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Introduction Pregnancy and physical activity are associated with oxidative stress and immune changes. We hypothesized that pregnant women physically more active in early pregnancy will display a better oxidative stress management and inflammatory response later in pregnancy compared with less active pregnant women. Material and methods Maternal physical activity using accelerometry monitors for 1 week and cardiorespiratory fitness (VO2 at anaerobic threshold) were assessed at 14–18 weeks in 58 pregnant women. Plasma and erythrocytes membrane samples were obtained from maternal blood samples at 14–18 and 34–37 weeks of pregnancy. Pro-inflammatory prostaglandin (PG) F2α and oxidative stress-derived F2-isoprostanes were measured by high-performance liquid chromatography coupled to tandem mass spectrometry. Results Higher physical activity levels at 14–18 weeks measured by mean counts per minute, >30 min/d of moderate to vigorous activity or >6500 steps/d at 14–18 weeks of pregnancy were associated with lower levels of total plasmatic PGF2α later in pregnancy. Concentrations of 5 F2-isomers in erythrocyte membranes in late pregnancy were significantly higher in the third (17.5–19.5 mL kg−1 min−1) and/or fourth (19.6–27.7 mL kg−1 min−1) quartiles of cardio-respiratory fitness compared to the first quartile (13.9–15.9 mL kg−1 min−1). Conclusions Overall, higher cardio-respiratory fitness in early pregnancy is associated with enhanced erythrocyte membranes oxidation at 34–37 weeks reflecting a higher oxygen transfer capacity. Also, the most active women experienced lower circulating levels of pro-inflammatory PGF2α in plasma at 34–37 weeks, a marker associated with adverse antenatal inflammation-associated conditions. These results support the practice of physical activity by pregnant women.

Background Diabetes and pregnancy are both associated with oxidative stress, characterized by an increase of F2-isoprostanes from the non-enzymatic oxidation of arachidonic acid, a n − 6 polyunsaturated fatty acid (PUFA). We hypothesized that pregnant women with pre-existing diabetes will be characterized by elevated levels of specific F2-isoPs isomers and altered PUFA composition in plasma early pregnancy when compared to normoglycemic controls. Methods Plasma samples from 23 women with uncomplicated pregnancies and 11 women with pre-existing diabetes in pregnancy were collected between 12 and 18 weeks of pregnancy (MIROS cohort). Six F2-isoprostanes isomers were measured by high-performance liquid chromatography coupled to tandem mass spectrometry. Fatty acids concentrations in plasmatic phospholipids were measured by gas chromatography coupled to a flame ionization detector. Results F2-isoprostanes, specifically the 8-iso-15(R)-PGF2α levels, were 67% higher in diabetic than normoglycemic pregnancies (p = 0.026). The total n − 6 PUFA and arachidonic acid level did not differ between study groups. In contrast, total n − 3 level was 32% lower in diabetic pregnancies than in controls (p = 0.002); EPA(20:5) and DHA(22:6) being specifically reduced (p = 0.035 and p = 0.003 respectively). Delta-6-desaturase (D6D) activity index, calculated using fatty acid ratios, was 9% lower in pre-existing diabetes than in controls (p = 0.042). Conclusions Pre-existing diabetes in early pregnancy displays a distinctive F2-isoprostanes profile when compared to other pathologies of pregnancy, such as preeclampsia, as previously assessed in the same cohort.

OBJECTIVES: We hypothesized that the intake of industrially originated trans-fatty acids (elaidic acid (EA trans 18: 1n-9)) and ruminant trans fatty acids (trans-palmitoleic acid (TPA t16:1 n-7)) will differentially modify gut microbiota and short-chain fatty acids (SCFA) profiles. The objective is to compare the long- and short-term effects of EA and TPA on the fecal microbiome and SCFAs profiles in mice. METHODS: Forty C57BL/6 mice were divided to 4 groups. Each group was given one of the following 4 formulations in the drinking water: lecithin nanovesicles, nanovesicles containing either lecithin with EA or TPA (86:14 (w/w)) or water alone (control) for 28 days with a normal fat diet. Fecal samples were collected at days 0, 7 and 28. Gut microbiota profiles were determined by 16S rRNA gene sequencing. SCFAs were measured by headspace gas chromatography coupled to a single quadrupole mass spectrometer. Baseline data (relative abundance of bacteria or levels of SCFAs) was pooled and then compared with data from day 7 or day 28 for each formulation. RESULTS: After 7 days of lecithin, 16S rRNA analysis revealed an increase in the relative abundance of Lactobacillus. After 28 days of lecithin, an increase in the relative abundance of Lactobacillus, Erysipelotrichaceae, and Enterobacteriaceae together with a decrease in Bacteroidaceae was observed. Further, a tendency to increase level of butyric acid (P = 0.053) was observed after 28 days of lecithin. After 7 days of EA, an increase in the relative abundance of Lactobacillus, whereas a decrease in the relative abundance of Parabacteroides, Bacteroides, Rumininococcaceae, Lachnospiraceae and Peptococcaceae was observed. After 7 days of TPA, results show a decreased level of isovaleric acid (P = 0.04) and valeric acid (P = 0.03). After 28 days of TPA, data demonstrates an increase in the level of butyric acid (P = 0.01) and propionic acid (P = 0.01). Water intake for 28 days decreased the level of valeric acid (P = 0.02). CONCLUSIONS: Consumption of industrial and ruminant trans-fatty acids modify differentially bacterial taxa present in the gut microbiome and SCFA profiles. FUNDING SOURCES: NSERC, CMDO.

This study aimed to determine whether dairy macronutrients alter markers of inflammation and oxidative stress in endothelial cells. Human endothelial cells (HUVEC) were treated with ruminant trans fatty acids (rTFA), either trans-vaccenic acid (tVA) or trans-palmitoleic acid (tPA), whey protein hydrolysate, leucine or combinations of rTFA and dairy protein compounds. Industrial TFA elaidic acid (EA) was also investigated and compared with rTFA. Inflammatory prostaglandins (PG) and F2-isoprostanes (F2-isoP) isomers, markers of oxidative stress, were assessed in cell supernatants by LC-MS/MS. Both tVA and tPA, as well as whey protein hydrolysate, decreased TNFα-induced PG excretion. Combinations of rTFA and dairy protein compounds decreased inflammation to a similar extent than rTFA alone. EA increased class VI F2-isoP isomers, whereas tVA mostly raised class III isomers. In summary, rTFA decreased inflammatory markers and increased oxidative stress markers in endothelial cells. Combinations of rTFA with whey proteins or leucine showed no additive effect.

Vasoactive eicosanoids such as thromboxane (TX) A2 and F2-isoprostanes (F2-isoPs) were shown to be increased in the preeclamptic placenta. Only one of the 64 possible isomers of F2-isoPs derived from the oxidation of arachidonic acid was investigated in the placenta so far. F2-isoPs are released from membrane phospholipids by phospholipases A2 (PLA2) and were shown to act on the TXA2 receptor (TBXA2R). However, the PLA2 deregulated in preeclampsia (PE) remains to be determined. In this study, we analyzed the concentrations of six isomers of F2-isoPs; 8-iso-PGF2α, 8-iso-15(R)-PGF2α, 15(R)-PGF2α, iPF2α-IV, iPF2α-VI, 5-iPF2α-VI and the concentrations of the stable metabolites of TXA2, TXB2, by high performance liquid chromatography coupled with tandem mass spectrometry in placentas of PE (n = 17) and normotensive (n = 15) pregnancies according to the biopsy site: peri-insertion or periphery. In the same biopsies, relative mRNA expression of PLA2G2A, PLA2G4A, PLA2G5, PLA2G7, the PLA2 receptor (PLA2R1), the TXA2 synthase and TBXAR2 were measured by quantitative RT-PCR. We observed similar concentrations of total F2-isoP isomers between groups whereas higher concentrations (>40%) of free F2-isoP were observed for all isomers (p ≤ 0.033) in PE than normotensive controls. As expected, we also observed higher placental concentrations of TXB2 in PE (p = 0.005). Interestingly, we concomitantly found higher mRNA expression of secretory PLA2G2A (p = 0.010), PLA2G5 (p = 0.038) and TBXA2R (p = 0.023) in PE than normotensive placentas. In sum, deregulated PLA2 could potentially be implicated in freeing F2-isoP which could participate in local hypertension observed in the PE placenta through the TX pathway.

International audience; Central to drug discovery is the correct characterization of the primary structures of compounds. In general, medicinal chemists make great synthetic and characterization efforts to deliver the intended compounds. However, there are occasions which incorrect compounds are presented, such as those reported for Bosutinib and TIC10. This may be due to a variety of reasons such as uncontrolled reaction schemes, reliance on limited characterization techniques (LC-MS and/or 1D 1H NMR spectra), or even the lack of availability or knowledge of characterization strategies. Here, we present practical NMR approaches that support medicinal chemist workflows for addressing compound characterization issues and allow for reliable primary structure determinations. These strategies serve to differentiate between regioisomers and geometric isomers, distinguish between N- versus O-alkyl analogues, and identify rotamers and atropisomers. Overall, awareness and application of these available NMR methods (e.g. HMBC/HSQC, ROESY and VT experiments, to name only a few) should help practicing chemists to reveal chemical phenomena and avoid mis-assignment of the primary structures of compounds.

We hypothesized that the mild physiological oxidative stress present during pregnancy could increase both, plasma F 2 -isoprostanes (F 2 -isoPs) by lipid oxidation and trans fatty acids (TFA) through cis-trans isomerization respectively. Plasma samples collected at 12–18 weeks (MIROS cohort; n =65) and 38–41 weeks of pregnancy (CHUL cohort; n =21) were subjected to alkaline hydrolysis followed by liquid–liquid extraction in order to extract total F 2 -isoPs for quantification by HPLC–MS/MS. Several positive correlations were found between F 2 -isoPs and TFA, measured by GC-FID in plasma phospholipids, such as 6 t -18:1, 9 t -18:1 and 9 t ,12 c -18:2 ( r >0.306; p t ,12 c -18:2 trans isomer, known to be associated to cardiovascular diseases, showed the most significant correlations with F 2 -isoPs. No correlation was observed between F 2 -isoPs and 9 t -16:1 or 11 t -18:1. In summary, this study suggests either a concomitant phenomenon or a competition between lipid peroxidation and cis - trans isomerisation of the cis precursor fatty acid in vivo during pregnancy.

Reports indicate that antioxidant enzymes like the glutathione peroxidases (GPx) can be regulated by sex steroids. The GPx, a major class of antioxidants involved in H(2)O(2) and lipid hydroperoxides neutralization, showed an age- and sex-specific expression in many adult organs including the lung. High levels of androgens in the male lung are known to delay the surge of surfactant synthesis during gestation in several species. However, the impact of male androgens on antioxidant GPx early in life remains to be determined. The objective was to study the lung sex-specific expression of GPx during BALB/c mouse perinatal development. The mRNA expression of four seleno-dependent Gpx (Gpx1 to 4) in the lung of both sexes was characterized by real-time PCR from gestational day 15 to postnatal day 30, covering the entire canalicular, saccular and alveolar stages. Immunohistochemistry of GPx-1, -3 and -4, and seleno-dependent GPx enzymatic assays were also performed in the lung. We found a transient lower Gpx1 mRNA level in male than in female lungs during the first 5 days after birth, corresponding to the saccular phase. This dimorphic expression was concomitant to a sex difference in GPx enzymatic activity corrected for blood. It is, to our knowledge, the first report of a sex dimorphism for murine lung enzymatic antioxidant defenses during the perinatal period.

Increased placental oxidative stress in preeclampsia (PE) has been associated in part to a decrease in glutathione peroxidase (GPX) antioxidant activity. However, it is not clear if GPX mRNA expression is affected in PE, and how the presence of labor may impair this expression. In this study, we characterized by quantitative PCR, in situ hybridization and immunohistochemistry, the expression of four GPX (GPX1 to 4) in the placenta of normotensive (NP; n = 23) and PE pregnancies (n = 25) according to mode of delivery: vaginal delivery (with labor) or cesarean (without labor); the tissue layer: amnion-chorion (AC) and villi; and the sampling site: peri-insertion or peripheral. Concomitantly, oxidative stress markers mRNA expression, HSP70 and HO-1 were measured. All GPX mRNA and protein were detected in all layers of the placenta and sampling sites. In absence of labor, GPX1 is more expressed near the umbilical cord than at the periphery of the villi (p = 0.037). At the periphery of AC membranes, GPX2 was more expressed in PE than in controls in presence of labor (p = 0.037). Interestingly, GPX4 mRNA level was clearly deficient in the PE villi in presence or absence of labor (p < 0.0473). Also, the GPX4 expression in PE was lower than controls in AC membranes in presence of labor (p = 0.0007). Oxidative stress markers, HSP70 and HO-1, were higher in PE placental membranes than in controls in absence of labor (p < 0.011). HSP70 was also upregulated in PE placental membranes in presence of labor (p = 0.034). Correlations between stress markers and GPX mRNA expression were mostly present in AC membranes in presence of labor in NP. Most of the latter correlations were lost in PE. In conclusion, our results suggest that the reported decrease in GPx activity and increased oxidative stress in PE placental villi may be attributed in part to GPX4 independently of the presence or absence of labor.

A decreased antioxidant activity for superoxide dismutases (SODs) in the placenta was reported in preeclampsia (PE). However, it is unclear if this reduced enzymatic activity can be attributed to a specific SOD isoform. Moreover, the specific spatial SOD expression in the placenta and the impact of the mode of delivery on the latter are still lacking. There are three known SOD isoforms: SOD1 (cytosolic), SOD2 (mitochondrial) and SOD3 (extracellular). Our main objective was to characterize by RT-PCR, western blot and immunolocalization, the expression of SOD1, SOD2, and SOD3 in placentas of normotensive (n = 23) and PE pregnancies (n = 25) according to the presence or absence of labor, the sampling site (peri-insertion, mid-disc and periphery) and the placental layer: amnion-chorion, villi, and maternal side layer (MS). In absence of labor (cesarean), SOD1 expression in the placental villi and MS was lower in PE than in controls (p0.049). In presence of labor (vaginal deliveries), SOD1 expression in the amnion-chorion only was higher in PE than controls (p = 0.014). Additionally, SOD2 and SOD3 expression in presence of labor were higher in all three layers in PE than controls, with a strong positive correlation between these two SODs (mRNA; r0.65, p0.008). The sampling site and gestational age had no effect on SOD expression within the placenta. In this study, we showed that the reported decrease for SOD activity in PE may be attributed to SOD1 in absence of labor. Also, this is the first study characterizing specific SOD isoforms according to the mode of delivery. We demonstrated in PE that labor upregulates SOD1 in fetal membranes as well as SOD2 and SOD3 in the whole placenta.

A new method for solid phase parallel synthesis of chemically and conformationally diverse macrocyclic peptidomimetics is reported. A key feature of the method is access to broad chemical and conformational diversity. Synthesis and mechanistic studies on the macrocyclization step are reported.

In models which allow for random jumps, statistical tests for jumps are typically non-standard and nuisance parameter-dependent. To handle these problems, we combine bounds and Monte-Carlo (MC) simulation techniques to derive nuisance-parameter-free bounds and obtain level-exact p-values for a wide class of processes with random jumps and time varying heteroskedasticity. When identified nuisance parameters are absent under the null, we show that MC p-values are finite sample, level-exact. To illustrate this easy-to-implement approach, we analyze the spot prices of four commodities (aluminium, copper, gold and lead) and the closing prices of four technology stocks (Intel, Microsoft, Oracle and Sun). We find significant jumps in these time series. Our approach can easily be extended to other nuisance-parameter dependent tests.

Preeclampsia is a leading cause of maternal and neonatal mortality and morbidity. It is a complex syndrome of undetermined etiologic origin, usually diagnosed during the second half of pregnancy, with clinical features of hypertension, proteinuria, and edema. No cure for preeclampsia exists, except premature delivery. There is increasing evidence that oxidative stress is an important contributing factor to the pathogenesis of preeclampsia. Oxidative stress is defined as an imbalance between reactive oxygen species (ROS), such as nitric oxide(NO · ), superoxide anion (O 2 ·↔ ), and hydrogen peroxide (H 2 O 2 ), and antioxidants, favouring an overabundance of ROS. The consequence of an overproduction of ROS can be observed as increased levels of markers of oxidative stress, such as lipid peroxides.Pregnant women affected by preeclampsia may have abnormal ROS production, particularly NO · and O 2 ·↔ , abnormal levels of antioxidant defences, and increased placental lipid peroxidation. Several observations suggest that decreased bioavailability of endothelium-derived NO · , due to oxidative destruction of NO · by ROS, might contribute to the impaired endothelium-dependent vasodilatory responses and multisystemie pathology of preeclampsia, a phenomenon in which antioxidant vitamins may play a beneficial role. This review focuses on the rationale for vitamins C and E supplementation toward prevention of preeclampsia, with an emphasis on the limit of our scientific knowledge concerning the deleterious oxidative events taking place in this pathology.

The synthesis of a 15-membered macrocyclic triene containing all the required substituents of ring A of (+)-aphidicolin (1) is reported. This compound underwent a thermal transannular cycloaddition followed by an intramolecular aldol reaction to yield tetracycle 32 containing 8 chiral centers which is considered a key intermediate for the synthesis of (+)-aphidicolin and related analogs.

Antioxidants may be beneficial additives to synthetic culture media because these well defined media lack serum or other macromolecules that serve as reactive oxygen species scavengers. In this study, three separate experiments were performed to determine the effects of antioxidants on the development of oocytes to the morula and blastocyst stage when added during in vitro maturation (IVM) of bovine oocytes, during in vitro fertilization (IVF), and during embryo culture for the first 72 h of the development period. Bovine oocytes were matured, fertilized (under 20% O(2)), and embryos were cultured (under 7% O(2)) in defined conditioned medium in vitro with or without supplementation with the antioxidant cysteine, N-acetyl-L-cysteine (NAC), catalase and superoxide dismutase (SOD). Significant improvements in the proportion of oocytes undergoing morula and blastocyst development (33.3% versus 20.3%, P

It is known that large amounts of leukocytes colonize the uterus, and that these leukocytes can produce considerable quantities of hydrogen peroxide (H2O2) and other reactive oxygen species that are toxic to sperm. It has been shown recently that oviductal fluid has a catalase that helps to maintain sperm motility. Therefore, the current experiment was performed to determine if a similar mechanism of protection exists against peroxides within uterine cells. Sperm motility and velocity were recorded after a 6h incubation in 1) conditioned media in the presence of endometrial cells, 2) conditioned media without endometrial cells, 3) control media (48h without cells) over endometrial cells, or 4) control media alone. All these treatments were performed in the presence or absence of added catalase. Conditioned media, endometrial cells and catalase had a significant positive effect on the maintenance of sperm motility and velocity. Addition of anti-catalase antibodies did not neutralize the beneficial effect of the conditioned media. However, the concentrations of aromatic amino acids, known substrates for sperm amino acid oxidase, were significantly lower in uterine conditioned media as compared to control medium. This reduction of aromatic amino acids was in correlation with reduced H2O2 production by sperm as estimated by chemiluminescence. These results suggest that epithelial and stromal uterine cells do not maintain sperm motility by secreting catalase in the conditioned media, but rather by reducing the levels of aromatic amino acids and thus of peroxides generated in the presence of spermatozoa.

Little is known about cell-cycle checkpoint activation by oxidative stress in mammalian cells. The effects of hyperoxia on cell-cycle progression were investigated in asynchronous human T47D-H3 cells, which contain mutated p53 and fail to arrest at G1/S in response to DNA damage. Hyperoxic exposure (95% O 2 , 40–64 h) induced an S-phase arrest associated with acute inhibition of Cdk2 activity and DNA synthesis. In contrast, exit from G2/M was not inhibited in these cells. After 40 h of hyperoxia, these effects were partially reversible during recovery under normoxic conditions. The inhibition of Cdk2 activity was not due to degradation of Cdk2, cyclin E or A, nor impairment of Cdk2 complex formation with cyclin A or E and p21 Cip1 . The loss of Cdk2 activity occurred in the absence of induction and recruitment of cdk inhibitor p21 Cip1 or p27 Kip1 in cyclin A/Cdk2 or cyclin E/Cdk2 complexes. In contrast, Cdk2 inhibition was associated with increased Cdk2-Tyr15 phosphorylation, increased E2F-1 recruitment, and decreased PCNA contents in Cdk2 complexes. The latter results indicate a p21 Cip1 /p27 Kip1 -independent mechanism of S-phase checkpoint activation in the hyperoxic T47D cell model investigated.