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Journal of Lipid Research Jul 2013Western diets are enriched in omega-6 vs. omega-3 fatty acids, and a shift in this balance toward omega-3 fatty acids may have health benefits. There is limited...
Western diets are enriched in omega-6 vs. omega-3 fatty acids, and a shift in this balance toward omega-3 fatty acids may have health benefits. There is limited information about the catabolism of 3-series prostaglandins (PG) formed from eicosapentaenoic acid (EPA), a fish oil omega-3 fatty acid that becomes elevated in tissues following fish oil consumption. Quantification of appropriate urinary 3-series PG metabolites could be used for noninvasive measurement of omega-3 fatty acid tone. Here we describe the preparation of tritium- and deuterium-labeled 6-keto-PGF2α and their use in identifying urinary metabolites in mice using LC-MS/MS. The major 6-keto-PGF2α urinary metabolites included dinor-6-keto-PGF2α (~10%) and dinor-13,14-dihydro-6,15-diketo-PGF1α (~10%). These metabolites can arise only from the enzymatic conversion of EPA to the 3-series PGH endoperoxide by cyclooxygenases, then PGI3 by prostacyclin synthase and, finally, nonenzymatic hydrolysis to 6-keto-PGF2α. The 6-keto-PGF derivatives are not formed by free radical mechanisms that generate isoprostanes, and thus, these metabolites provide an unbiased marker for utilization of EPA by cyclooxygenases.
Topics: Animals; Chromatography, Liquid; Deuterium; Dinoprost; Humans; Isotope Labeling; Male; Mice; Mice, Inbred C57BL; Molecular Structure; Tandem Mass Spectrometry; Tritium
PubMed: 23644380
DOI: 10.1194/jlr.M037192 -
Scientific Reports May 2020Heart failure (HF) is a cardiovascular disease affecting about 26 million people worldwide costing about $100 billons per year. HF activates several compensatory...
Heart failure (HF) is a cardiovascular disease affecting about 26 million people worldwide costing about $100 billons per year. HF activates several compensatory mechanisms and neurohormonal systems, so we hypothesized that the concomitant monitoring of a panel of potential biomarkers related to such conditions might help predicting HF evolution. Saliva analysis by point-of-care devices is expected to become an innovative and powerful monitoring approach since the chemical composition of saliva mirrors that of blood. The aims of this study were (i) to develop an innovative procedure combining MEPS with UHPLC-MS/MS for the simultaneous determination of 8-isoprostaglandin F and cortisol in saliva and (ii) to monitor lactate, uric acid, TNF-α, cortisol, α-amylase and 8-isoprostaglandin F concentrations in stimulated saliva samples collected from 44 HF patients during their hospitalisation due to acute HF. Limit of detection of 10 pg/mL, satisfactory recovery (95-110%), and good intra- and inter-day precisions (RSD ≤ 10%) were obtained for 8-isoprostaglandin F and cortisol. Salivary lactate and 8-isoprostaglandin F were strongly correlated with NT-proBNP. Most patients (about 70%) showed a significant decrease (a factor of 3 at least) of both lactate and 8-isoprostaglandin F levels at discharge, suggesting a relationship between salivary levels and improved clinical conditions during hospitalization.
Topics: Aged; Area Under Curve; Biomarkers; Dinoprost; Heart Failure; Hospitalization; Humans; Lactic Acid; Limit of Detection; Middle Aged; Pilot Projects; ROC Curve; Reproducibility of Results; Saliva
PubMed: 32366899
DOI: 10.1038/s41598-020-64112-2 -
Laboratory Investigation; a Journal of... Feb 2008F2-isoprostanes are considered as the most reliable markers of oxidative stress and can be used to evaluate the oxidative status in a number of human pathologies....
F2-isoprostanes are considered as the most reliable markers of oxidative stress and can be used to evaluate the oxidative status in a number of human pathologies. Besides being markers of oxidative stress, F2-isoprostanes proved to be mediators of important biological effects and would act through the activation of receptors analogous to those for thromboxane A2. In a previous work, we provided evidence that F(2)-isoprostanes, generated during carbon tetrachloride-induced hepatic fibrosis, mediate hepatic stellate cell (HSC) proliferation and collagen hyperproduction. To investigate whether TxA2 receptor (TxA2r or TPr) is involved in the effects of F2-isoprostanes on HSC, experiments on DNA synthesis were carried out in the presence of 8-epi-prostaglandin F(2alpha) (8-epi-PGF(2alpha)) or the TxA2r-specific agonist I-BOP ([1S-[1alpha,2alpha(Z),3beta(1E,3S*), 4alpha]]-7-[3-[3-hydroxy-4-(4-iodophenoxy)-1-butenyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid). Both agonists significantly stimulated DNA synthesis, which was almost completely inhibited by the TxA2r-specific antagonist SQ29548 ([1S-[1alpha,2alpha(Z),3alpha,4alpha]]-7-[3-[[2-[(phenylamino)carbonyl] hydrazino] methyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5-heptanoic acid), suggesting that much of the effect of 8-epi-PGF(2alpha) is mediated by the TxA2r. Further studies showed that increasing concentrations of SQ29548 progressively inhibit DNA synthesis, suggesting a possible competitive antagonism between the two molecules. In addition, we demonstrated that the stimulatory effect of 8-epi-PGF(2alpha) on collagen synthesis could be mediated by TxA2r. The occurrence of TxA2r on HSC was also investigated using western blotting analysis and immunocytochemistry, which reveals that TP is distributed both on plasma membranes and within the cells. Moreover, binding studies indicated the presence of a specific binding site for 3H-SQ29548 on HSC. Competition binding studies indicated that 8-epi-PGF(2alpha) and I-BOP were both able to displace 3H-SQ29548 binding with a very different affinity (K(i)=4.14+/-1.9 x 10(-6) M and K(i)=1.15+/-0.3 x 10(-9) M, respectively), suggesting the involvement of a modified form of isoprostane receptor, homologous to the classic thromboxane A2-binding site in F2-isoprostanes-evoked responses on HSC.
Topics: Binding, Competitive; Bridged Bicyclo Compounds, Heterocyclic; Cells, Cultured; Collagen; DNA; Dinoprost; F2-Isoprostanes; Fatty Acids, Unsaturated; Humans; Hydrazines; Liver; Pericytes; Receptors, Thromboxane A2, Prostaglandin H2; Tritium; Vasoconstrictor Agents
PubMed: 18158556
DOI: 10.1038/labinvest.3700712 -
Transfusion Jun 2014Stored red blood cells (RBCs) release hemoglobin (Hb) that leads to oxidative damage, which may contribute to thrombosis in susceptible transfusion recipients. Oxidative...
BACKGROUND
Stored red blood cells (RBCs) release hemoglobin (Hb) that leads to oxidative damage, which may contribute to thrombosis in susceptible transfusion recipients. Oxidative stress stimulates the generation of a new class of lipid mediators called F2 -isoprostanes (F2 -IsoPs) and isofurans (IsoFs) that influence cellular behavior. This study investigated RBC-derived F2 -IsoPs and IsoFs during storage and their influence on human platelets (PLTs).
STUDY DESIGN AND METHODS
F2 -IsoP and IsoF levels in RBC supernatants were measured by mass spectrometry during storage and after washing. The effects of stored supernatants, cell-free Hb, or a key F2 -IsoP, 8-iso-prostaglandin F2α (PGF2α ), on PLT function were examined in vitro.
RESULTS
F2 -IsoPs, IsoFs, and Hb accumulated in stored RBC supernatants. Prestorage leukoreduction reduced supernatant F2 -IsoPs and IsoFs levels, which increased again over storage time. Stored RBC supernatants and 8-iso-PGF2α induced PLT activation marker CD62P (P-selectin) expression and prothrombotic thromboxane A2 release. Cell-free Hb did not alter PLT mediator release, but did inhibit PLT spreading. Poststorage RBC washing reduced F2 -IsoP and IsoF levels up to 24 hours.
CONCLUSIONS
F2 -IsoPs and IsoFs are produced by stored RBCs and induce adverse effects on PLT function in vitro, supporting a potential novel role for bioactive lipids in adverse transfusion outcomes. F2 -IsoP and IsoF levels could be useful biomarkers for determining the suitability of blood components for transfusion. A novel finding is that cell-free Hb inhibits PLT spreading and could adversely influence wound healing. Poststorage RBC washing minimizes harmful lipid mediators, and its use could potentially reduce transfusion complications.
Topics: Blood Platelets; Dinoprost; Erythrocytes; F2-Isoprostanes; Furans; Humans; Immunoassay; Isoprostanes; Reactive Oxygen Species
PubMed: 24192515
DOI: 10.1111/trf.12485 -
Journal of Dental Research Dec 2015In a previous report, we demonstrated the inverse association of high serum 8-isoprostane levels, a marker for oxidative stress, with decreased serum IgG antibodies to...
In a previous report, we demonstrated the inverse association of high serum 8-isoprostane levels, a marker for oxidative stress, with decreased serum IgG antibodies to oral bacteria. The association between increased serum IgG with increased plaque and periodontitis (increased probing depths) was attenuated by high systemic oxidative stress. Other investigations have reported a role for systemic oxidative stress as a stimulus of hepatic C-reactive protein (CRP) response. These observations led us to hypothesize that the reported relationship of periodontitis to elevated serum CRP, a systemic inflammatory marker, may be modified by oxidative stress and that the levels of serum antibodies to oral bacteria might be an intermediary explanatory variable linking the association of systemic oxidative stress, periodontal disease, and levels of CRP. This hypothesis was explored as a secondary analysis of the Dental ARIC (Atherosclerosis Risk in Communities) study using serum levels of CRP, serum IgG levels to 16 oral organisms, serum levels of 8-isoprostane, and periodontal status. The findings indicate periodontitis is associated with high CRP in the presence of elevated oxidative stress that serves to suppress the IgG response. Only within the highest 8-isoprostane quartile was periodontitis (pocket depth) associated with increased serum CRP levels (P = 0.0003). Increased serum IgG antibody levels to oral bacteria were associated with lowered serum CRP levels. Thus, systemic oxidative stress, which has been demonstrated to be associated with increased levels of CRP in other studies, appears to be associated with the suppression of bacterial-specific IgG levels, which in the presence of periodontal disease can result in an enhanced systemic CRP response. Conversely, individuals with increased serum IgG antibodies to plaque bacteria exhibit lowered serum CRP levels. These 2 factors, oxidative stress and the serum IgG response, appear to function in opposing directions to modify serum levels of CRP and the association with periodontitis.
Topics: Biofilms; C-Reactive Protein; Dinoprost; Female; Humans; Immunoglobulin G; Male; Middle Aged; Mouth; Oxidative Stress; Periodontitis; Prospective Studies
PubMed: 26318589
DOI: 10.1177/0022034515602693 -
Journal of the American Veterinary... Apr 2002To assess the efficacy and safety of 2 protocols using bromocriptine mesylate and prostaglandins to terminate unwanted pregnancy in bitches. (Clinical Trial)
Clinical Trial Randomized Controlled Trial
OBJECTIVE
To assess the efficacy and safety of 2 protocols using bromocriptine mesylate and prostaglandins to terminate unwanted pregnancy in bitches.
DESIGN
Prospective randomized single-blind controlled study.
ANIMALS
34 crossbred and purebred bitches referred for possible pregnancy termination. Seven additional pregnant bitches were used as controls.
PROCEDURE
Pregnancy was assessed by ultrasonographic examination from day 25 after mating in all bitches. Of the 34 bitches, 25 were pregnant and were randomly allocated to a treatment group. Group-1 dogs (n = 12) received a combination of increasing amounts of bromocriptine mesylate (15 to 30 microg/kg [6.8 to 13.6 microg/lb], p.o., q 12 h) and dinoprost tromethamine (0.1 to 0.2 mg/kg [0.045 to 0.09 mg/lb], s.c., q 24 h). Group-2 dogs (n =13) received a combination of increasing amounts of bromocriptine mesylate (the same schedule as group-1 dogs) and cloprostenol sodium (1 microg/kg [0.45 microg/lb], s.c., q 48 h). Both groups were treated until pregnancy termination. Results-Treatment success was 100% in both groups. Days of treatment required for pregnancy termination did not significantly differ between groups (5.0 +/- 0.6 vs 3.7 +/- 0.6 days, group-1 and group-2 dogs, respectively) although adverse effects only developed in group-1 dogs. At the end of the protocols, pseudopregnancy was observed in 3 of 12 and 6 of 13 group-1 and group-2 dogs, respectively. Pregnancy termination was followed by a mucoid sanguineous vulvar discharge for 3 to 10 days.
CONCLUSIONS AND CLINICAL RELEVANCE
Results of this study indicate that protocols that combine the use of bromocriptine mesylate and prostaglandins for the termination of unwanted pregnancy in bitches are efficient and safe. The use of bromocriptine mesylate and cloprostenol had the best results and could be easily used on an outpatient basis.
Topics: Abortifacient Agents, Nonsteroidal; Abortion, Induced; Abortion, Veterinary; Animals; Bromocriptine; Cloprostenol; Dinoprost; Dogs; Female; Pregnancy; Prospective Studies; Prostaglandins; Safety; Single-Blind Method; Treatment Outcome
PubMed: 12420779
DOI: 10.2460/javma.2002.220.1017 -
Cells Nov 2021Mild hypoglycemia is common in clinical practice. Severe hypoglycemia results in heat shock protein and associate co-chaperone changes. Whether mild prolonged... (Clinical Trial)
Clinical Trial
Mild hypoglycemia is common in clinical practice. Severe hypoglycemia results in heat shock protein and associate co-chaperone changes. Whether mild prolonged hypoglycemia elicits a similar response with inflammatory and oxidative-stress responses compared with a severe hypoglycemic event is unclear; therefore, this pilot exploratory study was undertaken. We performed a case-control induced hypoglycemia clamp study, maintaining blood glucose at 2.8 mmol/L (50 mg/dL) for 1 h in 17 subjects (T2D ( = 10); controls ( = 7)). Blood sampling was performed at baseline, hypoglycemia, and 24 h; slow off-rate modified aptamer (SOMA)-scan plasma protein analysis of HSP-related proteins, inflammatory stress markers, and oxidative stress markers was performed. In total, 16 HSPs were analyzed. At baseline, TLR4:MD-2 complex was elevated ( = 0.01), whilst HSPA8 was lower ( < 0.05) in T2D. At hypoglycemia, UBE2N, STIP1, and UBE2L3 increased (all < 0.05), whilst TLR4:MD-2 and HSPA8 decreased ( < 0.05) in T2D versus baseline. In follow-up after hypoglycemia, HSPs normalized to baseline by 24 h, except UBE2L3 ( < 0.05), which was decreased in controls versus baseline. Correlation of altered inflammatory markers with HSPs revealed the following: at baseline, TLR4:MD-2 correlated with CXCL10 ( < 0.01) and SIGLEC1 ( < 0.05) in controls; HSPA8 negatively correlated with IL5 ( < 0.05) in T2D. A negative correlation between urinary isoprostane 8-iso PGF2α, a marker of oxidative stress, and HSPA1A was seen at 24 h in T2D only ( < 0.05). In conclusion, the HSP changes seen for mild prolonged hypoglycemia were similar to those previously reported for a severe event. However, mild prolonged hypoglycemia appeared to elicit an increased inflammatory response that was associated with heat shock and related proteins.
Topics: Adult; Biomarkers; Case-Control Studies; Diabetes Mellitus, Type 2; Dinoprost; Female; Heat-Shock Proteins; Heat-Shock Response; Humans; Hypoglycemia; Inflammation; Male; Middle Aged; Oxidative Stress; Protein Interaction Mapping; Proteins; Ubiquitin-Conjugating Enzymes
PubMed: 34831332
DOI: 10.3390/cells10113109 -
Kidney International Aug 1999Free-radical-generated F2-isoprostane stimulates DNA synthesis and endothelin-1 (ET-1) expression on endothelial cells. 8-Iso-prostaglandin F2alpha (8-iso-PGF2alpha) is...
BACKGROUND
Free-radical-generated F2-isoprostane stimulates DNA synthesis and endothelin-1 (ET-1) expression on endothelial cells. 8-Iso-prostaglandin F2alpha (8-iso-PGF2alpha) is a member of the recently discovered family of prostanoids, the F2-isoprostanes, produced in vivo by cyclooxygenase-independent, free-radical-catalyzed lipid peroxidation. The goal of our study is to establish the effect of isoprostane on ET-1 production by endothelial cells, as well to determine the receptors responsible for these effects.
METHODS
The proliferative effect of isoprostanes was measured as an increase of viable cell number and [3H]-thymidine uptake. ET-1 gene expression and protein synthesis were determined by Northern blot and radioimmunoassay, respectively. We also determined inositol 1,4,5-trisphosphate synthesis. Thromboxane A2 (TXA2) receptor antagonist SQ29,548 was used to establish the role of TXA2 receptor in isoprostane effect, as well as to determine the type of receptors involved in these effects.
RESULTS
Our results show that physiological concentrations of 8-iso-PGF2alpha stimulated cell proliferation, DNA synthesis, and ET-1 mRNA and protein expression in bovine aortic endothelial cells (BAECs). The proliferative effect was partially abolished by treatment with anti-endothelin antibody. 8-Iso-PGF2alpha also increased inositol 1, 4,5-trisphosphate formation in these cells. These effects were partially inhibited by SQ29,548. In competitive binding assays, two binding sites were recognized on BAECs with dissociation constants (Kd) and binding site densities at equilibrium similar to those previously described in smooth muscle cells and likely represent [3H]-8-iso-PGF2alpha binding to its own receptor (high-affinity binding site) and cross-recognition of the TXA2 receptor (low-affinity binding site).
CONCLUSION
These studies expand the potential scope of the pathophysiologic significance of F2-isoprostanes, released during oxidant injury, to include alteration of endothelial cell biology.
Topics: Animals; Aorta; Binding, Competitive; Cattle; Cell Count; Cell Division; Cell Survival; Cells, Cultured; DNA; Dinoprost; Endothelin-1; Endothelium, Vascular; F2-Isoprostanes; Free Radicals; Gene Expression; Inositol 1,4,5-Trisphosphate; Lipid Peroxidation; Oxidative Stress; RNA, Messenger; Receptors, Thromboxane; Signal Transduction; Tritium; Vasoconstrictor Agents
PubMed: 10432385
DOI: 10.1046/j.1523-1755.1999.00596.x -
The Journal of Reproduction and... Jun 2024The number of cows in estrus often influences estrus behavior; however, the effects of social order are not well documented. This study examined the effects of social...
The number of cows in estrus often influences estrus behavior; however, the effects of social order are not well documented. This study examined the effects of social order on the expression of behaviorally-scored and pedometer-detected estrus, combined with the effects of the number of cows in estrus. In a herd comprising 13 or 15 beef cattle, cows with orders 1st-7th were defined as dominant and the remaining cows as subordinate. Sole or simultaneous estrus was induced by prostaglandin F analog injection and/or intravaginal progesterone treatment. Ovulation timing was determined using ultrasonography at 6-hour intervals. Estrous signs and steps of the cows were recorded 49 h before ovulation using video monitoring and a pedometer, respectively. Among the 59 treated cows, 56 behaviorally-scored estruses (27 sole and 29 simultaneous) were detected. In the sole estrus, 61.5% of the dominant-rank cows had no zero-point period; however, 35.7% of the subordinate-rank cows had that period. The dominant-rank cows in estrus alone had a significantly shorter duration of scored estrus than those in simultaneous estrus (P < 0.05). Among the 50 pedometer-detected estruses (24 sole and 26 simultaneous), the subordinate-rank cows in sole estrus had a shorter interval from estrus onset to ovulation than the dominant-rank cows in simultaneous estrus (P < 0.05). The effects of social order varied in response to the number of cows in estrus, which might have influenced determining the optimal time for artificial insemination.
Topics: Animals; Cattle; Female; Estrus; Estrus Detection; Ovulation; Behavior, Animal; Progesterone; Social Behavior; Estrus Synchronization; Dinoprost
PubMed: 38508766
DOI: 10.1262/jrd.2024-005 -
Journal of Dairy Science Nov 2020The objective of this randomized controlled experiment was to evaluate reproductive performance and reproductive physiological outcomes of lactating Holstein cows...
Lactating dairy cows managed for second and greater artificial insemination services with the Short-Resynch or Day 25 Resynch program had similar reproductive performance.
The objective of this randomized controlled experiment was to evaluate reproductive performance and reproductive physiological outcomes of lactating Holstein cows managed for second and greater artificial insemination (AI) services with the Short-Resynch or Day 25 Resynch program. Cows from 2 commercial farms were randomly assigned after first service to the Short-Resynch (SR; n = 870) or Day 25 Resynch (D25R; n = 917) program in which they remained until 210 d after first service or left the herd. Cows in D25R received GnRH 25 ± 3 d after AI, whereas cows in SR did not. Cows not reinseminated at detected estrus (AIE) by 32 ± 3 d after AI underwent nonpregnancy diagnosis (NPD) through transrectal ultrasonography (TUS). Nonpregnant cows from both treatments with a corpus luteum (CL) ≥15 mm and an ovarian follicle ≥10 mm (hereafter, CL cows) received 2 PGF treatments 24 h apart, GnRH 32 h after the second PGF, and timed AI 16 to 18 h later. Cows that did not meet the criteria to be included in the CL group (NoCL cows) received a modified Ovsynch protocol with progesterone (P4) supplementation [P4-Ovsynch; GnRH and controlled internal drug-release device (CIDR) in, 7 d later CIDR removal and PGF, 24 h later PGF, 32 h later GnRH, and 16 to 18 h later timed AI]. In a subgroup of cows, blood samples were collected and TUS conducted at each treatment to evaluate ovarian responses to resynchronization. Binary data were analyzed with logistic regression, continuous data by ANOVA, and time-to-event data by Cox's proportional hazard regression. A greater proportion (mean; 95% CI) of cows were AIE before NPD in the SR (60.5%; 57.0-63.8; n = 3,416) than the D25R (50.1%; 46.5-53.7; n = 3,177) treatment, whereas pregnancy per AI (P/AI) at 32 d for AIE services before NPD was greater for the D25R (41.3%; 38.8-43.8; n = 1,560) than the SR (37.6%; 35.5-39.8; n = 1,961) treatment. At NPD, a greater proportion of cows in the D25R (84.3%; 82.2-86.2) than the SR (77.0%; 74.4-79.4) treatment were considered CL cows. Pregnancy per AI at 32 d was greater for the D25R than the SR treatment for all timed AI services (D25R = 43.0%; 40.2-45.9 vs. SR = 36.8%; 33.8-39.8) and for CL cows (D25R = 42.8%; 39.7-45.9 vs. SR = 33.8%; 30.6-37.2) but did not differ for NoCL cows (D25R = 39.4%; 32.1-47.3 vs. SR = 44.0%; 36.8-51.4). The hazard ratio for time to pregnancy (1.03; 0.93-1.14) and the proportion of cows not pregnant at the end of the observation period (D25R = 5.9%; 4.4-7.8 vs. SR = 6.7%; 5.0-8.7) did not differ between SR and D25R treatments. The GnRH treatment 25 d after AI resulted in more cows with P4 >1 ng/mL (D25R = 80.5%; 75.3-84.9 vs. SR = 63.6%; 57.3-69.4) and smaller follicle diameter at NPD 32 ± 3 d after AI for D25R (16.2 ± 0.4 mm) than for SR (17.5 ± 0.4 mm); however, it did not affect follicle diameter and luteal regression risk (CL cows only) before TAI. We concluded that the use of reproductive management programs including SR and D25R for CL cows and the P4-Ovsynch protocol for NoCL cows resulted in similar hazard of pregnancy and proportion of nonpregnant cows for up to 210 d after first service.
Topics: Animals; Cattle; Dinoprost; Drug Administration Schedule; Estrus Synchronization; Female; Gonadotropin-Releasing Hormone; Insemination, Artificial; Lactation; Pregnancy; Reproduction
PubMed: 32921468
DOI: 10.3168/jds.2020-18607