Possible Role of Bent Structure of Methylated Lithocholic Acid on Artificial and Plasma Membranes.

Bile acids form micelles that are essential for the absorption of dietary lipids. However, excessive bile acid micelles can disrupt the plasma membrane by removing phospholipids, resulting in cell death. We hypothesized that the bent geometrical structure of the steroid scaffold of bile acids decreases the lipid order (similar to unsaturated phospholipids with cis double bonds), disrupting the plasma membrane. Here, lithocholic acid (LCA), a bile acid, was methylated to prevent micellization. Methylated lithocholic acid (Me-LCA) was mixed with a thin phase-separated lipid bilayer comprising 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and cholesterol (Chol). Me-LCA was not localized in the DPPC-rich rigid phase but localized in the DOPC-rich fluid phase, and excess Me-LCA did not affect the phase separation. Me-LCA is distributed in the plasma and organelle membranes. However, Me-LCA with bent structure did not affect the membrane properties, membrane fluidity, and hydrophobicity of liposomes composed of DOPC, DPPC, and Chol and also did not affect the proliferation of cells.

Combination of UV and green light synergistically enhances the attractiveness of light to green stink bugs Nezara spp.

The southern green stink bug Nezara viridula and its congener N. antennata are important agricultural pests worldwide. These species show positive phototaxis and their compound eyes have high sensitivity to UV and green lights. The attractiveness of monochromatic UV, green lights and combined UV and green light to stink bugs was investigated under field conditions. The number of stink bugs caught increased with the number of UV LEDs, but very few bugs were caught using green light, irrespective of the number of LEDs. However, the most stink bugs were caught when both colors were combined. These results indicate that monochromatic green light is less attractive to Nezara bugs, but when mixed with UV light, it synergistically enhances the attractiveness of UV light. This finding contributes to the construction of reliable and highly specific light traps to monitor Nezara bugs. The addition of green light hardly affected the attractiveness of the UV light to other insects, such as Anomala beetles, which are often caught in light traps. We conclude that the spectral composition of light that is attractive to nocturnal insects depends on the species, hence it is possible to make ecologically friendly light traps that are target specific.

Novel Anti-Adhesive CMC-PE Hydrogel Significantly Enhanced Morphological and Physiological Recovery after Surgical Decompression in an Animal Model of Entrapment Neuropathy.

We developed a novel hydrogel derived from sodium carboxymethylcellulose (CMC) in which phosphatidylethanolamine (PE) was introduced into the carboxyl groups of CMC to prevent perineural adhesions. This hydrogel has previously shown excellent anti-adhesive effects even after aggressive internal neurolysis in a rat model. Here, we confirmed the effects of the hydrogel on morphological and physiological recovery after nerve decompression. We prepared a rat model of chronic sciatic nerve compression using silicone tubing. Morphological and physiological recovery was confirmed at one, two, and three months after nerve decompression by assessing motor conduction velocity (MCV), the wet weight of the tibialis anterior muscle and morphometric evaluations of nerves. Electrophysiology showed significantly quicker recovery in the CMC-PE group than in the control group (24.0 ± 3.1 vs. 21.0± 2.1 m/s (p < 0.05) at one months and MCV continued to be significantly faster thereafter. Wet muscle weight at one month significantly differed between the CMC-PE (BW) and control groups (0.148 ± 0.020 vs. 0.108 ± 0.019%BW). The mean wet muscle weight was constantly higher in the CMC-PE group than in the control group throughout the experimental period. The axon area at one month was twice as large in the CMC-PE group compared with the control group (24.1 ± 17.3 vs. 12.3 ± 9 µm2) due to the higher ratio of axons with a larger diameter. Although the trend continued throughout the experimental period, the difference decreased after two months and was not statistically significant at three months. Although anti-adhesives can reduce adhesion after nerve injury, their effects on morphological and physiological recovery after surgical decompression of chronic entrapment neuropathy have not been investigated in detail. The present study showed that the new anti-adhesive CMC-PE gel can accelerate morphological and physiological recovery of nerves after decompression surgery.

Multiple free-radical scavenging capacity in serum.

We have developed a method to determine serum scavenging-capacity profile against multiple free radical species, namely hydroxyl radical, superoxide radical, alkoxyl radical, alkylperoxyl radical, alkyl radical, and singlet oxygen. This method was applied to a cohort of chronic kidney disease patients. Each free radical species was produced with a common experimental procedure; i.e., uv/visible-light photolysis of free-radical precursor/sensitizer. The decrease in free-radical concentration by the presence of serum was quantified with electron spin resonance spin trapping method, from which the scavenging capacity was calculated. There was a significant capacity change in the disease group (n = 45) as compared with the healthy control group (n = 30). The percent values of disease's scavenging capacity with respect to control group indicated statistically significant differences in all free-radical species except alkylperoxyl radical, i.e., hydroxyl radical, 73 ± 12% (p = 0.001); superoxide radical, 158 ± 50% (p = 0.001); alkoxyl radical, 121 ± 30% (p = 0.005); alkylperoxyl radical, 123 ± 32% (p>0.1); alkyl radical, 26 ± 14% (p = 0.001); and singlet oxygen, 57 ± 18% (p = 0.001). The scavenging capacity profile was illustrated using a radar chart, clearly demonstrating the characteristic change in the disease group. Although the cause of the scavenging capacity change by the disease state is not completely understood, the profile of multiple radical scavenging capacities may become a useful diagnostic tool.

Insecticide susceptibility of Nezara viridula (Heteroptera: Pentatomidae) and three other stink bug species composing a soybean pest complex in Japan.

The susceptibility of the stink bug species Nezara viridula (L.), Nezara antennata Scott, Piezodorus hybneri (Gmelin), and Riptortus pedestris (F.) to insecticides was tested, establishing their 50% lethal dose (LD50) values as baseline data. Third instars and adults of the four species were treated by topical application with seven insecticides: fenitrothion, fenthion, etofenprox, silafluofen, dinotefuran, clothianidin, and ethiprole. The weight of the stink bug and weight of the insecticide applied to each bug were used as explanatory variables in the probit regression analysis. The effect of the body weight on the dose-response relationship, the proportional model, was not uniform among the tested insecticide-stink bug combinations. However, the basic model fit all combinations and could estimate LD50 values successfully. Therefore, LD50 values at the medium (average) weight estimated by the basic model were selected to describe the susceptibility of the stink bugs. The LD50 value of silafluofen for N. viridula adults, and that of silafluofen and etofenprox for N. antennata adults, was at least 2,338 ng greater than the other species exposed to each insecticide. Almost all of the LD50 values for adults were over 10 times greater than those of the same species' nymphs treated with the same insecticide. Thus monitoring of occurring species and their developmental stages is important for controlling effectively the stink bug pest complex by insecticides, especially by silafluofen or etofenprox. The estimated LD50 values can be used as baseline data to compare the susceptibility of the species collected in another year or location.

Edaravone directly reacts with singlet oxygen and protects cells from attack.

AIMS: Protective effects of edaravone, an approved medicine for acute brain infarction in Japan, on cell death induced by singlet oxygen (1O2) were examined. MAIN METHOD: The 1O2 scavenging activity was examined by direct analysis of near-infrared luminescence in a cell-free system and by fluorospectrometry in the presence of cells. The protective effects of edaravone on 1O2-induced cell death were examined, using rat neuronal B50 cells. Cell death was evaluated by mitochondrial respiration (MTT assay), confocal microscopy and time-lapse imaging. The chemical reaction of edaravone with 1O2 was examined by production analysis using high performance liquid chromatography (HPLC). KEY FINDINGS: When rose Bengal (RB) in D2O was irradiated by a 514nm laser beam, the signal of 1O2 was observed. Edaravone suppressed the 1O2 signal more potently than azide, a 1O2 scavenger. When B50 cells were irradiated by 525nm green light in the RB solution, production of 1O2 and induction of cell death were observed. The fluorospectrometric study and the MTT assay revealed that 100-400microM edaravone suppressed the 1O2 production and attenuated cell death in a concentration-dependent manner. Confocal microscopy and the time-lapse imaging revealed that edaravone prevented the impairment of membrane integrity and the progression of cell death induced by 1O2. The HPLC study revealed that edaravone chemically reacted with 1O2 and changed another compound. SIGNIFICANCE: Since 1O2 is possibly involved in post-ischemic neuronal damage, the clinically approved curative effects of edaravone on acute brain infarction might be attributed to its potent 1O2 scavenging activity.

Novel polysaccharide-derived hydrogel prevents perineural adhesions in a rat model of sciatic nerve adhesion.

We investigated the effects of a novel carboxymethylcellulose (CMC)-derived hydrogel, in which phosphatidylethanolamine (PE) was introduced into the carboxyl groups of CMC, for preventing perineural adhesion after extensive internal neurolysis of rat sciatic nerve. Sciatic nerves were randomly assigned to one of the following groups: the Control group, operated but no treatment; the HA group, operated and treated with 1% hyaluronan; the CMC-PE(L) group, operated and treated with low-viscosity CMC-PE hydrogel; and the CMC-PE(H) group, operated and treated with high-viscosity CMC-PE hydrogel. Perineural adhesions were evaluated at 6 weeks. Nerves were also subjected to biomechanical testing to assess ultimate breaking strength. Electrophysiological and wet muscle weight measurements were performed. Breaking strengths were significantly lower for the CMC-PE(L) group than for the Control and HA groups. Latency was significantly longer for the Control group than for the CMC-PE(L) group at 20 days. The mean percentage of wet muscle weight to body weight was significantly lower for the Control group than for the CMC-PE(L) group at 6 weeks. Low-viscosity CMC-PE hydrogel appears to prevent perineural adhesions and allow early restoration of nerve function.

Serum hydroxyl radical scavenging capacity as quantified with iron-free hydroxyl radical source.

We have developed a simple ESR spin trapping based method for hydroxyl (OH) radical scavenging-capacity determination, using iron-free OH radical source. Instead of the widely used Fenton reaction, a short (typically 5 seconds) in situ UV-photolysis of a dilute hydrogen peroxide aqueous solution was employed to generate reproducible amounts of OH radicals. ESR spin trapping was applied to quantify OH radicals; the decrease in the OH radical level due to the specimen's scavenging activity was converted into the OH radical scavenging capacity (rate). The validity of the method was confirmed in pure antioxidants, and the agreement with the previous data was satisfactory. In the second half of this work, the new method was applied to the sera of chronic renal failure (CRF) patients. We show for the first time that after hemodialysis, OH radical scavenging capacity of the CRF serum was restored to the level of healthy control. This method is simple and rapid, and the low concentration hydrogen peroxide is the only chemical added to the system, that could eliminate the complexity of iron-involved Fenton reactions or the use of the pulse-radiolysis system.

Alpha-phenyl-N-tert-butyl nitrone has scavenging activity against singlet oxygen ((1)O(2)) and attenuates (1)O(2)-induced neuronal cell death.

Scavenging activity of alpha-phenyl-N-tert-butyl nitrone (PBN) against singlet oxygen ((1)O(2)) and its effects on (1)O(2)-induced neuronal cell death were examined. PBN at 1 - 4 mM dose-dependently suppressed (1)O(2) released from activated human neutrophils. PBN did not react with hydrogen peroxide or hypochlorite and did not affect myeloperoxidase activity, which are involved in the (1)O(2) formation in neutrophils. PBN also suppressed chemically generated (1)O(2) in a cell-free system. These findings collectively indicated that PBN certainly has scavenging activity against (1)O(2). Furthermore, PBN attenuated (1)O(2)-induced neuronal cell death. The well-known neuroprotective effects of PBN might be attributed to its (1)O(2)-scavenging activity.

Ozone production by amino acids contributes to killing of bacteria.

Reactive oxygen species produced by phagocytosing neutrophils are essential for innate host defense against invading microbes. Previous observations revealed that antibody-catalyzed ozone formation by human neutrophils contributed to the killing of bacteria. In this study, we discovered that 4 amino acids themselves were able to catalyze the production of an oxidant with the chemical signature of ozone from singlet oxygen in the water-oxidation pathway, at comparable level to antibodies. The resultant oxidant with the chemical signature of ozone exhibited significant bactericidal activity in our distinct cell-free system and in human neutrophils. The results also suggest that an oxidant with the chemical signature of ozone produced by neutrophils might potentiate a host defense system, when the host is challenged by high doses of infectious agents. Our findings provide biological insights into the killing of bacteria by neutrophils.

Reactive oxygen species generation through NADH oxidation by pterin derivatives.

Pterin is an electron transfer compound in biological systems. Among the analogs, 6-formylpterin (6FP) has been demonstrated to have many marked physiological and pharmacological activities. In previous study, we have elucidated that 6FP derivatives in which the 3-position is modified possess reactive oxygen species (ROS), which are involved in the modulation of a variety of cell functions, generation activities through the oxidation of NADH to NAD(+) in the dark at neutral pH. In the present study, we have demonstrated that the ROS generation activity by 6FP derivative is enhanced in the presence of 3-methyl-1-phenyl-2-pyrazolin-5-one. In this reaction, 3-methyl-1-phenyl-2-pyrazolin-5-one is reacted with the formyl group on the 6-position of 6FP derivative to give the activated product. The present results would be helpful for designing pharmaceutical ROS generation system in vivo.

Effects of injectable propofol emulsion on singlet oxygen released from activated human neutrophils and that chemically generated.

Effects of an injectable emulsion of propofol and its emulsifier on singlet oxygen (1O2) were examined. 1O2 released from activated human neutrophils was detected by chemiluminescence, and chemically generated 1O2 was detected by electron paramagnetic resonance (EPR). Both the propofol emulsion and the emulsifier suppressed 1O2 release from neutrophils. However, the emulsifier did not quench chemically generated 1O2, while the propofol emulsion quenched it. These results indicated that the emulsifier did not scavenge 1O2 released from neutrophils but inhibited 1O2 generation. The suppressive effects of propofol emulsion on 1O2 release from neutrophils consist of 1O2 scavenging and inhibition of 1O2 generation.

The radical scavenger edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) reacts with a pterin derivative and produces a cytotoxic substance that induces intracellular reactive oxygen species generation and cell death.

Cytotoxic effects of the combined use of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one), a radical scavenger and an approved medicine for acute brain infarction in Japan, with a pterin derivative, were examined in vitro. When pancreatic cancer cell line Panc-1 cells were incubated with 50 to 400 microM of a pterin derivative, 2-(N,N-dimethylaminomethyleneamino)-6-formyl-3-pivaloylpteridine-4-one (DFP), and the equivalent dose of edaravone, reactive oxygen species (ROS), were generated, and cell death was induced. ROS generation and the loss of mitochondrial membrane potential (MMP) preceding cell death were simultaneously monitored using time-lapse microscopy with an ROS-sensitive dye and a probe to monitor MMP, respectively. Cell death was also estimated quantitatively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. ROS generation and cell death were prominent when more than 100 microM of each agent was used in combination, whereas the sole use of each agent did not show any effects even at the highest dose, 400 microM. Chemical analysis revealed that DFP and edaravone react immediately in aqueous solution and produce a new compound named DFP-E. DFP-E chemically reacted with NADH much faster than DFP and generated ROS, and biologically, it was much more cell-permeable than DFP. These findings collectively indicated that the combined use of DFP with edaravone produced DFP-E, which caused intracellular ROS generation and cell death. Cell death was observed in normal cells, and edaravone reacted with another pterin derivative to yield an ROS-generating compound. As a result, care should be taken with the clinical use of edaravone when pterin derivatives stay in the body.

Chemical natures of 6-formylpterin nucleoside analogs.

Pterin is an electron transfer compound in biological systems. Among the analogs, 6-formylpterin (6FP) has been demonstrated to have many marked physiological activities. In the present study, we have developed the synthetic procedure for nucleoside analogs of 6FP and prepared 1-(beta-D-ribofuranosyl)-2-(N,N-diethylaminomethyleneamino)-6-formylpteridin-4-one (RDEF) and 1-(beta-D-ribofuranosyl)-2-(piperidine-1-ylmethyleneamino)-6-formylpteridin-4-one (RPIF) in which the 1-position is glycosylated. By electron paramagnetic resonance analysis coupled with the DMPO spin trapping technique, it has been elucidated that O2 was converted to H2O2 by RDEF at pH 7.4 in the presence of NADH in the dark. This result indicates that marked reactive oxygen species (ROS) generation activities of 6FP occurring under light illumination, which gives rise to the particular physiological activities such as induction of apoptosis, can be reproduced in the cellular and living systems by such derivatives and gives suggestion for designing pharmaceutical compounds that generate appropriate and controllable amounts of ROS in vivo.

Synthesis of 6-formylpterin nucleoside analogs and their ROS generation activities in the presence of NADH in the dark.

We demonstrated previously that 3-position-modified 6-formylpterin (6FP) derivatives produce reactive oxygen species (ROS) such as hydrogen peroxide (H(2)O(2)) from oxygen in the presence of NADH in the dark. It has been shown that 6FP derivatives markedly generate ROS, which gives rise to their particular physiological activities, such as induction of apoptosis in cellular and living systems, suggesting that such compounds provide a hint for the design of a ROS controlling agent in vivo. However, it is not well understood why such unique activities appear on chemical modification. In the present study, in order to see the effect on ROS generation activity in the dark by the modification of the 1-position in 6FP, we have developed a new synthetic procedure for nucleoside analogs of 6FP and prepared 1-(beta-d-ribofuranosyl)-2-(N,N-diethylaminomethyleneamino)-6-formylpteridin-4-one (RDEF) and 1-(beta-d-ribofuranosyl)-2-(piperidine-1-ylmethyleneamino)-6-formylpteridin-4-one (RPIF) in which the 1-position of 6FP is glycosylated. At pH 7.4, NADH was spontaneously oxidized to NAD(+) in the presence of RDEF in the dark. Using electron paramagnetic resonance analysis coupled with the spin trapping technique, we show that O(2) was converted to H(2)O(2)via superoxide anion radical ( O(2)(-)) during this reaction. The modification of the 1-position of 6FP did not cancel ROS generation activities, which were demonstrated in 3-position-modified 6FPs. Since the 6FP derivatives developed in the present study have a ribose moiety, these compounds can be subjected to further derivatization, such as incorporation into oligonucleotides, oligosaccharides, proteins, or any other compounds that recognize and interact with specific biomolecules, and therefore would be useful in pharmaceutical investigations that need generation of appropriate and controllable amounts of ROS in vivo.

Reactive oxygen species generation through NADH oxidation by 6-formylpterin derivatives in the dark.

6-formylpterin (6FP) has been reported to produce reactive oxygen species (ROS) such as *O2- and H2O2 from O2 in the presence of NADH under light condition. In the present study, we prepared a variety of 6FP derivatives and found that 2-(N,N-dimethylaminomethyleneamino)-6-formyl-3-pivaloylpteridin-4-one and 2-(N,N-dimethylaminomethyleneamino)-6-formyl-3-methylpteridin-4-one, in which the 2-amino groups are modified by a dimethylaminomethylene group and the 3-positions by pivaloyl and methyl groups and 2-amino-6-formyl-3-methylpteridin-4-one in which the amino group at the 2-position is free and the 3-position is modified by a methyl group generated H2O2 from O2 on oxidation of NADH to NAD+ in the dark. However, 6FP and 2-(N,N-dimethylaminomethyleneamino)-6-formylpteridin-4-one, in which the 3-position is free did not yield H2O2. These results indicate that modification of the 3-position is essential to make the activities of 6FP available in the dark and would be suggestive for designing pharmaceutical compounds that generate appropriate and controllable amounts of ROS in vivo.

Chemical natures and application of 6-formylpterin derivatives.

Pterin, an analog of guanine, is an electron transfer compound in biological systems. Among the analogs, 6-formylpterin (6FP) has been demonstrated to have many marked physiological and pharmacological activities. In vitro, 6FP converts molecular oxygen to reactive oxygen species (ROS) in the presence of NADH or NADPH under light illumination, with the oxidation of NADH is to NAD(+). In the present study, it has been elucidated that some of 6FP derivatives in which the 3-position of 6FP is modified have such unique activities even in the dark where the most of in vivo biological events occur.

Interspecific pheromone cross-attraction among soybean bugs (Heteroptera): does Piezodorus hybneri (Pentatomidae) utilize the pheromone of Riptortus clavatus (Alydidae) as a kairomone?

The chemical and ecological function of cross-attraction of Piezodorus hybneri (Pentatomidae) to the Riptortus clavatus (Alydidae) pheromone (a mixture of three components) was studied. In a field attraction test using traps with synthetic pheromone components, P. hybneri was attracted to (E)-2-hexenyl (E)-2-hexenoate, a component of the R. clavatus pheromone. Other components had neither an additive nor a synergistic effect on the attraction of P. hybneri. Neither (E)-2-hexenyl (E)-2-hexenoate nor other components of the R. clavatus pheromone were detected in volatiles or whole-body extracts of P. hybneri adults by gas chromatographic analysis. In addition, (E)-2-hexenyl (E)-2-hexenoate could not be found in volatiles of soybean plants. Therefore, it appears that P. hybneri responds to a component of the R. clavatus pheromone that is not emitted by P. hybneri itself. We discuss this interspecific pheromone cross-attraction of the soybean bug and hypothesize that P. hybneri utilizes the pheromone of its competitor as a kairomone for host location.

Novel 6-formylpterin derivatives: chemical synthesis and O2 to ROS conversion activities.

6-Formylpterin (6FP) has been demonstrated to have strong neuroprotective effects against transient ischemia-reperfusion injury in gerbils. Also it has been shown that in rats, 6FP protected retinal neurons even when it was administered after the ischemic insult. Since there is a significant need for such a compound that effectively suppresses the events caused by the lack of oxygen supply, 6FP has attracted further investigation. Unfortunately, however, 6FP is hardly soluble in water at neutral pH and in organic solvents because of its self-assembling ability. Although a several mM solution of 6FP is available in alkaline water, it is unstable. In the present study, a novel chemical derivatization of 6FP has been developed which maintains the formyl group on the 6-position of 6FP, which is essential for the physiological activities of 6FP, and increases solubility in water and organic solvents. In the method, the 2- and 3-positions of 6FP were modified by a three component coupling reaction: 6FP was subjected to the reaction with acid chloride and N,N-dimethylformamide. The derivatives synthesized here, 2-(N,N-dimethylaminomethyleneamino)-6-formyl-3-pivaloylpteridine-4-one 1, 2-(N,N-dimethylaminomethyleneamino)-6-formyl-3-isobutyrylpteridine-4-one 2, and 2-(N,N-dimethylaminomethyleneamino)-6-formyl-3-o-toluoylpteridine-4-one 3, showed high solubility in water (1.0-5.6 mM) and organic solvents. The O(2) conversion property has also been determined for the derivative 1. Using an oxygen electrode, it has been found that O(2) is consumed in the presence of 1 and NADH at around pH 7.4 and that the rate of O(2) consumption is enhanced by UV-A irradiation. Electron paramagnetic resonance (EPR) analysis coupled with DMPO spin trapping has also revealed that in the presence of NADH, 1 converts O(2) to O(2)(-), which is further reduced to OH. By UV-A illumination in the analogous systems, (1)O(2) formation was observed. These results are similar to those reported previously for 6FP.

LPS-induced ROS generation and changes in glutathione level and their relation to the maturation of human monocyte-derived dendritic cells.

Lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) generation and the concomitant decline in the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) were demonstrated in human monocyte-derived dendritic cells (DC). Further, their relation to the maturation of DC, characterized by the production of cytokines, up-regulation of cell surface molecules and allo-stimulatory capacity, was examined. The LPS-induced ROS generation was demonstrated using electron paramagnetic resonance spectroscopy in intact cells, and was also confirmed using laser scanning confocal microscopy. The GSH/GSSG was assesed using a glutathione assay kit. When the DC were treated with alpha-phenyl-tert-butylnitrone, the ROS generation was attenuated, but the declined GSH/GSSG was not attenuated, and only cytokine production was suppressed among the above-mentioned maturation characteristics. When the DC were treated with glutathione monoethyl ester, both the ROS generation and the declined GSH/GSSG were attenuated, and the maturation characteristics were all suppressed. These findings suggest that the LPS-induced ROS generation and the concomitant decline in GSH/GSSG occur in human monocyte-derived DC and that the former is involved in cytokine production, while the latter is involved in the up-regulation of cell surface molecules and allo-stimulatory capacity. Since the cytokine production and the allo-stimulatory capacity of DC play an important role in inflammatory and immune responses, differential regulation of the ROS generation and the declined GSH/GSSG may be useful as therapeutic tools in diseases where both responses become entangled, such as sepsis and graft-versus-host disease.