The Effects of Omega‐3 Polyunsaturated Fatty Acid Consumption on Mammary Carcinogenesis

The consumption of omega‐3 polyunsaturated fatty acids (n‐3 PUFA) is associated with a reduced risk of breast cancer. Studies in animals and in vitro have demonstrated mechanisms that could explain this apparent effect, but clinical and epidemiological studies have returned conflicting results on the practical benefits of dietary n‐3 PUFA for prevention of breast cancer. Effects are often only significant within a population when comparing the highest n‐3 PUFA consumption group to the lowest n‐3 group or highest n‐6 group. The beneficial effects of n‐3 PUFA eicosapentaenoic and docosahexaenoic on the risk of breast cancer are dose dependent and are negatively affected by total n‐6 consumption. The majority of the world population, including the most highly developed regions, consumes insufficient n‐3 PUFA to significantly reduce breast cancer risk. This review discusses the physiological and dietary context in which reduction of breast cancer risk may occur, some proposed mechanisms of action and meaningful recommendations for consumption of n‐3 PUFA in the diet of developed regions.     

Increased Prostaglandin Response to Oxytocin in Ewes Fed a Diet High in Omega-6 Polyunsaturated Fatty Acids

Diets high in omega-6 polyunsaturated fatty acids (n-6) are associated with increased prostaglandin F_2α (PGF_2α) synthesis in cattle, however, the specific effects on the potential prostaglandin response to an oxytocin challenge in sheep have not been reported. The aim of the current study was to determine whether oxytocin-stimulated PGF_2α was significantly increased when ewes were fed a diet high in n-6 compared with a control diet low in n-6. Merino x Border Leicester ewes (n = 30) received one of two dietary treatments, either high in n-6 (70 % oat grain) or low in n-6 (control diet, 100 % cereal/legume silage). Ewes consumed the diets for 44 days prior to two consecutive oxytocin challenges. Plasma n-6 and PGF_2α metabolite (PGFM) concentrations following oxytocin challenge were greater (P < 0.05) when ewes were fed a diet high in n-6 compared with the control diet. A higher availability of n-6 may have lead to an increased in vivo synthesis of PGF_2α, however, further research is required to determine the exact mechanisms involved.     

Dietary supplement of isohumulones inhibits the formation of aberrant crypt foci with a concomitant decrease in prostaglandin E2 level in rat colon

Male Fischer 344 rats were subcutaneously injected with azoxymethane (AOM) twice weekly at a dose of 15 mg/kg and were fed with freeze-dried (FD) samples of beer brewed without hops (non-hops beer), beer with hops at 4 times the amount of regular lager beer (x 4-hops beer), and isomerized hop extract (IHE) for the whole experimental period (I/PI) or for the post-initiation period (PI) only. Feeding FD beer samples at a dose of 1% significantly decreased the number of aberrant cryp foci (ACF) in the PI protocol over five weeks.x4-hops beer showed stronger inhibitory effects on the development of the numbers of aberrant crypts per focus and large ACF with four or more crypts than non-hops beer. Feeding IHE to rats at a dose of 0.01% or 0.05% in either the I/PI or PI experiment significantly reduced the numbers of ACF. Prostaglandin E2 (PGE2) levels in colonic mucosa of AOM-treated rats were significantly reduced by feeding of IHE. PGE2 production induced by lipopolysaccharide/interferon-gamma (LPS/IFN-gamma) in RAW264.7 cells was also reduced by treatment with IHE and isohumulone in a dose-dependent manner. These observations suggest that isohumulones show chemopreventive effects on ACF formation in rat colon by inhibiting the production of PGE2.     

Women and Smokers Have Elevated Urinary F2-Isoprostane Metabolites: A Novel Extraction and LC–MS Methodology

F(2)-Isoprostanes (F(2)-IsoPs), regio- and stereoisomers of prostaglandin F(2alpha) (PGF(2alpha)), and urinary F(2)-IsoP metabolites including 2,3-dinor-5,6-dihydro-8-iso-PGF(2alpha) [2,3-dinor-8-iso-PGF(1alpha) (2,3-dinor-F1)] and 2,3 dinor-8-iso-PGF(2alpha) (2,3-dinor-F2), have all been used as biomarkers of oxidative stress. A novel method was developed to measure these biomarkers using a single solid phase extraction (SPE) cartridge, separation by HPLC, and detection by negative mode selected reaction monitoring (SRM) mass spectrometry (MS), using authentic standards of PGF(2alpha); 8-iso-PGF(2alpha); 2,3-dinor-F1 and 2,3-dinor-F2 to identify specific chromatographic peaks. The method was validated in a population of healthy, college-aged nonsmokers (n = 6 M/8F) and smokers (n = 6 M/5F). Urinary F(2)-IsoP concentrations were approximately 0.2-1.5 microg/g creatinine, 2,3-dinor-F1 was approximately1-3 microg/g and 2,3-dinor-F2 was approximately 3-5 microg/g. Additional F(2)-IsoPs metabolites were identified using SRM. The sum of all urinary F(2)-IsoP metabolites was 50-100 microg/g creatinine indicating their greater abundance than F(2)-IsoPs. Women had higher F(2)-IsoP metabolite concentrations than did men (MANOVA, main effect P = 0.003); cigarette smokers had higher concentrations than did nonsmokers (main effect P = 0.036). For men or women, respectively, smokers had higher metabolite concentrations than did nonsmokers (P < 0.05). Thus, our method simultaneously allows measurement of urinary F(2)-IsoPs and their metabolites for the determination of oxidative stress.     

CLA Reduces Inflammatory Mediators from A427 Human Lung Cancer Cells and A427 Conditioned Medium Promotes Differentiation of C2C12 Murine Muscle Cells

Conjugated linoleic acid (CLA) is thought to have anti-proliferative and anti-inflammatory properties, but its effect on cancer cachexia is unknown. Two effects were here investigated: that of CLA on inflammatory mediator production in human lung cancer cells, and that of reduced mediators on the myogenic differentiation of murine muscle C2C12 cells. The latter cells were grown in medium conditioned by human lung cancer A427 cells, with or without CLA, to mimic only the effect of molecules released from the tumor “in vivo”, excluding the effect of host-produced cachectic factors. The results obtained show that CLA was found to reduce the production of tumor necrosis factor-α, interleukin (IL)-1β and prostaglandin E2 (PGE2), but had no effect on IL-6 production. The mechanisms underlying the effect of CLA on cytokine or PGE2 release in A427 cells are probably mediated by activation of peroxisome proliferator-activated receptor (PPAR)α, which increased at 24 h CLA treatment. In turn, the reduced content of inflammatory mediators in medium conditioned by A427 cells, in the presence of CLA, allowed muscle cells to proliferate, again by inducing PPAR. The involvement of PPARα was demonstrated by treatment with the antagonist MK-886. The findings demonstrate the anti-inflammatory and myogenic action of CLA and point to its possible application as a novel dietary supplement and therapeutic agent in inflammatory disease states, such as cachexia.     

Development and applications of quaternary ammonium (QA) membrane electrodes in pharmaceutical preparation and in bioavailability of Prostaglandin E1 and Deoxycholate

The two novel ion-pairs (PB-TPB and NB-TPB) of quaternary ammonium drugs; propantheline bromide (PB), N,N-Diisopropyl-N-methyl-N-[2-(xanthen-9ylcarbonyloxy)ethyl] ammonium bromide and neostigmine bromide (NB), 3-(dimethylcarbamoyloxy) phenyl]-trimethylazanium have been synthesized, respectively and incorporated in poly (vinyl chloride)-based membrane electrodes for the quantification of propantheline bromide and neostigmine bromide in different pharmaceutical preparations. The influences of membrane compositions on the potentiometric responses of membrane electrodes have been found to substantially improve the performance characteristics. The best performance was reported with membranes having composition (w/w) of PB-TPB or NB-TPB (6%): PVC (34%): o-NPOE (60%). The proposed electrodes exhibit nernstian response in the concentration ranges of 2.1 × 10⁻⁷ M to 1.0 × 10⁻² M and 4.4 × 10⁻⁷ M to 1.0 × 10⁻² M with detection limit of 1.5 × 10⁻⁷ M and 3.3 × 10⁻⁷ M, respectively. Both the membrane electrodes perform satisfactorily over pH ranges of (3.5–7.5 and 4.0–7.0) with fast response times (11 s and 13 s), respectively. These drugs (PB and NB) were further utilized as different ion-pairs of Prostaglandin E₁ (PGE₁) and Deoxycholate (DOC) in poly (vinyl chloride)-based membrane electrodes for the determination of bioavailability of Prostaglandin E₁ and Deoxycholate in plasma of different patients.     

丹参饮促进胃溃疡愈合作用及其机理研究

目的: 观察丹参饮对大鼠乙酸性胃溃疡的影响并探讨其作用机制。方法: 应用大鼠乙酸性胃溃疡模型, 给予丹参饮治疗14 d 后, 观察胃粘膜损伤程度(UI), 检测血清NO含量和血浆PGE₂水平, 测定胃壁结合粘液的含量、溃疡边缘粘膜细胞凋亡(AI)、凋亡相关基因Bcl-2 以及表皮生长因子受体(EGFR) 的表达。结果: 与模型组相比, 丹参饮可明显促进大鼠溃疡愈合(P <0.01), 提高血清NO 、血浆PGE₂ 和胃壁结合粘液含量(P < 0.05), 并显著抑制胃溃疡边缘粘膜细胞凋亡(P <0.01), 同时促进EGFR 和凋亡抑制基因Bcl-2 的表达, 其中7、3.5g·kg^-1剂量组作用更为显著(P < 0.01) 。结论: 通过胃粘膜防御因子及抑制凋亡途径来促进溃疡愈合可能是丹参饮治疗胃溃疡的作用机制之一。     

Lipocalin‐Type Prostaglandin D Synthase Is a Novel Phytocannabinoid‐Binding Protein

Lipocalin‐type prostaglandin D synthase (L‐PGDS; EC:5.3.99.2) is an enzyme with dual functional roles as a prostaglandin D₂‐synthesizing enzyme and as an extracellular transporter for diverse lipophilic compounds in the cerebrospinal fluid (CSF). Transport of hydrophobic endocannabinoids is mediated by serum albumin in the blood and intracellularly by the fatty acid binding proteins, but no analogous transport mechanism has yet been described in CSF. L‐PGDS has been reported to promiscuously bind a wide variety of lipophilic ligands and is among the most abundant proteins found in the CSF. Here, we examine the binding of several classes of endogenous and synthetic ligands to L‐PGDS. Endocannabinoids exhibited low affinity toward L‐PGDS, while cannabinoid metabolites and synthetic cannabinoids displayed higher affinities for L‐PGDS. These results indicate that L‐PGDS is unlikely to function as a carrier for endocannabinoids in the CSF, but it may bind and transport a subset of cannabinoids.     

A Fast One-Step Extraction and UPLC–MS/MS Analysis for E2/D2 Series Prostaglandins and Isoprostanes

Prostaglandins (PG) and isoprostanes (iso-PG) may be derived through cyclooxygenase or free radical pathways and are important signaling molecules that are also robust biomarkers of oxidative stress. Their quantification is important for understanding many biological processes where PG, iso-PG, or oxidative stress are involved. One of the common methods for PG and iso-PG quantifications is LC–MS/MS that allows a highly selective, sensitive, simultaneous analysis for prostanoids without derivatization. However, the currently used LC–MS/MS methods require a multi-step extraction and a long (within an hour) LC separation to achieve simultaneous separation and analysis of the major iso-PG. The developed and validated for brain tissue analysis one-step extraction protocol and UPLC–MS/MS method significantly increases the recovery of the PG extraction up to 95 %, and allows for a much faster (within 4 min) major iso-PGE₂ and -PGD₂ separation with 5 times narrower chromatographic peaks as compared to previously used methods. In addition, it decreases the time and cost of analysis due to the one-step extraction approach performed in disposable centrifuge tubes. All together, this significantly increases the sensitivity, and the time and cost efficiency of the PG and iso-PG analysis.