Uteroglobin gene expression in the rabbit uterus throughout gestation and in the fetal lung. Relationship between uteroglobin and eicosanoid levels in the developing fetal lung.

Uteroglobin (UG) gene encodes a cytokine-like, multifunctional, antiinflammatory protein, with potent phospholipase A2-inhibitory activity. It has been suggested that during implantation this protein protects the embryos from maternal immunological assault, facilitates the maintenance of quiescence in the uterus throughout pregnancy, prevents the onset of premature labor, and helps maintain an inflammation-free respiratory organ. This latter function of UG is suggested to be accomplished by preventing hydrolysis of surfactant phospholipids by a lung-specific phospholipase A2. Using reverse transcription polymerase chain reaction, in situ hybridization, immunofluorescence, and radioimmunoassay, we studied UG gene expression in the rabbit uterus throughout gestation and in the fetal lung. Here, we report that: (a) contrary to previous reports, UG gene expression in the rabbit uterus occurs throughout gestation with a precipitous decline just before parturition; (b) this gene expression is dramatically increased in the fetal lung with increasing gestational age; and (c) while there is an inverse relationship between the levels of UG, PGE2, and PGF2 alpha, a positive correlation was found in that of UG and leukotriene C4 in the fetal lung. Our results raise the possibility that dysregulation of UG gene expression, at least in part, may contribute to the onset of premature labor and the development of inflammatory lung disease in premature neonates.

A prospective assessment of carpal tunnel surgery with respect to age.

Six hundred and thirty five carpal tunnel decompressions in 490 patients were studied prospectively in two hand surgery centres to assess the effect of increasing age on the outcome after surgery. The outcome was assessed using the Levine-Katz carpal tunnel questionnaire, Tinel's sign, Phalen's test, Semmes-Weinstein monofilaments and pinch and grip strengths. Assessments were made pre-operatively, at 2 weeks and 6 months postoperatively. Information was also sought concerning co-morbid conditions. Cases were divided into four groups (less than 40 years of age, 40 to 60, 61 to 80, and over 80 years of age). Patients improved significantly in all age groups after carpal tunnel surgery. Despite a relatively high number of co-morbidities, older patients had an acceptable complication rate and their improvement was comparable to all other age groups.

Digital anaesthesia: comparison of the efficacy and pain associated with three digital nerve block techniques.

There are three commonly used methods of digital block anaesthesia: viz. subcutaneous, metacarpal and transthecal. A randomized, single-blinded study on 50 healthy volunteers was performed to determine time to onset, pain level and preference. Volunteers each received all three blocks, serving as their own controls. Time to onset was significantly longer (P<0.001) for the metacarpal block than for the subcutaneous or transthecal blocks. There was no significant difference in average pain level between the methods, as measured on a scale from 1 to 10. Volunteers chose the subcutaneous block (43%) as their first choice over the metacarpal block (33%) or the transthecal block (25%). The transthecal block had prolonged discomfort lasting 24 to 72 hours after injection in 20 subjects (40%). These findings suggest that subcutaneous block is effective and preferred by healthy volunteers for digital anaesthesia.

Angiotensin II and [Sar1, Ile5, Ala8]angiotensin II effect on contractile activity and prostaglandin production of in vitro pregnant rat uteri.

The effect of angiotensin II and [Sar1,Ile5,Ala8]-angiotensin II on uterine contractions and the relationship of uterine prostaglandins to these effects were studied. Uterine segments from pregnant rats were monitored in vitro for isometric contractile activity in Krebs-Ringer medium (95% O2-5% CO2; 37 C). The medium was sampled periodically and assayed for prostaglandin E2, prostaglandin F2 alpha, and 13,14-dihydro-15-keto-prostaglandin F2 alpha by RIA. Angiotensin II increased frequency of contractions and integrated contractile force in a dose-related fashion. Angiotensin II (1 microgram) resulted in increased prostaglandin (PG) production, but there was no clear dose-related effect. Indomethacin significantly reduced PG production (P < 0.001); however, the contractile response to angiotensin II was not affected. [Sar1,Ile5,Ala8]Angiotensin II had no effect on spontaneous contractile activity or PG production in uteri from 18 or 21 days of pregnancy, nor did [Sar1,Ile5,Ala8]angiotensin II affect oxytocin-stimulated uterine contractions. [Sar1,Ile5,Ala8]Angiotensin II (2.5 microgram) did inhibit (P < 0.05) uterine contractions induced by angiotensin II (0.5 microgram), but PG production was not affected. In conclusion, the studies described provide evidence that angiotensin II-induced uterine contractions of in vitro pregnant rat uteri are not dependent upon increased PG production.

Progesterone and prostaglandin secretion by ovulated rat cumulus cell-oocyte complexes.

Hormone production (progesterone and prostaglandin) by oocyte-cumulus cell complexes was studied in the postovulatory period. Cumulus-oocyte complexes were collected from oviducts of prepuberal rats after the synchronization of follicle development and ovulation with PMS gonadotropin and hCG injections. Cumulus masses from individual animals were cultured in roller vessels containing Eagles' Minimal Essential Media with or without added heat-inactivated rat sera (5% CO2-air; 37 C). Culture media were collected after 4.5 h of incubation and analyzed by RIA for prostaglandin E2 (PGE), PGF, and progesterone. Extensive progesterone secretion occurred during the culture period and was markedly stimulated by the addition of rat serum; however, cumulus masses did secrete some progesterone in the absence of rat serum and without exogenous gonadotropins. The addition of aminoglutethimide, a steroidogenesis inhibitor, suppressed progesterone secretion but did not alter the secretion of PGF or PGE. Both PGF and PGE were detected in the culture medium, and in all cases, PGE was the predominant PG detected. Indomethacin, an inhibitor of PG synthesis, suppressed PGE and PGF secretion but had no effect on progesterone synthesis. Small amounts of PGs were also detected in nonincubated cumulus-oocyte masses. Hormone secretion in culture was measured after cumulus mass dissociation by enzymatic digestion. Somatic cumulus cells, rather than germ cells, were the primary cellular source of progesterone and PGs. The results demonstrate that cumulus-oocyte complexes can synthesize and secrete steroid and PG hormones subsequent to ovulation, and thus, these masses should be considered functional endocrine tissues. The physiological significance of the endocrine activities of the cumulus-oocyte complex are discussed in terms of oviductal and gametic processes. (Endocrinology 108: 457, 1981)

The effects of bradykinin on ovulation and prostaglandin production by the perfused rabbit ovary.

Involvement of the vasoactive peptide bradykinin (BK) in ovulation, oocyte maturation, and prostaglandin (PG) production was assessed using an in vitro perfused rabbit ovary preparation. In the first experiment, BK at a concentration of 0.033, 0.33, or 3.3 micrograms/ml was added to the perfusate of one ovary at hourly intervals for the first 10 h of perfusion. The contralateral control ovary was treated with medium alone parallel to the experimental ovary. Ovaries were perfused for a total of 12 h. BK induced ovulation in the absence of gonadotropin in a dose-related fashion, but did not induce maturation of ovulated ova or follicular oocytes. BK significantly stimulated PG production at all concentrations tested, but the effect was not dose related. Prostacyclin, as reflected by the concentration of 6-keto-PGF1 alpha in the perfusate, was the major PG produced. Smaller quantities of PGE2 and PGF2 alpha were present in the perfusate. After a single injection of BK (3.3 micrograms/ml), 6-keto-PGF1 alpha and PGF2 alpha production increased within 15 min and reached a maximum at 60-90 min. PGE2 did not change significantly over this time period. The addition of 1 microgram/ml indomethacin to the perfusate completely inhibited BK-stimulated PG production. However, indomethacin did not significantly affect the ovulatory efficiency of BK-treated ovaries. Neither BK nor indomethacin induced any degenerative changes in ovulated ova or follicular oocytes. The addition of a BK antagonist at 1 microgram/ml every 30 min to the perfusate resulted in an effective blockade of hCG-induced ovulation. These results suggest that BK is involved in the process of follicle rupture, but BK may induce ovulation by a mechanism(s) other than through PG stimulation.

Prostaglandins and progesterone secretion in the in vitro perfused pseudopregnant rabbit ovary.

The relationship between progesterone and prostaglandin (PG) secretion in the pseudopregnant rabbit corpus luteum was investigated using isolated in vitro perfused ovaries. Progesterone and PG secretion were measured on days 1, 11, and 18 of hCG-induced pseudopregnancy. The mean progesterone secretion increased significantly from days 1 to 11, and then decreased significantly by day 18. PG secretion was inversely correlated with progesterone secretion, suggesting that PG might inhibit progesterone secretion. To test this hypothesis, indomethacin, an inhibitor of PG secretion, was administered to intact rabbits from days 11-18 of pseudopregnancy and/or on day 18 ovaries were perfused in vitro with indomethacin. Indomethacin administered in vivo, in vitro, or both in vivo and in vitro significantly reduced PG secretion compared to that in controls, but did not affect progesterone secretion. In addition, perfusion of ovaries in vitro with PGF2 alpha did not alter progesterone secretion on either day 11 or day 18. Thus, although there is an inverse relationship between progesterone and PG secretion during pseudopregnancy, PGF2 alpha alone had no effect on progesterone secretion. These results question the hypothesis that PGF2 alpha alone is the luteolytic factor.

Prostaglandins as mediators of vasotonia in the immature retina.

In vivo observations of the immature beagle puppy retina indicates that in general O2-breathing induces vasoconstriction whereas CO2 or CO2 combined with O2 induces vasodilation. It was the goal of this study to determine whether a specific correlation could be found between this pattern of vascular response to changes in blood gas content and production of prostaglandins in the eye. Prostaglandin production by puppy choroidal and retinal tissues and platelets exposed in vitro to these gas combinations was such that it appears platelet-derived thromboxane could have a significant influence in determining retinal vasotonia.

Serum and urine markers of bone metabolism during the year after hip fracture.

OBJECTIVE: As part of a larger study to describe indices of recovery during the year after hip fracture, the current prospective study investigated longitudinal changes in serum and urine markers of bone metabolism for the year after hip fracture and related them to bone mineral density (BMD). DESIGN: A representative subset of participants provided serum and urine samples and had bone density measured at 3, 10, 60, 180, and 365 days postfracture. SETTING: Two Baltimore hospitals. PARTICIPANTS: The subjects were 205 community-dwelling, white women age 65 and older with fresh proximal femur fractures. MEASUREMENTS: Samples were assayed for specific bone-related proteins and bone turnover markers, including serum osteocalcin (OC), procollagen type 1 carboxy-terminal extension peptide (PICP), bone-specific alkaline phosphatase (BAP), and urinary deoxypyridinoline (DPD) cross-links. Selected hormonal regulators of bone metabolism, including parathyroid hormone (PTH), calcitonin (CT), 1,25-dihydroxy vitamin D(3) (1,25 (OH)(2)D), and estrone (E(1)) were measured from serum samples. Repeated measures analyses were used to evaluate postfracture changes in each of the markers. RESULTS: BAP, OC, and PICP were most active during the early postfracture period (3-60 days). BAP and OC remained elevated at 365 days compared with 3 days. DPD rose 48% from 3 days to 60 days, but this difference was not statistically significant. PTH and 1,25 (OH)(2)D increased steadily and significantly from 3 to 365 days. E(1) was highest at baseline and decreased at each time point, whereas CT showed no significant changes. When subjects were stratified into high-, medium-, and low-BMD groups based on their measurement at 3 days, both osteoclastic and osteoblastic markers in the low-BMD group displayed exaggerated and different patterns over time compared with the other groups. CONCLUSION: Currently, the standard treatment of care for hip fractures still results in high morbidity and mortality and failure to regain prefracture quality of life. Gaining an understanding of bone cell activity in these patients after hip fracture, derived by measuring markers longitudinally during recovery, provides a baseline by which to measure the effectiveness of new interventions to improve recovery from hip fracture.

Serum concentrations of steroids, parathyroid hormone, and calcitonin in postmenopausal women during the year following hip fracture: effect of location of fracture and age.

BACKGROUND: Hip fracture in the aged is a major health problem, especially considering the increasing proportion of the elderly in the population. This study examines changes in circulating levels of hormones, which are purported to affect bone metabolism, in response to hip fracture in postmenopausal women. METHODS: Patients consisted of women ages 65 and older who had surgery within 2 days of fracture. Serum samples were obtained at 3, 10, 60, 180, and 360 days postfracture. Healthy women without hip fractures from the same age range served as a control group (n = 17). Hormones were determined by radioimmunoassay. Subjects with fractures in the neck region of the femur (n = 78) were compared to subjects with fractures in the trochanteric region (n = 88). RESULTS: Estrone concentration (47.6 +/- 5.7 pg/mL; mean +/- SEM) at 3 days postfracture was elevated (p < .001) compared to control levels of 20.7 +/- 4.6 pg/mL. By 2 months, levels had declined to control levels. Androstenedione and the adrenal hormones, DHEAS and cortisol, displayed similar responses. Parathyroid hormone (PTH) levels were not significantly different from the control concentration at 3 days following fracture, but increased (p < .001) during the year following fracture. Calcitonin concentrations were much higher (p < .001) 3 days postfracture (42.1 +/- 3.7 pg/mL) compared to controls without fracture (9.8 +/- 3 pg/mL). Except for testosterone, no differences could be attributed to fracture location. Only PTH, with concentrations higher in the older age groups (p < .001), showed an age-related response. CONCLUSIONS: Following hip fracture, there are some dramatic responses in hormones that purportedly are mechanistically important in bone metabolism. These changes include transient increases in steroid hormones, chronic elevations in calcitonin, and rising levels of PTH during the year after fracture.

Placental immaturity associated with anencephaly.

The placenta associated with anencephalic infants demonstrates striking histologic immaturity near term. This consists of large numbers of prominent cytotrophoblastic cells with frequent mitoses.

Changes in uterine venous prostaglandin F metabolites following intravenous infusion of ritodrine in sheep.

The effectiveness of the beta-adrenergic receptor agonist ritodrine as a tocolytic agent is limited by the tachyphylaxis that occurs with its sustained usage. In order to understand the nature of this tachyphylaxis, we investigated the effect of ritodrine on uteroplacental prostaglandin (PG)F2 alpha production in pregnant sheep. Using general anesthesia in five pregnant sheep, we placed catheters in the aorta and vena cava and in the uterine vein from the uterine horn. In random order on different days, we infused ritodrine (4 micrograms/kg/minute) or physiologic saline into the maternal vena cava at a rate of 0.184 mL/minute. Uterine venous and maternal arterial blood was sampled 60 minutes before and immediately before the infusion and then at 60, 120, 180, and 240 minutes during the infusion. After centrifugation, the serum was frozen and then assayed for the metabolite of PGF2 alpha (PGFM). Uterine venous PGFM increased significantly after 60 minutes of ritodrine infusion (mean increase 1.164 ng/mL; P less than .05), and this increase was sustained during the 4-hour infusion. The PGFM gradient across the uteroplacental bed (uterine vein - aorta) was also significantly elevated during the infusion, suggesting a uteroplacental or fetal membrane source of the PG. Saline had no effect on uterine venous PGFM or the PGFM gradient. These results suggest that ritodrine stimulates PGF2 alpha production, and this may contribute to the tachyphylaxis that occurs with ritodrine and limits its long-term effectiveness as a tocolytic agent.

Prostaglandin F metabolite concentration as a prognostic factor in preterm labor.

Concentration of the 13,14-dihydro,15 keto-metabolite of prostaglandin F2 alpha (PGFM) was measured in women being observed for preterm labor. The mean initial PGFM level was significantly higher in patients who delivered preterm (65.9 +/- 9.7 pg/mL; N = 14) than in patients not in preterm labor (32.1 +/- 4.3 pg/mL; N = 11; P less than .01). Plasma PGFM concentrations decreased significantly during ritodrine therapy only in successfully treated patients (P less than .05). All patients with initial PGFM concentrations greater than or equal to 55 pg/mL delivered preterm. Two of four patients not considered to be in preterm labor but who delivered prematurely (within five days of initial evaluation) had initial PGFM concentrations of greater than 55 pg/mL. Concentration determinations of PGFM might be a useful adjunct in identifying early preterm labor and in predicting success of tocolytic therapy.

Plasma prostaglandin in pregnant women with term and preterm deliveries.

Because of the involvement of prostaglandins in the labor mechanism, it was hypothesized that elevated prostaglandin levels would presage preterm deliveries. A comparison of plasma 13,14-dihydro-15-ketoprostaglandin F2 alpha (PGFM) levels was made between pregnant patients who eventually delivered at term (N = 22) and those who delivered preterm (N = 17). Maternal blood was collected from each patient at approximately 1-week intervals between 16 and 40 weeks' gestation and during labor. Mixed cord blood was also obtained at delivery. PGFM was measured in plasma by radioimmunoassay. Plasma PGFM concentrations did not vary throughout pregnancy. PGFM concentrations did increase during labor, and the highest levels were found in cord blood at delivery. Mean PGFM levels during labor and in cord blood were not significantly different between the preterm and term groups. In 1 preterm delivery patient the PGFM concentration during labor was higher than that of all other patients, including those who delivered at term. Excluding this patient, preterm labor PGFM concentrations were significantly lower (P less than .05) than those at term. In general, the study provides evidence that most cases of preterm labor are not characterized by elevated prostaglandin levels.

Stimulation of follicular growth with "pure" FSH in patients with anovulation and elevated LH levels.

Ovarian follicular growth was stimulated in 4 patients with clomiphene resistant polycystic ovary (PCO)-like disease, with "pure" human pituitary FSH during 10 cycles. In 8 cycles additional hCG was given to induce ovulation. Serum LH and FSH and plasma estradiol and progesterone were determined daily. From the response patterns of steroidogenesis, four functional stages of follicular development can be distinguished, each subsequent stage being characterized by an increasing spontaneous estradiol production and a decreasing capacity to produce extra estradiol in response to stimulation with additional hCG. It is concluded that the unpredictable response to gonadotropin stimulation of patients with PCO-like disease is due to the varying state of development of the follicles at the start of the stimulation. The suggestion is made, based on the different responses in 3 cycles of the same patient (EW), that the lack of positive estrogen feedback in patients with PCO-like disease may be a hyposensitivity to estrogens rather than an absolute insensitivity.

Cul-de-sac fluid in women with endometriosis: fluid volume, protein and prostanoid concentration during the periovulatory period--days 13 to 18.

Cul-de-sac fluid from women with histologically confirmed endometriosis (n = 45) or from infertile women without evidence of endometriosis (n = 28) was collected at the time of laparoscopy during the periovulatory period (days 13 to 18). This fluid was analyzed for prostaglandin E2 (PGE2), prostaglandin F2a (PGF2a), 13,14-dihydro-15 keto-PGF2a (PGFM), and thromboxane B2 (TXB2) by radioimmunoassay (RIA). Protein content of the fluid also was determined. No difference (P greater than 0.05) in cul-de-sac fluid volume was found between women with and without endometriosis, nor were differences detected in the level of any of the prostanoids measured in fluid from infertile control patients compared with those with endometriosis. This was true regardless of whether the prostanoids were expressed as a concentration, total amount in fluid, or as a ratio of prostanoid to protein content. The present study does not support the theory that cul-de-sac fluid prostanoids provide a useful diagnostic index of endometriosis.

The rate at which human sperm are immobilized and killed by mild acidity.

OBJECTIVE: To determine the rate at which mild acidity immobilizes and kills human sperm and to evaluate an acidic microbicide, BufferGel, for sperm immobilization. DESIGN: Controlled in vitro study. SETTING: An academic research university and hospital andrology lab. PATIENT(S): Eight volunteer male sperm donors. INTERVENTION(S): Semen samples were treated with hydrochloric acid (HCl) or BufferGel. MAIN OUTCOME MEASURE(S): Sperm motility was measured by using a computerized automated semen analyzer and video microscopy. Sperm membrane permeability and intracellular pH were measured by using fluorescent techniques. RESULT(S): In semen acidified with HCl to pH 4.0, sperm were rapidly immobilized (within 1 min) and were irreversibly immobilized (killed) within 10 minutes. The speed of immobilization and of killing were both linearly proportional to hydrogen ion activity over a pH range of 7.5-4.0. Across the same range, the intracellular pH of human sperm equilibrated to within 0.5 pH units of extracellular pH within 1-2 minutes. BufferGel immobilized sperm significantly faster than HCl from pH 4.0-6.0. CONCLUSION(S): Exposure to mild acidity rapidly acidifies the intracellular pH of human sperm and is rapidly spermicidal. BufferGel accelerates acid immobilization of sperm.

Pharmacokinetic studies on quinacrine following intrauterine administration to cynomolgus monkeys.

Recent efforts have been made to develop a chemical oviductal occluding agent. Intrauterine quinacrine has been used in certain areas of the world with moderate success in effected tubal closure. This report presents the pharmacokinetics of a quinacrine solution (30 mg) as administered to cynomolgus monkeys via the intrauterine route, compared with intravascular injection. The data show rapid transfer of the drug from the uterine to the vascular compartment and uptake by almost all tissues examined. Although plasma concentrations disappear within 24 hours, levels can be detected in most tissues for at least 1 week following intrauterine injection. After 28 days, however, tissue levels of the drug are absent or near the limit of detection.

Hormonal considerations in early normal pregnancy and blighted ovum syndrome.

The hormonal profiles of six pregnancies which terminated in miscarriage with the blighted ovum syndrome have been studied and compared with those of a group of patients similarly studied who had clinically normal pregnancies terminating in live birth at term. The serum chorionic gonadotropin (HCG) values were below normal or at the lowest limit of normal in five of six patients. Three patients had progesterone values within 1 SD of the normal, with normal serum estradiol values. It was concluded that the hormonal profile of early pregnancy is characterized by rising serum HCG and estradiol levels and a declining serum progesterone level from the 5th to the 8th week. The theoretical explanation for the dichotomy seems to be that the fetal adrenal anlagen, even at this early embryonic stage, can produce steroid precursors which are aromatized to estradiol. The production of progesterone, however, does not seem to be possible. Abnormal serum estradiol levels strongly suggest the absence of fetal development and a blighted ovum. However, no single hormonal level will distinguish between blighted ovum and potentially salvagable threatened abortion.

Effect of silver nitrate on pregnancy termination in cynomolgus monkeys.

The effect of intrauterine injection of 1% silver nitrate on pregnancy was studied in 13 cynomolgus monkeys. The monkeys received, in a random manner, intrauterine injections of 1 ml of either 1% silver nitrate (n = 7) or 0.9% NaCl solution (n = 6) between days 27 and 43 of pregnancy. Silver nitrate injection resulted in vaginal bleeding, beginning 1 or 2 days after treatment, that lasted for an average of 5.3 days. In all of these cases, pregnancy was terminated. Injection of normal saline had variable sequelae, but four of six monkeys were delivered of healthy offspring. All of the silver nitrate-treated animals recycled and, after remating, two of the seven subsequently became pregnant again and were delivered of normal healthy infants. This study demonstrates the efficacy of intrauterine injection of 1% silver nitrate in terminating early pregnancy.