Paeonia ; Quality Control ; Thiocyanates

Oxidative damage is thought to be a major cause of the progression of dopamine (DA)rgic neurodegeneration as in Parkinson's disease. We have previously reported that tetrahydrobiopterin (BH4), an endogenous molecule required for DA synthesis, exerts oxidative stress to DA-producing cells and facilitates the production of DA quinone. It is known that aconitase, present in both mitochondrial and cytosolic forms, act as an reactive oxygen species (ROS) sensor, and that their inactivation leads to further generation of ROS. In the present study we investigated whether the BH4-associated vulnerability of DA cells might involve aconitase. In DArgic cell line CATH.a, BH4 treatment caused reduction of activity of both mitochondrial and cytosolic aconitases, and this appeared to be due to direct inactivation of the pre-existing enzyme molecules. Although most of the activity reduced by BH4 was increased upon reactivation reaction under a reducing condition, the restoration was not complete, suggesting that irreversible and covalent modification has occurred. The aconitase inactivation was exacerbated in the presence of DA and attenuated in the presence of tyrosine hydroxylase inhibitor a-methyl-p-tyrosine, suggesting the involvement of DA. The degree of inactivation increased when the cells were treated with the quinone reductase inhibitor dicoumarol and decreased in the presence of quinone reductase inducer sulforaphane. Taken together, BH4 appeared to lead to both reversible and irreversible inactivation of aconitase and that this is facilitated by the presence of DA and accumulation of DA quinone.
Aconitate Hydratase ; Benzoquinones ; Biopterin ; Cell Line ; Cytosol ; Dicumarol ; Dopamine ; NAD(P)H Dehydrogenase (Quinone) ; Oxidative Stress ; Parkinson Disease ; Reactive Oxygen Species ; Thiocyanates ; Tyrosine 3-Monooxygenase

OBJECTIVETo explore material basis of the pharmacological differences between the roasting and pro-roasting Raphani Semen.

METHODThe two new sulfur-containing compounds (A209 and B221) were found changed after processing in the water decoction. The common precursor-C3 of A209 and B211 and the precursor of C3 were seperated and purified. Their transforming relationship was proved.

RESULTThe result showed that glucosinolates could decompose into sulforaphane and transform into A209 and B221 further in the boiling process.

CONCLUSIONThis study provides some experimental evidences for revealing the mechanism of Raphani Semen processing.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glucosinolates ; chemistry ; isolation & purification ; metabolism ; Isothiocyanates ; Seeds ; chemistry ; enzymology ; Sulfur Compounds ; chemistry ; Thiocyanates ; chemistry