Longitudinal ultrasound data were collected for 18 structures in 37 pigtailed macaque (Macaca nemestrina) fetuses to 1) generate standards of normal fetal growth, 2) measure operator reliability, 3) assess the accuracy of linear and nonlinear regression models to estimate gestational age and dates of delivery, and 4) evaluate the portability of equations and absolute values derived from data at one facility (Seattle) to describe independently collected data on the same species at another facility (Medical Lake Breeding Colony). Femur length, biparietal diameter, and head area were found to be the best predictors of gestational age, as judged by maximum "explained" variance (R2) with minimum error estimates. Days to delivery could not be adequately predicted by any single fetal growth parameter or by the best combination of parameters. Operator reliability was very good: error was 5 5 % over all parameters studied. Although the form of growth curves from each facility was generally the same, variability was much greater for most parameters at Medical Lake, and both femur and humerus lengths were overestimated relative to the Seattle data. The same result was obtained for femur length when Seattle data on pigtailed macaques were compared with published data on rhesus macaques. We believe that these differences in facilities and populations may be due to the practices of the ultrasound operators. We suggest that portability of regression coefficients and absolute values for structures at a given gestational age can be accomplished only if operators are trained to use the same standard methods and receive periodic reliability checks. 0 1995 Wiley-Liss, Inc.

The determination of urinary 17-ketosteroids has limited applications in the understanding and diagnosis of the nature of endocrine disorders associated with mild hirsutism and virilism in the female, because a very large number of these patients excretes 17-ketosteroids well within the normal range. Recent experience of Finkelstein, Forchielli and Dorfman (1) suggests that in these cases the blood levels and nature of the circulating androgens may be very important. Earlier indirect evidence (2-12) and the more recent isolation and partial identification of sulfates of dehydroisoandrosterone, androsterone, and etiocholanolone by Baulieu (13) indicate that 17-ketosteroids are present in plasma mainly in the form of esters. Direct estimation of these conjugated steroids in plasma offers distinct advantages: the alteration of the steroid molecule which occurs with certain types of hydrolysis as well as possible problems of incomplete hydrolysis can be circumvented. Also, information concerning the physiological significance of the steroid conjugating mechanisms may be gained through direct qualitative and quantitative estimation of these conjugates. The present paper describes a convenient method, using quantitative paper chromatography, for the estimation of dehydroisoandrosterone and androsterone sulfates (DHIA-SO and Andro-SO4) in plasma. MATERIALS AND METHODAll reagents and solvents used were of analytical grade, and all solvents, except absolute ethanol, were redistilled prior to use. Florisil was purified as described earlier (14). M-dinitrobenzene was purified by sublimation, using a modification of the "cold finger" technique, and stored in the dark at room temperature. Collection of plasm1ia. Blood from fasting subjects was drawn into heparinized syringes between 8:30 and 9: 30 a.m. The blood was then transferred to a glass centrifuge bottle containing 1 ml of heparin per 100 ml of blood and centrifuged at 2,500 rpm for 30 minutes. The plasma was removed with a syringe equipped with a long spinal needle and transferred to a clean glass vessel.Extraction of steroid con jngates. All analyses were run in triplicate. Fifteen-ml aliquots 1 of the plasma were placed in three 250-ml Erlenmeyer flasks and 45 ml of absolute ethanol was added to each flask. The flasks were swirled several times and placed in an ice bath for 10 to 15 minutes. After filtering the mixture, under vacuum, through two discs of Whatman no. 1 filter paper, the flasks were rinsed three times with 10 ml of absolute ethanol, and these washings were then used to wash the precipitate. The filtrate was transferred quantitatively to a 1 L round-bottomed flask with further washings of absolute ethanol and evaporated to near dryness (1 to 2 drops) on a flash evaporator (Laboratory Glass and Instruments Corp., model FE-2) under vacuum, using dry ice in ethylene glycol for refrigeration. Secondary butanol (15 ml) was then added to effect a partial separation of steroid conjugates from inorganic salts. The steroid conjugates, which dissolve quantita...

The toxicity of azidothymidine (AZT) was studied in monkey dams and fetuses that were exposed to the drug over the entire gestational period. Fourteen virus-free female macaques (Macaca nemestrina) were randomly assigned to AZT or control groups. AZT animals received the drug through a gastric catheter at a dose of 1.5 mg/kg every 4 hours, which produced plasma concentrations similar to those in humans taking 500 to 600 mg/day of AZT. Control animals received water placebo, also through gastric catheter. Some animals participated in both groups. All females were mated with the same male; 41 matings produced 20 pregnancies, of which 16 were carried to term (9 in AZT females; 7 in control females). The AZT animals developed an asymptomatic macrocytic anemia, but hematologic parameters returned to normal when AZT was discontinued. Total leukocyte count decreased during pregnancy and was further affected by AZT administration. AZT-exposed infants were mildly anemic at birth. AZT caused deficits in growth, rooting and snouting reflexes, and the ability to fixate and follow near stimuli visually, but the deficits disappeared over time. These data indicate that early exposure to AZT in utero should have no irreversible adverse effects on the fetus.

Monkeys with excellent reproductive histories were immunized with the laminin peptides YIGSR, RGD, IKVAV, and YD, a control sequence with no known biological function. Sera from the YIGSR-immunized monkey became toxic, causing neural tube defects in whole rat embryo cultures, and this monkey experienced fetal loss after immunization. Sera from the RGD-immunized monkey also became embryotoxic in culture after immunization, but this monkey appeared to become infertile as she failed to initiate a pregnancy for at least 2 years after immunization. In contrast, embryos cultured on sera from the IKVAV-or YD-immunized monkeys were predominantly normal and both monkeys completed successful pregnancies. Antibody levels to the respective peptides or to laminin were not predictive of embryotoxicity, but antibody binding to homogenized yolk sacs as well as to yolk sacs of cultured embryos was associated with sera embryotoxicity and reproductive outcomes in vivo. These observations suggested that the laminin sequences YIGSR and RGD may play a role in immune-mediated reproductive failure by reacting directly with embryonic tissue and could provide a basis for identifying individuals at risk for both spontaneous abortion and infertility.Immune dysfunction has been implicated in recurrent pregnancy loss (1). For example, isolated T cells from recurrent aborters were found to secrete embryotoxic cytokines in response to trophoblast cells (2), while a direct role of autoantibodies in fetal loss has been implicated for women with autoimmune diseases, such as systemic lupus erythematosus (3) and herpes gestationis (4). Anti-phospholipid autoantibodies particularly have been associated with spontaneous abortions (5), but their presence alone could not predict poor pregnancy outcomes (6). Therefore, there has been a need to identify specific autoantibodies responsible for reproductive failure and to establish their mechanisms of pathogenicity (7).Anti-laminin autoantibodies were detected in the sera of some monkeys with histories of reproductive failure, and sera from these monkeys caused neural tube defects in cultures of whole rat embryos (8). When monkeys with excellent reproductive histories were immunized with intact murine laminin, their sera became embryotoxic, again causing neural tube defects in rat embryo cultures, and these monkeys subsequently failed to reproduce (9). When pregnant mice were injected with heterologous anti-laminin antibodies, they spontaneously aborted (10). However, sera from lamininimmunized rats were not consistently embryotoxic and sera anti-laminin antibody levels were not predictive of embryotoxicity (11). These toxic and nontoxic sera were found to recognize different epitopes on Western blots of enzymedigested laminin, suggesting that antibodies to only certainThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. laminin epitopes were r...

The inhibition of the pregnenolone Δ⁵–3Β-hydroxysteroid dehydrogenase-Δ^5-4 isomerase systems in microsomes of human placenta or corpus luteum of pregnancy has been investigated using three steroidal substances: 2Α-cyano- 4,4,17Β-trimethylandrost-5-en-17Α-ol-3-one (cyanoketone), 2- hydroxymethylene, 17Α-methyl-5Α-androstan-17Β-ol-3-one (oxymetholone), and estradiol 17Β. The inhibition is of the competitive type with the synthetic steroids. The apparent K_i’s are approximately 0.3xl0^-5 M and 0.2xI0^-5 m for cyanoketone in the placental and ovarian systems, respectively, and the apparent K_i produced by oxymetholone is 0.6x10^-6 m in the ovarian system, however, that for the placental system is much smaller, 5.5xl0^-8 M. Approximately 50% inhibition was obtained in both systems with 2–5xl0^-5 M estradiol-17Β. Oxymetholone may be useful for the inhibition of corpora luteal function in women.