Ellagic Acid Prevents L-NAME-Induced Hypertension via Restoration of eNOS and p47phox Expression in Rats

The effect of ellagic acid on oxidative stress and hypertension induced by N^ω-Nitro-l-arginine methyl ester hydrochloride (L-NAME) was investigated. Male Sprague-Dawley rats were administrated with L-NAME (40 mg/kg/day) for five weeks. L-NAME induced high systolic blood pressure (SBP) and increased heart rate (HR), hindlimb vascular resistance (HVR) and oxidative stress. Concurrent treatment with ellagic acid (7.5 or 15 mg/kg) prevented these alterations. Co-treatment with ellagic acid was associated with up-regulation of endothelial nitric oxide synthase (eNOS) protein production and alleviation of oxidative stress as indicated by decreased superoxide production in the vascular tissue, reduced plasma malondialdehyde levels, reduced NADPH oxidase subunit p47^phox expression and increased plasma nitrate/nitrite levels. Our results indicate that ellagic acid attenuates hypertension by reducing NADPH oxidase subunit p47^phox expression, which prevents oxidative stress and restores NO bioavailability.     

One-week high-fat diet leads to reduced toll-like receptor 2 expression and function in young healthy men

Toll-like receptor 2 (TLR2) is implicated in inflammatory responses to high-fat diet (HFD)–induced obesity in rodents, but human HFD studies examining TLR2-mediated immune responses are lacking. Our aim was to determine whether HFD affected TLR2 function in humans. We hypothesized that a short-term HFD in humans would impair TLR2-mediated immune function. Fasting blood samples were obtained from healthy young men (N = 9) before and after a 7-day HFD. Toll-like receptor 2 function was assessed in ex vivo whole blood cultures stimulated with the TLR2 agonist N-palmitoyl-S-[2,3-bis[palmitoyloxy]-[2RS]-propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine (Pam3-Cys-SK4). Peripheral blood mononuclear cells (PBMCs) were isolated to examine TLR2, TLR4, and p47 subunit of nicotinamide adenine dinucleotide phosphate oxidase (p47^phox) protein expression via Western blotting. Pam3-Cys-SK4–stimulated secretion of interleukin-1β (−35%, P = .005), interleukin-6 (−32%, P = .01), and tumor necrosis factor–α (−33%, P = .06) was reduced following the HFD. High-fat diet resulted in decreased TLR2 (P = .049) and p47^phox (P = .037) protein expression from PBMCs. To mimic lipid overload ex vivo, follow-up experiments were performed in whole blood cultures exposed to a mixture of free fatty acids for 24 hours; and surface protein expression of TLR2 and TLR4 on CD14+ monocytes was measured by flow cytometry. Free fatty acid exposure for 24 hours ex vivo reduced monocyte TLR2 levels by about 20% (P = .028). A 7-day HFD in young healthy men resulted in impaired TLR2 function. Decreased TLR2 and p47^phox protein expression in PBMCs, possibly due to excess free fatty acids, may mediate this response. Our current findings indicate that impaired TLR2 response after HFD might be partially responsible for increased risk of infection in diet-induced obesity.     

Effects of Whole-Grain Consumption on Selected Biomarkers of Systematic Inflammation: A Systematic Review and Meta-analysis of Randomized Controlled Trials

Whole grains have potential benefits in preventing cardiovascular diseases and diabetes; nevertheless, results from randomized clinical trials (RCTs) on inflammatory markers are controversial. The aim of this meta-analysis of RCTs was to examine the effect of whole grains on inflammatory markers A systematic literature search was conducted by using the online database of PubMed, Embase, Google Scholar, and Scopus for relevant studies up to November 2017, using Medical Subject Headings and other related keywords. Only studies that compared the effects of whole grains on inflammatory markers with refined grains were included. From more than 2278 articles, 17 RCTs met the inclusion criteria and were systematically reviewed. Weighted mean differences were estimated and pooled effect size was calculated by random effects model. Thirteen RCTs with 466 participants were included in the meta-analysis. Whole-grain consumption had a significant effect on serum concentration of high-sensitivity C-reactive protein (hs-CRP; Hedges’ g: ?0.22; 95% confidence interval (CI): ?0.45, 0.00; p?=?0.047), interleukin-6 (IL-6; ?0.28?pg/mL; 95% CI: ?0.55, ?0.02; p?=?0.037) but did not result in a significant decline in serum concentration of tumor necrosis factor alpha (TNF-α; ?0.12?pg/mL; 95% CI: ?0.39, 0.15; p?=?0.396). Significant heterogeneity was observed between studies for hs-CRP (I²?=?69.0%, p?<?0.0001), IL-6 (I²?=?96.6%, p?<?0.001), and TNF-α (I²?= 95.4%, p?<?0.001). In sensitivity analysis, the effect of whole grain intake on hs-CRP, IL-6, and TNF-α was not substantially modified by the result of a single study. Meta-regression for duration showed no significant association between the duration of study and changes in serum C-reactive protein levels (β coefficient?=?0.006, standard error?=?0.036; p?=?0.870).

Conclusions: This meta-analysis of RCTs suggested that whole grains might affect health status via improving systematic inflammation.

• Key teaching points:

• We conducted a systematic review and meta-analysis of randomized controlled trials.

• Whole grains consumption was associated with lower serum levels of hs-CRP and IL-6.

• Whole grains cannot significantly decrease serum levels of TNF-α.

• We could not found any source for heterogeneity.

• The effect of whole grains on serum inflammatory biomarkers was independent from duration.

     

Sesamin Ameliorates Advanced Glycation End Products-Induced Pancreatic β-Cell Dysfunction and Apoptosis

Advanced glycation end products (AGEs), the direct modulators of β-cells, have been shown to cause insulin-producing β-cell dysfunction and apoptosis through increase of intracellular reactive oxygen species (ROS) production. Sesamin has been demonstrated to possess antioxidative activity. This study was designed to investigate whether sesamin protects against AGEs-evoked β-cell damage via its antioxidant property. The effects of sesamin were examined in C57BL/6J mice and MIN6 cell line. In in vivo studies, mice were intraperitoneally injected with AGEs (120 mg/kg) and orally treated with sesamin (160 mg/kg) for four weeks. Intraperitoneal glucose tolerance and insulin releasing tests were performed. Insulin content, ROS generation and β-cell apoptosis in pancreatic islets were also measured. In in vitro studies, MIN6 cells were pretreated with sesamin (50 or 100 μM) and then exposed to AGEs (200 mg/L) for 24 h. Insulin secretion, β-cell death, ROS production as well as expression and activity of NADPH oxidase were determined. Sesamin treatment obviously ameliorated AGE-induced β-cell dysfunction and apoptosis both in vivo and in vitro. These effects were associated with decreased ROS production, down-regulated expression of p67^phox and p22^phox, and reduced NADPH oxidase activity. These results suggest that sesamin protects β-cells from damage caused by AGEs through suppressing NADPH oxidase-mediated oxidative stress.     

Leishmania infantum sterol 24-c-methyltransferase formulated with MPL-SE induces cross-protection against L. major infection

The enzyme sterol 24-c-methyltranferase (SMT) is required for the biosynthesis of ergosterol, the major membrane sterol in Leishmania parasites. SMT and ergosterol are not found in mammals, so this protein may be an attractive target for anti-leishmanial vaccines and drugs. We have previously demonstrated that SMT from L. infantum, which causes visceral leishmaniasis, is a protective antigen against this parasite. Because this protein is highly conserved among Leishmania species, we evaluated the potential of SMT to cross-protect against a different form of leishmaniasis. Here, we show that immunization with L. infantum SMT, formulated with monophosphoryl lipid A in stable emulsion (MPL-SE), protects mice from cutaneous leishmaniasis caused by L. major. In BALB/c mice the vaccine preparation induced antigen-specific multi-functional CD4⁺ T cells capable of producing IFN-γ, IL-2, and/or TNF-α upon antigen re-exposure, and MPL-SE was indispensable to direct immune responses to SMT towards Th1. Mice immunized with the SMT/MPL-SE vaccine developed significantly smaller lesions following ear challenge with L. major. These results suggest that SMT is a promising vaccine antigen for multiple forms of leishmaniasis.     

葡萄籽提取物改善砷暴露致大鼠肝损伤的作用及机制

目的研究葡萄籽提取物(GSE)对砷诱导肝损伤大鼠抗氧化防御系统和NADPH氧化酶的影响。方法40只健康雄性SD大鼠随机分为4组:正常对照组、砷暴露组(As,NaAsO230 mg/L)、GSE干预组(GSE100 mg/kg隔日灌胃)、砷暴露+GSE干预组(As+GSE)。12个月后,检测谷丙转氨酶(ALT)、谷草转氨酶(AST)和白蛋白,作肝功能分析;检测谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、超氧化物歧化酶(SOD)血清抗氧化酶的活性、检测组织活性氧(ROS)水平、检测脂质过氧化水平;Western blot检测肝脏中烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶亚单位Nox2、Nox4、p47phox蛋白的表达。结果与对照组相比,砷干预组的肝功能明显受损,抗氧化酶活性降低,组织ROS水平、脂质过氧化水平、Nox2、Nox4、p47phox蛋白表达水平升高(P<0.01);与As组相比,联合干预组的各项指标均得到明显改善(P<0.01)。结论 GSE可抑制慢性砷暴露引起大鼠的氧化应激及肝损伤,其机制可能与抑制NADPH氧化酶活性有关。[营养学报,2012,34(4):388-391]     

Fucoxanthin in association with Vitamin c acts as modulators of human neutrophil function

Introduction

Neutrophils provide the first line of defense of the innate immune system by phagocytosing, killing and digesting bacteria and fungi. During this process, neutrophils produce reactive oxygen species (ROS), which in excess, can damage the cells themselves and surrounding tissues. The carotenoid fucoxanthin (Fc) has been studied concerning its antioxidant and anti-inflammatory actions. Vitamin c (Vc) also demonstrates potent antioxidant action. This study aimed to evaluate the effect of Fc (2 μM) in association with Vc (100 μM) on functional parameters of human neutrophils in vitro.

Materials and methods

We evaluated the migration and phagocytic capacity, intracellular calcium mobilization, ROS production (O ₂ ^·? , H₂O₂, HOCl), myeloperoxidase activity, profile of antioxidant enzymes, phosphorylation of p38 MAPK and p65 NFκB subunit, GSH/GSSG ratio and release of pro-inflammatory cytokines (TNF-α and IL-6) in neutrophils under different stimuli.

Results

We verified an increase in phagocytic capacity for all treatments, together with an increase in intracellular calcium only in cells treated with Fc and Fc + Vc. ROS production was reduced by all treatments, although Vc was a better antioxidant than Fc. Phosphorylation of the p-65 subunit of NFκB was reduced in cells treated with Fc + Vc and release of TNF-α and IL-6 was reduced by all treatments. These findings indicate that the regulation of inflammatory cytokines by neutrophils is not exclusively under the control of the NFκB pathway. Fc reduced the activity of some antioxidant enzymes, whereas Vc increased GR activity and the GSH/GSSG ratio.

Conclusion

In conclusion, the results presented in this study clearly show an immunomodulatory effect of the carotenoid fc alone or in combination with Vc on the function of human neutrophils.     

Effects of Vitamin D and Omega-3 Fatty Acids Co-Supplementation on Inflammatory Factors and Tumor Marker CEA in Colorectal Cancer Patients Undergoing Chemotherapy: A Randomized,Double-Blind,Placebo-Controlled Clinical Trial

Abstract

Objectives: This study aimed to investigate the effects of vitamin D and omega-3 fatty acids co-supplementation on inflammatory factors and tumor marker CEA in colorectal cancer patients undergoing chemotherapy.

Methods: In this study, 81 patients with stage ?? or ??? colorectal cancer were randomly assigned into four groups: (1) control: receiving a vitamin D placebo, weekly?+?two omega-3 fatty acid placebo capsules, daily; (2) omega-3 fatty acid, receiving two omega-3 fatty acid capsules (each capsule containing 330?mg of omega-3 fatty acids), daily?+?a vitamin D placebo, weekly; (3) vitamin D, receiving a 50,000?IU vitamin D soft gel, weekly?+?two omega-3 fatty acid placebo capsules, daily; (4) co-supplementation, receiving a 50,000?IU vitamin D soft gel, weekly?+?two omega-3 fatty acids capsules, for 8?weeks. Before and after the intervention, serum levels of 25(OH)D, TNF-α, IL-1β, IL-6, IL-8, NF-kB activity, and tumor marker CEA, were measured.

Results: After 8?weeks of intervention, patients who received combined vitamin D and omega-3 fatty acids supplements compared with omega-3, vitamin D, and placebo had significantly decreased TNF-α, and IL-1β (P?<?.05). In addition, serum levels of TNF-α, IL-1β, IL-6, IL-8, and tumor marker CEA were decreased significantly in omega-3, vitamin D, and co-supplementation of them, compared with baseline. NF-kB activity was decreased significantly in vitamin D and co-supplementation groups, compared with baseline. Regarding CEA, there was no significant difference between the four groups at the end of intervention (P?>?.05).

Conclusion: Results show that co-supplementation of vitamin D and omega-3 fatty acids co-supplementation, in colorectal cancer patients have beneficial impacts on inflammation and tumor marker CEA.     

Anti-inflammatory effects of daidzein on primary astroglial cell culture

Abstract

Introduction: Alzheimer's disease is the common cause of dementia in old people. The pathological hallmarks of Alzheimer's disease include neuronal loss, deposition of amyloid-β, and presence of neurofibrillary tangles. The endogenous steroid estrogen has been shown to affect neuronal growth, differentiation and survival, while isoflavones also have a neuroprotective effect on human cortical neurons. Daidzein, however, has a superior neuron-protective effect to other isoflavones. The present study is to determine whether daidzein is able to inhibit the production of pro-inflammatory mediators under amyloid-β and lipopolysaccharide stimulation.

Materials and methods: Astrocyte cells were stimulated with amyloid-β or lipopolysaccharide in the absence and presence of diadzein. Nitric oxide released into the culture media was determined using the Griess reaction, and concentrations of IL-1, IL-6, TNF-α and estrogen receptor gene expression were measured by semi-quantitative real-time polymerase chain reaction assay.

Results: Diadzein-treatment increases astrocyte cell counts and attains its maximal effect at the 10^?12M concentration. The addition of 20 μM amyloid-β or 10^?6 g/ml LPS can significantly decrease the viability of astrocytes, up-regulated IL-1, IL-6, TNF-α mRNA and estrogen receptor expression; in addition, 1-h daidzein pre-treatment can restore the decreased viability of astrocytes induced by amyloid-β or lipopolysaccharide as well as down-regulate their mRNA expression.

Conclusions: It seems that this response is estrogen receptor-mediated. These results further increase the possibility that daidzein may have potential to ameliorate the inflammatory process and also alleviate the risk of Alzheimer's disease progression.     

Gp91phox NADPH oxidase modulates litter size by regulating mucin1 in the uterus of mice

Active oxygen derived from gp91phox is critical for gestation. However, no reports have evaluated the relationship between reactive oxygen species (ROS) and the number of births in a given pregnancy. In this study, we examined the influence of ROS produced by gp91phox activity on the number of births using C57BL/6j (control) and gp91phox-knockout (gp91phox^?/-) mice. The number of births in gp91phox^?/- mice was found to be lower than that in control mice. We observed sequential increases in gp91phox, ROS, nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3), caspase-1, and interleukin-18 (IL-18), followed by increased expression of mucin1 (MUC1), in control mice. However, none of these markers were upregulated in gp91phox^?/- mice. In addition, in control mice administered IL-18 or MUC1 inhibitors, the number of births decreased to a number similar to that of gp91phox^?/- mice. These results suggest that ROS derived from gp91phox activity altered the inflammatory system and produced IL-18, which subsequently increased the expression of MUC1, thereby modulating fetal development.

Abbreviations: IL-1 β: interleukin-1β; IL-18: interleukin-18; NLRP3: nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3; IgA: immunoglobulin A; MUC1: mucin1     

Soy protein inhibits inflammation-induced VCAM-1 and inflammatory cytokine induction by inhibiting the NF-κB and AKT signaling pathway in apolipoprotein E–deficient mice

Purpose

Inflammation is a hallmark of many diseases, such as atherosclerosis, autoimmune diseases, obesity, and cancer. Isoflavone-free soy protein diet (SPI^?) has been shown to reduce atherosclerotic lesions in a hyperlipidemic mouse model compared to casein (CAS)-fed mice, despite unchanged serum lipid levels. However, possible mechanisms contributing to the athero-protective effect of soy protein remain unknown. Therefore, we investigated whether and how SPI^? diet inhibits inflammatory responses associated with atherosclerosis.

Methods

Apolipoprotein E knockout (apoE?/?) mice (5-week) were fed CAS or SPI^? diet for 1 or 5 week to determine LPS- and hyperlipidemia-induced acute and chronic inflammatory responses, respectively. Expression of NF-κB-dependent inflammation mediators such as VCAM-1, TNF-α, and MCP-1 were determined in aorta and liver. NF-κB, MAP kinase, and AKT activation was determined to address mechanisms contributing to the anti-inflammatory properties of soy protein/peptides.

Results

Isoflavone-free soy protein diet significantly reduced LPS-induced VCAM-1 mRNA and protein expression in aorta compared to CAS-fed mice. Reduced VCAM-1 expression in SPI^?-fed mice also paralleled attenuated monocyte adhesion to vascular endothelium, a critical and primary processes during inflammation. Notably, VCAM-1 mRNA and protein expression in lesion-prone aortic arch was significantly reduced in apoE?/? mice fed SPI^? for 5 weeks compared with CAS-fed mice. Moreover, dietary SPI^? potently inhibited LPS-induced NF-κB activation and the subsequent upregulation of pro-inflammatory cytokines, including TNF-α, IL-6, IL-1β, and MCP-1. Interestingly, SPI^? inhibited NF-κB-dependent inflammatory responses by targeting I-κB phosphorylation and AKT activation with no effect on MAP kinase pathway. Of the five putative soy peptides, four of the soy peptides inhibited LPS-induced VCAM-1, IL-6, IL-8, and MCP-1 protein expression in human vascular endothelial cells in vitro.

Conclusions

Collectively, our findings suggest that anti-inflammatory properties of component(s) of soy protein/peptides may be a possible mechanism for the prevention of chronic inflammatory diseases such as atherosclerosis.     

Effect of High‐Dose Cysteine Supplementation on Erythrocyte Glutathione

Background: This study's objective was to determine if parenteral cysteine when compared with isonitrogenous noncysteine supplementation increases erythrocyte reduced glutathione (GSH) in neonates at high risk for inflammatory injury. Material and Methods: Neonates with a score for neonatal acute physiology >10 requiring mechanical ventilation and parenteral nutrition (PN) were randomized in a double‐blinded, placebo‐controlled study to receive parenteral cysteine‐HCl (CYS group) or additional PN amino acids (ISO group) at 121 mg/kg/d for ≥7 days. A 6‐hour [¹³C₂] glycine IV infusion was administered at study week 1 to determine the fractional synthetic rate of GSH (FSR‐GSH). Results: Baseline characteristics were similar between the CYS (n = 17) and ISO groups (n = 21). Erythrocyte GSH and total glutathione concentrations, GSH:oxidized GSH (GSSG), and FSR‐GSH after treatment were not different between groups. However, the CYS group had a larger individual positive change in GSH and total glutathione (infusion day – baseline) compared with the ISO group (P = .02 for each). After adjusting for treatment, a lower enrollment weight and rate of red blood cell transfusion were associated with a decreased change in total glutathione and GSH (P < .05 for each). Conclusion: When compared with isonitrogenous noncysteine supplementation, high‐dose cysteine supplementation for at least 1 week in critically ill neonates resulted in a larger and more positive individual change in GSH. Smaller infants and those who received transfused blood demonstrated less effective change in GSH with cysteine supplementation. The benefit of cysteine remains promising and deserves further investigation.     

Biochemical Response of the Copepod <Emphasis Type="Italic">Tigriopus japonicus</Emphasis> Mori Experimentally Exposed to Cadmium

The response of the copepod (Tigriopus japonicus Mori) to cadmium (Cd) additions was investigated under laboratory-controlled conditions in a 12-day exposure. Superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione-S-transferase (GST), acetylcholinesterase (AchE), reduced glutathione (GSH), the ratio of reduced to oxidized glutathione (GSH/GSSG), and metallothionein (MT) were analyzed for Cd treatments (0, 10, 20, 40, and 100 μg/L) after exposure for 1, 4, 7, and 12 days. Additionally, thiobarbituric reactive species assay was used to evaluate lipid peroxidation (LPO) of the copepod after the 12-day exposure. The results indicated that Cd treatments significantly influenced the biochemical indexes (SOD, GPx, GST, AchE, GSH, and GSH/GSSG) after certain exposure times. Exposure to Cd induced LPO in the treated copepods, hinting that the copepods had suffered from oxidative damage. During exposure, the Cd initiated an induced MT synthesis in the copepods by day 7, which peaked at day 12 and which was probably responsible for Cd detoxification. Thus, Cd exposure significantly affected the detoxification process and antioxidant system of this copepod, and T. japonicus could be used as a suitable bioindicator of exposure to Cd using SOD, GPx, GST, LPO, and GSH/GSSG as biomarkers.     

Human dendritic cell maturation and activation by a heat-killed recombinant yeast (Saccharomyces cerevisiae) vector encoding carcinoembryonic antigen

Tumor-associated antigens are weakly immunogenic. Human carcinoembryonic antigen (CEA) is overexpressed on a wide range of human carcinomas and represents an attractive target for cancer immunotherapy. This study analyzes the ability of a Saccharomyces cerevisiae vector containing the transgene encoding CEA (yeast-CEA) to activate human dendritic cells (DCs) and stimulate CEA-specific T-cell responses. We demonstrate for the first time that treatment with yeast-CEA can activate human DCs, resulting in increases in surface expression of CD80, CD83, CD54, CD58, and MHC class II, and increased production by DCs of IL-12p70, TNF-α, IFN-γ, IL-8, IL-2, IL-13, IL-10, and IL-1β. We also show that human DCs treated with yeast-CEA can activate CEA-specific T-cell lines and can act as antigen-presenting cells (APCs) to generate CEA-specific T-cell lines capable of lysing CEA⁺ human tumor cells. Gene expression profiles of human DCs treated with yeast-CEA show increased expression of numerous genes involved in the production of chemokines and cytokines and their receptors, and genes related to antigen uptake, antigen presentation, and signal transduction.     

A phase I trial evaluating the safety and immunogenicity of a candidate tuberculosis vaccination regimen,ChAdOx1 85A prime – MVA85A boost in healthy UK adults

BackgroundThis phase I trial evaluated the safety and immunogenicity of a candidate tuberculosis vaccination regimen, ChAdOx1 85A prime-MVA85A boost, previously demonstrated to be protective in animal studies, in healthy UK adults.MethodsWe enrolled 42 healthy, BCG-vaccinated adults into 4 groups: low dose Starter Group (n = 6; ChAdOx1 85A alone), high dose groups; Group A (n = 12; ChAdOx1 85A), Group B (n = 12; ChAdOx1 85A prime – MVA85A boost) or Group C (n = 12; ChAdOx1 85A – ChAdOx1 85A prime – MVA85A boost). Safety was determined by collection of solicited and unsolicited vaccine-related adverse events (AEs). Immunogenicity was measured by antigen-specific ex-vivo IFN-γ ELISpot, IgG serum ELISA, and antigen-specific intracellular IFN-γ, TNF-α, IL-2 and IL-17.ResultsAEs were mostly mild/moderate, with no Serious Adverse Events. ChAdOx1 85A induced Ag85A-specific ELISpot and intracellular cytokine CD4+ and CD8+ T cell responses, which were not boosted by a second dose, but were boosted with MVA85A. Polyfunctional CD4+ T cells (IFN-γ, TNF-α and IL-2) and IFN-γ+, TNF-α+ CD8+ T cells were induced by ChAdOx1 85A and boosted by MVA85A. ChAdOx1 85A induced serum Ag85A IgG responses which were boosted by MVA85A.ConclusionA ChAdOx1 85A prime – MVA85A boost is well tolerated and immunogenic in healthy UK adults.     

Enhanced antigen uptake by dendritic cells induced by the B pentamer of the type II heat-labile enterotoxin LT-IIa requires engagement of TLR2

The potent mucosal adjuvant properties of the type II heat-labile enterotoxin LT-IIa of Escherichia coli are dependent upon binding of the B pentamer of the enterotoxin (LT-IIa-B₅) to ganglioside receptors on immunocompetent cells. To evaluate the immunomodulatory activities of LT-IIa-B₅, in vitro experiments employing bone marrow-derived dendritic cells (BMDC) were performed. Uptake of OVA-FITC, a model antigen (Ag), was enhanced by treatment of BMDC with LT-IIa-B5, but not by treatment of cells with the B pentamer of cholera toxin (CTB). Expression of co-stimulatory molecules (CD40, CD80, CD86, and MHC-II) and cytokines (IL-12p40, TNF-α, and IFN-γ) was increased in BMDC treated with LT-IIa-B₅. The capacity of LT-IIa-B₅ to enhance Ag uptake and to induce expression of co-stimulatory receptors and cytokines by BMDC was dependent upon expression of TLR2 by the cell. Increased Ag uptake induced by LT-IIa-B₅ was correlated with increased Ag-specific proliferation of CD4⁺ T cells in an in vitro syngeneic DO11.10 CD4⁺ T cell proliferation assay. These experiments confirm that LT-IIa-B₅ exhibits potent immunomodulatory properties which may be exploitable as a non-toxic mucosal adjuvant.     

Glutamine attenuates lipopolysaccharide-induced acute lung injury

ObjectivesIt has been reported that glutamine (GLN) can attenuate acute lung injury after sepsis. GLN is also thought to be a precursor of glutathione (GSH) synthesis. Using the GSH synthesis blocker, L-buthionine-(S,R)-sulfoximine (BSO), we investigated the role of GSH synthesis in the protective effect of GLN on acute lung injury.MethodsIn this study, we used an acute lung injury model induced by intratracheal injection of lipopolysaccharide (1 mg · mL^?1 · kg^?1). GLN (0.75 g/kg, intravenous) and BSO (2 mmol/kg, intraperitoneal) were administrated simultaneously. At 2 and 18 h after the injections, the rats were sacrificed by right ventricular puncture and bronchoalveolar lavage was done. The lower right lung was excised for histologic examination. Total protein concentration and total cell and neutrophil counts in the bronchoalveolar lavage fluid were determined. CD11b expression in the blood was determined by flow cytometry. We also analyzed myeloperoxidase activity, and GSH and interleukin-8 levels in lung tissues.ResultsGLN supplementation reduced the total protein concentration and total cell and neutrophils counts in bronchoalveolar lavage fluid after lipopolysaccharide challenge. GLN enhanced GSH synthesis and attenuated interleukin-8 release and myeloperoxidase activity in lung tissues. GLN also decreased CD11b expression in blood neutrophils and prevented lung histologic changes. BSO abolished the effects of GLN and attenuated its protection on acute lung injury.ConclusionThese results indicate that GLN could prevent neutrophil recruitment and infiltration, protect the alveolar barrier, and attenuate inflammatory injury during sepsis. This effect may be related to enhanced GSH synthesis.     

Impact of hydrosalpinx fluid on early human embryos

This study aimed to investigate the impact of hydrosalpinx fluid (HF) on early human embryonic development. A total of 33 patients who underwent laparoscopic surgery for hydrosalpinx were selected, and the HF specimens obtained from these patients were subjected to bacterial culture, Chlamydia antigen detection, biochemical analysis, and cytokine detection. Meanwhile, human embryos derived from three pronuclei (3PN) were cultured in various HF concentrations. There was no significant difference in the chemical components and physical characteristics between colorless and colored HF specimens, apart from the glucose concentration which was significantly higher in colorless HF. K⁺ and HCO₃^? were significantly increased (P < 0.05 and P < 0.01), and Ca²⁺, Mg²⁺, and glucose were significantly decreased (P < 0.05, P = 0.006, and P = 0.007) in the two HF specimens, compared to blastocyst culture medium (G-2 medium); no phosphates were detected in the HF specimens. Compared to colorless HF, the concentrations of tumor necrosis factor α (TNF-α) and interleukin 2 (IL-2) in the colored HF specimens were significantly increased (P < 0.05). There were no differences in the Chlamydia antigen-positive rate between the HF groups (62.5% vs. 70.6%), and no bacterial growth occurred in the HF specimens. There were no significant differences in the development of the 3PN embryos between the two HF groups (P > 0.05). High-concentration HF (75%) significantly affected the rates of blastulation, blastocyst hatching, and high-quality blastocyst formation (P < 0.05). HF is related to chlamydial infection. Embryonic development may be significantly affected only in high-concentration HF, possibly due to the deficiency of essential elements required for embryonic development. TNF-α and IL-2 concentrations were found to vary between the clear and colored HF specimens; however, TNF-α and IL-2 in HF do not appear to exert adverse effects on embryonic development.