Plasma concentrations of 13,14-dihydro-15-keto-prostaglandin E2 in rabbits bearing the VX2 carcinoma: Effects of hydrocortisone and indomethacin

In rabbits bearing the prostaglandin-producing VX₂ carcinoma, the plasma concentration of 13,14-dihydro-15-keto-PGE₂ (PGE₂-M) was elevated within one week after tumor implantation and preceded the development of hypercalcemia. Both the rate of rise and magnitude of the increase were greater for the metabolite than for PGE₂; at the time of peak hypercalcemia (about 4 to 5 weeks after tumor implantation), the increase over basal in plasma PGE₂-M was about 75 fold whereas it was previously shown that the increase in PGE₂ was less than 2 fold. Indomethacin, which inhibits PGE₂ synthesis in VX₂ cells in culture, lowered in parallel plasma calcium and PGE₂-M in tumor-bearing rabbits. Administration of hydrocortisone to rabbits bearing the VX₂ tumor prevented the development of hypercalcemia when given at the time of tumor implantation and reversed the elevated plasma calcium in previously untreated animals; the steroid hormone also lowered plasma concentrations of PGE₂-M. These findings are consistent with our hypothesis that the hypercalcemic syndrome in VX₂ tumor-bearing rabbits is due to the secretion of PGE₂ by the tumor.     

Effect of partial fetectomy on plasma 13,14-dihydro-15-keto-prostaglandin F2α in late pregnancy of sows

The aim of this study was to ascertain if partial fetectomy in late pregnancy affected prostaglandin F_2α levels, thereby influencing the time of delivery of the remaining fetuses in the sow. Sham fetectomy or surgical removal of one, two, three or four fetal piglets was performed on each of ten sows during the last 3 weeks of pregnancy. Removal of no fetuses (two sows) and in one sow a single fetus was followed by a continuation of pregnancy to the expected time of parturition. Plasma values for oestrone, progesterone and 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM) were similar to those reported previously for normal sows. Removal of one (two sows), two (two sows), three (two sows) or four (one sow) fetuses was followed by premature parturition, within 42–144 h of surgery. Labour lasted 24 to 30 h. Almost immediately after fetectomy, PGFM levels in plasma increased and were accompanied by a decline in progesterone concentrations. High PGFM values (13–60 ng/ml) were present at parturition. Oestrone concentrations were variable or rose slightly at this time. The results suggest that all fetuses in a litter must be present to maintain pregnancy to term. Pregnancy may depend upon fetal suppression of prostaglandin F_2α production and release until the appropriate time for parturition.     

The origin of circulating 13,14-dihydro-15-keto-prostaglandin F2α during delivery

All uterine tissues as well as the fetal membranes and the placenta can form prostaglandins from endogenous precursors but it is not clear which of the tissues is the main site for the increase in PGF_2α production during human parturition. To examine this question, we measured plasma prostaglandin levels before and at intervals after expulsion of the fetus, placenta, and membranes. The concentration of PGFM at the beginning of the second stage of labor was significantly higher than before the onset of labor. Five minutes after the birth of the infant, the concentration had doubled. Thirty minutes after the expulsion of placenta and membranes, plasma PGFM had fallen to the levels at full dilatation; two hours postpartum it was still significantly raised over levels before labor. Since the halflife of PGFM in the circulation is about 7 minutes, these findings indicate that the uterine tissues are important sources of PGFM during labor. In contrast, endogenous oxytocin levels, which were significantly raised over control levels at the second stage of labor, did not change during the third stage, and decline postpartum to control levels. Oxytocin infusion did not influence PGFM levels at 5 and at 30 minutes postpartum, but raised them at 2 hours.     

The stability of 13,14-dihydro-15 keto-PGE2

13,14-Dihydro-15 keto-PGE₂ decomposes by first order reaction kinetics, dependent on pH, temperature and albumin concentration. Under common experimental conditions at or near neutrality in the absence of albumin decomposition is suspended with the formation of 13,14-dihydro-15 keto-PGA₂. Cyclization into 11-deoxy-13,14-dihydro-15 keto-11,16-bicyclo-PGE₂ occurs at elevated pH in purely aqueous buffers, and also at or near neutrality in the presence of albumin. Albumin accelerates, quantitatively, the decomposition of 13,14-dihydro-15 keto-PGE₂ and promotes, qualitatively, the formation of the bicyclo rearrangement product. High performance liquid chromatographic analysis indicates that the cyclization product exists in at least three epimerically distinct forms. The two major epimers have been synthesized and isolated in pure form. They differ, mainly, by their optical rotation.     

Peripheral plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F2α and progesterone around luteolysis and during early pregnancy in the goat

Peripheral plasma concentrations of 13,14-dihdyro-15-keto-prostaglandin F_2α (PGFM) and progesterone were determined during both luteolysis in the oestrous cycle and early pregnancy in four goats. Luteal regression, characterised by decreasing progesterone concentrations, began on day 12 or 13. PGFM concentrations showed a pulsatile pattern around this time, with peak concentrations increasingly markedly as progesterone levels fell and oestrus approached. During early pregnancy progesterone concentrations did not fall after day 12 and no marked elevation of PGFM above basal values of 50–150 pg/ml was detected.     

Radioimmunoassay of 15-keto-prostaglandin E2 in peripheral plasma after oral administration of prostaglandin E2 tablets used for induction of labour

A radioimmunoassay of the 15-keto-metabolite of prostaglandin E₂ has been developed. The details of the assay are described. Using unextracted plama, serial measurements of 15-keto-PGE₂ have been carried out every 15 minutes for 3 hours in 9 women ingesting 0.5 mg PGE₂ oral tablets used for the induction of labour.     

Plasma levels of 13,14-dihydro-15-keto PGE2 after vaginal application of a new PGE2 film

To determine the release and absorption profile of prostaglandin E₂ from a new vaginal film formulation containing 850 μg PGE₂, serial plasma levels of 13,14-dihydro-15-keto PGE₂ were measured by radioimmunoassay in pregnant women between 16 and 18 weeks gestation. A control group, using placebo vaginal film was included in the study. There was a somewhat uniform increase in the plasma levels of the PGE₂ metabolite, reaching peak levels between 4 and 6 hours after application of the film. The findings suggest that this drug formulation could be used clinically when slow constant release of the prostaglandin is required over a period of hours such as in pre-induction cervical ripening of term pregnancy.     

Conditions of the cervix for the increase of plasma levels of 13,14-dihydro-15-keto-prostaglandin F2 alpha after amniotomy at term

Amniotomy was performed in 12 multiparas at term but not in labor. In 6 of these patients (group I), the fetal head and cervix condition were favorable for amniotomy, and in the other 6 (group II), they were not favorable. In all group I patients, a sudden and progressive descent of the fetal head, and onset and progress of labor were noted within 5 hours. Plasma 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) levels increased significantly (P less than 0.05) in 4 of these cases with time. In group II patients, descent of the head was less than that in group I patients (P less than 0.05), and neither strong labor nor rise of PGFM levels was noted within 5 hours. These data support our view that amniotomy at an appropriate time results in the onset and progress of labor, and the rise of plasma PGFM in virtue of the sudden and exponential increase of the head to cervix force, but amniotomy at an inappropriate time does not, because this force is unchanged.     

Plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F during pregnancy in sheep

A specific and sensitive radioimmunoassay is described for 13,14-dihydro-15-keto-prostaglandin F in ovine plasma. Using this assay it has been shown that, in sheep, jugular venous 13,14-dihydro-15-keto-prostaglandin F concentrations increase at parturition and correlate well with concentrations of prostaglandin F in the utero-ovarian vein. It is suggested that uterine prostaglandin F production under these conditions may be assessed by measuring peripheral venous 13,14-dihydro-15-keto-prostaglandin F, thereby avoiding the need for chronic utero-ovarian venous catheters.     

Effects of corticotropin-releasing hormone on ACTH, cortisol and 13,14-dihydro-15-keto prostaglandin E2 in patients with diabetes insipidus before and after captopril treatment

A corticotropin-releasing hormone (CRH) test was performed on 7 patients with central diabetes insipidus (DI) and on 7 healthy subjects. The test was repeated on the patients with DI after 3 days of oral treatment with captopril at a dose of 100 mg daily. No significant difference in the responses of plasma ACTH and cortisol to CRH between the patients and the controls was found. The short-term captopril treatment resulted in a significant decrease of both basal and CRH-stimulated ACTH and cortisol levels in the patients with DI. CRH did not induce any changes in the stable metabolite of prostaglandin E₂ 13,14-dihydro-15-keto-prostaglandin E₂ (PGE₂-M) in the patients with DI before or after the captopril treatment. The results obtained suggest that vasopressin is not an obligatory factor for a normal ACTH response to CRH. Angiotensin II (A II) is involved in the regulation of ACTH. This study confirmed our previous data showing the lack of any specific effect of CRH on PGE₂ production.     

Jugular levels of 13,14-dihydro-15-keto-prostaglandin F and progesterone around luteolysis and early pregnancy in the ewe

Six non-pregnant ewes at day 12 of the estrous cycle each had a day-12 embryo transferred into the uterine horn ipsilateral to the corpus luteum, and 4 non-pregnant ewes at day 13 each had a day-13 embryo similarly transferred. Four control ewes, 2 at day 12 and 2 at day 13 received sheep serum into the uterine horn ipsilateral to the corpus luteum. Jugular blood samples were taken at 2-hourly intervals for 3 days post-surgery, then twice-daily for a further 4 days, and the plasma radioimmunoassayed for progesterone and 13,14-dihydro-15-keto-prostaglandin F. All control ewes exhibited estrus within the expected time range and pulsatile peaks of 13,14-dihydro-15-keto-prostaglandin F occurred coincident with declining progesterone levels. With one exception, the recipient ewes had prolonged cycles and those ewes found pregnant at necropsy, 30 days after transfer, showed no progesterone decline and no pulsatile peaks of prostaglandin during days 12 to 16 after estrus. These observations suggest that the presence of the embryo at a critical stage after mating suppresses the release of uterine prostaglandin F_2α.     

Temporal relationship between plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F and neurophysin I/II around luteolysis in sheep

Plasma concentrations of neurophysin I/II (N-I/II), 13,14-dihydro-15-keto-prostaglandin F (PGFM) and progesterone were measured by radioimmunoassay in plasma samples collected from four sheep at hourly intervals between 0700 and 1900 h from Days 12–17 of the estrous cycle. Plasma samples were also collected from a fifth sheep at 2-hourly intervals during Days 12–16 of the cycle. In all sheep, intermittent surges in the plasma concentrations of PGFM and N-I/II occurred during the period of luteal regression. On at least one occasion in each sheep a surge in the plasma concentration of N-I/II was observed coincident with a rise in PGFM concentrations. In general, the highest levels of N-I/II were observed early in luteolysis (Days 13–14 of the cycle) while the corresponding levels of PGFM in plasma were maximal around Day 15 when luteolysis was well advanced.It is suggested from this temporal data that oxytocin, which is considered to be released in association with N-I/II, may play an important role in ovine luteolysis by stimulating the secretion of prostaglandin F from the uterus during Days 13–15 of the estrous cycle.     

Phosphatidylethanolamide derivatives of prostaglandins E1 and E2

Prostaglandins E₁ (PGE₁) and E₂ (PGE₂) have been coupled with the amine group of phosphatidylethanolamine (PE) by means of dicyclohexylcarbodiimide. These complexes basically mimic the relaxant and contractile effects of the corresponding free prostaglandins (PGs) on various smooth muscle preparations, but exhibit a delayed onset of action and a lower affinity for the PG receptors. The complexes are comparable with the free, parent PGs, in their intrinsic activities. The same holds true for the effects on blood pressure and on the motility of the uterus . The PGE₂-PE complex is hydrolysed to release obviously free PGE₂ by cell-free homogenates prepared from various tissues, but not by blood plasma. The PGE₂-PE complex is immunologically indistinguishable from the free PGE₂.     

Effects of dibuturyl cAMP, LHRH, and aromatase inhibitor on simultaneous outputs of prostaglandin F2α, and 13,14-dihydro-15-keto-prostaglandin F2α by term placental explants

Explants from term human placentas were maintained in culture with daily changes of medium. Daily output of PGF_2α and PGFM¹ decreased during the course of the incubation. Addition of 4 μg/ml DHEAS or 67 μg/ml LDL cholesterol had no effect on output of PGF_2α or PGFM. Addition of 1.6, 3.2, or 6.4 μg/ml of LHRH to the culture plates had no effect on output of PGFM or PGF_2α, but LHRH increased hCG output. Dibutyryl cAMP (1mM, 2mM, and 4mM) increased output of PGF_2α, PGFM, and hCG. Aromatase inhibitor decreased hCG output, but it was without effect on output of PGF_2α, or PGFM. Significant correlations were demonstrated between progesterone, PGFM, PGF_2α, and hCG, suggesting that PGF_2α originates in the syncytiotrophoblast cell. The ability of LHRH to stimulate output of hCG but not PGF_2α while dbcAMP stimulated both suggests that either PGF_2α and hCG arise in different cells or that LHRH does not act through cAMP.     

Effects of prostaglandins E1 and E2 on activity in laryngeal and pharyngeal afferent fibers

Prostaglandins (PG) E₁ and E₂ were applied topically to the receptive fields of feline laryngeal and pharyngeal sensory receptors, while action potentials were recorded from single - or few-fiber preparations of the superior laryngeal nerve. When initially dissolved in ethanol, PGs stimulated these sensory receptors. If ethanol was not used as a solvent for the PGs, they did not stimulate the sensory receptors. Similarly, local application of dilute (0.025%, v/v) solutions of ethanol alone excited the receptors, whereas phosphate buffer alone did not. Thus PGE₁ and PGE₂ do not themselves stimulate sensory receptors in the larynx and pharynx. These findings suggest that irritant properties of PGEs on upper airways are attributable to the ethanol used as a solvent.     

Radioimmunoassay of 13, 14-dihydro-15-keto-prostaglandin F2a

A simple method is described for the determination of 13, 14-dihydro-15-keto-prostaglandin F_2a, (PGFM), using a highly specific antiserum raised in New Zealand rabbits. It involves extraction of human peripheral venous plasma with diethyl ether after addition of tritiated PGFM and HCl. Radioimmunoassay is performed on appropriate aliquots; after 2 hours or overnight equilibration the bound and free metabolite are separated using dextran-coated charcoal. The mean values ± S.D. obtained are as follows: healthy males 32 ± 16 pg/ml, females during follicular phase 48 ± 18 pg/ml, luteal phase 37 ± 8 pg/ml, first trimester of pregnancy 66 ± 33 pg/ml, second trimester 67 ± 42 pg/ml and third trimester 72 ± 26 pg/ml.     

Levels of 13,14-dihydro-15-keto-PGE2 in some biological fluids as measured by radioimmunoassay

A rabbit antiserum directed toward the prostaglandin E₂ metabolite 13,14-dihydro-15-keto-prostaglandin E₂ (KH₂PGE₂) was produced by immunization with a human albumin-KH₂PGE₂ conjugate. The antiserum recognized the 15-keto-group (it cross reacts with 13,14-dihydro-prostaglandin E₂ 0.2%); the saturated 13,14-bond (it cross reacts with 15-keto-prostaglandin E₂ 7%); the 9-keto group (it cross reacts with 13,14-dihydro-15-keto-prostaglandin F_2α 5%); and the 11-hydroxy group (it cross reacts with 13,14-dihydro-15-keto-PGA₂ 0.4%).By subjecting the antiserum to preparative isoelectric electrofocusing, populations of antibodies that varied in their cross reaction with 13,14-dihydro-15-keto-prostaglandin F_2α (KH₂PGF_2α) from 20% to 1% were obtained. The levels of KH₂PGE₂ in plasma of rat and mouse as measured by radioimmunoassay of the unfractionated plasma were 0.39 ± 0.07 ng/ml and 0.41 ± 0.13 ng/ml, respectively. Recovery of exogenously added KH₂PGE₂ from human plasma was 100%. Radioimmunoassay with two antisera; an antiserum directed toward KH₂PGF_2α that cross reacts with KH₂PGE₂ 1% and the antiserum to KH₂PGE₂, demonstrated that KH₂PGE₂, not KH₂PGF_2α, was being measured with the anti-KH₂PGE₂. The levels of KH₂PGE₂ in rat plasma did not vary with sex. In rats, the levels of KH₂PGE₂ markedly increased after exercise stress.In mice carrying a spindle-cell sarcoma (SAI) and a fibrosarcoma (SaD2), the levels of KH₂PGE₂ in the plasma increased with time after transplantation. The increase was not observed in the plasma of mice carrying a transplantable anaplastic carcinoma (15091AK), a lymphatic leukemia (AW5147), two mammary adenocarcinomas (CADI, CAD2), a myeloid leukemia (C1498), and a hepatoma (BW7756).     

Conditions of the cervix for the increase of plasma levels of 13, 14-dihydro-15-keto-prostaglandin F2α after amniotomy at term

Amniotomy was performed in 12 multiparas at term but not in labor. In 6 of these patients (group I), the fetal head and cervix condition were favorable for amniotomy, and in the other 6 (group II), they were not favorable. In all group I patients, a sudden and progressive descent of the fetal head, and onset and progress of labor were noted within 5 hours. Plasma 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM) levels increased significantly (P < 0.05)_in 4 of these cases with time. In group II patients, descent of the head was less than that in group I patients (P < 0.05), and neither strong labor nor rise of PGFM levels was noted within 5 hours. These data support our view that amniotomy at an appropriate time results in the onset and progress of labor, and the rise of plasma PGFM in virtue of the sudden and exponential increase of the head to cervix force, but amniotomy at an inappropriate time does not, because this force is unchanged.     

The effect of endocervical PGE2-gel (Prepidil™) gel on plasma levels of 13,14-dihydro-15-keto-PGE2(PGEM) in women at term

Term pregnant patients undergoing preinduction cervical softening were randomized to receive no treatment (controls) or 0.5 mg PGE₂-triacetin gel (Prepidil™ gel) endocervically. Plasma samples containing PGEM collected 4 hrs post-treatment and converted to the stable bicyclo degradation product (bicyclo-PGEM) were assayed by RIA. Positive clinical effect (responders) during 12 hrs after treatment (Bishop score increase≥3, in labor or delivered) were assessed. All evaluations were blind.In nonresponders (n=35), the means of the bicyclo-PGEM variables (mean, maximum, area under the curve) were all about 18% higher in gel-treated patients (n=6) than controls (n.s.). In responders (n=38), the variables were all about 80% higher in gel-treated women (n=32) than controls (p<.01). In controls (n=35), the responders (n=6) had 50% higher levels than nonresponders (n.s.). In the gel-treated women (n=38), responders (n=32) had about 140% higher levels than nonresponders (<.01).The results suggest that both exogenous and endogenous bicyclo-PGEM were measured. Differences in pairwise comparisons suggest that there may be substantially less exogenous bicyclo-PGEM in the gel nonresponders than in gel responders or substantially more endogenous bicyclo-PGEM in gel responders than in control-responders.     

Effect of oxytocin on plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F2 alpha during the oestrous cycle and early pregnancy in the goat.

The effect of subcutaneous oxytocin on plasma concentrations of 13,14-dihydro-15-keto-prostaglandin (PG) F2 alpha (PGFM) was examined in the goat at various periods during the oestrous cycle and early pregnancy. 100 i.u. oxytocin was administered daily for 4 day, the dose being divided and given at 0900 and 2100 h; PGFM concentrations were assessed after the first treatment of each day. On days 3-6 (oestrus, day 0) PGFM concentrations increased significantly (P less than 0.001) within 15 minutes and both non-pregnant and mated goats exhibited oestrus behaviour by day 7. Significant (P less than 0.01) increases in PGFM were also produced on days 7-10, in both non-pregnant and pregnant goats, but the responses diminished from day 7 to day 10; only one goat (non-pregnant) came into oestrus. There was a marked difference in response between groups, however, during days 12-15. In non-pregnant goats significant (P less than 0.05) increases in PGFM were detected on days 13-15, but in pregnant animals oxytocin was without effect. Similarly, oxytocin did not increase PGFM concentrations on days 17-20 of pregnancy. However, uterine responsiveness reappeared in pregnant goats with significant (P less than 0.01) increases in PGFM on days 24 and 25.