OBJECTIVETo evaluate the effect of Nigella sativa (NS) extract on memory performance and its possible mechanisms in scopolamine (Sco)-induced spatial memory impairment model using Morris water maze test.

METHODSThirty-two male Wistar rats were randomly divided into four groups. The control group received saline instead of both NS extract and Sco. The Sco group was treated by saline for two weeks, and was injected by Sco (2 mg/kg, intraperitoneally) 30 min before each trail in Morris water maze test. Sco+NS 200 and Sco+NS 400 groups were daily treated by 200 or 400 mg/kg of NS (intraperitoneally) for two weeks, respectively, and were finally injected by Sco 30 min before Morris water maze test. The brains of animals were removed to determine the acetylcholinesterase (AChE) activity and oxidative stress criteria in cortical tissues.

RESULTSTime latency and path length in the Sco group were significantly higher than in the control group (P<0.01), while the Sco+NS 400 group showed a significantly shorter traveled path length and time latency compared with the Sco group (P<0.01). AChE activity in the cortical tissues of the Sco group was significantly higher than the control group (P<0.01), while AChE activity in the Sco+NS 200 and Sco+NS 400 groups was lower than the Sco group (P<0.01). Following Sco administration, malondialdehyde (MDA) concentrations were increased (P<0.01) in comparison with the control group, while cortical total thiol content decreased (P<0.01). Pretreatment with extracts caused a significant elevation in cortical total thiol content (P<0.01) and reduction in cortical MDA concentration (P<0.01) compared with the Sco group.

CONCLUSIONSHydro-alcoholic extract of NS prevents Sco-induced spatial memory deficits and decreases the AChE activity as well as oxidative stress of brain tissues in rats. Our results support the traditional belief about the beneficial effects of NS in nervous system. Moreover, further investigations are needed for better understanding of this protective effect.

Acetylcholinesterase ; metabolism ; Animals ; Ethanol ; chemistry ; Male ; Malondialdehyde ; metabolism ; Maze Learning ; drug effects ; Memory Disorders ; drug therapy ; physiopathology ; Nigella sativa ; chemistry ; Plant Extracts ; pharmacology ; therapeutic use ; Rats, Wistar ; Reaction Time ; drug effects ; Scopolamine Hydrobromide ; Spatial Memory ; drug effects ; Sulfhydryl Compounds ; metabolism ; Water ; chemistry

The aimed of this study was to prepare stabilized thiomers to overcome the poor stability character of traditional thiomers. Poly(acrylic acid)-cysteine (PAA-Cys) was synthesized by conjugating cysteine with poly(acrylic acid) and poly(acrylic acid)-cysteine-6-mercaptonicotinic acid (PAA-Cys-6MNA, stabilized thiomers) was synthesized by grafting a protecting group 6-mercaptonicotinic acid (6MNA) with PAA-Cys. The free thiol of PAA-Cys was determined by Ellmann's reagent method and the ratio of 6MNA coupled was determined by glutathione reduction method. The study of permeation enhancement and stabilized function was conducted by using Franz diffusion cell method, with fluorescein isothiocyanate dextran (FD4) used as model drug. The influence of polymers on tight junctions of Caco-2 cell monolayer was detected with laser scanning confocal fluorescence microscope. The results indicated that both PAA-Cys and PAA-Cys-6MNA could promote the permeation of FD4 across excised rat intestine, and the permeation function of PAA-Cys-6MNA was not influence by the pH of the storage environment and the oxidation of air after the protecting group 6MNA was grafted. The distribution of tight junction protein of Caco-2 cell monolayer F-actin was influenced after incubation with PAA-Cys and PAA-Cys-6MNA. In conclusion, stabilized thiomers (PAA-Cys-6MNA) maintained the permeation function compared with the traditional thiomers (PAA-Cys) and its stability was improved. The mechanism of the permeation enhancement function of the polymers might be related to their influence on tight junction relating proteins of cells.
Acrylic Resins ; chemistry ; Actins ; metabolism ; Animals ; Caco-2 Cells ; Cysteine ; chemistry ; Dextrans ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Glutathione ; Humans ; Intestinal Absorption ; Intestinal Mucosa ; drug effects ; Nicotinic Acids ; chemistry ; Rats ; Sulfhydryl Compounds ; chemistry

OBJECTIVEFlavonoids are present in foods such as fruits and vegetables. Several studies have demonstrated a relationship between the consumption of flavonoid-rich foods and prevention of human disease, including neurodegenerative disorders. We assessed the effect of rutin (quercetin-3-O-rutinoside) on oxidative stress in kainic acid (KA)-induced seizure.

METHODSThirty-six BALB/c mice were randomly divided into three groups. In the control group, saline (intra-peritoneal, i.p.) was administered for 7 d, and on the last day, KA (10 mg/kg, i.p.) was injected 30 min after administration of saline. In rutin groups, mice were pretreated with rutin (100 and 200 mg/kg, i.p.) for 7 d, and on the last day, KA (10 mg/kg, i.p.) was injected 30 min after administration of rutin. Subsequently, behavioural changes were observed in mice. Lipid peroxidation and oxidative stress were measured respectively in the early and late phases after KA-induced seizures.

RESULTSSeizure scores in the rutin groups were significantly lower than those in the control group (P < 0.01). Furthermore, rutin dose-dependently inhibited the number of wet-dog shakes (WDS) (P < 0.05). Malondialdehyde level in the hippocampus of the rutin groups was significantly lower than that in the hippocampus of the control group on days 1 and 21 after KA administration. In the rutin groups, the thiol levels observed on day 1 after KA administration were higher than that in the control group (P < 0.01).

CONCLUSIONThese results indicate that rutin has potential anticonvulsant and antioxidative activities against oxidative stress in KA-induced seizure in mice.

Animals ; Dose-Response Relationship, Drug ; Kainic Acid ; toxicity ; Lipid Peroxidation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Oxidative Stress ; drug effects ; Rutin ; pharmacology ; Seizures ; chemically induced ; metabolism ; Sulfhydryl Compounds ; analysis

Redox signal transduction, especially the oxidative modification of proein thiols, correlates with many diseases and becomes an expanding research area. However, there was rare method for quick and specific detection of protein thiols and their oxidative modification in living cells. In this article, we review the current chemical strategies for the detection and quantification of protein thiols and related cysteine oxidation. We also look into the future of the development of fluorescent probes for protein thiols and their potential application in the research of reactive cysteine proteomes and early detection of redox-related diseases.
Animals ; Cysteine ; metabolism ; Fluorescent Dyes ; Humans ; Nitrosation ; Oxidation-Reduction ; Proteins ; chemistry ; metabolism ; Reactive Nitrogen Species ; metabolism ; Reactive Oxygen Species ; metabolism ; Sulfenic Acids ; analysis ; Sulfhydryl Compounds ; analysis ; chemistry ; metabolism

In order to explore the potential influences of the disulfide bridge on the physical and chemical properties of PrP protein, the expressed recombinant human wild-type PrP protein was purified for using in an established redox process for the reduction and oxidation of the ethanethiol group within PrP. Sedimentation tests illustrated that redox process remarkably promoted the aggregation of recombinant PrP. Thioflavin T binding assay revealed an enhanced fibrillization of the recombinant human PrP after redox process. Far-UV circular dichroism demonstrated that the PrP treated with redox process showed a significant p-sheet rich structure. Furthermore, PrP-specific Western blot identified that the recombinant PrP after redox possessed stronger proteinase K-resistance. Those data indicates that the formation of the disulfide bridge induces the alteration of the secondary structure and enhances the progresses of aggregation and fibrillization of PrP protein.
Amyloid ; chemistry ; Endopeptidase K ; metabolism ; Humans ; Oxidation-Reduction ; Prions ; chemistry ; metabolism ; Protein Multimerization ; Protein Structure, Secondary ; Proteolysis ; Sulfhydryl Compounds ; chemistry

BACKGROUND: The aim of this study was to evaluate oxidative stress in various clinical forms of hepatitis B infection and to investigate its role in the development of the chronic form of the disease. METHODS: Ninety-three patients with inactive hepatitis B surface antigen (HbsAg) carrier state (IHBCS), 65 patients with chronic hepatitis B infection (CHB), and 42 healthy adults were included in the study. The following values were measured and compared in patient groups: total antioxidant status (TAS), total oxidative stress (TOS), oxidative stress index (OSI), sulfhydryl (SH), lipid peroxidation (LOOH), catalase (CAT), and ceruloplasmin. In patients with chronic hepatitis B, these values were compared with HBV DNA and fibrosis levels. RESULTS: ALT, TOS, LOOH, and OSI levels were higher in the CHB group compared to the other groups (P<0.001). Catalase levels increased in the CHB and IHBCS groups compared to the control group (P<0.001). Total aminooxidant and ceruloplasmin levels were found to be lowest in the CHB group and highest in the control group (P<0.001). Sulfhyrdyl was higher in the control group compared to the other groups (P<0.001). In the CHB group, there was no correlation between the HBV DNA and OSI (P>0.05). CONCLUSIONS: These finding suggested that oxidative stress is associated with hepatitis B activity.
Adolescent ; Adult ; Aged ; Alanine Transaminase/blood ; Antioxidants/metabolism ; Carrier State ; Catalase/blood ; DNA, Viral/*analysis ; Female ; Fibrosis ; Hepatitis B/*metabolism/pathology ; Hepatitis B Surface Antigens/blood ; Hepatitis B virus/*genetics ; Hepatitis B, Chronic/metabolism/pathology ; Humans ; Lipid Peroxidation ; Male ; Middle Aged ; *Oxidative Stress ; Sulfhydryl Compounds/blood ; Young Adult

Holo glutaredoxin (Grx) is a homo-dimer that bridges a [2Fe-2S] cluster with two glutathione (GSH) ligands. In this study, both monothiol and dithiol holo Grxs are found capable of transferring their iron-sulfur (FeS) cluster to an apo ferredoxin (Fdx) through direct interaction, regardless of FeS cluster stability in holo Grxs. The ligand GSH molecules in holo Grxs are unstable and can be exchanged with free GSH, which inhibits the FeS cluster transfer from holo Grxs to apo Fdx. This phenomenon suggests a novel role of GSH in FeS cluster trafficking.
Circular Dichroism ; Dimerization ; Ferredoxins ; chemistry ; metabolism ; Glutaredoxins ; chemistry ; metabolism ; Glutathione ; metabolism ; Iron ; chemistry ; Ligands ; Magnetic Resonance Spectroscopy ; Sulfhydryl Compounds ; chemistry ; Sulfur ; chemistry ; Toluene ; analogs & derivatives ; chemistry

OBJECTIVETo study the relationship of renal aquaporin -1, -2, -3, and -4 (AQP1-4) expression with renal parenchymal thickness and glomerular filtration rate (GFR) in children with congenital hydronephrotis.

METHODSRenal tissue samples were obtained from 10 kidneys of 10 children (age: 62.3±18.3 months) with hydronephrosis and who underwent Anderson-Hynes pyeloplasty. Renal control samples were obtained from 6 children (age: 62.7±17.1 months) undergoing nephrectomy for nephroblastoma and were confirmed histologically as normal renal tissues. Renal parenchymal thickness of the hydronephrotic kidneys was measured by ultrasound preoperatively and was verified at operation. Renal GFR was assessed using 99mTc-DTPA scintigraphy preoperatively. Western blot was used to examine the expression of AQP1-4 in the renal tissues. The correlations of renal AQP1-4 expression with the renal parenchymal thickness and GFR were assessed by Pearson correlation analysis.

RESULTSThe expression of AQP1-4 in the hydronephrotis group was markedly reduced compared to that in the control group (P<0.05). The mean renal parenchymal thickness of the hydronephrotic kidney was 4.59±2.25 mm measured by ultrasound preoperatively. The mean GFR of the obstructed kidney was significantly lower than that of the contralateral kidney in the hydronephrosis group (40±12 mL/min vs 105±20 mL/min; P<0.05). The expression of AQP1, 2, 3 and 4 was positively correlated with preoperative renal GFR and renal parenchymal thickness in the hydronephrosis group (P<0.05). Renal parenchymal thickness was positively correlated with renal GFR (P<0.05).

CONCLUSIONSThe expression of renal AQP1-4 is reduced in children with congenital hydronephrosis. The expression levels of AQP1-4 are positively correlated with renal parenchymal thickness and GFR.

Aquaporins ; analysis ; Child ; Child, Preschool ; Female ; Glomerular Filtration Rate ; Humans ; Hydronephrosis ; congenital ; metabolism ; pathology ; Kidney ; pathology ; Male ; Sulfhydryl Compounds ; Technetium Tc 99m Aggregated Albumin ; Technetium Tc 99m Pentetate

Genetic modification of barley variety can be an efficient way to improve beer quality. The objective of this study was to understand the effect of trxS gene on hydrolases activities in transgenic and non-transgenic barley seeds. The results showed that alpha-amylase, free beta-amylase and limit dextrinase activity were increased in transgenic seeds in comparison with non-transgenic seeds. Sulfhydryl content of protein in transgenic seeds was also higher than that in non-transgenic seeds, suggesting that trxS gene could express in barley seeds, which opens a new way for breeding new barley varieties to improve beer quality.
Germination ; genetics ; Glucosyltransferases ; metabolism ; Hordeum ; enzymology ; genetics ; Plants, Genetically Modified ; enzymology ; genetics ; Seeds ; enzymology ; genetics ; Sulfhydryl Compounds ; metabolism ; Thioredoxins ; genetics ; alpha-Amylases ; metabolism ; beta-Amylase ; metabolism

Ionizing radiation produces reactive oxygen species, which exert diverse biological effects on cells and animals. We investigated alterations of heme oxygenase (HO) and non-protein thiols (NPSH), which are known as two major anti-oxidant enzymes, in female and male C57BL/6 mice in the lung, liver, and brain after whole-body gamma-irradiation with 10 Gy (1-7 days) as well as in the lung after whole-thorax gamma-irradiation (WTI) with 12.5 Gy (1-26 weeks). Most significant alteration of HO activity was observed in the liver, which elevated 250% in males. NPSH level in female liver was increased on the 5th-7th days but decreased in males on the 3rd day. In the lung, the elevation of HO activity in both sexes and the pattern of NPSH change were similar to that of the liver. On the other hand, the increase of HO activity on the 16th week and the decrease of NPSH level on the 2nd week were observed only in male lung after WTI. This study shows that the liver is the most sensitive tissue to gamma-irradiation-induced alterations of HO activity in both female and male mice. In addition, there exists significant differential effect of gamma-irradiation on anti-oxidant system in female and male mice.
Animals ; Brain/*enzymology/metabolism/radiation effects ; Comparative Study ; Female ; Gamma Rays ; Gene Expression Regulation, Enzymologic/radiation effects ; Liver/*enzymology/metabolism/radiation effects ; Lung/*enzymology/metabolism/radiation effects ; Male ; Mice ; Mice, Inbred C57BL ; RNA, Messenger/genetics/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sex Factors ; Sulfhydryl Compounds/metabolism ; Time Factors ; Whole-Body Irradiation