Uterine catecholamines and prostaglandins during the estrous cycle of the rat.

The uterine contents of norepinephrine, epinephrine, dopamine, and prostaglandins F and E were determined on each day of the rat estrous cycle. Catecholamines were determined by high performance liquid chromatography with electrochemical detection (HPLC-EC) as well as by a radioenzymatic technique; prostaglandins were quantitated by RIA. The norepinephrine and dopamine values obtained by the radioenzymatic assay were approximately 1.5 times as high as the values obtained by HPLC-EC (norepinephrine, 285 vs. 188 ng/g; dopamine, 11.0 vs. 7.5 ng/g). Despite this difference in levels, both analytic techniques showed a decline in uterine norepinephrine from diestrus to estrus, followed by a significant (P less than 0.01) increase in norepinephrine on the day of metestrus. A cyclic pattern was also revealed for uterine dopamine concentration. There was a decline in dopamine concentration from diestrus to proestrus (radioenzymatic, P less than 0.01), followed by a return to high levels at metestrus (HPLC-EC). Epinephrine levels were low (undetectable by radioenzymatic assay; 24 ng/g by HPLC-EC) and showed no variation during the estrous cycle. Prostaglandin F was uniformly higher than prostaglandin E (10 vs. 2.5 ng/uterus). Significant increases in the uterine contents of both prostaglandins were shown on the day of proestrus.

Plasma prostaglandin levels in radiation-induced enteritis.

Increased prostaglandin synthesis has been implicated as a causative factor in the production of radiation induced enteritis. Seventeen patients selected to begin pelvic irradiation for treatment of gynecological cancer had plasma Prostaglandin E, Prostaglandin F, and 13, 14 dihydro 15 keto PGF2 alpha metabolite determined by radioimmunoassay, prior to initiation of radiotherapy, at weekly intervals during treatment and at six weeks following completion of radiotherapy. A total of 362 prostaglandin determinations were performed. Thirteen patients (76%) developed significant diarrhea consisting of three or more watery bowel movements per day. Nine patients (53%) had intermittent colicky pain and six patients (35%) had nausea and vomiting during treatment. Statistical evaluation revealed no significant elevation of plasma prostaglandins during radiotherapy.

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.

A modification of the unlabeled antibody enzyme method using heterologous antisera for the light microscopic and ultrastructural localization of insulin, glucagon and growth hormone.

The requirement of using homologous antisera (primary antiserum and peroxidase-antiperoxidase (PAP) complex raised in the same species) in the unlabeled antibody enzyme method has been investigated at the light and electron microscopic level using the localization of insulin, glucagon and growth hormone as model systems. Optimum immunocytochemical staining for all three antigens was observed when sheep or goat antirabbit gamma-globulin (S-ARgammaG or G-ARgammaG) were used to couple rabbit peroxidase-antiperoxidase complex with either guinea pig antisera to insulin (GP-AIS) or glucagon (GP-AGS), or monkey antisera to rat growth hormone (M-ARGH). The cross-reactivity between S-ARgammaG or G-ARgammaG and immunoglobulins in these primary antisera were substantiated by immunoelectrophoresis and radioimmunoassay. S-ARgammaG was shown to produce precipitation arcs with GP-AIS and M-ARGH that were similar to those seen when the latter were reacted with rabbit antiguinea pig gamma-globulin antiserum and goat antimonkey gamma-globulin antiserum, respectively. Radioimmunoassay results revealed that immunoprecipitation of 6-10% as compared to homologous antisera controls yielded excellent staining localization when S-ARgammaG was used for immunocytochemistry. Thus, heterologous antisera (primary antiserum and PAP complex raised in different species) may be used in the unlabeled antibody enzyme method as long as the coupling antiserum shows cross-reactivity with immunoglobulins of the primary antiserum and the PAP complex.

Plasma prostaglandins in pregnancy.

Plasma prostaglandins were determined by radioimmunoassay in 92 pregnant and 14 nonpregnant women. There was significant elevation of PGA-like material in the first trimester of pregnancy (1744 pg/ml) over that seen in nonpregnant women (576 pg/ml) with continuation of that elevation in the second and third trimesters. No significant difference existed among PGE levels of the nonpregnant group (251 pg/ml) and the first two trimesters of pregnancy (384 pg/ml and 294 pg/ml); the PGE level of the third trimester group (443 pg/ml) was significantly elevated over that of the nonpregnant group. PGF levels remained constant during all trimesters (135 pg/ml, 144 pg/ml, and 130 pg/ml) but exhibited plasma concentrations significantly higher than the nonpregnant group (78 pg/ml). Potential role (s) of prostaglandins as mediators of cardiovascular and renal changes of pregnancy are discussed.

Plasma prostaglandins in the normal menstrual cycle.

Plasma prostaglandins were studied by radioimmunoassay on alternate days during the menstrual cycle in fourteen normal women. No cyclic patterns were found for A-like prostaglandins using an assay which measured 13,14-dihydro-PGA. Mean subject values of PGA-like prostaglandin ranged from 367 to 904 pg/ml and varied significantly among women. Prostaglandin E determinations showed an upward trend beginning 8 days before the LH peak. PGE subject means varied significantly and ranged from 182 to 362 pg/ml. Prostaglandin F did not exhibit a cyclic pattern. The average concentration of PGF for individual women ranged from 58 to 153 pg/ml, showing significant variance. The physiologic implications of the results are discussed as well as recommendations for the design of future studies.

Dysmenorrhea: treatment with an antiprostaglandin.

Painful menses, one of the most frequent gynecologic complaints, is incapacitating for many women. It has recently been proposed that increased endometrial prostaglandin production and prostaglandin-induced myometrial contractility may be responsible for dysmenorrhea. In this prospective, double-blind, 3-way, crossover study, relief of pain by an antiprostaglanding drug, ibuprofen (400 mg), was compared with propoxyphene (64 mg) and placebo in 22 women with severe primary dysmenorrhea. Ibuprofen was significantly more effective in 18 patients when compared to the other 2 treatment regimens (P less than 0.001), while propoxyphene was superior to placebo in 13 patients (P less than 0.05). Prostaglandin E and F synthesis rates in endometrial biopsy specimens taken on the second day of treatment in 2 patients during each treatment cycle were lowest during ibuprofen in one case but showed no definite pattern in the second.

Structural and mechanical changes of uterine arteries during pregnancy in the pig.

A dramatic 15-fold increase in uterine blood flow in pigs occurs during pregnancy in association with marked increases in uterine arterial (UA) diameter. This study was conducted to determine UA collagen and elastin content, diameter, alpha 1- and alpha 2-adrenergic receptor (AR) numbers, norepinephrine (NE) and in vitro reactivity to phenylephrine on d 0, 20, 50, 80 and 110 of pregnancy in the pig. Uterine arterial collagen content declined progressively throughout pregnancy (P less than .01), whereas the content of elastin remained constant from d 0 to d 80, then increased (P less than .05) from d 80 to d 110. The UA collagen to elastin ratio was correlated with UA diameter (r = -.69; P less than .01), which increased from 4.5 mm on d 0 to 9.0 mm on d 110. Uterine arterial alpha 1-AR numbers remained low and constant throughout pregnancy, consistent with its retained ability to contract in response to phenylephrine. Uterine arterial NE content declined (P less than .05) from d 20 to d 80 before increasing slightly to d 110. Uterine arterial alpha 2-AR numbers remained high from d 0 to d 80 before decreasing (P less than .05) to low values on d 110. These data are consistent with a reduced adrenergic neuronal control and increased elasticity of the UA during pregnancy in the pig.

Changing adrenergic sensitivity of the caruncular arterial vasculature supplying the bovine placentome.

With the advancement of gestation, blood flow increases preferentially to the caruncular bed of the gravid uterus in association with a decreasing sensitivity of the uterus to the vasoconstrictive effects of circulating catecholamines. This study directly compared the sensitivity of the caruncular artery (CA) of the isolated bovine placentome to phenylephrine (PE), a specific alpha 1-adrenergic receptor (AR) agonist, with that to norepinephrine (NE) and epinephrine (E), both of which are alpha 1-, alpha 2-, and beta-AR agonists, at two stages of gestation (140 to 170 d, mid-pregnant; 210 to 270 d, late pregnant). The CA of each placentome was perfused with oxygenated Krebs Ringer solution into which PE, NE or E were administered; increases in intra-arterial pressure were recorded. Further, NE content and numbers of alpha 1- and alpha 2-AR in the CA, intercaruncular arteries (ICA) and uterine arteries (UA) were quantitated. The CA from mid-pregnant cows exhibited greater (P less than .05) contractile responses to NE and E than did the CA from late pregnant cows, whereas responsiveness to PE remained constant. No difference in NE content, alpha 1-AR or alpha 2-AR numbers were observed in the UA, ICA or CA between mid-pregnant or late pregnant cows. Alpha 1-AR numbers were similar in CA, ICA and UA. However, CA contained threefold greater alpha 2-AR numbers than either the ICA or UA (50.2 +/- 6.1 vs 14.6 +/- 1.6 and 14.8 +/- 2.4 fmol/mg protein, respectively; P less than .01).(ABSTRACT TRUNCATED AT 250 WORDS)

Local and systemic effects of intrauterine estradiol-17 beta on luteal function of nonpregnant sows.

This study was conducted to determine if intrauterine injections of estradiol-17 beta (E2 beta) could maintain luteal function in nonpregnant sows. Eight sows were assigned to surgery on d 8 or 9 of the estrous cycle (first day of estrus = d 0). At surgery, cannulas were inserted bilaterally into the uterine artery (UA) and common utero-ovarian vein (UOV), as well as into the lumen of each isolated uterine horn. Electromagnetic blood flow transducers were placed around the middle uterine artery supplying each horn of four sows. After surgery, sows were assigned randomly to receive either intrauterine injections of vehicle (.9% NaCl) into both uterine horns (control sows) or E2 beta into one uterine horn (375 ng/injection) and vehicle into the other (treated sows) every 6 h from 1200 h on d 11 to 1200 h on d 15. Uterine blood flow (UBF) was quantified, and blood was sampled from the UA and UOV, periodically, from d 11 to 18. On d 18, sows were ovariectomized and corpora lutea (CL) were weighed. Blood plasma was subsequently analyzed for progesterone (P4) and prostaglandin F (PGF) by radioimmunoassay. Control sows had smaller (P less than .05) CL than treated sows on d 18 (3,046 +/- 614 vs 4,451 +/- 324 mg). Progesterone concentrations in UOV blood of treated sows tended to increase from d 11 (469 +/- 110 ng/ml) to 18 (626 +/- 209 ng/ml) while P4 in UOV blood of control sows decreased markedly (P less than .01) from d 11 (579 +/- 79 ng/ml) to 18 (14 +/- 5 ng/ml). In addition, UOV P4 concentrations on the E2-beta-injected side of treated sows were higher (P less than .05) than those on the vehicle-injected side from d 14 to 18. The UBF of two treated sows increased eightfold to 10-fold within 12 h of the first E2 beta injection and remained elevated through d 17, while UBF of two control sows remained constant. Prostaglandin F concentrations in UOV blood of treated sows were lower (P less than .05) than in UOV blood of control sows on d 14 and 15. There was no effect of side of E2 beta injection on PGF concentrations, which were similar in UOV blood draining both uterine horns of treated sows. Thus, the local effect of E2 beta in stimulating P4 secretion by the ipsilateral ovary is not due to reduced PGF concentrations in UOV blood draining the E2 beta-injected horn.

Prostaglandin F2 radioimmunoassay utilizing polyethylene glycol separation technique.

PIP: A radioimmunoassay for Prostaglandin F which utilizes polyethylene glycol to separate antibody-bound and free prostaglandin is discribed. The detailed procedure for extraction and chromatographic fractionalization of the "E" and "F" series is presented. The recoveries of labeled prostaglandin obtained at each step of the procedure compared favorably with the 68% overall recovery figure for the complete assay. Data on the sensitivity, specificity, accuracy and reproducibility of the assay are presented. PgF levels in plasmas obtained from normal males and females, pregnant females in the third trimester and females in labor have been determined. Ovarian and uterine tissue levels of Prostaglandin F are also included.^ieng

Effect of prostacyclin inhibition by tranylcypromine on uterine 6-keto-pgf 1 alpha levels during estrogen hyperemia in rats.

A role for prostacyclin (PGI2) as a mediator of estrogen-induced increases in uterine blood volume (UBV) was investigated by measuring uterine tissue levels of 6-keto-prostaglandin F1 alpha (6-keto-PGF 1 alpha), and testing estrogen responses in rats pretreated with the PGI2 synthesis inhibitor, tranylcypromine (TCP). Uterine 6-keto-PGF1 alpha content was determined by radioimmunoassay of tissue extracts purified through the use of high-performance liquid chromatography (HPLC). Estrogen treatment of castrate rats resulted in a significant increase of uterine 6-keto-PGF 1 alpha was compared to saline treated controls (9.3 ng/uterine horn vs 6.7 ng/uterine horn, p=0.01). Pretreatment with TCP (20 mg/kg) markedly reduced the uterine content of 6-keto-PGF 1 alpha (2.5 ng/uterine horn). The typical 50% increase in UBV observed after estrogen was unaffected by tranylcypromine pretreatment. It was concluded that the increased PGI2 synthesis, as indicated by elevated levels of 6-keto-PGF1 alpha, may function as an amplifying mechanism for the uterine vasodilation-induced by estrogen in castrate rats, but that production of this prostanoid is not essential for the estrogen response.

Effect of parasympathetic decentralization and paracervical ganglion excision on reproductive function in the rat.

The uterine paracervical ganglion (Frankenhauser's ganglion) contains the terminal ganglion cells of the sacral parasympathetic system and, in some species, the short adrenergic system. Histochemical studies also show numerous chromaffin cells with morphologic attributes of interneurons. The present study explores the function of cell types present in this ganglion, by seeking changes in reproductive function following either parasympathetic decentralization (transection of the cauda equina) or excision of the ganglion itself. Two well-known reproductive phenomena were observed after each surgical intervention, namely, induction of uterine hyperemia by estrogen administration, and maintenance of normal vaginal cycles. Estrogen-induced uterine hyperemia was not affected by parasympathetic decentralization or ganglion excision. Therefore, nerves originating in, or passing through this structure may be eliminated as components of the vascular control mechanism. In contrast, compared to sham-operated controls, ganglionectomy caused a significant reduction in the proportion of animals exhibiting normal vaginal cycles postoperatively (P less than 0.05). Cycle distribution was more evident in animals ganglionectomized on metestrus (P less than 0.01) and proestrus (P less than 0.05) than in animals ganglionectomized on diestrus or estrus. Since parasympathetic decentralization did not produce cycle disruption similar to ganglionectomy, one may conclude that the cycle-modulating effect does not involve preganglionic fibers of the sacral parasympathetic nerves.

A technique for monitoring blood flow changes with miniaturized Doppler flow probes.

A method for constructing miniaturized Doppler blood flow probes is presented. Since these probes weigh less than 100 mg and have crystal heads less than 0.5 mm in size, they are suitable for chronic placement on vessels as small as 200 micron. The probes are positioned under the vessel and rotated to optimize the Doppler signal. While held in that position, the crystal head is attached to the adventitia of the vessel with cyanoacrylate glue. A cuff holding the vessel and probe in the chosen position is then formed in situ by the application of a drop of silicone polymer. Data are presented showing the linearity of a flow probe response with the volume blood flow at rates as low as 14 microliter/min. With the use of the uterine artery of the conscious, unrestrained rat as an example, the probe was demonstrated to detect a dynamic change in flow.

Effects of indomethacin on the vascular abnormalities of Bartter's syndrome.

We examined the hypothesis that the vascular abnormalities of Bartter's syndrome are due to excess production of prostaglandin. Balance studies and vascular reactivity studies were performed before and after indomethacin (200 mg/day) in a patient with well-documented Bartter's syndrome. During indomethacin, potassium balance became positive, serum potassium rose from 2.1--3 mEq/1 in the absence of potassium supplementation, plasma renin activity decreased from 55--3.2 ng/day and peripheral plasma PGA-like activity fell from 1460 +/- 220 to 456 +/- 71 pg/ml. Before indomethacin, forearm vasoconstrictor responses to brachial arterial infusions of angiotensin II, norepinephrine and to neurogenic reflex stimulation elicited by lower body suction were greatly depressed compared to those of normal subjects. During indomethacin these responses were restored to normal. The dose of intravenous angiotensin II required to increase diastolic blood pressure 20 mm Hg decreased from 160--30 ng/kg/min. These data support the hypothesis that the vascular insensitivity to exogenous angiotensin II, norepinephrine and to neurogenic reflex stimulation observed in this patient with Bartter's syndrome is due to excess prostaglandin. Moreover, stimulation of the renin-angiotensin-aldosterone system in this syndrome appears to be a compensatory adaptation to excess prostaglandin production.