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The content of prostaglandin E and prostaglandin F2alpha in the exudate of carrageenin granuloma of rats.

1. Granuloma was made by the subcutaneous injection of 2% carrageenin solution on the dorsum of male rats. Eight, 16, 24 and 72 h after the injection. the exudate from each rat granuloma was withdrawn and extracted for rpstaglandins. 2. Extracted prostaglandins were separated prostaglandin E and prostaglandin F group by silicic acid mini-column chromatography. Then the amount of prostaglandin E and prostaglandin F2alpha were determined by the radioimmunoassay method. 3. The levels of prostaglandin E in the granuloma exudates were 4.6 ng/ml at 8 h after the carrageenin injection, then decreased 3.6 ng/ml and to 1.1 ng/ml at 16 h and 24 h, respectively. Seventy-two h after the injection, prostaglandin E level was increased to 8.1 ng/ml. 4. The levels of prostaglandin F2alpha in the exudate were as follows: At 8 h after the carrageenin injection, the level was 9.4 ng/ml, then decreased to 1.3 ng/ml and to 0.8 ng/ml at 16 h and 24 h, respectively. Seventy-two h after the carrageenin injection, it was again elevated to 4.7 ng/ml. 5. The exudate of granuloma, 24 and 72 h after the carrageenin injection, was incubated with [3H]prostaglandin E1 at 37 degrees C for 30 min. Then the acidic ether extract was subjected to reversed phase partition chromatography. It was found that the exudate of 24 h and 72 h granuloma had little activity of prostaglandin 15alpha-hydroxy dehydrogenase.

Animals

A technique for the immunohistochemical localization of prostaglandin E.

Prostaglandin E (PGE) has been localized via the unlabeled antibody technique in freeze-dried and ethanol-fixed cryostat sections. Discrete perivascular and stromal localization was present in the uterus prepared by the method presented, but not in classically fixed specimens. Absorption of the anti-PGE by addition of free PGE was ineffective; whereas, removal of PGE-reactive antibodies from the anti-serum was effectively accomplished with an Affigel-101-PGE immunoadsorbant column.

Animals

Purification and regulatory properties of chicken heart prostaglandin E 9-ketoreductase.

Prostaglandin E 9-ketoreductase was purified from chicken heart by ammonium sulfate fractionation, and DEAE-Sephadex, hydroxylapatite and phosphocellulose chromatography. Two peaks of activity were resolved during the phosphocellulose chromatographic step. Both peaks were stimulated by a substance that was not bound to the phosphocellulose column. This stimulatory substance was destroyed by treatment with phosphodiesterase and 0.1 M NaOH. It was heat-stable (100 degrees, 2 min), nondialyzable, and resistant to treatment with pronase, ribonuclease, and deoxyribonuclease; but it was dialyzable after heating or digestion with pronase. Sodium pyrophosphate also enhanced the activities of the prostaglandin E 9-ketoreductases as did angiotensin I; but not angiotensin II. In the presence of 3':5'-cyclic AMP, AMP, or several other ribonucleotides, the enhancing effects of the natural stimulatory substance, sodium pyrophosphate or angiotensin I were blocked, but these ribonucleotides themselves had little effect on the enzymes activity. The substrate specificities of the two prostaglandin E 9-ketoreductases were also studied. Both the 9-keto group and the 15-keto group of 15-ketoprostaglandin F2 alpha could be converted to the corresponding hydroxyl group; the 15-keto group was reduced faster than the 9-keto group. Prostaglandin D2, a prostaglandin with a 9-hydroxyl and an 11-keto group, could not be converted to prostaglandin F2 alpha nor could cyclohexanone be converted to cyclohexanol by the prostaglandin E 9-ketoreductase.

Alcohol Oxidoreductases

Variability in rates of urine prostaglandin E excretion.

Urine prostaglandin E excretion rates were determined by hepatic receptor assay in three groups of conscious animals. Although 24-hour urine collections gave reproducibly similar levels of prostaglandin E excretion rates, levels obtained with consecutive 20-minute urine collections were extremely variable despite no obvious changes in renal function. Since short urine collection periods are used frequently in physiologic studies, the significance of variations in prostaglandin excretion rates in these studies may have to be re-evaluated.

Animals

Zero extracellular K+ and prostaglandin E release in the guinea-pig taenia coli.

Prostaglandin E release rates from isolated strips of guinea-pig taenia coli increased during exposure to zero K+ bathing fluid, from control values of 0.78 +/- 0.11 ng/g per min to levels as high as 29.2 ng/per min. Release rates increased for 40-50 min and then remained constant or fell despite progressive increases in intracellular sodium [Nai+] or fall in intracellular potassium [Ki+]. Readmittance of K+ to the bathing solution resulted in rapid reversal of elevated prostaglandin E release rates. [Nai+] and [Ki+] were markedly more abnormal in strips exposed to zero K+ for 70-201 min compared to 30-min exposures. Upon the readdition of K+ after long zero K+ exposure, the rate of prostaglandin E release fell long before [Nai+] and [Ki+] returned to control levels. After K+ was readded to the bathing solution, the ion concentration of tissues exposed to zero K+ for 30 min returned to normal much more quickly than did those of tissues exposed for the longer time periods, yet the exponential rate constants for fall of prostaglandin E release rate after K+ was added were not significantly different after short or long zero K+ exposure. Thus there was a dissociation between the return of [Nai+] and [Ki+] and the fall of prostaglandin E release rate to control levels. Ouabain augmented prostaglandin E release under conditions where [Ki+] could not fall. Addition of known neurotransmitters present in this tissue to the bathing fluid did not augment prostaglandin E release. Guinea-pig taenia coli strips that had been incubated with [3H]arachidonic acid, constantly released [3H]arachidonic acid and [3H]prostaglandin E and a prostaglandin which cochromatographed with prostaglandin E but could not be converted to prostaglandin B by alkali and was shown to be 6-ketoprostaglandin F1 alpha. Release of [3H]arachidonic acid and [3H]prostaglandin E plus 6-[3H]ketoprostaglandin F1 alpha was increased when strips were exposed to zero K+. Data obtained in this study suggest the augmented prostaglandin E release seen during zero K+ or ouabain is related to increased availability of unbound arachidonic acid at the site of cyclooxygenase in the cell. Augmented prostaglandin E release is apparently not related to alterations in intracellular electrolyte concentrations or release of known neurotransmitters.

Acetylcholine

Prostaglandin E, thromboxane B2 and 6-oxo-prostaglandin F1 alpha in amniotic fluid and maternal plasma of rhesus monkeys (Macaca mulatta) during the latter third of gestation.

The concentrations of prostaglandin E (PGE), thromboxane B2 (TXB2) and 6-oxo-prostaglandin F1 alpha (6-oxo-PGF1 alpha) were measured by radioimmunoassay in serial samples of amniotic fluid and maternal peripheral plasma in the latter third of pregnancy in rhesus monkeys (Macaca mulatta). The samples were collected under ketamine-induced anaesthesia. The concentration of PGE was undetectable in amniotic fluid until a few days before delivery when a large increase was observed in three of the five animals. There were small increases of TXB2 and 6-oxo-PGF1 alpha in amniotic fluid before delivery. In maternal plasma the concentrations of PGE, TXB2 and 6-oxo-PGF1 alpha were generally higher and more variable than in amniotic fluid and did not increase with advancing gestation. It is suggested that increased production of primary prostaglandins occurs before, and is involved in, the onset of parturition in the rhesus monkey.

Amniotic Fluid

Effect of triamterene on urinary excretion of immunoreactive prostaglandin E in essential hypertension.

The effect of triamterene on urinary excretion of prostaglandin E was studied, and the results were compared with those obtained with spironolactone. Urinary excretion of immunoreactive prostaglandin E was measured radioimmunologically. Triamterene was administered in a dose of 100 mg/day for 8 days to 7 patients with essential hypertension. Following the administration of triamterene, urinary prostaglandin E tended to increase. However, the increment was not significant. The lack of significant increase in urinary prostaglandin E excretion during the administration of triamterene contrasted with our previous finding with spironolactone, in which a significant increase in prostaglandin E excretion was observed on the first day of spironolactone administration. Urinary Na excretion and urinary Na/K ratio were significantly increased and urine volume also tended to increase following the administration of triamterene. Plasma renin activity and plasma aldosterone concentration were increased in all cases. However, there was no significant correlation between these parameters and urinary prostaglandin E. These results suggest that the effect of triamterene on renal prostaglandin E synthesis is different from that of spironolactone and that the change in urinary prostaglandin E after the administration of triamterene is not the reflection of the change in the renin-angiotensin-aldosterone system.

Adolescent

The concentrations of the prostaglandins E and F, 13 14-dihydro-15-oxo-prostaglandin F and thromboxane B2. In tissues obtained from women with and without pre-eclampsia.

The concentrations of prostaglandins E (PGE) and F (PGF), 13, 14-dihydro-15-oxo-prostaglandin F (PGFM) and thromboxane B2 (TXB2) were measured by specific radioimmunoassays in tissues obtained from women with and without pre-eclampsia. The concentrations of PGE in the amnion, chorion, decidua and placenta obtained from subjects with pre-eclampsia were significantly lower than those from subjects without pre-eclampsia. The concentration of PGE in these tissues increased significantly with gestational age and correlated with urinary oestrogen excretion. PGF concentrations were lower in the amnion and placenta of the pre-eclamptics compared to those without pre-eclampsia. The concentrations of PGFM in the amnion, decidua, and myometrium were lower in the pre-eclamptics. No significant difference in the TXB2 concentrations between the two groups of subjects were found. It is suggested that the altered tissue concentrations of prostaglandins in pre-eclamptics are due to the effects of gestational age and oestrogens and may or may not be involved in the pathogenesis of pre-eclampsia.

Adolescent

[Development of estradiol, progesterone, and prostaglandins E and F2 alpha levels during pregnancy in mice].

17 beta estradiol, progesterone, prostaglandin E and prostaglandin F2 alpha were determined in the plasma of mice at different times of pregnancy. Estradiol displays a preimplantation peak on day 4. Other peaks are visible at the end of the implantation period (day 6) and on day 10. Progesterone increases when implantation begins (day 5). Prostaglandins increase at the beginning of pregnancy and decrease temporarily before implantation. Other peaks are also apparent for prostaglandin E on days 5.5 and 11.

Animals

Effect of prostaglandin E on the adenyl cyclase-cyclic AMP system and gluconeogenesis in rat renal cortical slices.

Prostaglandin E was found to increase the formation of cyclic acdenosine 3',5'-monophosphate (cyclic AMP) by renal cortical slices. This increased release of cyclic AMP was not influenced by the absence of Ca2+ in the incubating media. The enhanced production of cyclic AMP was probably mediated by stimulation of membrane-bound adenylate cyclase activity. An increase in adenyl cyclase activity was observed with increasing concentrations of prostaglandin E. Furthermore, prostaglandin E augmented glucose production from alpha-ketoglutarate. This effect on gluconeogenesis was abolished by the removal of Ca2+ from the incubating medium. These effects are similar to those described for parathyroid hormone and suggest that the renal cortex is a prostaglandin-dependent system. Prostaglandin E decreased cyclic AMP production and glucose production (from alpha-ketoglutarate) in response to submaximal doses of parathyroid hormone, suggesting that prostaglandin may be important in modulating the intracelluar action of parathyroid hormone in the kidney cortex.

Adenylyl Cyclases

Prostaglandin E and hypercalcemia in breast carcinoma: only a tumor marker? A need for perspective.

Prostaglandin E concentrations were measured in a patiet with breast carcinoma, hypercalcemia, undetectable parathyroid hormone (PTH) and no evidence of bone metastases. Catheterization of the drainage bed of her tumor documented production of E series prostaglandins. Treatment with the largest recommended doses of indomethacin for 10 days failed to lower her plasma prostaglandin E (PGE) concentrations or to correct the hypercalcemia, but it normalized urinary excretion of PGE. Subsequent chemotherapy reduced prostaglandin concentrations toward normal values concomitant with a reduction of clinically estimated tumor burden. During this period of time, serum calcium concentrations had no consistent relationship to the plasma PGE levels. We suggest that PGE merely reflected the tumor burden of this patient and did not directly contribute to the genesis of her hypercalcemia. The pertinent literature relating PGE and hypercalcemia is reviewed.

Antineoplastic Agents

Prostaglandin E-2-9-ketoreductase in ovarian tissues.

The existence of the enzyme prostaglandin E-2-9-ketoreductase which can convert prostaglandin E-2 to prostaglandin F-2 alpha was indicated in experiments with pig and human ovarian tissues in vitro, using radioimmunoassay methods and a superfusion technique. Further studies involving radiotracer techniques demonstrated that the enzyme was localized in the high-speed (105 000 g) supernatant fraction of human, pig and rat luteal tissue and human stromal tissue. The enzyme was shown to be NADPH-dependent and its activity in luteal tissue increased in the order : pig less than human less than rat.

Animals

Changes in blood level of kininogen, prostaglandin E and hemodynamics during experimental acute myocardial ischemia with and without FOY-007.

Changes in kininogen, prostaglandin E and hemodynamics were studied during 2 hours after left anterior descending artery ligation in 19 anesthetized dogs with and without FOY-007, an inhibitor of kinin forming enzyme. Significant decrease in kininogen in aorta and great cardiac vein (aorta greater than great cardiac vein) and increase in prostaglandin E in great cardiac vein were observed after ligation, indicating release of kinin and prostaglandin E from ischemic area. Both kininogen and prostaglandin E changes were inhibited by FOY-007 but further decrease in cardiac output and increase in systemic vascular resistance were observed. In summary, myocardial release of both kinin and prostaglandin E was inhibited by FOY-007, with increase of afterload of the heart.

Animals

Prostaglandins E and F in the uterine tissue of cyclic rats.

Prostaglandins E and F (PGE and PGF) levels and the biosynthetic ability of rat uterine tissue have been estimated in the various phases of oestrous cycle by radioimmunoassay technique. Significantly high PGF concentration was found during metaoestrous phase. In comparison, PGE levels did not show as high elevation. The metaoestrous phase also showed significantly higher biosynthesis of prostaglandins E and F. Our experimental findings support the role of prstaglandins in ovulation and luteolysis of the corpus luteum.

Animals

Selective stimulation of venous prostaglandin E 9-ketoreductase by bradykinin.

The effects of bradykin on prostaglandin metabolism in canine mesenteric vessels were examined. Bradykinin stimulated microsomal prostaglandin synthesis in both artery and vein; this stimulation was more pronounced when [14C] hosphatidylcholine rather than [14C] arachidonate was used as the substrate for prostaglandin synthetase. This suggested that bradykinin enhanced a membrane phospholipase. In addition, bradykinin selectively stimulated prostaglandin E 9-ketoreductase activity from veins but not arteries. This may explain the finding that bradykinin induces the release of prostaglandin E compounds from arteries but prostaglandin F compounds from veins.

Alcohol Oxidoreductases

Relationship between concentration of prostaglandins E and F in the regulation of ovum transport in rabbits.

Prostaglandin E and F levels were measured by a radioimmunoassay technique in different parts of the rabbit fallopian tube treated with indomethacin and in untreated rabbit at 0, 24, 48 and 72 hours after Human Chorionic Gonadotrophin (HCG) administration. The results indicate a significant rise in the levels of PGE and fall in the levels of PGF throughout the isthmus at 48 hours and in the distal isthmus at 72 hours in the untreated rabbits. In the indomethacin treated rabbits, PGF levels were reduced relatively more than PGE levels in different parts of the rabbit fallopian tube at various time intervals after HCG administration. Prostaglandins E and F may have a role in regulating ovum transport.

Animals