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Objective: To observe the clinical significance of nitric oxide (NO) and 8-isoprostane (8-isoPG) changes in exhaled breath condensate ( EBC) of acute respiratory distress syndrome (ARDS) patients after treated by Qingfei Decoction (QD)., Methods: Totally 48 ARDS patients receiving mechanical ventilation were equally assigned to the QD treatment group and the control group by random digit table. EBC specimens were collected by modified Ecoscreen breath condensate collector (German JAEGER Company) on the first day and the fifth day after confirmed diagnosis of ARDS. Concentrations of NO and 8-isoPG in EBC were measured by ELISA. The oxygenation index and APACHE II scores were recorded at the same time., Results: (1) The fatality rate in the QD treatment group was lower than that in the control group (8.3% vs 37.5%, P < 0.05). (2) After treatment NO and 8-isoPG concentrations in EBC were lower in the QD treatment group (34.49 ± 5.67 µmol/L, 30.09 ± 7.89 ng/L) than in the control group (39.78 ± 9.27 µmol/L, 35.65 ± 8.90 ng/L; P < 0.05). (3) After treatment improved oxygenation index value was higher in the QD treatment group than in the control group (120.88 ± 35.16 vs 101.50 ± 37.70, P < 0.05). After treatment APACHEII scores was lower in the QD treatment group than in the control group (6.21 ± 3.51 vs 10. 26 ± 4.33, P < 0.05)., Conclusion: Treatment of ARDS patients by QD was favorable in controlling inflammation, alleviating lung injury, and improving clinical efficacy.

Fertilization is an indispensable step for formation of a zygote in sexual reproduction, leading to species survival. When mating occurs, sperm is transported to the female reproductive tracts via the seminal plasma (SP). SP is derived from male accessory sex glands and it plays pivotal roles for fertilization in animals. However, molecular mechanisms of SP or a fluid derived from male accessory sex glands for successful fertilization remain unclear. Here, we report that in male quail the cloacal gland (CG) produces prostaglandin F2α (PGF2α) that contributes to successful fertilization. PGF2α, as well as the secretion of CG (CGS), induced vaginal contractions and caused the opening of the entrance of the sperm storage tubules, the structures responsible for the long-term sperm storage and fertilization. The removal of CGS from the male before mating reduced the fertility, but the supplementation of CGS or PGF2α rescued the subfertility. We further showed that male CG contains glucose that is utilized as energy source for the intrinsic sperm mobility after transportation to female vagina. This mechanism, in concert with the excitatory effects of PGF2α enables successful fertilization in the domestic bird.

Aim: In patients with renal failure or chronic inflammation, the accumulation of fetuin-A-containing calciprotein particles (CPP) in the extracellular fluid has been implicated in driving inflammatory pathways and extraosseous mineral deposition. We aimed to discover whether CPP are present in the peritoneal dialysis fluid effluent (PDF) of stable peritoneal dialysis (PD) patients, and if so, how these particles might be formed., Methods: Serum and PDF were sampled from 20 stable PD patients. CPP were quantified by the reduction in fetuin-A concentration after high speed centrifugation. 8-iso-PGF2α in PDF was measured as a marker of oxidative stress. Fetuin-A and phosphate were added to commercially available dialysis fluids to assess their ability to support CPP formation ex vivo., Results: We report that the major protein component of these mineral-containing nanoparticles, fetuin-A, is relatively abundant in PDF and that CPP were present in the PDF of 17/20 PD patients. PDF CPP levels were strongly correlated with 8-iso-PGF2α concentrations. In vitro experiments suggested that commonly used peritoneal dialysate fluids, irrespective of composition, could not sustain appreciable de novo CPP formation ex vivo., Conclusion: Fetuin-A is either actively transported or locally produced by the peritoneal membrane in PD patients. The association between fetuin-A-containing CPP and markers of oxidative stress warrants further mechanistic studies., (© 2014 Asian Pacific Society of Nephrology.)

Background: The current study aimed at assessing the associations between black carbon (BC) exposure and markers for airway inflammation and oxidative stress in primary school children in a Western European urban area., Methods: In 130 children aged 6-12 years old, the fraction of exhaled nitric oxide (FeNO), exhaled breath condensate (EBC) pH, 8-isoprostane and interleukin (IL)-1β were measured in two seasons. BC concentrations on the sampling day (2-h average, 8:00-10:00 AM) and on the day before (24-h average) were assessed using measurements at a central monitoring site. Land use regression (LUR) models were applied to estimate weekly average BC exposure integrated for the time spent at home and at school, and seasonal average BC exposure at the home address. Associations between exposure and biomarkers were tested using linear mixed effect regression models. Next to single exposure models, models combining different BC exposure metrics were used., Results: In single exposure models, an interquartile range (IQR) increase in 2-h BC (3.10 μg/m(3)) was linked with a 5.9% (95% CI: 0.1 to 12.0%) increase in 8-isoprostane. FeNO increased by 16.7% (95% CI: 2.2 to 33.2%) per IQR increase in 24-h average BC (4.50 μg/m(3)) and by 12.1% (95% CI: 2.5 to 22.8%) per IQR increase in weekly BC (1.73 μg/m(3)). IL-1β was associated with weekly and seasonal (IQR=1.70 μg/m(3)) BC with respective changes of 38.4% (95% CI: 9.0 to 75.4%) and 61.8% (95% CI: 3.5 to 153.9%) per IQR increase in BC. An IQR increase in weekly BC was linked with a lowering in EBC pH of 0.05 (95% CI: -0.10 to -0.01). All associations were observed independent of sex, age, allergy status, parental education level and meteorological conditions on the sampling day. Most of the associations remained when different BC exposure metrics were combined in multiple exposure models, after additional correction for sampling period or after exclusion of children with airway allergies. In additional analyses, FeNO was linked with 24-h PM10 levels, but the effect size was smaller than for BC. 8-Isoprostane was not linked with either 2-h or 24-h concentrations of PM2.5 or PM10., Conclusion: BC exposure on the morning of sampling was associated with airway oxidative stress while 24-h and weekly exposures were linked with airway inflammation., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: In vitro fertilisation (IVF) treatment provides an opportunity to study early developmental responses to periconceptional dietary interventions. Retrospective studies have suggested links between preconception diet and fertility, and more recently, a "Mediterranean" diet has been reported to increase pregnancy rates by up to 40%. In addition, a prospective study examining increased intake of omega-3 polyunsaturated fats demonstrated a quickened rate of embryo development after IVF. However, up to now, few prospective randomised controlled trials have investigated the impact of periconceptional dietary interventions on fertility outcomes., Methods and Design: The study is a randomised controlled trial of a dietary intervention consisting of olive oil for cooking, an olive oil based spread, and a daily supplement drink enriched with Vitamin D (10 microgram daily) and marine omega-3 fatty acids (2 g daily) for 6 weeks preconception versus a control diet of sunflower seed oil for cooking, a sunflower oil based spread, and a daily supplement drink without added Vitamin D or marine omega-3 fatty acids. Couples undergoing IVF will be randomised to either the intervention or control group (55 in each arm). The primary endpoint is embryo developmental competency in vitro, measured by validated morphokinetic markers. Secondary outcomes will include the effect of the dietary intervention on the nutritional content of the intrauterine environment., Discussion: This approach will enable rigorous examination of the impact of the dietary intervention on early embryo development, together with the influence of the peri-implantation intra-uterine nutritional environment., Trial Registration: ISRCTN50956936.

F2-isoprostanes such as 8-iso-prostaglandin F2 (8-iso-PGF2α) are formed by free radical-catalyzed mechanisms from membrane phospholipids and from low density lipoproteins through peroxidation of arachidonic acid. Esterified 8-iso-PGF2α is cleaved by phospholipases, circulates in blood and is excreted as putatively harmful oxidatively modified lipid via the kidney into urine. In this study we demonstrate that 8-iso-PGF2α concentrations in plasma samples from heifers are higher (p<0.005) compared to those from first-lactating dairy cows at 71 days postpartum. Furthermore, plasma 8-iso-PGF2α concentrations vary with ovarian activity and differ in response to luteolytic initiation as well as activation of the hypothalamic-pituitary-gonadal axis between heifers and first-lactating cows. Sustainable concentrations of 8-iso-PGF2α (50-150 pg/ml) are detectable in the phospholipid fraction of milk, suggesting milk as an additional excretion route for 8-isoprostanes. Plasma levels largely paralleled levels in milk (p<0.001). Plasma phospholipid 8-iso-PGF2α concentrations in cyclic cows decreased (p<0.05) from day 38 to day 71 postpartum, whereas milk phospholipid 8-iso-PGF2α rather increased (p<0.05). Cyclic cows tend to have higher 8-isoprostane levels compared to acyclic animals. In contrast to lipohydroperoxides, concentration of 8-iso-PGF2α were not correlated with milk yield (p>0.05). Our data indicate 8-iso-PGF2α may be a novel biomarker of oxidative stress in dairy cow, which is detectable in blood as well as in milk., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The roles of oxidative stress on nuclear factor (NF)‑κB activity and cardiomyocyte apoptosis during heart failure were examined using the antioxidant N‑acetylcysteine (NAC). Heart failure was established in Japanese white rabbits with intravenous injections of doxorubicin, with ten rabbits serving as a control group. Of the rabbits with heart failure, 12 were not treated (HF group) and 13 received NAC (NAC group). Cardiac function was assessed using echocardiography and hemodynamic analysis. Myocardial cell apoptosis, apoptosis‑related protein expression, NF‑κBp65 expression and activity, total anti‑oxidative capacity (tAOC), 8‑iso‑prostaglandin F2α (8‑iso‑PGF2α) expression and glutathione (GSH) expression levels were determined. In the HF group, reduced tAOC, GSH levels and Bcl‑2/Bax ratios as well as increased 8‑iso‑PGF2α levels and apoptosis were observed (all P<0.05), which were effects that were attenuated by the treatment with NAC. NF‑κBp65 and iNOS levels were significantly higher and the P‑IκB‑α levels were significantly lower in the HF group; expression of all three proteins returned to pre‑HF levels following treatment with NAC. Myocardial cell apoptosis was positively correlated with left ventricular end-diastolic pressure (LVEDP), NF‑κBp65 expression and 8‑iso‑PGF2α levels, but negatively correlated with the maximal and minimal rates of increase in left ventricular pressure (+dp/dtmax and ‑dp/dtmin, respectively) and the Bcl‑2/Bax ratio (all P<0.001). The 8‑iso‑PGF2α levels were positively correlated with LVEDP and negatively correlated with +dp/dtmax and ‑dp/dtmin (all P<0.001). The present study demonstrated that NAC increased the antioxidant capacity, decreased the NF‑κB activation and reduced myocardial cell apoptosis in an in vivo heart failure model.

Unlabelled: The aim of this study was to evaluate the role of oxidative and nitrosative stress in autogenous bone grafts to the mandible based on immunohistochemical analysis., Material and Methods: Using a well-established sheep model autogenous bone grafts were harvested form the iliac bone. A combination of a Collagen Membrane (CM) and Deproteinized Bovine Bone Material (DBBM) was used to cover the bone graft (Experiment 2). This modification was compared with simple onlay bone grafts (Experiment 1). Immunohistochemically, the expression of specific stable degradation products of oxidative and nitrosative stress was compared between the two experimental groups., Results: Specific markers for oxidative and nitrosative stress showed statistically significant differences in expression in the different experimental groups. The influence of oxidative and nitrosative stress on osteoblasts (OB), osteoclasts (OC), and osteocytes (OCy) was analysed. Experiment 2 showed increased expression of markers in OB and decreased expression in OC., Conclusions: Taking the result of this study and reports from the literature into consideration grafts in Experiment 2 showed less resorption and atrophy, higher activity of OB and inhibition of OC, and less expression of Reactive Oxygen and Nitrogen Species (RONS) as markers of oxidative stress within the graft. These data illustrate the improved remodelling processes in grafts using CM and DBBM., (Copyright © 2013 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.)

The bovine corpus luteum (CL) is hypothesized to utilize a local auto-amplification system for prostaglandin (PG) F2α production. The objective of the present study was to determine if such a PGF2α auto-amplification system exists in the bovine CL, and if so, which factors regulate it. PGF2α significantly stimulated intra-luteal PGF2α production in all luteal phases, but did not affect PGE2 production. The stimulatory effect of exogenous PGF2α on CL PGF2α production was lower at the early luteal phase. Indomethacin, an inhibitor of prostaglandin-endoperoxide synthase (PTGS), significantly suppressed the PGF2α-stimulated PGF2α production by luteal tissue, indicating that the PGF2α in the medium was of luteal origin. Consistent with these secreted-PGF2α profiles, PGF2α receptor (PTGFR) protein expression was higher during the mid and late luteal phases than at early and developing luteal phases. Treatment of cultured bovine luteal cells obtained from the mid-luteal phase with PGF2α (1 µM) significantly increased the expressions of PTGS2, PGF synthase (PGFS), and carbonyl reductase1 (CBR1) at 24 hr post-treatment. Together, these results suggest the presence of a local auto-amplification system for PGF2α mediated by PTGS2, PGFS, and CBR1 in the bovine CL, which may play an important role in luteolysis., (© 2014 Wiley Periodicals, Inc.)

Interferon-τ (IFN-τ) has been recognized as the primary embryonic signal responsible for maternal recognition of pregnancy. Uterine endometrium produces both prostaglandin F2α (PGF2α ) and prostaglandin E2 (PGE2 ). PGF2α is responsible for the luteolysis; however, PGE2 favours establishment of pregnancy by its luteoprotective action. In this study, the dose-response effect of recombinant bovine IFN-τ (rbIFN-τ) on prostaglandin (PG) production by buffalo endometrial stromal cells cultured in vitro was studied. Buffalo endometrial stromal cells were isolated by double enzymatic digestion, initially with trypsin III followed by a cocktail of trypsin III, collagenase type II and DNase I and subsequently cultured till confluence. Further, cells were treated with different doses of rbIFN-τ (0.001, 0.01, 0.1, 1.0 and 10 μg/ml) and keeping a separate set of control. Culture supernatant was collected after 6, 12 and 24 h of treatment. PG levels in the culture supernatant were measured by enzyme immune assay (EIA) and total cellular protein estimated by Bradford method. Results indicated that buffalo endometrial stromal cells following rbIFN-τ treatment enhanced the secretion of both PGE2 and PGF2α , and also its ratio in a strict dose-dependent manner with a significant increase (p < 0.01) in PGE2 production at 1 μg/ml dose of rbIFN-τ and maximal stimulation for both PG was observed at 10 μg/ml. Further, both PG production and its ratio were increased significantly (p < 0.01) in a time-dependent fashion in all the groups at 6, 12 and 24 h post-treatment with highest level achieved at 24 h as compared with control. Absolute levels of PGE2 remained higher than PGF2α indicating PGE2 as the major PG produced by endometrial stromal cells. The dose-dependent response of rbIFN-τ signifies the importance of optimum concentration of IFN-τ for the embryonic development especially during the critical period to establish successful pregnancy., (© 2014 Blackwell Verlag GmbH.)

Objectives: To search for optimal markers in the exhaled breath condensate (EBC), plasma and urine that would reflect the activity/severity of occupational asthma (OA) after the withdrawal from the exposure to the allergen., Material and Methods: Markers of oxidative stress: 8-iso-prostaglandin F2α (8-isoprostane, 8-ISO), malondialdehyde (MDA), 4-hydroxy-trans-2-nonenale (HNE), cysteinyl leukotrienes (LT) and LTB4 were determined using liquid chromatography and mass spectrometry in 43 subjects with immunological OA (49.3 ± 11.8 years), removed from the exposure to the sensitizing agent 10.5 ± 6.5 years ago; and in 20 healthy subjects (49.0 ± 14.9 years). EBC was harvested both before and after the methacholine challenge test. In parallel, identical markers were collected in plasma and urine. The results were analyzed together with forced expiratory volume in one second (FEV1), blood eosinophils, immunoglobulin E (IgE) and eosinophilic cationic protein (ECP) and statistically evaluated (Spearman rank correlation rS, two- or one-sample t tests and alternatively Kruskal Wallis or pair Wilcoxon tests)., Results: Several parameters of lung functions were lower in the patients (FEV1% predicted, MEF25% and MEF50%, Rtot%, p < 0.001). Shorter time interval since the removal from the allergen exposure correlated with higher ECP (rS = 0.375) and lower FEV1%, MEF25% and MEF50% after methacholine challenge (rS = -0.404, -0.425 and -0.532, respectively). In the patients, IgE (p < 0.001) and ECP (p = 0.009) was increased compared to controls. In EBC, 8-ISO and cysteinyl LTs were elevated in the asthmatics initially and after the challenge. Initial 8-ISO in plasma correlated negatively with FEV1 (rS = -0.409) and with methacholine PD20 (rS = -0.474). 8-ISO in plasma after the challenge correlated with IgE (rS = 0.396)., Conclusions: The improvement in OA is very slow and objective impairments persist years after removal from the exposure. Cysteinyl LTs and 8-ISO in EBC and 8-ISO in plasma might enrich the spectrum of useful objective tests for the follow-up of OA.

Methods for monitoring endocrine activities are useful tools for reproduction management. In particular, captive breeding of endangered felid species is considered to be an important part of the species conservation efforts. Within breeding programs, reliable methods for pregnancy diagnosis are highly demanded to prevent peri- and postpartal losses, but pregnancy diagnosis based on gestagen metabolites in felids is hampered by pseudopregnancies. Recently, we described fecal PGFM as an indicator for pregnancy in several feline species, but peak levels of PGFM secretion differed dramatically between species. It is believed that prostaglandin composition and metabolism pathways may differ as well. Therefore, a study was devised to both compare various fecal immunoreactive PGFM metabolites and to identify prostaglandins in fecal extracts by liquid chromatography-mass spectrometry (LCMS). Our results confirmed that fecal metabolite patterns differ between feline species. The identity of PGFM was confirmed in six of eight felids. In Iberian lynx and the Sumatran tiger, PGFM did not exceed 5% of all immunoreactivities. The total number of immunoreactivities varied between two (e.g., domestic cat) and four (e.g., oncilla). Several prostaglandins were identified by LCMS; apart from PGFM, all LCMS-identified prostaglandins, including tetranor-PGFM, did not show any cross-reactivity with our PGFM-specific antibody. This indicates the existence of still unknown eicosanoids and further studies are needed to clarify the origin of the different metabolites. Although differing stages of pregnancy did not reveal significant differences in the composition of metabolites, we could not exclude the possibility that metabolites from other prostaglandins (e.g. PGE2) contributed to the fecal metabolite patterns., (Copyright © 2014 Elsevier Inc. All rights reserved.)

This study assessed the influence of cigarette smoke condensate (CSC) and benzo(a)pyrene [B(a)P] on the levels of two oxidative stress biomarkers [8-isoprostane (8-IsoP) and 8-hydroxy-2-deoxy Guanosine (8-OH-dG)], in in-vitro spent media of follicle cells. Follicles (100-130 µm) isolated from ovaries of F₁ hybrid (C57Bl/6j × CBA/Ca) mice were cultured for 13 days in media exposed to B(a)P [0 ng ml⁻¹ (control) to 45 ng ml⁻¹] or CSC [0 µg ml⁻¹ (control) to 130 µg ml⁻¹]. The concentrations of oxidative stress biomarkers in spent media were quantified by enzyme-linked immune sorbent assays (ELISA). CSC and B(a)P treatment induced a significant, dose-dependent increase in the concentrations of 8-IsoP and 8-OH-dG in the spent media. We conclude that CSC and B(a)P exposure can induce oxidative stress in ovarian follicles, an effect that may contribute to the previously documented decline in follicle development and premature ovarian failure in women who smoke., (© Her Majesty the Queen in Right of Canada 2013.)

The Rho-associated coiled-coil containing protein kinase, (Rho-kinase or ROCK) undergoes activation by oxidative stress. ROCK-II, which is an isoform of ROCK, is activated in a murine model of lung fibrosis. The present study evaluated the level of oxidative stress and activation of ROCK-II in patients with idiopathic pulmonary fibrosis (IPF). The ROCK-II level and the phosphorylation of myosin phosphatase subunit-1 (p-MYPT-1), a hallmark of ROCK activation, were examined by immunohistochemistry of lung tissue sections. The 8-iso prostaglandin-F2alpha (8-isoPGF2alpha) level, as a marker of oxidative stress, of exhaled breath condensate was significantly higher in IPF patients than in control patients. In IPF lungs, ROCK-II was predominantly expressed by bronchial epithelial cells, as well as at a lower level by airway smooth muscle cells, vascular smooth muscle cells, and the fibroblasts of fibroblastic foci (FF). In addition, there was moderate p-MYPT-1 expression in these cells of IPF lungs. In control lungs, ROCK-II was expressed by these cells. p-MYPT-1 was weakly expressed by the bronchial epithelial cells. In conclusion, ROCK-II was activated in various lung cells of IPF patients along with oxidative stress detected by 8-isoPGF2alpha elevation. The ROCK pathway may play a role in the development of IPF oxidative stress.

Aim: Muscle injuries and extensive exercise are associated with cyclo-oxygenase dependent formation of inflammatory prostaglandins. Since the effect of topical administration of non-steroidal anti-inflammatory drugs (NSAIDs) on local cyclo-oxygenase is unknown, the present exploratory, open label, non-randomized study set out to measure exercise induced release of prostaglandins before and after epicutaneous administration of diclofenac., Methods: Microdialysis was used to determine the local interstitial concentration of PGE2 and 8-iso-PGF2α as well as diclofenac concentrations in the vastus lateralis under rest, dynamic exercise and during recovery in 12 healthy subjects at baseline and after a treatment phase applying a total of seven plasters medicated with 180 mg of diclofenac epolamine over 4 days., Results: At baseline PGE2 concentrations were 1169 ± 780 pg ml(-1) at rest and 1287 ± 459 pg ml(-1) during dynamic exercise and increased to 2005 ± 1126 pg ml(-1) during recovery. After treatment average PGE2 concentrations were 997 ± 588 pg ml(-1) at rest and 1339 ± 892 pg ml(-1) during exercise. In contrast with the baseline phase no increase in PGE2 concentrations was recorded during the recovery period after treatment (PGE2 1134 ± 874 pg ml(-1)). 8-iso-PGF2α was neither affected by exercise nor by treatment with diclofenac. Local and systemic concentrations of diclofenac were highly variable but comparable with previous clinical pharmacokinetic studies., Conclusions: We can hypothesize an effect of topical diclofenac epolamine plaster on limiting the increase of local concentrations of the pro-inflammatory prostaglandin PGE2 induced in the muscle of healthy human subjects following standardized physical exercise. No effect of diclofenac treatment on 8-iso-PGF2α concentrations was observed, mainly since isoprostane is produced by a free radical-catalyzed lipid peroxidation mechanism independent of cyclo-oxygenases., (© 2013 The Authors. British Journal of Clinical Pharmacology © 2013 The British Pharmacological Society.)

Introduction: Studies conducted so far on the effect of hyperthyroidism on oxidative stress (OS) have employed blood and urine samples. Exhaled Breath Condensate (EBC) is a non-invasive technique used to take sample from lungs to determine many biological indications. The aim of the present study was determine the possibility of using 8- isoprostane levels in EBC as an indicator of OS in hyperthyroid patients., Methods: The present study was performed on 42 patients with hyperthyroidism and 42 healthy control subjects. Hyperthyroid patients included patients with newly diagnosed Graves' disease, toxic multinodular goiter and toxic adenoma. Exhaled breath condensates were collected from patients in each group using a condensing device. 8- isoprostane levels as an indicator of OS in EBC were detected via immunoassay method., Results: Hyperthyroid patients and control groups had 8-isoprostane levels of 6.08±6.31 and 1.56±0.88 pg/ml, respectively. The difference between patient and control groups was statistically significant (p<0.001). Of the hyperthyroid patients, eleven had Graves', 21 multinodular goiter, and 10 toxic adenoma diagnosis. There were no significant differences among patients of different diagnoses for 8-isoprostane levels (p=0.541). No significant correlations were found between 8-isoprostane and free thyroxine (fT4) or thyroid stimulating hormone (TSH) levels., Conclusion: In the present study, 8-isoprostane levels in EBC of hyperthyroid patients were found to be significantly higher than that in healthy control group. This study is important in that it is the first to evaluate the effects on respiratory system of elevated OS of hyperthyroidism in EBC.

Background: Cysteinyl leukotrienes are important mediators of airway inflammation, whereas 8-isoprostane is a biomarker of oxidative stress. This study evaluated the distributions of cysteinyl leukotriene and 8-isoprostane concentrations in exhaled breath condensates (EBCs) of children. The relationship between cysteinyl leukotriene and 8-isoprostane concentrations in the EBCs was also evaluated., Methods: The EBCs were collected from 34 children with allergic respiratory diseases and 24 healthy children. All recruited children underwent pulmonary function testing every season. The severity of allergic respiratory diseases and medication status were assessed every month in children with allergic respiratory diseases., Results: The EBC cysteinyl leukotriene and 8-isoprostane levels were higher in children with asthma and allergic rhinitis than in those with asthma only and healthy children. In asthmatic children, cysteinyl leukotriene and 8-isoprostane levels peaked in the summer. All children showed a clear association between EBC cysteinyl leukotriene and EBC 8-isoprostane levels., Conclusion: The cysteinyl leukotriene and 8-isoprostane concentrations in the EBCs of children significantly varied by season. Oxidative stress correlated with airway inflammation in children.

Hypoxic ischemic encephalopathy is a serious condition due to inadequate oxygen supply to the brain. Regeneration of neural cells is a critical process for repairing the damaged brain. Nogo has been identified as an inhibitor of neurite outgrowth that is specific to the brain. In the present study, the Nogo-A receptor (NgR) antagonist NEP1-40 was used to study the effects of inhibition of NgR on the regeneration of neural cells and the related Wnt signaling pathway in newborn rats. The investigation focused on the transcription factors regulated in the Wnt signaling pathway during the repair process, together with the proliferation of neural cells. The results indicated that c-Jun and c-Myc were the main transcription factors involved in the Wnt signaling pathway, while neural cell proliferation in the subventricular zone was increased during this process.

Introduction: Exacerbations in chronic obstructive pulmonary disease (COPD) reduce quality of life and are associated with a more rapid deterioration of the disease. It is desirable to predict an oncoming exacerbation before it occurs. The aim of the present study was to identify biomarkers that may predict a forthcoming exacerbation., Materials and Methods: Forty-three patients with COPD in their stable state were included and followed up monthly until exacerbation, or for a maximum of 6 months. The patients come for an extra visit (prior to a scheduled visit) when exacerbated. The patients completed the questionnaires CCQ and MRC. Exhaled breath condensate (EBC) was collected followed by spirometry, impulse oscillometry, and sputum induction., Results: Twenty-five patients had an exacerbation within the 6-month period. Leukotriene B4 in sputum was the only biomarker that was increased at the visit prior to exacerbation compared to at the stable phase (p = 0.05). There also was a tendency for a similar but not significant increase in the sputum levels of 8-isoprostane, myeloperoxidase activity, and interleukin-8, as well as additional increases during exacerbation. Sputum purulence was not increased until exacerbation (p = 0.02). In contrast, none of the inflammatory biomarkers in EBC, the quality-of-life questionnaire score, CRP, spirometric parameters, or impulse oscillometry parameters were increased at the visit prior to exacerbation compared to the values at the stable phase., Conclusion: Sputum biomarkers, especially leukotriene B4, could be used as predictors of a forthcoming exacerbation and worsening of COPD. This would be of great value for the patient, who may be a subject for early treatment and thereby avoid a progression of the disease.

Ozone exposure effect on free radical-catalyzed oxidation products of lipids, proteins, and DNA in the plasma and urine of rats was studied as a continuation of the international Biomarker of Oxidative Stress Study (BOSS) sponsored by NIEHS/NIH. The goal was to identify a biomarker for ozone-induced oxidative stress and to assess whether inconsistent results often reported in the literature might be due to the limitations of the available methods for measuring the various types of oxidative products. The time- and dose-dependent effects of ozone exposure on rat plasma lipid hydroperoxides, malondialdehyde, F2-isoprostanes, protein carbonyls, methionine oxidation, and tyrosine- and phenylalanine oxidation products, as well as urinary malondialdehyde and F2-isoprostanes were investigated with various techniques. The criterion used to recognize a marker in the model of ozone exposure was that a significant effect could be identified and measured in a biological fluid seen at both doses at more than one time point. No statistically significant differences between the experimental and the control groups at either ozone dose and time point studied could be identified in this study. Tissue samples were not included. Despite all the work accomplished in the BOSS study of ozone, no available product of oxidation in biological fluid has yet met the required criteria of being a biomarker. The current negative findings as a consequence of ozone exposure are of great importance, because they document that in complex systems, as the present in vivo experiment, the assays used may not provide meaningful data of ozone oxidation, especially in human studies., (Published by Elsevier Inc.)

Polycyclic aromatic hydrocarbons (PAHs) are known to induce reactive oxygen species and oxidative stress, but the dose-response relationships between exposure to PAHs and oxidative stress levels have not been established. In this study, we recruited 1333 male coke oven workers, monitored the levels of environmental PAHs, and measured internal PAH exposure biomarkers including 12 urinary PAH metabolites and plasma benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydotetrol-albumin (BPDE-Alb) adducts, as well as the two oxidative biomarkers urinary 8-hydroxydeoxyguanosine (8-OHdG) and 8-iso-prostaglandin-F2α (8-iso-PGF2α). We found that the total concentration of urinary PAH metabolites and plasma BPDE-Alb adducts were both significantly associated with increased 8-OHdG and 8-iso-PGF2α in both smokers and nonsmokers (all p < 0.05). This exposure-response effect was also observed for most PAH metabolites (all p(trend) < 0.01), except for 4-hydroxyphenanthrene and 8-OHdG (p(trend) = 0.108). Furthermore, it was shown that only urinary 1-hydroxypyrene has a significant positive association with both 8-OHdG and 8-iso-PGF2α after a Bonferroni correction (p < 0.005). Our results indicated that urinary ΣOH-PAHs and plasma BPDE-Alb adducts can result in significant dose-related increases in oxidative damage to DNA and lipids. Furthermore, when a multianalyte method is unavailable, our findings demonstrate that urinary 1-hydroxypyrene is a useful biomarker for evaluating total PAHs exposure and assessing oxidative damage in coke oven workers.

The purpose of this study was to evaluate the seminal 8-Isoprostane, reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase, total antioxidant capacity (TAC) in subjects with different level of physical fitness. A total of 161 semen samples were obtained from three groups of healthy males, including elite athletes (23.8 ± 5.2 years, n = 56) who had regular training (4-5 days per week), recreationally active men (24.2 ± 4.9 years, n = 52) who participated in educational or recreational physical activities for 4-5 h per week and non-active men (23.9 ± 5.0 years, n = 53) who did not participate in any exercise programmes for at least 6 months prior to the study. The results showed significantly higher levels of SOD, catalase and TAC as well as lower levels of 8-Isoprostane, ROS and MDA in recreationally active men compared with either elite athletes or non-active men (p < 0.001). Also, elite athletes revealed significantly higher seminal 8-Isoprostane, ROS and MDA as well as lower SOD, catalase and TAC levels compared with recreationally active and non-active men (p < 0.001). In conclusion, the results of the present study indicate that there are differences in seminal oxidants and antioxidants of elite athletes, recreationally active and non-active men. These differences are more likely related to indices that favour decrement of oxidative stress-induced peroxidative damage in spermatozoa from recreationally active men. Hence, recreationally active men seem to have a healthier semen production. The physiological significance of this observation is worthy of further investigation., (© 2012 American Society of Andrology and European Academy of Andrology.)

Background: Urinary 8-iso-PGF2α, a marker of oxidative stress, is influenced by the activation of NOX2. It is unclear if platelets 8-iso-PGF2α contribute to urinary 8-iso-PGF2α., Methods and Results: In a cross-sectional study, platelet, urinary, and serum 8-iso-PGF2α were determined in subjects with downregulation (X-linked chronic granulomatous disease [X-CGD], n=25) and upregulation (type II diabetic patients [T2D], n=121) of NOX2 and 153 controls matched for sex and age. In diabetic patients (n=18), the above variables were repeated before and after 7 days treatment with 100 mg/day aspirin or 100 mg/day aspirin plus 40 mg/day atorvastatin. In vitro study was performed to see the contribution of blood cells to serum 8-iso-PGF2α. Compared with controls, X-CGD patients had lower platelet, serum, and urinary 8-iso-PGF2α values; conversely, diabetic patients had higher values of 8-iso-PGF2α compared with controls. Urinary 8-iso-PGF2α significantly correlated with both platelet and serum 8-iso-PGF2α in the 2 cohorts. A parallel increase of platelet, serum, and urinary 8-iso-PGF2α by aspirin and a parallel decrease by aspirin plus atorvastatin were detected in the interventional study. In vitro study demonstrated that platelets contribute to 37% of serum 8-iso-PGF2α and that only 13% of it is of extravascular origin., Conclusions: The study suggests that NOX2 contributes to the formation of 8-iso-PGF2α in both platelets and urine. The direct correlation between platelet and urinary 8-iso-PGF2α suggests that, at least partly, urinary 8-iso-PGF2α reflects platelet 8-iso-PGF2α production. Analysis of serum 8-iso-PGF2α may represent a novel tool to investigate the production of 8-iso-PGF2α by blood cells including platelets., Clinical Trial Registration: URL: ClinicalTrials.gov. Unique Identifier: NCT01250340.

Objective: Exhaled breath condensate (EBC) 8-isoprostane concentrations are increased in asthma, but it is not known if they acutely change following bronchoprovocation. The objective of this study was to evaluate EBC 8-isoprostane concentrations following allergen-induced bronchoprovocation in asthma., Methods: This comparison study included eight mild atopic asthmatics and six controls. Asthmatics were challenged with inhaled specific allergen, methacholine, and irrelevant allergen in random order. Controls were challenged with irrelevant allergen. EBCs collected at 0, 3, 6, 9, and 23 hours by the R-tube method were measured for 8-isoprostanes by ELISA technique. Repeated measures ANOVA technique was used for analysis., Results: EBC 8-isoprostane concentrations did not change following any inhalational challenge, as compared to baseline, in either asthmatics or controls., Conclusions: EBC 8-isoprostane concentrations do not acutely change following bronchoprovocation in subjects with mild asthma.

Objective: The idea that reactive oxygen species (ROS) are associated with the pathogenesis of inflammatory periodontal diseases and have a role (direct or indirect) in tissue damage has become a major area of research over the last decade. The purpose of this study is to determine, presence of 8-isoprostane in gingival crevicular fluid (GCF) in healthy, gingivitis and chronic periodontitis (CP) subjects and to find an association, if any between GCF 8-isoprostane levels and clinical periodontal parameters., Materials and Methods: 78 subjects (40 males and 38 females) were selected based on their clinical parameters into three groups: Group 1 (26 healthy), Group 2 (26 gingivitis subjects) and Group 3 (26 CP subjects). GCF 8-isoprostane levels were estimated by ELISA., Results: The 8-isoprostane concentration in GCF was highest in subjects with chronic periodontitis as compared to gingivitis and healthy subjects and a significant association was observed between GCF 8-isoprostane levels and all periodontal parameters., Conclusions: There was increase in 8-isoprostane levels in GCF as the disease process progressed from health to gingivitis and chronic periodontitis, suggesting a role for increased oxidative stress in CP., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The study was designed to examine the aglepristone (RU534) mechanisms affecting the corpora lutea (CL) lifespan in pseudopregnant rabbits. Aglepristone (10 mg/kg b.w.) was injected subcutaneously twice at either early- or mid-luteal phase (Days 3 and 4, or Days 8 and 9, respectively) after induction of ovulation with GnRH (Day 0). Corpora lutea and uteri, explanted at days 6 and 11, were evaluated for immunohistochemistry and Western blotting of progesterone (PR) and estrogen (ER) receptors, cyclooxygenase 1 (COX1), COX2, and PGE2-9-ketoreductase (PGE2-9-K) enzymatic activities, and progesterone, PGF2α, and PGE2 in vitro synthesis. Independent of luteal stage, aglepristone prolonged the functional luteal phase by 3 Days over that of controls as assessed by blood progesterone profiles. Aglepristone decreased protein for ER during both luteal-stages in CL and uteri. Progesterone receptor protein was decreased by RU354 at Days 6 in the uterus and at Days 11 in CL, whereas RU534 increased PR at Days 11 in uteri. In the CL, RU534 enhanced progesterone production at Days 6 and 11, whereas it decreased PGF2α and increased PGE2 at Day 11. In the uteri, RU534 decreased PGF2α and increased PGE2 synthesis at both days. COX2 and PGE2-9K activities were decreased by RU534 in the CL at Day 11, whereas in the uteri COX2 increased and PGE2-9-K decreased at Days 6 and 11. In conclusion, these data on aglepristone effects suggest that progesterone has a regulatory role on luteal function through direct and uterine-mediated mechanisms in pseudopregnant rabbits., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Purpose: To investigate renal ischemia injury during renal hilar clamping (artery alone versus clamping artery/vein together) by evaluating ischemic damage via two different modalities in animal models-near-infrared tissue oximetry and 8-isoprostane levels., Methods: Near-infrared renal oximetry measurements of Yorkshire swines (n = 4; 8 renal units) subject to hilar clamping were obtained at baseline, during warm ischemia (15- and 30-min trials) and after unclamping. Quantitation of 8-isoprostane levels is the second technique of quantitating interstitial fluid collected from a dialysis catheter placed through renal parenchyma of male Sprague-Dawley rats (n = 50) subject to hilar clamping during preclamp, clamp (either 15 or 30 min of hilum clamping), and post-clamp., Results: N ear-infrared tissue oximetry. In the 15-min trial, oxygen saturation decreased 6× faster with artery alone compared to artery/vein clamped together. In the 30-min trial, the decrease was 5× faster. Recovery of oxygen saturation with only artery clamped occurred more than 2× faster in the 15- and 30-min periods. Isoprostane. For 15-min clamp times, 8-isoprostane levels in the artery alone group demonstrated a 1.54 decrease in the artery clamped alone group (p = 0.006) versus artery/vein together: preclamp (11.47 and 11.63 pg/mL/g), clamp (14.61 and 17.70 pg/mL/g), and post-clamp (14.26 and 22.04 pg/mL/g)., Conclusions: Renal ischemia injury from clamping the renal artery alone was significantly less than clamping artery/vein together demonstrated in two different techniques. Recovery of oxygen saturation was twofold faster, and mean post-clamp 8-isoprostane levels demonstrated a 1.54-fold decrease with clamping renal artery alone compared to clamping artery/vein together.

Glucagon-like peptide-1 (GLP-1) has protective effects in the heart. We hypothesized that GLP-1 would mitigate coronary microvascular and left ventricular (LV) dysfunction if administered after cardiac arrest and resuscitation (CAR). Eighteen swine were subjected to ventricular fibrillation followed by resuscitation. Swine surviving to return of spontaneous circulation (ROSC) were randomized to receive an intravenous infusion of either human rGLP-1 (10 pmol·kg(-1)·min(-1); n = 8) or 0.9% saline (n = 8) for 4 h, beginning 1 min after ROSC. CAR caused a decline in coronary flow reserve (CFR) in control animals (pre-arrest, 1.86 ± 0.20; 1 h post-ROSC, 1.3 ± 0.05; 4 h post-ROSC, 1.25 ± 0.06; P < 0.05). GLP-1 preserved CFR for up to 4 h after ROSC (pre-arrest, 1.31 ± 0.17; 1 h post-ROSC, 1.5 ± 0.01; 4 h post-ROSC, 1.55 ± 0.22). Although there was a trend toward improvement in LV relaxation in the GLP-1-treated animals, overall LV function was not consistently different between groups. 8-iso-PGF(2α), a measure of reactive oxygen species load, was decreased in post-ROSC GLP-1-treated animals [placebo, control (NS): 38.1 ± 1.54 pg/ml; GLP-1: 26.59 ± 1.56 pg/ml; P < 0.05]. Infusion of GLP-1 after CAR preserved coronary microvascular and LV diastolic function. These effects may be mediated through a reduction in oxidative stress.

Exposures to ambient diesel exhaust particles have been associated with respiratory symptoms and asthma exacerbations in children; however, epidemiologic evidence linking short-term exposure to ambient diesel exhaust particles with airway inflammation is limited. We conducted a panel study with asthmatic and nonasthmatic adolescents to characterize associations between ambient diesel exhaust particle exposures and exhaled biological markers of airway inflammation and oxidative stress. Over four weeks, exhaled breath condensate was collected twice a week from 18 asthmatics and 18 nonasthmatics (ages 14-19 years) attending two New York City schools and analyzed for pH and 8-isoprostane as indicators of airway inflammation and oxidative stress, respectively. Air concentrations of black carbon, a diesel exhaust particle indicator, were measured outside schools. Air measurements of nitrogen dioxide, ozone, and fine particulate matter were obtained for the closest central monitoring sites. Relationships between ambient pollutants and exhaled biomarkers were characterized using mixed effects models. Among all subjects, increases in 1- to 5-day averages of black carbon were associated with decreases in exhaled breath condensate pH, indicating increased airway inflammation, and increases in 8-isoprostane, indicating increased oxidative stress. Increases in 1- to 5-day averages of nitrogen dioxide were associated with increases in 8-isoprostane. Ozone and fine particulate matter were inconsistently associated with exhaled biomarkers. Associations did not differ between asthmatics and nonasthmatics. The findings indicate that short-term exposure to traffic-related air pollutants may increase airway inflammation and/or oxidative stress in urban youth and provide mechanistic support for associations documented between traffic-related pollutant exposures and respiratory morbidity., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Purpose: Our research aimed to evaluate the effect of endometriosis on folliculogenesis and pregnancy, and to assess the involvement of inflammatory factors (IL1b, PGE2, PGF2α, and TGFβ2) in follicular fluid., Methods: A total of 65 follicular fluid aspirates were collected. Concentrations of inflammatory factors (IL1b, PGE2, PGF2α, and TGFβ2) and steroid hormones (E2, progesterone, FSH, and LH) within follicular fluid as well as serum E2 and LH concentrations were measured. The mRNA expression of IL1b, Ptgs2, aromatase, and PPARγ in granulosa cells was determined. The outcome of ART was monitored and recorded., Results: The oocyte retrieval, rate of metaphase II oocyte, cleavage rate, effective embryo rate, and pregnancy rates of patients with endometriosis were all significantly lower than those of the control patients. In those with endometriosis, serum E2 concentrations were lower than those observed in controls. Aromatase levels in the granulosa cells of the endometriosis group were lower while concentrations of PGE2 in follicular fluid were higher than in the control group. Concentrations of PGE2, PGF2α, TGFβ2, and IL1b were significantly correlated with each other., Conclusions: These results suggest that the outcomes of ART, in relation to serum E2 concentration, were adversely affected by the presence of endometriosis. Furthermore, the results supported that, among the endocrine and inflammatory factors, PGE2 within the follicular fluid impairs the number and quality of oocytes.

Prematurity and its main respiratory complication, bronchopulmonary dysplasia (BPD), are potentially associated with lifelong respiratory morbidities and/or lung function abnormalities. The mechanisms behind these long-term respiratory problems are still unclear. We assessed airway oxidative stress in adolescents born very pre-term (≤ 32 gestational weeks) by measuring 8-isoprostane concentration in exhaled breath condensate (EBC). In addition, the study protocol included spirometry and measurement of exhaled nitric oxide fraction (F(eNO)). The study groups included 34 ex-pre-term adolescents with BPD, 18 ex-pre-term adolescents without BPD and 34 healthy controls born at term. Regardless of a history of BPD, the ex-premature adolescents had higher EBC 8-isoprostane levels (median (interquartile range) BPD 9.5 (7.3-12.2) pg·mL(-1); pre-term non-BPD 10 (8.1-16) pg·mL(-1)) than the controls (3.2 (1.9-6.5) pg·mL(-1)) (p<0.001). Forced expiratory volume in 1 s was lower in the BPD group (mean ± sd Z-score -2.1 ± 1.58) than in the pre-term non-BPD individuals (-1.13 ± 1.15), who showed in turn significantly lower values than the controls (0.18 ± 0.83; p<0.001). F(eNO) was similar in the three groups (p=0.55). Our data show that, after premature birth, evidence of oxidative stress in the airways may be detected into adolescence, suggesting that long-term respiratory abnormalities after pre-term birth may be associated with an ongoing airway disease and not just a stabilised structural lung damage.

A simple extraction method for the analysis of PGE(2) and PGF(2α) in gonad samples from Atlantic cod and further quantification by using liquid chromatography-tandem mass spectrometry is proposed. The evaluation of the best solvent extraction conditions and the analytical performance parameters are reported. The method was highly selective for both prostaglandins and the calibration curves, based on the internal standard method, were linear between 5 and 1000 ng mL(-1) for PGE(2) and PGF(2α), with limits of detection of 1 ng mL(-1) and 1.5 ng mL(-1) and recovery values of 99.999±0.002 and 99.967±0.023 respectively. The homogenization of samples using liquid nitrogen combined with the developed extraction protocol can be implemented in different types of biological tissues., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The effects of early relief of heavy bilateral carotid stenosis and ischemic postconditioning on hippocampus CA1 neurons are still unclear. In this study, we used a rat model to imitate severe bilateral carotid stenosis in humans. The rats were divided into sham group, carotid stenosis group, stenosis relief group and ischemic postconditioning group. Ischemic postconditioning consisted of three cycles of 30 s ischemia and 30 s reperfusion. The cerebral blood flow was measured with a laser Doppler flowmeter. Neuronal death in the CA1 region was observed by hematoxylin-eosin staining, and the number of live neurons was assessed by cell counting under a light microscope. The levels of oxidative products MDA and 8-iso-PGF2α, inflammatory factors IL-1β and TNF-α, and the activities of anti-oxidative enzymes SOD and CAT were assayed by specific enzyme-linked immunosorbent assay (ELISA) kits, respectively. We found that relief of carotid stenosis and ischemic postconditioning could increase cerebral blood flow. When stenosis was relieved, the percentage of live neurons was 66.6% ± 6.2% on day 3 and 62.3% ± 9.8% on day 27, which was significantly higher than 55.5% ± 4.8% in stenosis group. Ischemic postconditioning markedly improved the live neurons to 92.5% ± 6.7% on day 3 and 88.6% ± 9.1% on day 27. Further study showed that, neuronal death caused by relief of stenosis is associated with increased oxidative stress and enhanced inflammatory response, and the protection of ischemic postconditioning is related to inhibition of oxidative stress and suppression of inflammatory response.

Aim: To evaluate the effect of TEGDMA on cell cycle progression as well as alterations of cell cycle-related gene and protein expression., Methodology: Human dental pulp cells were exposed to 0-5 mmol L(-1) TEGDMA for 24 h. Cytotoxicity was evaluated by 3-(4, 5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. Cell cycle progression was analysed by propidium iodide (PI) flow cytometry. Cell death pathway was surveyed by annexin V/PI dual-staining flow cytometry. The mRNA expression of cell cycle-related genes (cdc2, cyclinB1 and p21) and COX-2 was evaluated by reverse transcriptase-polymerase chain reaction, and their protein expression was evaluated by Western blotting. The production of PGE(2) and PGF(2α) in the culture medium was determined by enzyme-linked immunosorbent assay., Results: Triethylene glycol dimethacrylate inhibited cellular growth and induced cell cycle deregulation in dental pulp cells. High-dose exposure provoked both necrotic and apoptotic cell death. The gene and protein expression of cdc2, cyclin B1 and cdc25C declined obviously whilst cells treated with 2.5 mmol L(-1) TEGDMA concurrent with the elevated expression of p21. The mRNA and protein expression of COX-2, along with production of PGE(2) and PGF(2α), are drastically raised by 2.5-5 mmol L(-1) TEGDMA., Conclusions: Triethylene glycol dimethacrylate induced cytotoxicity, cell cycle arrest and apoptosis in dental pulp cells, which was associated with the decline of cdc2, cyclin B1, cdc25C expression and elevation of p21 expression. Concomitantly, COX-2 expression, PGE(2) and PGF(2α) production increased. These effects may contribute to explain the pulpal damage and inflammation induced by TEGDMA after operative procedures., (© 2012 International Endodontic Journal.)

Background: Mechanisms linking behavioral stress and inflammation are poorly understood, mainly in distal lung tissue., Objective: We have investigated whether the forced swim stress (FS) could modulate lung tissue mechanics, iNOS, cytokines, oxidative stress activation, eosinophilic recruitment, and remodeling in guinea pigs (GP) with chronic pulmonary inflammation., Methods: The GP were exposed to ovalbumin or saline aerosols (2×/wk/4wks, OVA, and SAL). Twenty-four hours after the 4th inhalation, the GP were submitted to the FS protocol (5×/wk/2wks, SAL-S, and OVA-S). Seventy-two hours after the 7th inhalation, lung strips were cut and tissue resistance (Rt) and elastance (Et) were obtained (at baseline and after OVA and Ach challenge). Strips were submitted to histopathological evaluation., Results: The adrenals' weight, the serum cortisol, and the catecholamines were measured. There was an increase in IL-2, IL-5, IL-13, IFN-γ, iNOS, 8-iso-PGF2α, and in %Rt and %Et after Ach challenge in the SAL-S group compared to the SAL one. The OVA-S group has had an increase in %Rt and %Et after the OVA challenge, in %Et after the Ach and in IL-4, 8-iso-PGF2α, and actin compared to the OVA. Adrenal weight and cortisol serum were increased in stressed animals compared to nonstressed ones, and the catecholamines were unaltered., Conclusion & Clinical Relevance: Repeated stress has increased distal lung constriction, which was associated with an increase of actin, IL-4, and 8-iso-PGF2α levels. Stress has also induced an activation of iNOS, cytokines, and oxidative stress pathways.

The link between cell death and increased cyclooxygenases-2 (COX-2) activity has not been clearly established. In this study, we examined whether COX-2 activation contributed to the mechanism of neurotoxicity produced by an organophosphorous nerve agent in cultured rat cortical neurons. Exposure of neuronal cells to the nerve agent, VX resulted in an increase in COX enzyme activity in the culture media. A concentration dependent increase in the activity levels of COX-2 enzyme was observed while there was little to no effect on COX-1. In addition, COX-2 mRNA and protein levels increased several hours post-VX exposure. Pre-treatment of the cortical cells with the COX-2 selective inhibitor, NS 398 resulted in a decrease in both the enzyme activity and prostaglandin (PGE(2) and PGF(2α)) release, as well as in a reduction in cell death. These findings indicate that the increase in COX-2 activity may contribute to the mechanism of VX-induced neurotoxicity in cultured rat cortical neuron., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Oxidative stress is a major contributor to the secondary injury process after experimental traumatic brain injury (TBI). The importance of oxidative stress in the pathobiology of human TBI is largely unknown. The F(2)-isoprostane 8-iso-prostaglandin F(2α) (8-iso-PGF(2α)), synthesized in vivo through non-enzymatic free radical catalyzed peroxidation of arachidonic acid, is a widely used biomarker of oxidative stress in multiple disease states, including TBI and cerebral ischemia/reperfusion. Our hypothesis is that harvesting of biomarkers directly in the injured brain by cerebral microdialysis (MD) is advantageous because of its high spatial and temporal resolution compared to blood or cerebrospinal fluid sampling. The aim of this study was to test the feasibility of measuring 8-iso-PGF(2α) in MD, ventricular cerebrospinal fluid (vCSF), and plasma samples collected from patients with severe TBI, and to compare the MD signals with MD-glycerol, implicated as a biomarker of oxidative stress, as well as MD-glutamate, a biomarker of excitotoxicity. Six patients (4 men, 2 women) were included in the study, three of whom had a focal/mixed TBI, and three a diffuse axonal injury (DAI). Following the bedside analysis of routine MD biomarkers (glucose, lactate:pyruvate ratio, glycerol, and glutamate), two 12-h MD samples per day were used to analyze 8-iso-PGF(2α) from 24 h up to 8 days post-injury. The interstitial levels of 8-iso-PGF(2α) were markedly higher than the levels obtained from plasma and vCSF (p<0.05), supporting our hypothesis. The MD-8-iso-PGF(2α) levels correlated strongly (p<0.05) with MD-glycerol and MD-glutamate, which are widely used biomarkers of membrane phospholipid degradation/oxidative stress and excitotoxicity, respectively. This study demonstrates the feasibility of analyzing 8-iso-PGF(2α) in MD samples from the human brain. Our results support a close relationship between oxidative stress and excitotoxicity following human TBI. MD-8-iso-PGF(2α) in combination with MD-glycerol may be useful biomarkers of oxidative stress in the neurointensive care setting.

Background: Oxidative stress has been implicated in the pathogenesis of diabetes mellitus (DM). Levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), 8-epi-prostaglandin-F(2α) (8-epi-PGF2α), and total protein carbonyls were measured to assess whether DM is associated with altered salivary redox homeostasis., Methods: A total of 215 patients with diabetes and 481 healthy controls were recruited from the Department of Endocrinology at the Jewish General Hospital in Montreal. Levels of oxidative biomarkers were assayed using enzyme-linked immunosorbent assay (ELISA) in whole unstimulated saliva. Associations of the redox data with exposure to insulin, metformin and dietary control were assessed by logistic regression analyses., Results: We observed (i) significantly higher mean levels of 8-OHdG and protein carbonyls in whole unstimulated saliva of patients with diabetes compared to controls, (ii) higher mean levels of protein carbonyls in type 1 diabetes as well as higher mean levels of 8-OHdG and protein carbonyls in type 2 diabetes compared to controls, (iii) elevated levels of protein carbonyls in diet-controlled patients and in patients with diabetes on insulin and metformin, (iv) elevated levels of 8-OHdG in patients on metformin, and (v) significant associations between subjects with DM and salivary 8-OHdG and protein carbonyls., Conclusion: DM is associated with increased oxidative modification of salivary DNA and proteins. Salivary redox homeostasis is perturbed in DM and may inform on the presence of the disease and efficacy of therapeutic interventions., (© 2011 John Wiley & Sons A/S.)

Reasons for Performing the Study: Ascending placentitis results in premature birth and high foal mortality. By understanding how placentitis induces premature delivery, it may be possible to develop diagnostic markers and to delay premature delivery pharmacologically, thereby decreasing perinatal foal mortality., Objective: To identify relationships between bacterial infection, inflammation and premature parturition in mares with experimentally induced placentitis., Materials and Methods: Experiment 1: Concentrations of allantoic fluid prostaglandins (PGs) F2alpha and E2 were measured in 8 mares after intracervical inoculation with Streptococcus equi ssp. zooepidemicus (at Days 285-291 of gestation) until parturition and compared with controls (n = 4). Experiment 2: mRNA expression of interleukin (IL)-1beta, IL-6, tumour necrosis factor (TNF)-alpha and IL-8 in the chorioallantois from inoculated mares in Experiment 1 were compared with 7 mares that foaled normally., Results: Bacterial inoculation resulted in 7 aborted fetuses and birth of one premature, viable foal. Infection was associated with inflammation of the chorioallantois in the region of the cervical star, isolation of bacteria and high concentrations of PGE2 and PGF2alpha in allantoic fluid obtained within 48 h of delivery (P = 0.04). Chorioallantois from all mares expressed mRNA for IL-8, TNF-alpha, IL-6 and IL-1beta. Experimentally infected mares expressed more mRNA for IL-6 (P = 0.003) and IL-8 (P = 0.009) in the cervical star region and more mRNA for IL-6 (P = 0.004) in tissues from placental horns than control mares., Conclusions and Clinical Relevance: Bacterial placentitis may result in liberation of cytokines from the chorioallantois and prostaglandin formation leading to abortion or birth of a precociously mature foal.

Introduction: Discriminating between active and inactive sarcoidosis may be problematic in everyday clinical practice. There are numerous biochemical markers used in the diagnosis and monitoring of sarcoidosis. Somatostatin receptor (SR) scintigraphy with the use of 99mTc-octreotide may be used to estimate disease activity., Objectives: The aim of the paper was to assess the value of traditional biomarkers (serum angiotensin-converting enzyme [SACE], C-reactive protein, markers of calcium metabolism, bronchoalveolar lavage fluid [BALF] lymphocytes) and a novel biomarker, 8-isoprostane (8-IP) in exhaled breath condensate (EBC), in the assessment of sarcoidosis activity in relation to somatostatin receptor scintigraphy., Patients and Methods: The study included 32 patients with sarcoidosis. Scintigraphy was performed using somatostatin analogue, 99mTc-HYNIC-TOC; planar and SPECT/CT images were recorded. The study group was divided into a subgroup with positive radiotracer uptake (n = 20) and without a visible uptake (n = 12). 8-IP levels were measured in EBC by an immunoenzymatic assay. RESULTS We observed a significantly higher EBC 8-IP levels in the subgroup with positive uptake compared with those with negative uptake (19.1 ± 19.8 vs. 5.4 ± 3.5 pg/ml, P = 0.02). The levels of SACE and the percentage of BALF lymphocytes were also nonsignificantly elevated. In the group of patients with positive scintigraphy results, a positive correlation was observed between the uptake ratio and SACE (r = 0.44, P = 0.041)., Conclusions: The results indicate low value of biochemical markers in the assessment of disease activity. SR scintigraphy may have practical usefulness in the monitoring of sarcoidosis.

Background: There is increasing evidence that oxidative stress is involved in the development and severity of bronchiolitis obliterans occurring in post-transplant patients. In developing countries, the most common form of bronchiolitis obliterans occurs after severe lung infection, mainly caused by adenovirus. However, the oxidative status in the lungs of children with post infectious bronchiolitis obliterans is unknown., Methods: The aim of this study was to measure the oxidant (8-isoprostane and protein carbonyls) and antioxidant (catalase and glutathione peroxidase) activity in the bronchoalveolar lavage fluid of 21 children with post-infectious bronchiolitis obliterans, and to correlate oxidant/antioxidant level with lung function. Lung function was assessed by spirometry and plethysmography, one week prior to fiberbronchoscopy., Results: There was a markedly increased oxidative stress (lipid and protein oxidation) in the bronchoalveolar lavage fluid, and a notorious impairment of lung function demonstrating moderate-severe distal airway narrowing. There was not a significant correlation between the level of oxidants or antioxidants and lung function. There was a consistent antioxidants/oxidants pattern characterised by markedly increased 8-isoprostane and carbonyls, increased GPx and normal catalase activity., Conclusion: The present study shows for the first time that children with post-infectious bronchiolitis obliterans have a markedly increased oxidative stress in their lungs., (Copyright © 2010 SEICAP. Published by Elsevier Espana. All rights reserved.)

Many species of teleost fish detect and release F prostaglandins (PGFs), but the specific identities of these compounds and how they function as species-specific pheromones have yet to be resolved. This study addressed these questions in the common carp. An initial set of experiments established that mature male common carp were attracted to chemicals released by ovulated conspecifics, whereas the odor of female goldfish, a close relative, was less attractive. Tests of fractionated holding water from ovulated carp revealed that only the non-polar fraction was attractive on its own. Mass spectrometry and immunoassay next demonstrated that the non-polar fraction contained large quantities of prostaglandin F(2α) (PGF(2α)), 15keto-prostaglandinF(2α), and 13,14-dihydro-15keto-prostaglandin F(2α) (100 g fish released over 1 μg of all 3 PGFs per h at a ratio of 1.0: 1.7: 0.7). Ovulated goldfish released the same three PGFs but at a slightly greater rate and in a different ratio. Tests of synthetic mixtures of these PGFs revealed that the carp-specific mixture attracted male carp but was no better than the goldfish-specific mixture or PGF(2α) alone and that PGF(2α) was just as attractive as mixture of all three PGFs. A final set of attraction tests revealed that although PGF(2α) could explain all of the activity of the non-polar portion of female carp holding water, it could not explain the entire activity of female water but that a mixture of PGFs and the polar fraction could. We conclude that ovulated female carp release a multi-component sex pheromone complex that is comprised of PGF(2α) and unknown species-specific polar compound(s) that synergize the activity of the former. The pheromone also might be useful in controlling this invasive species. The observation that a fish hormonal sex pheromone incorporates bodily metabolites in addition to relatively common hormonal products demonstrates a mechanism by which species specificity may be conferred to this common type of sex pheromone.

Reactive oxygen species (ROS) are involved in both bone and cartilage physiology and play an important role in the pathogenesis of osteoporosis and osteoarthritis. The present study investigated the effect of running exercise on bone and cartilage in heterozygous manganese superoxide dismutase (SOD2)-deficient mice. It was hypothesized that exercise might induce an increased production of ROS in these tissues. Heterozygous SOD2-deficient mice should exhibit an impaired capability to compensate, resulting in an increased oxidative stress in cartilage and bone. Thirteen female wild type and 20 SOD2(+/-) mice (aged 16 weeks) were randomly assigned to a non-active wild type (SOD2(+/+)Con, n = 7), a trained wild type (SOD2(+/+)Run, n = 6), a non-active SOD2(+/-) (SOD2(+/-)Con, n = 9) and a trained SOD2(+/-) (SOD2(+/-)Run, n = 11) group. Training groups underwent running exercise on a treadmill for 8 weeks. In SOD2(+/-) mice elevated levels of 15-F(2t)-isoprostane and nitrotyrosine were detected in bone and articular cartilage compared to wild type littermates. In osteocytes the elevated levels of these molecules were found to be reduced after exercise while in chondrocytes they were increased by aerobic running exercise. The observed changes in oxidative and nitrosative stress did neither affect morphological, structural nor mechanical properties of both tissues. These results demonstrate that exercise might protect bone against oxidative stress in heterozygous SOD2-deficient mice.

Background: The imbalance between oxidants and antioxidants is referred to as oxidative stress and has been associated with various respiratory disorders. The aim of this study was the assessment of 8-isoprostane (8-iso-PGF(2α)) and Cu/Zn superoxide dismutase (Cu/Zn SOD) in exudative pleural effusions in order to examine the diagnostic accuracy of these markers in the differentiation between complicated and uncomplicated parapneumonic effusions., Methods: The study included 214 consecutive patients with pleural effusions [68 parapneumonic (31 uncomplicated parapneumonic, 20 complicated parapneumonic, 17 empyemas), 24 tuberculous, 88 malignant and 34 transudates]. 8-Isoprostane and Cu/Zn SOD were determined by ELISA in pleural fluid and serum., Results: Parapneumonic effusions were characterized by higher pleural fluid 8-isoprostane levels compared to transudative, malignant and tuberculous effusions. Pleural fluid Cu/Zn SOD levels were lower in transudates, while serum levels were higher in transudative compared to all exudative pleural effusions. Both pleural fluid 8-isoprostane and Cu/Zn SOD were higher in complicated parapneumonic effusions and empyemas compared to uncomplicated parapneumonic effusions. Pleural fluid 8-isoprostane was the most accurate test to differentiate between complicated and uncomplicated parapneumonic pleural effusions with a sensitivity of 100% and a specificity of 58·1% at a cut-off point of 35·1 (AUC = 0·848)., Conclusions: Pleural fluid 8-isoprostane and Cu/Zn SOD may provide useful information for the differentiation between uncomplicated and complicated parapneumonic effusions and empyemas., (© 2010 The Authors. European Journal of Clinical Investigation © 2010 Stichting European Society for Clinical Investigation Journal Foundation.)

Background: As new methods are developed to increase efficiency and higher analytical performance, it is necessary to evaluate their quality in comparison to standard methods. To understand how the analytical performance changes between methods, it is common to compare the validation parameters; sensitivity, linearity, accuracy and precision. Here, we compare an UHPLC-UV method to the HPLC-UV method (reference method) for the simultaneous determination of seven prostanoids. Though the basic chromatography theory is the same for HPLC and UHPLC, the instrumentation has been modified to accommodate higher pressures, lower flow rates and smaller sample size. The differences in analytical instrumentation and procedures can give rise to method inequivalencies. Our approach evaluates the UHPLC and HPLC methods and poses the question: are the methods equivalent? To answer this question a statistical comparison of the analytical performance and method parameters is necessary., Results: Statistical comparisons were performed using the t-test, F-test, regression analyses (ordinary linear regression and Deming regression) and Bland-Altman analyses. Statistical comparison of the results, suggested that the precision (amount of variability) is different (p < 0.05) for the HPLC and UHPLC methods. Whereas, the accuracy (method bias and the means) is similar (p > 0.05) for 8-isoprostane, 11-dehydro TXB₂, PGE₂ PGF(2α), PGD₂ and 15-deoxy Δ¹²,¹⁴ PGJ₂., Discussion: Ordinary linear regression shows that the methods are well correlated for all compounds. The Deming regression, which assumes error in both the methods, suggests the existence of a proportional and constant bias for 11-dehydro TXB₂ and only proportional bias for 8-isoprostane, PGF(2α), PGD₂ and 15-deoxy Δ(12,14) PGJ₂ between the two methods. According to Deming regression, the two methods are statistically similar for 6-keto PGF(1α) and PGE₂. The Bland-Altman analyses indicate the two methods are commutable.

Prostaglandins (PGs), important modulators in bone biology, may also contribute to tumor formation and progression in human osteosarcoma. 15-Deoxy-Δ(12,14)-PGJ(2) (15d-PGJ(2)), a metabolite of PGD(2) and PPARγ-ligand, exerts a panel of biological activities via receptor-dependent and -independent mechanisms. As inducible cyclooxygenase-2 (Cox-2) is a candidate inflammatory marker in human osteosarcoma and a rate-limiting enzyme in PG biosynthesis, this study aimed at investigating intracellular redox status and signaling cascades leading to Cox-2 induction in human MG-63 osteosarcoma cells. 15d-PGJ(2) induced the accumulation of reactive oxygen species (ROS) that in turn may lead to upregulation of Cox-2 via two different routes in a PPARγ-independent manner. First, phosphorylation of p38 MAPK directly enhances Cox-2 expression by promoting mRNA stability. Second, 15d-PGJ(2) induces activation of epidermal growth factor receptors and downstream activation of Cox-2 via phosphorylation of p42/44 MAPK. Glutathione precursor molecules reversed enhanced ROS levels and Cox-2 expression. Functional activity of Cox-2 expression was tested by measurement of PGE(2) and PGF(2α). The synthetic compound 9,10-dihydro-15d-PGJ(2) lacking the α,β-unsaturated carbonyl group in the cyclopentenone ring did not exhibit the cellular responses observed with 15d-PGJ(2). We conclude that the electrophilic carbon atom of 15d-PGJ(2) is responsible for alterations in intracellular redox status and Cox-2 expression., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Acute chorioamnionitis of infectious origin and chronic chorioamnionitis of immunological origin are two major placental lesions of spontaneous preterm birth with elevated amniotic fluid interleukin-6 and CXCL10 concentrations, respectively. The changes in the amniotic fluid proteome associated with intra-amniotic infection and acute chorioamnionitis are well defined, yet alterations unique to chronic chorioamnionitis remain to be elucidated. This study was conducted to determine those amniotic fluid proteins changing specifically in the presence of chronic chorioamnionitis. Amniotic fluid obtained from acute chorioamnionitis, chronic chorioamnionitis and gestational age-matched controls were analysed by two-dimensional (2D) difference in gel electrophoresis and MALDI-TOF analyses. The type of histological inflammation was used to define each condition in preterm labour cases (n = 125) and term not in labour cases (n = 22), and the amniotic fluid concentrations of interleukin-6, CXCL8, CXCL10 and prostaglandin F(2α) were also measured by specific immunoassays. Among preterm labour cases, 31 differentially expressed proteins were identified in chronic chorioamnionitis cases as compared to both acute chorioamnionitis and control cases. Importantly, glycodelin-A, which maintains maternal tolerance against an allogeneic fetus, was decreased in chronic chorioamnionitis, while haptoglobin was increased. We report the amniotic fluid proteome of chronic chorioamnionitis for the first time, and the findings herein strongly suggest that there is a pathophysiological association between the changes of immunomodulatory proteins in the amniotic fluid and chronic chorioamnionitis, a histological manifestation of maternal anti-fetal allograft rejection., (Copyright © 2010 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.)

Lipid peroxidation products, malondialdehyde (MDA), 4-hydroxynonenal (4-HNE) and [Formula: see text], were determined in the plasma and urine of patients with Lyme arthritis and healthy people. The group consisted of 19 patients with Lyme arthritis (mean age 47 years) and the control group consisted of 16 healthy individuals (mean age 38 years). Diagnosis of Lyme disease was confirmed by epidemiological anamnesis, clinical manifestation of arthritis and serological examinations. Lipid peroxidation was estimated by the measurement of aldehydes (MDA and 4-HNE, determined by high-performance liquid chromatography [HPLC]) and prostaglandin derivatives (8 - isoPGF(2a), determined by liquid chromatography/mass spectrometry [LC/MS]). MDA and 4-HNE levels were increased about 2-4-fold in the plasma, while in the urine, the increases were about 2-fold. More significant increases were noted for the 8 - isoPGF(2a) total plasma level, which was enhanced over 4-fold, and for the urine 8 - isoPGF(2a) level, which was increased over 8-fold. The 8 - isoPGF(2a) total plasma level consists of free and esterified form. During infection, the ratio of free to esterified form is significantly smaller compared to healthy people. The ratio of free to esterified form of 8 - isoPGF(2a) may be a useful indicator of Lyme arthritis. Moreover, the complementarities of three lipid peroxidation product levels may be helpful in the diagnosis of Lyme arthritis.

To study oxidative stress differences between women with normal vaginal deliveries (VD) and those with elective cesarean sections without labor (CS), total antioxidant capacity (TAC), erythrocyte glutathione peroxidase (GPX) and superoxide dismutase (SOD) activity, 8-isoprostane, nitrotyrosine and 8-hydroxydeoxyguanosine (8-OHdG) in blood, urine, and placental samples were assessed. The VD group had significantly higher placental 8-isoprostane and 8-OHdG levels as well as greater plasma TAC and 8-OHdG levels and lower erythrocyte SOD activity in umbilical venous blood. Women with VD exhibited differential changes in maternal oxidative stress before and after delivery compared with women with CS. Furthermore, we found that repetitive hypoxia-reoxygenation increased the 8-isoprostane and 8-OHdG levels in villous explants compared with the normoxic controls. Together, these results indicate that labor is associated with increased placental oxidative stress and has an influence on maternal oxidative stress. Therefore, women with VD exhibit different oxidative stress indicators than do those with CS., (Copyright © 2010 Elsevier Inc. All rights reserved.)