Malnutrition promotes prostaglandin over leukotriene production and dysregulates eicosanoid-cytokine crosstalk in activated resident macrophages

We previously described a murine model of malnutrition that mimicked features of moderate human malnutrition, and led to increased dissemination of Leishmania donovani. In this study, we investigated the effect of malnutrition on macrophage production of cytokines, prostaglandins (PGs), and leukotrienes (LTs). Using either LPS or calcium ionophore A23187 as a stimulus, macrophages from the malnourished mice produced a 3-fold higher PG/LT ((PGE₂+6-keto-PGF_1α)/(LTB₄+cysteinyl leukotrienes)) ratio than macrophages from well-nourished mice. LPS-stimulated macrophages from the malnourished mice produced decreased levels of TNF-α, GM-CSF, and IL-10, but similar levels of IL-6 and NO compared to well-nourished mice. A complex crosstalk between the eicosanoids and cytokines in the LPS-stimulated macrophages from the malnourished mice was evident by the following: (1) high levels of PG secretion despite low levels of TNF-α; (2) supplemental IL-10 modulated the excessive PG production; (3) GM-CSF rectified the PG/LT ratio, but did not correct the abnormal cytokine profile; and (4) inhibitors of cyclooxygenase decreased the PG/LT ratio, but did not affect TNF-α. Thus, in this model of malnutrition, there is a relative increase in anti-inflammatory PGs compared to pro-inflammatory LTs, which may contribute to immunodeficiency.     

Stable isotope dilution gas chromatography/mass spectrometry of prostaglandins and leukotrienes

The group of arachidonic acid metabolites comprising the prostaglandins, thromboxanes, and leukotrienes (eicosanoids) are extremely potent, biologically active compounds. Their properties include proaggregatory anti-aggregatory activity for platelets, chemotactic activity for neutrophils, vasoactive activity, and contractile activity to smooth muscle. In order to determine the role of these substances in pathophysiological conditions, it is essential to have highly sensitive methods available for their analysis. It is generally accepted that combined gas chromatography/mass spectrometry is the most specific technique available for the quantitative analysis of eicosanoids. However, methods based on electron impact ionization and positive ion chemical ionization are relatively insensitive, and many investigators have preferred the use of less specific but more sensitive methods based on radioimmunoassay. We have explored the use of negative ion chemical ionization mass spectrometry to improve sensitivity coupled with capillary column chromatography to maximize specificity. Conversion of the terminal carboxyl group (present in all eicosanoids) to the pentafluorobenzyl ester derivative confers excellent electron capturing properties to the molecule. The derivative undergoes highly efficient thermal electron capture in the gas phase, and any fragmentation that occurs subsequently is directed almost entirely away from the analyte molecule. The stabilized carboxylate anion that results carries at least 30% of the total ion current. Using selected ion monitoring techniques it is possible to detect eicosanoids in the range 1–8 pg on column. This methodology has been applied to the development of stable isotope dilution assays for plasma 6-oxo-prostaglandin (PG) F_1α (1) and for the simultaneous analysis of six biologically important PGs in biological fluids (2). In addition, stable isotope dilution techniques have been developed for the analysis of serum thromboxane B₂ and serum leukotriene B₄ (3). The application of this technology to understanding the role of arachidonic acid metabolism in humans will be discussed.     

Biosynthesis and functions of eicosanoids generated by the coelomocytes of the starfish, Asterias rubens

Eicosanoids are a group of oxygenated fatty acid derivatives formed from C20 polyunsaturated fatty acids, including arachidonic and eicosapentaenoic acids. The potential of the coelomocytes of the starfish, Asterias rubens, to generate eicosanoids through the cyclooxygenase (COX) and lipoxygenase (LOX) pathways was investigated using reverse-phase high performance liquid chromatography, enzyme immunoassay and gas chromatography–mass spectrometry. The principal LOX product was identified as 8-hydroxyeicosatetraenoic acid (8-HETE) with 8-hydroxyeicosapentaenoic acid (8-HEPE) synthesised at significantly lower levels. No classical prostaglandins (PG), such as PGE₂ or PGD₂, were found to be generated by ionophore-challenged coelomocytes. Incubation of coelomocytes with lipopolysaccharides from either Escherichia coli or Salmonella abortus failed to induce an increase in generation of LOX products and the presence of 8-HETE (0–25 μM) had no significant effect on the in vitro phagocytic activity of Asterias coelomocytes. Neither indomethacin (a COX inhibitor) or esculetin (a LOX inhibitor) had any effect on the clearance of the bacterium, Vibrio splendidus, from the coelomic cavity of starfish suggesting that products of these enzymes are not involved in such coelomocyte responses to foreign particles.     

Phospholipase A2 in hemocytes of the tobacco hornworm,Manduca sexta

On the hypothesis that prostaglandins and other eicosanoids mediate nodulation responses to bacterial infections in insects, we describe an intracellular phospholipase A₂ (PLA₂) in homogenates prepared from hemocytes collected from the tobacco hornworm, Manduca sexta. PLA₂ hydrolyzes fatty acids from the sn-2 position of phospholipids. Some PLA₂s are thought to be the first and rate-limiting step in biosynthesis of prostaglandins and other eicosanoids. The hemocyte PLA₂ activity was sensitive to hemocyte homogenate protein concentration (up to 250 μg protein/reaction), pH (optimal activity at pH 8.0), and the presence of a Ca²⁺ chelator. Like PLA₂s from mammalian sources, the hemocyte PLA₂ was inhibited by the phospholipid analog oleyoxyethyl phosphorylcholine. Whereas most intracellular PLA₂s require Ca²⁺ for catalytic activity, some PLA₂s, including the hemocyte enzyme, are Ca²⁺-independent. The hemocyte PLA₂ exhibited a preference for arachidonyl-associated substrate over palmitoyl-associated substrate. These findings show that M. sexta hemocytes express a PLA₂ that shows a marked preference for hydrolyzing arachidonic acid from phospholipids. The biological significance of this enzyme relates to cellular immune responses to bacterial infections. The hemocyte PLA₂ may be the first biochemical step in synthesis of the eicosanoids that mediate cellular immunity in insects. © 1996 Wiley-Liss, Inc.     

Eicosanoids mediate control of thermoregulatory sweating in the cicada,Tibicen dealbatus (Insecta: Homoptera)

Cicadas prevent body temperature from exceeding tolerable levels by a combination of behavioral responses and sweating. Sweating is activated when body temperature reaches a critical set-point temperature. We investigated control of sweating in the cicada, Tibicen dealbatus, by chemically manipulating biosynthesis of prostaglandins and other eicosanoids. Injecting prostaglandins in amounts equal to those that induce behavioral fever in scorpions and crustaceans resulted in only a small increase in set-point temperature. Blocking prostaglandin biosynthesis with cyclo-oxygenase inhibitors such as aspirin produced significant changes in set-point temperature, confirming that prostaglandins are involved in control of sweating. However, the effect of cyclo-oxygenase inhibitors was not the opposite of the effect of prostaglandins. Instead, the effect of cyclo-oxygenase inhibitors depended strongly on the value of setpoint temperature prior to treatment. Results of biochemical manipulations of other steps in eicosanoid biosynthetic pathways corroborated the results of cyclo-oxygenase inhibition and indicated that eicosanoids other than prostaglandins may be involved in control of body temperature in normothermic T. dealbatus. The effect of cyclo-oxygenase inhibitors on a given set-point temperature depended on the ambient temperature experienced by cicadas during the experiment. Surprisingly, cicadas exposed to ambient temperatures 40°C delayed activation of sweating until body temperature exceeded values normally recorded from T. dealbatus in the field. Control of body temperature in normothermic cicadas is thus complex, involving inputs from body temperature sensors, ambient temperature sensors, and at least two cyclo-oxygenase-dependent regulatory pathways.Abbreviations PUFA polyunsatured fatty acid(s) - T _a ambient temperature - T _b body temperature - T _set set-point of body temperature for activation of sweating     

大肠杆菌细胞壁肽聚糖的化学修饰及荧光标记

【目的】发展一种活细菌细胞壁荧光标记方法,为后期研究细菌肽聚糖的生物合成和代谢规律以及其与细菌感染致病的关系提供新的工具。【方法】对细菌肽聚糖的生物合成前体N-乙酰葡萄糖胺-1-磷酸(GlcNAc-1-P)进行化学修饰,设计并合成含有叠氮基的GlcNAc-1-P类似物(化合物5:Ac3GlcNAz-1-P)。将该类似物与细菌共同孵育,使其作为探针进入细菌肽聚糖天然合成途径。之后提取并酶解肽聚糖组分,用红外光谱(FTIR)和液质联用(LC/MS)检测探针是否通过代谢进入细菌肽聚糖结构中。同时用外源荧光素对代谢掺入细菌肽聚糖中的探针进行染色。在激光共聚焦显微镜下观察对活细菌的荧光标记效果。【结果】通过四步有机合成反应,以79%的总收率成功获得了化合物5。将大肠杆菌(Escherichia coli BL21)作为模式菌株与化合物5共孵育后,其肽聚糖组分的LC/MS和FTIR分析结果均显示探针可以被细菌利用并被代谢掺入到肽聚糖结构中。激光共聚焦显微镜观察结果显示,荧光素可以高效标记表面携带有生物正交探针的大肠杆菌。【结论】设计合成了一种新型探针,可用于活细菌成像,为深入研究细菌肽聚糖的生物学功能及其与细菌感染致病的关系提供了一种简便的方法。     

Stimulated release of arachidonate and prostaglandins is vectorial in MDCK epithelial cells

The receptor mediated activation of phospholipase A₂ by appropriate ligands results in the synthesis and release of eicosanoids, a class of potent bioregulatory molecules. Madin-Darby canine kidney cells (MDCK) are polarized epithelial cells, with structurally and functionally distinct plasma membrane domains separated by tight junctions. Using MDCK cells grown in dual sided chambers, we show in this report, that a) the receptor mediated release of prostaglandins and arachidonate into the extracellular medium is predominantly unidirectional, b) the direction of release is agonist specific, and c) the magnitude of the response due to a given agonist is cell-domain specific. These characteristics, if operative in vivo, would contribute towards the optimal function of trans-cellular metabolism of eicosanoids already demonstrated.     

The distribution and taxonomic value of fatty acids and eicosanoids in the Lipomycetaceae and Dipodascaceae

Using radioimmunoassay, blood platelet aggregation studies and GC-MS the existence of prostaglandins in the endomycetalean yeastDipodascopsis uninucleata was confirmed by our group. These findings triggered the search for similar eicosanoids in the rest of the Endomycetales. We commenced by scanning for the easily detectable precursors of eicosanoids, linoleic- and linolenic acid. We selected two families (i.e. Lipomycetaceae and Dipodascaceae), both producing these precursors, for further investigation.Representative strains of the two families were tested for their ability to grow in the presence of 1mM aspirin, a specific inhibitor of prostaglandin biosynthesis. In contrast to the lipomycetaceous species the dipodascaceous species were insensitive to this drug. These results were verified when representative strains of both families were investigated for their ability to produce eicosanoids from externally fed radio-labeled arachidonic acid along an aspirin sensitive pathway. Thin layer chromatography of culture extracts, followed by autoradiography, showed that while none of the Dipodascaceae produced aspirin sensitive arachidonic acid metabolites, the members of the Lipomycetaceae tested positive for these metabolites. These findings supported the separation of the lipomycetaceous yeastDipodascopsis from the Dipodascaceae. The findings also correlate with the delimitation of these yeasts in two families (i.e. Dipodascaceae and Lipomycetaceae).Further investigation indicated that prostaglandin production by the genusDipodascopsis is mainly associated with ascosporogenesis. Thin layer chromatography of cell extracts fromDipodascopsis tóthii, followed by scintillation counting, indicated the presence of PGF₂ and PGE₂ during ascosporogenesis.     

Comitogenicity of eicosanoids and the peroxisome proliferator ciprofibrate in cultured rat hepatocytes

Several hypolipidemic drugs and environmental contaminants induce hepatic peroxisome proliferation and hepatic tumors when administered to rodents. These chemicals increase the expression of the peroxisomal β-oxidation pathway and the cytochrome P-450 4A family, which metabolize lipids, including eicosanoids and their precursor fatty acids. We previously found that the peroxisome proliferator ciprofibrate decreases the level of eicosanoids in the liver and in cultured hepatocytes. In this study, we examined the effect of prostaglandins E₂ and F_2α (PGE₂ and PGF_2α), leukotriene C₄ (LTC₄) and the peroxisome proliferator ciprofibrate on DNA synthesis in cultured hepatocytes. Primary rat hepatocytes were cultured on collagen gels in serum-free L-15 medium with varying concentrations of eicosanoids and ciprofibrate, and the absence or presence of growth factors. Ciprofibrate lowered hepatocyte eicosanoid concentrations; the addition of eicosanoids restored their levels. After a 48-h exposure with [³H]-thymidine, DNA synthesis was determined by measuring [³H]-thymidine incorporation into DNA. The addition of PGE₂, PGF_2α, and LTC₄ to cultures along with ciprofibrate increased DNA synthesis, whereas treatment with ciprofibrate or eicosanoids alone resulted in a much smaller increase. The addition of epidermal growth factor (EGF) to the eicosanoid-ciprofibrate combination increased DNA synthesis more than EGF or the eicosanoid-ciprofibrate combination alone. The PGF_2α-ciprofibrate combination also was comitogenic with transforming growth factor-α and hepatocyte growth factor. The addition of both ciprofibrate and prostaglandins also blocked the growth inhibitory effect of transforming growth factor-β on DNA synthesis induced by EGF. These results show that the eicosanoids PGE₂, PGF_2α, and LTC₄ are comitogenic with the peroxisome proliferator ciprofibrate in cultured rat hepatocytes. © 1996 Wiley-Liss, Inc.     

Eicosanoid Signaling in Insects: from Discovery to Plant Protection

Prostaglandins (PGs) and related eicosanoids are signal moieties derived from arachidonic acid and two other C20 polyunsaturated fatty acids. They were discovered in the 1930s in the context of mammalian reproductive physiology; PGs were associated with the prostate gland, hence their name, and they stimulate uterine smooth muscle contraction. Determining PG chemical structures in the early 1960s and demonstrating that they mediate many human pathophysiological events in the 1970s stimulated intensive research over the following decades in universities, governments and the private sector. Interest in the biological significance of PGs in insects arose in the 1970s and 1980s, which opened a new research frontier. PGs act in reproduction, releasing egg-laying behaviors in some species and signaling egg-maturation events in the Drosophila and silk moth models. They act in insect immunity, mediating and coordinating cellular and humoral responses to wounds, infection and invasion. PGs act in ion transport physiology in insect Malpighian tubules and recta. These compounds also mediate physiological trade-offs between insect immunity and reproduction. Finally, they are central players in the molecular ecology of interactions between blood-feeding insects and their vertebrate hosts. Some PG functions are critical at specific, crucial moments in insect lives, moments we consider ‘emergencies,’ such as the immediate reactions to infection. Certain microbial species have keyed into insect PG signaling and they evolved mechanisms to disable insect immune reactions to infection by inhibiting key enzymes in PG biosynthesis. We provide proof-of-principle that RNA interference treatments designed to silence genes in PG signaling disrupts insect immunity. In this review we describe the history, chemistry and biology of PGs. We use this background to argue that because PGs and other eicosanoids act in emergency situations, they are visible targets for development and deployment of novel insect pest management technologies.     

Degradation of Differently Substituted Xylogalacturonans by Endoxylogalacturonan Hydrolse and Endopolygalacturonases

Abstract

A method was developed to make xylogalacturonans (XGAs) with different degrees of xyloslyation from gum tragacanth (XGA-25, XGA-29, XGA-35 and XGA-47), using alkali treatment at 4°C and acid treatment at 100°C. Ester linkages as well as fucose and arabinose substituents could selectively be removed by this procedure. Galactosyl- and xylosyl-linkages appeared to be more stable, while some backbone degradation of the galacturonan took place upon prolonged acid treatment. Using XGA-35, endoxylogalacturonan hydrolase (XGH) from Aspergillus tubingensis, expressed in Kluyveromyces lactis, was characterised with respect to kinetic parameters, temperature and pH effects.

XGA-25 and XGA-47 were degraded with endopolygalacturonases (PGs) from Aspergillus niger (PG1, PG2), from A. tubingensis (PF-arf), from Kluyveromyces fragilis (PG-kluyv) and XGH from A. tubingensis. The activity of the different PGs decreased with increasing degrees of xylosylation. However, for each PG a different tolerance for the presence of side chains was observed. PG-arf and PG1 were hindered most by xylosyl branching, whereas XGH appeared to have a requirement for xylosylation and was almost not active towards polygalacturonic acid. The degradability of xylogalacturonans by XGH increased with higher degrees of xylosylation. Typically, a highly substituted xylogalacturonan from pea was almost resistant to XGH treatment. XGH produces a distinctive set of oligosaccharides from XGA, which is different from the hydrolysis products of PG action.

Saponified modified hairy regions from apple (MHR-S) containing xylogalacturonan, were partially degraded by XGH. A combination of XGH and rhamnogalacturonan hydrolase was able to fully degrade the high molecular weight fraction of MHR-S. The two enzymes acted additively, no synergy being observed.     

Prostaglandins and functional specificity of central neurons of Helix pomatia

The effect of extra- and intracellularly injected prostaglandins (PG) E₂ and F₂ on electrical activity and responses to acetylcholine and serotonin were studied in experiments on identified neurons ofHelix pomatia. As a rule prostaglandins modified the typical electrical activity of the identified neurons: PG E₂ enhanced and PG F₂ depressed it. These substances mainly weakened responses of the nerve cells to mediators: PG E₂ caused a greater change in the response to serotonin and PG F₂ in the response to acetylcholine. Effects of the prostaglandins when injected extracellularly and intracellularly differed. The possible molecular-cellular mechanisms of the central action of prostaglandins are discussed in the light of their functional connections with other universal regulators of cellular metabolism and with proteins specific for nerve tissues.P. K. Anokhin Research Institute of Normal Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 13, No. 6, pp. 580–588, November–December, 1981.     

Direct actions of prostaglandins E1, A1, and F2α on myocardial contraction

The inotropic and chronotropic actions of prostaglandin (PG) types PGE₁, PGA₁, and PGF_2α were studied in isolated guinea pig right and left atria, and papillary muscles; rabbit atria; and toad ventricular strips in order to more completely define the cardiac contractile properties of PG. All three prostaglandins, in muscle bath concentrations of 10μg/ml, exerted positive inotropic and chronotropic actions on guinea pig atrium. These contractile effects were persistent after removal of PG from the muscle bath and appeared to limit the relative response to a subsequent dose of PG. The inotropic action of PGE₁ was present over a wide range of bath calcium concentrations (1.1 to 4.4 mM/L). Beta adrenergic receptor blockade, histamine blockade, and pretreatment with reserpine failed to significantly affect the inotropic actions of PG. Norepinephrine and histamine produced more potent inotropic and chronotropic effects on guinea pig atria than did PG and these contractile effects did not exhibit persistence or tachyphylaxis. The prostaglandins did not significantly affect dose response curves for norepinephrine inotropic and chronotropic actions. The prostaglandins had no effect on the force or frequency of contraction in rabbit atria. PGE₁ exerted a positive inotropic effect on toad ventricular myocardium whereas PGA₁ had no effect and PGF_2α had a negative inotropic action.     

Fluorescent/ultraviolet absorbing ester derivative formation and analysis of eicosanoids by high-pressure liquid chromatography

A method is described for the quantitative analysis of eicosanoids (arachidonic acid metabolites, nee, prostaglandins) by reverse-phase high-pressure liquid chromatography following formation of the ester derivative with p-(9-anthroyloxy)phenacyl bromide. The lower limit of detection of the eicosanoid ester is 280 pg (ultraviolet—254 nm) and approximately 50 pg (fluorescence 249 emission, 413-nm cutoff). We separated the esters of seven common eicosanoids by reverse-phase chromatography with acetonitrile and water. Thromboxane B₂ chromatographs as two species and coelutes with PGF_2α. Separation of all others is adequate, including the three metabolites of prostacyclin (6-keto-PGF_1α, 6-keto-PGE₁, 13,14-dihydro-6,15-diketo-PGF_1α). We obtained good correlation between radioimmunoassay and derivative analysis of standard 6-keto-PGF_1α extracted from lactated Ringer's solution with standard technique, as well as 6-keto-PGF_1α quantitation from tissue culture medium that had contained pulmonary endothelial cells. This method should be applicable to analysis of eicosanoids extracted from biological matrices.     

Interaction of prostaglandins with adrenal microsomal cytochrome P-450 in the guinea pig

Studies were carried out to investigate the effects of prostaglandins (PG) $\text{in vitro}$ on adrenal microsomal steroid and drug metabolism in the guinea pig. The addition of PGE₁, PGE₂, PGA₁, PGF_1α or PGF_2α to isolated adrenal microsomes produced typical type I difference spectra. The sizes of the spectra (ΔA_385–420) produced by prostaglandins were smaller than those produced by various steroids including progesterone, 17-hydroxyprogesterone and 11β-hydroxyprogesterone. However, the affinities of prostaglandins and steroids for adrenal microsomal cytochrome P-450, as estimated by the spectral dissociation constants, were similar. Prior addition of prostaglandins to isolated adrenal microsomes did not affect steroid binding to cytochrome P-450 or the rate of steroid 21-hydroxylation. In contrast, prostaglandins inhibited adrenal metabolism of ethylmorphine and diminished the magnitude of the ethylmorphine-induced spectral change in adrenal microsomes. The results indicate that prostaglandins inhibit adrenal drug metabolism by interfering with substrate binding to cytochrome P-450. Since 21-hydroxylation was unaffected by PG, different cytochrome P-450 moieties are probably involved in adrenal drug and steroid metabolism.     

A deep learning approach for converting prompt gamma images to proton dose distributions: A Monte Carlo simulation study

PurposeIn proton therapy, imaging prompt gamma (PG) rays has the potential to verify proton dose (PD) distribution. Despite the fact that there is a strong correlation between the gamma-ray emission and PD, they are still different in terms of the distribution and the Bragg peak (BP) position. In this work, we investigated the feasibility of using a deep learning approach to convert PG images to PD distributions.MethodsWe designed the Monte Carlo simulations using 20 digital brain phantoms irradiated with a 100-MeV proton pencil beam. Each phantom was used to simulate 200 pairs of PG images and PD distributions. A convolutional neural network based on the U-net architecture was trained to predict PD distributions from PG images.ResultsOur simulation results show that the pseudo PD distributions derived from the corresponding PG images agree well with the simulated ground truths. The mean of the BP position errors from each phantom was less than 0.4 mm. We also found that 2000 pairs of PG images and dose distributions would be sufficient to train the U-net. Moreover, the trained network could be deployed on the unseen data (i.e. different beam sizes, proton energies and real patient CT data).ConclusionsOur simulation study has shown the feasibility of predicting PD distributions from PG images using a deep learning approach, but the reliable prediction of PD distributions requires high-quality PG images. Image-degrading factors such as low counts and limited spatial resolution need to be considered in order to obtain high-quality PG images.     

Prothoracicotropic hormone bioassay: Bombyx larva assay

Summary

An assay for the prothoracicotropic hormone (PTTH) has been established using in situ activation of the prothoracic glands (PG) of Bombyx mori in its larva-to-larva development. The timing of PTTH release was estimated by examining developmental response of 4th instar larvae to brain removal and neck ligation, and changes in the haemolymph ecdysteroid titer and ecdysone-releasing activity of PG in vitro during the development. Injection of Bombyx brain extracts into 4th instar larvae neck-ligated shortly before full activation of PG elicited larval moulting rather than precocious pupation in headless larvae. This developmental shift was regarded as due to the action of PTTH, and the PTTH unit has been defined from a linear log dose-response relationship. Materials chromatographically fractionated from Bombyx brain extracts were examined for the presence of stage- and species-specific PTTH molecules by using this Bombyx larva assay and Bombyx and Samia pupa assays previously developed. The same fractions were active when assayed by Bombyx larva and pupa assays.     

Studies of prostaglandins and prostaglandin antagonists on bovine pulmonary vein in vitro

Acetylsalicylic acid (ASA), indomethacin, sodium meclofenamate (FEN), phenylbutazone (PB), phloretin phosphates (PP), SC-19220, and diethylcarbamazine citrate (DECC) were screened against histamine, 5-hydroxytryptamine (5-HT), bradykinin, acetylcholine, and prostaglandins (PG) E₁, E₂, and F_2α to determine their specificity in antagonizing PG's on the bovine pulmonary vein. PG E₂ relaxed the smooth muscle preparation at low concentrations and induced contraction at higher concentrations. PG E₁ consistently evoked dose-related relaxations, whereas PG F_2α contracted the bovine pulmonary vein. Studies with inhibitors suggest that the different actions of prostaglandins could be mediated through different receptors. Sodium meclofenamate and PP dimer blocked PG E₂-induced contractions, whereas relaxations were not blocked. DECC inhibited the relaxant effect of PG E₂. DECC also antagonized histamine, 5-HT, and PG F_2α, suggesting the drug is rather non-specific. Phenylbutazone antagonized the actions of both PG E₂ and PG F_2α on the bovine pulmonary vein. By classifying receptors by antagonism the bovine pulmonary vein appears to contain PG E₂ (PP-type), PG E₂ (FEN-type), PG E₂ (PB-type), and PG F_2α (PB-type) receptors. An absence of SC-type PG-receptors is noted.     

Characterization of prostaglandin synthesis in the housefly,Musca domestica (L.)

The in vitro formation of prostaglandins (PG) was examined in the housefly Musca domestica. PG synthetase activity was detected in homogenates of whole insects and in head and thorax, abdomen, ovary and male reproductive tissues. Studies to determine the sub-cellular localization of PG synthetase indicated that the microsomal fraction contained the highest activity. Products obtained from radiolabeled arachidonic acid (20:4) and 8,11,14-eicosatrienoic acid (20:3, n-6) were PGE₂ and PGE₁, respectively, with lower amounts of the PGF series also present. In microsomal preparations from whole insects and reproductive tissues from both males and females, 20:3(n-6) was 2–2.5 times more efficiently converted to PG than was 20:4. Non-steroidal anti-inflammatory drugs (NSAID) fed to houseflies did not inhibit PG production from 20:4, whereas when they were included in microsomal preparations at high levels, they inhibited PG synthesis.