Identification of human NK cells that are deficient for signaling adaptor FcRγ and specialized for antibody-dependent immune functions.

NK cells respond to tumor and virus-infected cells directly through several activation receptors, including natural cytotoxicity receptors, or indirectly through the activating Fc receptor CD16 for antibody-coated cells. Triggering of NK-cell effector functions through these receptors depends on physically associated transmembrane signaling adaptors, such as FcRγ (also known as FcεRIγ) and CD3ζ, both of which have been traditionally believed to be expressed by all mature NK cells. However, we have identified a distinct subset of human NK cells that are deficient for FcRγ expression but express normal levels of CD3ζ. FcRγ-deficient NK cells were readily detectable in about one-third of the healthy individuals examined. The deficiency was confined to the CD56(dim) population and was due to low FcRγ mRNA. FcRγ-deficient NK cells displayed dramatically reduced expression of the natural cytotoxicity receptors NKp46 and NKp30 but still expressed substantial levels of CD16. Compared to FcRγ-expressing NK cells, FcRγ-deficient NK cells showed poor direct reactivity toward tumor targets as measured by cytokine production and degranulation. Unexpectedly, however, FcRγ-deficient NK cells exhibited significantly more robust responsiveness upon stimulation through CD16, particularly for cytokine production, compared to FcRγ-expressing NK cells. Thus, our study reveals FcRγ-deficient NK cells as a novel subset of human NK cells that have remarkably potent responses toward antibody-coated targets. These findings also illustrate a differential contribution of FcRγ and CD3ζ for the expression and functional activity of their associated receptors.

Akt Cys-310-targeted inhibition by hydroxylated benzene derivatives is tightly linked to their immunosuppressive effects.

The hydroxylated benzene metabolite hydroquinone (HQ) is mainly generated from benzene, an important industrial chemical, and is also a common dietary component. Although numerous reports have addressed the tumorigenesis-inducing effects of HQ, few papers have explored its molecular regulatory mechanism in immunological responses. In this study we characterized Akt (protein kinase B)-targeted regulation by HQ and its derivatives, in suppressing inflammatory responses using cellular, molecular, biochemical, and immunopharmacological approaches. HQ down-regulated inflammatory responses such as NO production, surface levels of pattern recognition receptors, and cytokine gene expression with IC(50) values that ranged from 5 to 10 microm. HQ inhibition was mediated by blocking NF-kappaB activation via suppression of its translocation pathway, which is composed of Akt, I kappaB alpha kinase beta, and I kappaB alpha. Of the targets in this pathway, HQ directly targeted and bound to the sulfhydryl group of Cys-310 of Akt and sequentially interrupted the phosphorylation of both Thr-308 and Ser-473 by mediation of beta-mercaptoethanol, according to the liquid chromatography/mass spectroscopy analysis of the interaction of HQ with an Akt-derived peptide. Therefore, our data suggest that Akt and its target site Cys-310 can be considered as a prime molecular target of HQ-mediated immunosuppression and for novel anti-Akt-targeted immunosuppressive drugs.

Inhibition of cytokine expression by a butanol extract from Cordyceps bassiana.

Cordyceps species have been known since long as a multi-utility ethnomedicinal herbal in Korea, China and Japan. It has been reported to exhibit a number of properties such as anti-oxidative, anti-cancer, antiinflammatory, anti-diabetic, and anti-obesity effects. In a previously conducted study, we had demonstrated that the ethanol extract of Cordyceps bassiana was able to suppress the production of interleukin (IL)-12 and interferon (IFN)-gamma in macrophages and T lymphocytes. In this study, we were able to further explore the molecular basis of its inhibitory mechanism using a butanol fraction of this herbal (Cb-BF) preparation. Similarly, this fraction also blocked the expression of cytokines such as IL-12 and tumor necrosis factor (TNF)-alpha as well as the proliferation of splenic lymphocytes and their production of IFN-gamma but not IL-4. Cb-BF suppressed the luciferase activities that are mediated by nuclear factor (NF)-kappaB, activator protein (AP)-1, and signal transducers and activators of transcription (STAT)-1. In agreement with this, these fractions diminished the translocation of the transcription factors into the nucleus. The study also demonstrated that the upstream signaling events for the activation of these factors such as spleen tyrosine kinase (Syk), janus kinase (JAK)-2, and extracellular signal-regulated kinase (ERK) were suppressed. Therefore, these results suggest that the butanol extract of Cordyceps bassiana may contain more than one active component capable of inhibiting the inflammatory signaling cascade and this can be considered as a potential candidate for treatment of diseases that require suppression of immune system.

Effect of ferrule on the fracture resistance of mandibular premolars with prefabricated posts and cores.

PURPOSE: This study evaluated fracture resistance with regard to ferrule lengths and post reinforcement on endodontically treated mandibular premolars incorporating a prefabricated post and resin core. MATERIALS AND METHODS: One hundred extracted mandibular premolars were randomly divided into 5 groups (n=20): intact teeth (NR); endodontically treated teeth (ETT) without post (NP); ETT restored with a prefabricated post with ferrule lengths of either 0 mm (F0), 1 mm (F1), or 2 mm (F2). Prepared teeth were restored with metal crowns. A thermal cycling test was performed for 1,000 cycles. Loading was applied at an angle of 135 degrees to the axis of the tooth using a universal testing machine with a crosshead speed of 2.54 mm/min. Fracture loads were analyzed by one-way ANOVA and Tukey HSD test using a statistical program (α=.05). RESULTS: There were statistical differences in fracture loads among groups (P<.001). The fracture load of F2 (237.7 ± 83.4) was significantly higher than those of NP (155.6 ± 74.3 N), F0 (98.8 ± 43.3 N), and F1 (152.8 ± 78.5 N) (P=.011, P<.001, and P=.008, respectively). CONCLUSION: Fracture resistance of ETT depends on the length of the ferrule, as shown by the significantly increased fracture resistance in the 2 mm ferrule group (F2) compared to the groups with shorter ferrule lengths (F0, F1) and without post (NP).

Protective effect of quercetin, a natural flavonoid against neuronal damage after transient global cerebral ischemia.

Previous studies have demonstrated that quercetin, a bioflavonoid shows the inhibitory effect against ischemia and reperfusion-induced injury in various tissues including neural tissue. Quercetin is also reported to have an inhibitory effect against matrix metalloproteinases (MMPs). Because MMPs are known to play a main role in the pathophysiology of brain ischemic insult, their mechanisms of possible protective effect of quercetin against brain ischemia remain to be clarified. In the present study, C57BL/6 mice were subjected to 20 min transient global brain ischemia. Cerebral blood flow was monitored by laser doppler flowmeter. Animals were sacrificed 72 h after ischemia. Quercetin (50 mg/kg, dissolved in saline) was intraperitoneally administered to mice at 30 min before and immediately after ischemia and from the second day, quercetin was then administered once daily until sacrifice. The present study was undertaken to test the effect of quercetin on neuronal damage after transient cerebral ischemia. Neuronal damages were remarkable in the medial portion of CA1 and CA2 areas after ischemic insult. In quercetin-treated mice, delayed neuronal damage was significantly decreased compared with vehicle-treated mice. Mice treated with quercetin showed attenuated brain MMP-9 activity. Gelatin gel zymography showed an induction of MMP-9 protein after ischemia. Quercetin significantly inhibited ischemia-induced elevation of MMP-9. In situ zymography showed elevations in gelatinase activities after brain ischemia. Quercetin also inhibited TdT-mediated dUTP nick end labeling (TUNEL) staining in CA1 and CA2 areas. These results demonstrate that quercetin, a natural flavonoid reduces global ischemia-induced neuronal damage through inhibition of MMP-9 activity.

Hydroquinone modulates reactivity of peroxynitrite and nitric oxide production.

Peroxynitrite (ONOO(-)), a potent cytotoxic oxidant formed by the reaction of nitric oxide ((.-)NO) and superoxide radical ((.-)O(2)(-)), may be rapidly lethal in a cellular milieu due to oxidization and nitration processes. In the present study, hydroquinone displayed strong ONOO(-) scavenging activity and inhibitory effect on NO production in murine macrophage RAW264.7 cells. Hydroquinone strongly scavenged ONOO(-)induced dihydrorhodamine 123 oxidation in a dose-dependent manner compared with other reactive species such as (.-)O(2)(-) and (.-)NO. Hydroquinone also decreased levels of ONOO(-) induced nitrotyrosine of glutathione reductase and consequently prevented the enzyme from ONOO(-) induced damage. Furthermore, hydroquinone suppressed NO production, a cellular pathway for ONOO(-) formation, in lipopolysaccharide-activated RAW264.7 cells via inhibition of inducible NO synthase expression. The inhibitory effect by hydroquinone seems to be mediated by interruption of lipopolysaccharide-induced signalling such as activation of nuclear factor-kappaB and extracellular signalrelated kinases 1 and 2. The results suggest that hydroquinone may potently modulate reactivity of ONOO(-) and may therefore be a useful agent against ONOO(-) mediated diseases.

Flavonoids differentially modulate nitric oxide production pathways in lipopolysaccharide-activated RAW264.7 cells.

Naturally occurring flavonoids are known to modulate various inflammatory and immune processes. Based on structural property, in this study, molecular mechanism of flavonoids in modulating nitric oxide (NO) production and its signaling pathway were investigated using lipopolysaccharide (LPS)-activated RAW264.7 cells. Although flavonol-typed flavonoids (kaempferol and quercetin) more potently scavenged reactivity of nitric oxide (*NO) as well as peroxynitrite (ONOO-) than isoflavones (genistein and genistin), kaempferol, quercetin and genistein showed a little difference in inhibition of both inducible NO synthase expression and NO production, with IC50 values of 13.9, 20.1 and 26.8 microM. However, there was a striking pattern related to structural feature in modulation of LPS-mediated signaling pathways. Thus, flavonols only inhibited transcription factor AP-1 activation, whereas isoflavones suppressed the DNA binding activation of NF-kappaB and C/EBPbeta. Therefore, these data suggest that structural feature may be linked to decide drugs target molecule in LPS-mediated signaling pathways, rather than its potency.

The Feasibility of a Modified Method of Laparoscopic Transabdominal Cervicoisthmic Cerclage During Pregnancy.

OBJECTIVE: To evaluate a modified laparoscopic transabdominal cervicoisthmic cerclage (LTCC) technique after failure of transvaginal cerclage during pregnancy in women with cervical weakness. MATERIALS AND METHODS: Eighty women in whom transvaginal cerclage was unsuccessful or who were anatomically unsuitable for the procedure underwent modified LTCC between January 2003 and December 2008 at Keimyung University, Dongsan Medical Center, Daegu, South Korea. The modified LTCC was performed using a polyfilament polyester double-armed needle that was sutured laterally to the uterine vessels at the level of the internal cervical os. Survival of the fetus was used to calculate the successful pregnancy rate of this modified LTCC. The relationship between successful pregnancy rate and clinical variables was evaluated using a chi-squared test and a Mann-Whitney U test. RESULTS: The mean gestational age was 12.1 weeks (range, 11-15 weeks). The operation time was 52 minutes (range, 25-100 minutes). The successful pregnancy rate was 90% (72/80 pregnancies), with a mean gestational age of 36.3±2.7 weeks. The mean newborn weight was 2690 g (range, 1860-3750 g). Eight pregnancies were lost in the first and second trimesters due to spontaneous abortion, premature rupture of the membrane, and termination due to anomaly; no other complications occurred. No statistical difference was found between the successful pregnancy rate and the measured clinical variables. CONCLUSIONS: The modified LTCC is feasible and safer than traditional LTCC.

Cytotoxic and pro-apoptotic activities of cynaropicrin, a sesquiterpene lactone, on the viability of leukocyte cancer cell lines.

Cynaropicrin, a sesquiterpene lactone from Saussurea lappa, has been reported to possess immunomodulatory effects on cytokine release, nitric oxide production and immunosuppressive effects. In this study, we have examined cytotoxic effect of cynaropicrin against several types of cell lines such as macrophages, eosinophils, fibroblasts and lymphocytes. Cynaropicrin potently inhibited the proliferation of leukocyte cancer cell lines, such as U937, Eol-1 and Jurkat T cells, but some other cells such as Chang liver cells and human fibroblast cell lines were not strongly suppressed by cynaropicrin treatment. The cytotoxic effect of cynaropicrin was due to inducing apoptosis and cell cycle arrest at G1/S phase, according to flow-cytometric, DNA fragmentation and morphological analyses using U937 cells. Evidence that combination treatment with l-cysteine and N-acetyl-l-cysteine, reactive oxygen species scavengers, or rottlerin (1-[6-[(3-acetyl-2,4,6-trihydroxy-5-methylphenyl)methyl]-5,7-dihydroxy-2, 2-dimethyl-2H-1-benzopyran-8-yl]-3-phenyl-2-propen-1-one), a specific protein kinase (PK) Cdelta inhibitor, abolished cynaropicrin-mediated cytotoxicity and morphological change, and that cynaropicrin-induced proteolytic cleavage of PKCdelta suggests that reactive oxygen species and PKCdelta may play an important role in mediating pro-apoptotic activity by cynaropicrin. Taken together, these results indicate that cynaropicrin may be a potential anticancer agent against some leukocyte cancer cells such as lymphoma or leukemia, through pro-apoptotic activity.

In vitro anti-inflammatory and pro-aggregative effects of a lipid compound, petrocortyne A, from marine sponges.

(3 S,14 S)-Petrocortyne A, a lipid compound (a C(46) polyacetylenic alcohol), from marine sponges ( Petrosia sp.) is potently cytotoxic against several solid tumour cells. In this study, we investigated in vitro anti-inflammatory and pro-aggregative effects of petrocortyne A at non-cytotoxic concentrations on various cellular inflammatory phenomena using the macrophage and monocytic cell lines RAW264.7 and U937. Petrocortyne A blocked tumour necrosis factor-alpha (TNF-alpha) production strongly and concentration-dependently in lipopolysaccharide (LPS)-activated RAW264.7 cells and phorbol 12-myristate 13-acetate (PMA)/LPS-treated U937 cells. It also blocked NO production concentration-dependently in LPS- or interferon (IFN)-gamma-treated RAW264.7 cells. Among the migration factors tested, the compound selectively blocked the expression of hepatocyte growth factor/scatter factor (HGF/SF). On the other hand, as assessed by a cell-cell adhesion assay, petrocortyne A did not block the activation of adhesion molecules induced by aggregative antibodies to adhesion molecules, but suppressed PMA-induced cell-cell adhesion significantly. Intriguingly, petrocortyne A induced U937 homotypic aggregation following long exposure (2 and 3 days), accompanied by weak induction of pro-aggregative signals such as tyrosine phosphorylation of p132 and phosphorylation of extracellular signal-related kinase 1 and 2 (ERK 1/2). Petrocortyne A may thus inhibit cellular inflammatory processes and immune cell migration to inflamed tissue.

Peroxynitrite scavenging activity of sinapic acid (3,5-dimethoxy-4-hydroxycinnamic acid) isolated from Brassica juncea.

Peroxynitrite (ONOO(-)), formed from a reaction of superoxide and nitric oxide, is one of the most potent cytotoxic species that are known to oxidize cellular constituents including essential proteins, lipids, and DNA. In this study, the ability of sinapic acid (3,5-dimethoxy-4-hydroxycinnamic acid), isolated from Brassica juncea, to scavenge ONOO(-) was investigated. The data obtained show that sinapic acid can efficiently scavenge native ONOO(-) as well as ONOO(-) derived from the peroxynitrite donor 3-morpholinosydnonimine hydrochloride (SIN-1). Spectrophotometric analyses revealed that sinapic acid suppressed the formation of ONOO(-)-mediated tyrosine nitration through an electron donation mechanism. In further studies, sinapic acid also showed a significant ability of inhibiting nitration of bovine serum albumin and low-density lipoprotein (LDL) in a dose-dependent manner. Sinapic acid decreased the LDL peroxidation induced by SIN-1-derived ONOO(-). The present study suggests that sinapic acid has an efficient ONOO(-) scavenging ability, which may well be a potent ONOO(-) oxidant scavenger for the protection of the cellular defense activity against the ONOO(-)-involved diseases.

Effects of dialkoxylphenyl compounds with oxime group on macrophage function and the proliferation of lymphocytes.

Dialkoxyphenyl compounds have been reported to possess anti-inflammatory activity through inhibition of phosphodieseterase (PDE) type IV. In this study, a series of derivatives of dialkoxyphenyl compounds with an oxime group, which is generally known to be one of the biologically active functional groups, were prepared and evaluated for their ability to inhibit the production of inflammatory mediators in activated macrophages and the proliferation of lymphocytes. The structure-activity relationship (SAR) study with 12 compounds on tumour necrosis factor (TNF)-alpha inhibition, analysed by the oxime geometry and different size of spacers between the oxime and phenyl group, indicated that there might be at least three possible hydrogen bonding sites in the inhibitor binding pocket of PDE IV. Of them, compound 6 clearly displayed the highest inhibitory effect on in-vitro TNF-alpha production from lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Compound 6 also suppressed in-vivo TNF-alpha release from LPS-primed mice, a level comparable with that of the standard PDE IV inhibitor, rolipram. In addition, oxime compounds also significantly inhibited both nitric oxide production from activated RAW264.7 cells and T lymphocyte proliferation elicited by concanavalin A but not IL-2. The data suggest that the oxime group may act as a functional group, capable of interacting with the inhibitor-binding pocket of target PDE IV. Therefore, it is conceivable that compound 6 may have the potential either to be developed as a new anti-inflammatory drug or to be used to develop more potent analogues.

Selective peroxynitrite scavenging activity of 3-methyl-1,2-cyclopentanedione from coffee extract.

It has been known that reactive oxygen and nitrogen species such as nitric oxide (NO), superoxide radical (*O2-) and their byproduct peroxynitrite (ONOO-) induce cellular and tissue injury, ultimately resulting in several human diseases. In this study, we examined scavenging effects of 3-methyl-1,2-cyclopentanedione (MCP) from coffee extract on the reactivity of those toxic molecules. MCP significantly inhibited both the oxidation of 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA) by reactive oxygen species (ROS) (mainly *O2-) from kidney homogenate (41% at 100 microM) and the generation of fluorescent 4,5-diaminofluorescein (DAF-2) by NO from sodium nitroprusside (IC50 (concn producing 50% inhibition), 63.8 microM). More potently, however, MCP suppressed the oxidation of dihydrorhodamine 123 (DHR 123) to fluorescent rhodamine 123 mediated by authentic ONOO- with an IC50 value of 3.3 microM. The neutralizing effect of the reactivity of ONOO- by MCP was due to electron donation, not nitration of the compound. Additionally, MCP also decreased ONOO- formation of nitrotyrosine adducts of glutathione (GSH) reductase, and consequently protected the enzyme activity of GSH reductase against decreasing by ONOO-, indicating that MCP may prevent ONOO- -induced damage of GSH reductase. Furthermore, MCP only weakly suppressed NO production, which is one of the upstream sources of ONOO- in-vivo, suggesting that NO production may be not a pharmacological target for MCP. Taken together, our results suggest that MCP may be regarded as a selective regulator of ONOO- -mediated diseases via direct scavenging activity of ONOO-.

In vitro antioxidant activity of some selected Prunus species in Korea.

In the course of the investigations of natural antioxidants, we examined the antioxidant activities of the methanol (MeOH) extracts of some selected Prunus species, including P buergeriana, P davidiana, P padus, P pendula for. ascendens, P. sargentii, P. serrulata var. spontanea and P. yedoensis by three methods as represented by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, total ROS (reactive oxygen species) and the peroxynitrite (ONOO-) scavenging activity tests. We also evaluated the activities of the organic solvent-soluble fractions, including the dichloromethane (CH2Cl2), ethyl acetate (EtOAc), n-butanol (n-BuOH) fractions and the water (H2O) layer of P serrulata var. spontanea leaves. By means of bioassay-directed fractionation, we isolated eleven known flavonoids (1-11) from the EtOAc soluble fraction of the MeOH extract of the Prunus serrulata var. spontanea leaves, exhibiting strong antioxidant activity and characterized as prunetin (1), genistein (2), quercetin (3), prunetin 4'-O-beta-glucopyranoside (4), kaempferol 3-O-alpha-arabinofuranoside (5), prunetin 5-O-beta-glucopyranoside (6), kaempferol 3-O-beta-xylopyranoside (7), genistin (8), kaempferol 3-O-beta-glucopyranoside (9), quercetin 3-O-beta-glucopyranoside (10) and kaempferol 3-O-beta-xylopyranosyl-(1-->2)-beta-glucopyranoside (11). Compounds 3 and 10 showed good activities in all tested model systems. Compounds 2 and 8 showed scavenging activities in the DPPH and ONOO- tests, while compounds 5, 7, 9 and 11 were active in the ONOO- and ROS tests. On the other hand, compounds 1, 4 and 6 did not show any activities in the tested model systems.

Incidence of intravascular insertion in thoracic epidural catheterization by using real time fluoroscopy.

BACKGROUND: Epidural analgesia is commonly used to provide several types of pain relief. Although this technique has been widely used with many advantages, currently the complications appear to be increasing. Especially, inadvertent intravascular cannulation and intravascular local anesthetic administration can lead to fatal consequences. METHODS: Data was collected on 296 patients undergoing elective thoracic or abdominal surgery. Two detection methods were utilized to confirm the epidural intravascular cannulation; flashback and aspiration of indwelling catheter, and injection of a contrast agent through the catheter under fluoroscopy were used to guide the placement of the catheter and to examine the intravascular cannulation. RESULTS: Epidural intravascular cannulation was reported in 4 out of 296 cases (1.4%), and 1 patient underwent subdural cannulation. Among the 4 cases of epidural intravascular cannulation, two were confirmed by the flashback and aspiration methods, while the remaining cases were only detected by real time fluoroscopy. CONCLUSIONS: In this study, inadvertent epidural intravascular cannulation occurred by as much as 1.4% of thoracic epidural catheterization. Utilizing real time fluoroscopy in addition to flashback and aspiration can enhance the sensitivity of detection.

Modest inhibition of the growth hormone axis does not affect mitochondrial reactive oxygen species generation or redox state, unlike calorie restriction.

AIM: Modest inhibition of the growth hormone (GH) axis by overexpression of the antisense GH gene in male Wistar rats reduced food intake and body weight, and lengthened the lifespan, even if fed ad libitum (AL). These findings were comparable with those induced by 30% calorie restriction (CR) in wild-type (WT) rats, suggesting importance of the GH signal pathway in the effect of CR. The present study evaluated the effects of GH inhibition and CR on mitochondrial oxidative stress and redox state in the liver. METHODS: Transgenic and WT rats were fed AL or 30% CR diets from 6weeks of age. Liver tissues were collected at 6 and 24months of age. The mitochondria fraction was prepared from liver tissue homogenates. The total reactive oxygen species (ROS) generation, the protein levels of glutathione (GSH) and oxidized GSH (GSSG), and the superoxide dismutase 2 activity were measured. RESULTS: The results revealed that CR, but not modest inhibition of GH, decreased mitochondrial ROS generation and increased the mitochondrial GSH redox potential. CONCLUSION: The present study suggests that CR affects mitochondrial function and redox state through a pathway distinct from GH signaling.

Anti-inflammatory activity of Sorbus commixta water extract and its molecular inhibitory mechanism.

ETHNOPHARMACOLOGICAL SIGNIFICANCE: Sorbus commixta Hedl. (Rosaceae) is a well known traditionally valuable medicinal plant in Korea, China and Japan. This plant has been prescribed for long time for various inflammatory symptoms such as asthma, bronchitis, gastritis and dropsy. AIM OF STUDY: Although a number of pharmacological properties have already been demonstrated, the anti-inflammatory effect of this plant and its associated molecular mechanisms has not yet been fully investigated. MATERIALS AND METHODS: In order to address the anti-inflammatory activity of S. commixta water extract (Sc-WE), lipopolysaccharide (LPS)-stimulated macrophages were employed and production of inflammatory mediators by these cells were evaluated. RESULTS: Sc-WE significantly suppressed the production of nitric oxide (NO) and prostaglandin (PG)E(2) in a dose-dependent manner and blocked ear edema formation induced by arachidonic acid in mouse. In addition, this extract effectively diminished the mRNA levels of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2, indicating that the inhibition occurs at the transcriptional level. Interestingly, Sc-WE remarkably blocked NF-κB translocation and its upstream signaling events by inhibition of κBα (IκBα), IκBα kinase (IKK), Akt (protein kinase B), phosphoinositide-dependent kinase 1 (PDK1), p85/phosphoinositide-3-kinase (PI3K), as per the results obtained from the reporter gene assay and immunoblotting analysis. More intriguingly, Sc-WE suppressed activities of Src and Syk kinases as well as their phosphorylation levels without altering molecular complex formation between them and toll like receptor (TLR)4 or MyD88, an adaptor protein of TLR4-mediated signaling. CONCLUSION: Therefore, our results suggest that Sc-WE can be developed as a potent anti-inflammatory remedy, acting by suppressing the inflammatory signaling cascade composed of Src, Syk, and NF-κB.

Is the rapid sequence induction possible with 0.6 mg/kg rocuronium in pediatric patient?

BACKGROUND: We have investigated the possibility of rocuronium 0.6 mg/kg and timing principle application with the same dose for rapid sequence induction (RSI) in 65 children, aged 4-8 yr. METHODS: Sixty five patients were randomly assigned to one of two groups; Group A (n = 31, timing principle application) received rocuronium (0.6 mg/kg) followed by administration of propofol (2.5 mg/kg), and group B (n = 36) received rocuronium (0.6 mg/kg) after administration of propofol. Intubation was assessed at 60 seconds just after administration of last injectants. Intubating conditions (jaw relaxation, vocal cord movement, and response to tracheal intubation) were evaluated as excellent, good, fair and poor. RESULTS: Excellent intubation conditions were obtained in 87% in group A and 61% in group B. However, clinically acceptable intubation conditions which means excellent and good did not show any significant difference as 100% (group A) and 99% (group B). CONCLUSIONS: In cases of pediatiric patients undergoing elective surgery, RSI was possible irrespective of the use of timing principle.

Mitogen activated protein kinases are prime signalling enzymes in nitric oxide production induced by soluble ß-glucan from Sparassis crispa.

Sparassis crispa (SC) is an edible mushroom that harbours ß-glucans reported to possess immunostimulatory and anticancer properties. The role of SC in regulating the functional activation of macrophages is yet to be fully elucidated. The objective of this study was to investigate the molecular mechanism underlying the immune-stimulatory function of Sparassis crispa soluble ß-glucan (Sc-SG) on macrophages. According to this study, Sc-SG was able to stimulate nitric oxide (NO) production as well as enhance the expression of inducible NO synthase (iNOS) from macrophage-like RAW264.7 cells. NO production was strongly suppressed by mitogen-activated protein kinase (MAPK) inhibitors such as U0126, extracellular signal-regulated kinase, SB203580, a p38 inhibitor, and SP600125, a c-Jun N-terminal kinase inhibitor. Thus, indicating that Sc-SG-induced NO release is possibly mediated by MAPK. Sc-SG induced phosphorylation of extracellular signal-regulated kinase, p38, and JNK in a time-dependent manner. Moreover, Sc-SG triggered the phosphorylation and translocation of c-Jun and c-Fos, components of the transcription factor AP-1, activated by MAPK. The results of this study suggest that MAPK may be a major signaling enzyme that regulates the Sc-SG-mediated NO production in macrophages.

Metabolic inhibition and kinetics of raloxifene by pharmaceutical excipients in human liver microsomes.

This study was originally undertaken to establish the in vitro metabolic conditions and then evaluate the effect of pharmaceutical excipients (PEs) on drug metabolism in uridine diphosphoglucuronic acid-supplemented human liver microsomes. Poorly bioavailable raloxifene was chosen as a model drug. Intact drug and its two glucuronide metabolites were successfully isolated using gradient HPLC analysis and LC/MS analysis. Formation of raloxifene metabolites was affected by buffer compositions, incubation time and initial raloxifene concentrations. Under optimized metabolic conditions, 41.0% of raloxifene was converted to its metabolites after 2h incubation. This metabolic inhibition of raloxifene by the PEs occurred in a dose-dependent manner and accordingly formed two glucuronide metabolites. In the metabolic kinetics using Lineweaver-Burk analyses, Cremophor EL competitively inhibited formation of metabolites while sodium lauryl sulfate (SLS), polyvinylpyrrolidone K30 (PVP) and Tween 80 significantly inhibited in a mixed competition. Although some PEs showed inhibition on glucuronidation of raloxifene in vitro, their effects on in vivo bioavailability of raloxifene need to be confirmed directly due to the dilution factors and other complicated situations influencing the bioavailability.