PubMed HealthSearch

Biomedical subjects

B L Lasley

Publications and source records attributed to B L Lasley.

At least 19 recordsLinked to original sources

The role of relaxin in glycodelin secretion.

Glycodelin is a glycoprotein named for its unique carbohydrate structure. Glycodelin is produced by the secretory endometrium during the late luteal phase and returns to baseline during menses of the ensuing cycle, whereas in conceptive cycles it rapidly increases. Although progesterone and possibly estradiol are required for glycodelin production, they are not directly involved in the synthesis and release of this protein. Their role may be development of the endometrial secretory glandular elements, whereas other factors are required to initiate and maintain glycodelin secretion. The pattern of relaxin secretion during the luteal phase and early pregnancy is similar to that of glycodelin, but their profiles have not been determined simultaneously. To investigate the relationship of relaxin and glycodelin, two studies were conducted. In the first study, relaxin, glycodelin, and ovarian steroids were measured in daily serum samples from nonconceptive and conceptive natural cycles. Profiles of relaxin and glycodelin were closely associated, with the onset of relaxin preceding glycodelin secretion by 1-2 days in nonconceptive cycles, and the pregnancy-associated increases in each hormone differing by about 2 days. The second study tested the hypothesis that relaxin stimulates glycodelin secretion. Samples were obtained from patients injected with human relaxin for 28 days. In subjects demonstrating ovarian cyclicity, glycodelin secretion was elevated, but it was not detected in subjects without ovarian cyclicity or in placebo-treated control subjects. This study reveals a close temporal and quantitative relationship between relaxin and glycodelin profiles in the late luteal phase and early pregnancy. It also demonstrates that relaxin administration can stimulate glycodelin production from a developed endometrium. This is the first report of a nonsteroidal ovarian factor that controls glycodelin secretion, and these results suggest a function for relaxin during early pregnancy. Glycodelin is a potent inhibitor of sperm zona pellucida binding by virtue of its extensive carbohydrate structure, but it is normally at a nadir in the periovulatory period. The data demonstrate that relaxin can stimulate glycodelin secretion throughout the menstrual cycle, including the periovulatory period, when relaxin-induced glycodelin secretion could have a contraceptive effect.

Adolescent

Hormonal indicators of pregnancy in llamas and alpacas.

OBJECTIVE: To determine concentrations of estrone sulfate in serum, estrone sulfate in urine, relaxin in serum, and progesterone in serum in pregnant llamas and alpacas and to assess the potential of these hormones as pregnancy indicators. DESIGN: Prospective study. ANIMALS: 19 parous pregnant camelids (8 llamas and 11 alpacas). PROCEDURE: Estrone sulfate concentrations (in serum and in urine) and progesterone concentrations (in serum) were determined by enzyme immunoassay. Relaxin concentrations (in serum) were measured by radioimmunoassay. Serum and urine samples were collected daily for the first 30 days after breeding and, thereafter, once weekly until parturition. RESULTS: Estrone sulfate concentrations (in serum and in urine) peaked twice during pregnancy. The first took place 21 days after breeding and the second during the last month of pregnancy. Relaxin concentrations increased at 3 months of gestation to > 20 ng/mL, decreased at 5 months to 5 ng/mL, then increased from 8 months of pregnancy until parturition. Progesterone concentrations were detectable 4 days after breeding and were maintained > 2 ng/mL throughout pregnancy. CLINICAL IMPLICATIONS: The first increase in estrone sulfate concentration over basal values may indicate early interaction between mother and embryo, whereas the second increase may reflect fetal viability. Use of estrone sulfate concentration to diagnose pregnancy in llamas and alpacas is highly dependent on time of sampling. Relaxin concentration in serum is a superior indicator of pregnancy after the second month in the Ilama and alpaca because its existence is suggestive of interaction between mother and fetus, and concentrations are greater than basal values for a long period of pregnancy. Progesterone is not a direct product of the embryo or fetus and only indirectly confirms a diagnosis of pregnancy.

Animals

An enzyme immunoassay for serum and urinary levonorgestrel in human and non-human primates.

A microtiter plate enzyme immunoassay (EIA) is reported for the measurement of levonorgestrel (LNG) in serum and urine samples of human and non-human primates, and the results are compared to data obtained by radioimmunoassay (RIA). Rabbit polyclonal antibodies were raised against the bovine serum albumin conjugate of the 3-O-carboxymethyl oxime (CMO) derivative of LNG. The enzyme label was produced by the conjugation of horseradish peroxidase to LNG at the 3-position by the same CMO bridge used for the immunogen. The assay requires 2.5 hours to perform using 2.2-azino-di-(3-ethylbenzthiazoline sulfonic acid) diammonium salt as the chromogenic substrate. Serum (100 microliters) is extracted with petroleum ether prior to assay, whereas urine samples (25 microliters) are diluted and measured directly. The sensitivity of the assay is 0.25 pg/well with a 50% displacement of label at 7.5-9.5 pg and a linear response through 250 pg/well. Minimum levels of 8.7 and 10.0 pg/ml can be detected in serum and urine samples, respectively. Changes in serum LNG concentrations were measured in women and non-human primates following LNG implantation or injection. In the non-human primate study, serum LNG concentrations began to rise rapidly following i.m. injection of LNG, with peak levels occurring on days 3 to 5, then decreasing to approximately 25-35% of peak levels for the duration of the study. Circulating concentrations of 1.86 +/- 0.18 ng/ml LNG were reached in women the first week post-insertion of Norplant implants and decreased by 50% at 7-10 days, 75% after 14-21 days, followed by a steady decrease during the next 60-70 days to constant low levels that exhibited a high individual variation. Correlation coefficients of EIA and RIA results were 0.988 for human serum, 0.926 for human urine, and 0.972 for non-human primate serum.

Adolescent

Effects of two antiprogestins on early pregnancy in the long-tailed macaque (Macaca fascicularis).

The abortifacient effects of mifepristone and HRP 2000 were compared in gravid long-tailed macaques. Thirty-six animals were studied with treatment administered either by the oral (0.5 or 5.0 mg/kg; N = 5 per antiprogestin per dose) or intramuscular (i.m.) routes (0.5 mg/kg; N = 5 per antiprogestin) on gestational days (GD) 23-26; six vehicle controls were included. Blood samples were collected for assay of progesterone (P4) and each of the antiprogestins (pre-treatment, daily GD 23-28, every other day GD 30-40), and animals were monitored sonographically throughout gestation. Results of these studies indicated high rates of abortion with i.m. administration (3/5 mifepristone, 4/5 HRP 2000) and 5.0 mg/kg oral route (4/5, 2/5, respectively), with less effects noted at oral doses of 0.5 mg/kg (2/5, 0/5, respectively). No early abortions were observed in the control groups. Following daily i.m. treatment, peak levels of 8-16 ng/ml mifepristone were detected whereas 6-10 ng/ ml of HRP 2000 were noted (GD 26-27). No serum levels of mifepristone were detected following either of the oral doses whereas serum levels of 2-6 ng/ml HRP 2000 were noted with high dose oral administratation. Results of these studies suggest: (1) both antiprogestins are roughly comparable in terminating early pregnancy although HRP 2000 may be more efficacious when administered i.m. whereas mifepristone may be more effective when administered orally; (2) similar levels of biological activity are seen with the i.m. and high dose oral dosing regimens, with little or no activity with the oral low dose; and (3) infants resulting from surviving pregnancies were not affected by early gestation exposure.

Abortifacient Agents, Steroidal

Bone density and cyclic ovarian function in trained runners and active controls.

This study was conducted to determine whether rigorous exercise training adversely affects ovarian hormone levels and bone health in cyclically menstruating trained runners. Ovarian hormones, bone mineral density (BMD), body composition, 3-d diet records, 3-d estimated energy expenditure, and menstrual histories were evaluated in 10 trained collegiate runners and 10 moderately active controls. The trained runners had lower total body calcium per kg of soft lean tissue measured by DEXA (P = 0.045). Half of the trained runners had experienced stress fractures compared with only one of the moderately active controls. The trained runners' lumbar (L2-L4) BMD (1.178 g.cm-2) was not significantly different from that of the active controls (1.283 g.cm-2) (P = 0.074) but, for all subjects combined, there wasa significant inverse relation between L2-L4 BMD and distance run per week (P = 0.036). Further, adding age, body weight, percent body fat, daily energy intake, and daily calcium intake to a stepwise multiple regression analysis did not significantly improve predictive precision. The trained runners consumed nearly twice the amount of calcium (1089 mg.d-1 vs 641 mg.d-1, respectively; P = 0.036), while intake of other nutrients did not differ significantly between groups. Urinary estrone conjugates (E1C) were lower in the trained runners during the early follicular phase (P = 0.028), while pregnanediol-3-glucuronide (PdG) was not significantly different between groups during the luteal phase (P = 0.213). Thus, it appears that lower estrogen production, especially during the early follicular phase, and not progesterone, is associated with lower whole body calcium per kg of soft lean tissue and, probably, L2-L4 BMD. Results of this study also suggest that regular menstrual cycles do not imply normal ovarian hormone function in young women who are engaged in either recreational or competitive running.

Bone Density

Identification of anovulation and transient luteal function using a urinary pregnanediol-3-glucuronide ratio algorithm.

The sensitivity and specificity of a urinary pregnanediol-3-glucuronide (PdG) ratio algorithm to identify anovulatory cycles was studied prospectively in two independent populations of women. Urinary hormone data from the first group was used to develop the algorithm, and data from the second group was used for its validation. PdG ratios were calculated by a cycles method in which daily PdG concentrations indexed by creatinine (CR) from cycle day 11 onward were divided by a baseline PdG (average PdG/Cr concentration for cycle days 6-10). In the interval method, daily PdG/CR concentrations from day 1 onward were divided by baseline PdG (lowest 5-day average of PdG/CR values throughout the collection period). Evaluation of the first study population (n = 6) resulted in cycles with PdG ratios > or = 3 for > or = 3 consecutive days being classified as ovulatory; otherwise they were anovulatory. The sensitivity and specificity of the PdG ratio algorithm to identify anovulatory cycles in the second population were 75% and 89.5%, respectively, for all cycles (n = 88); 50% and 88.3% for first cycles (n = 40) using the cycles method; 75% and 92.2%, respectively, for all cycles (n = 89); and 50% and 94.1% for first cycles (n = 40) using the interval method. The "gold standard" for anovulation was weekly serum samples < or = 2 ng/ml progesterone. The sensitivity values for all cycles and for the first cycle using both methods were underestimated because of apparent misclassification of cycles using serum progesterone due to infrequent blood collection. Blood collection more than once a week would have greatly improved the sensitivity and modestly improved the specificity of the algorithm. The PdG ratio algorithm provides an efficient approach for screening urine samples collected in epidemiologic studies of reproductive health in women.

Adult

Association of spontaneous abortion and other reproductive effects with work in the semiconductor industry.

This study tested the hypothesis that fabrication room (fab) work in the silicon-based semiconductor industry is associated with an increased risk of spontaneous abortion (SAB). The study was conducted nationwide at 14 companies representing a spectrum of large to small manufacturers. A small increase in risk of SAB was observed among fab workers compared with nonfabrication room (nonfab) workers in two cohorts, historical (adjusted RR = 1.43, 95% confidence interval [CI] 0.95-2.09) and prospective (adjusted RR = 1.25, 95% CI = 0.63-1.76). Analysis of specific fab exposures in the historical cohort showed a consistent, dose-response association of SAB with photoresist and developer solvents, whose major component was ethylene-based glycol ethers. The consistency of our findings and the toxicological data for these agents suggest that this is a causal association. Independent associations of SAB with self-reported stress and with etching fluorides were observed and require further research. No significant decrease in fertility was observed among men or women working in fabs, but reduced fecundability was suggested for some women fab workers.

Abortion, Spontaneous

Laboratory methods for evaluating early pregnancy loss in an industry-based population.

Laboratory methods were adapted or developed to analyze approximately 70,000 daily urine samples collected during more than 2,500 menstrual cycles from 448 women working in the semiconductor industry. An immunoenzymometric assay (IEMA) for human chorionic gonadotropin (hCG) was employed for screening cycles in order to optimize laboratory resources and to reduce the number of samples requiring analysis by less efficient methods. The presence of hCG in urine was confirmed by the definitive immunoradiometric assay (IRMA). The screening assay eliminated 78% of cycles from further analysis because there was no evidence of conception. Thirty-eight of 448 cycles identified as having significant levels of hCG with the IEMA were confirmed as hCG positive with the IRMA. HCG-positive cycles were further evaluated by examination of daily diary data and by laboratory assays for ovarian and pituitary hormones. As a result of these evaluations, 17 of the 38 cycles identified by the IRMA as positive for hCG were found to be nonconceptive cycles. These results demonstrate the effectiveness of screening assays for hCG, as well as the importance of using multiple urinary biomarkers for the detection of early fetal loss with daily urine samples.

Abortion, Spontaneous

Epidemiologic methods for prospective assessment of menstrual cycle and reproductive characteristics in female semiconductor workers.

Methods were developed to assess potential adverse effects of semiconductor fabrication work in a prospective study of reproductive health. All women aged 18-44 years who worked in seven silicon-wafer fabrication sites in five companies and a frequency-matched sample of women in nonfabrication jobs were included. Among 3,480 selected for screening, 2,639 (75%) completed a self-administered questionnaire to identify women at risk of pregnancy. Among the 739 (28%) eligible women, 481 (65%) completed baseline interviews and 402 (83.6%) completed at least one menstrual cycle of follow-up by providing daily diaries and daily urine samples. Menstrual cycle characteristics were assessed from questionnaires and diaries. Urine samples were assayed for reproductive hormones to identify conceptions. The usual cycle length recalled at baseline was 28 days. The mean cycle length (MCL) recorded in diaries was 29 days, with greater dispersion than at baseline. The median of the MCL from diary data was 28 days for women reporting regular cycles at baseline but 34 days for women reporting irregular cycles at baseline, and the median standard deviation in cycle length per woman was 2.5 days and 7.5 days, respectively. The prospective method, while expensive and labor intensive, showed good compliance. Nevertheless, recall also provided reasonably accurate estimates and distinguished women with regular and irregular cycles.

Adolescent

Prospectively assessed menstrual cycle characteristics in female wafer-fabrication and nonfabrication semiconductor employees.

Women aged 18-44 years in silicon-wafer fabrication-room (fab) jobs and frequency-matched women in nonfab jobs were screened for a prospective study of reproductive health (n = 2,639). Among the 739 (28%) eligible women, 481 (65%) completed a baseline interview; 402 completed at least one menstrual cycle of follow-up with daily diaries and urinary assays to exclude conceptive cycles. Adjusted mean cycle lengths (MCL) did not differ between fab and nonfab women (p = 0.97). Women working in thin film and ion implantation (TFII) had the highest adjusted MCL (34.8 +/- 1.7 days) compared with nonfab workers (32.5 +/- 1.4 days, p = 0.07). Among women working exclusively in one group, TFII women had significantly higher MCL (36.1 +/- 2.04 days) than nonfab women (32.0 +/- 1.38 days, p = 0.017). TFII women were also more likely to have all cycles > 35 days (adjusted relative risk [RR] = 2.45; 95% CI = 0.85-6.06). Variability was assessed by logarithmic transformation of the mean standard deviation (MLSD) in cycle length per woman and adjusted for age and ethnicity (4.5 days for fab vs. 4.0 days for nonfab, p = 0.16). Women working exclusively in TFII or photolithography (PHOTO) had significantly higher adjusted MLSD in cycle length (6.68 +/- 1.28 and 5.72 +/- 1.24 days, respectively) than women in nonfab (4.1 +/- 1.16 days, p = 0.013 and 0.019, respectively). Fab and nonfab women did not differ significantly in mean days of bleeding or risk of having cycles > 35 or < 24 days. However, elevated risks of having cycles < 24 days were seen in supervisor engineers (adjusted RR = 2.46, 95% CI = 1.19-3.63) and PHOTO women (adjusted RR = 1.83, 95% CI = 0.94-2.88).

Adolescent

Prospective assessment of fecundability of female semiconductor workers.

To investigate a possible effect of reduced fecundability (probability of conception per menstrual cycle) among women who fabricate silicon wafers, 152 fabrication-room (fab) and 251 nonfab workers were followed for an average of five menstrual cycles. Daily urine samples were analyzed to confirm clinical spontaneous abortions (SABs) and early fetal losses (EFLs). Adjusted fecundability odds ratios (FRs) for fab workers ranged from 0.59 to 0.72 (p = 0.09-0.28 vs. nonfab). For clinical pregnancies only, the adjusted FR ranged from 0.43-0.50 (p = 0.04-0.09 vs. nonfab). This lower fecundability was most pronounced among dopants and thin-film workers [adjusted FR = 0.61, 95% confidence interval (CI) = 0.27-1.40 for all pregnancies; adjusted FR = 0.22, 95% CI = 0.05-0.96 for clinical pregnancies] and in workers exposed to ethylene-based glycol ethers (adjusted FR = 0.37, 95% CI = 0.11-1.19).

Abortion, Spontaneous

Prospective monitoring of early fetal loss and clinical spontaneous abortion among female semiconductor workers.

Women who work in silicon wafer fabrication rooms (fabs) have been reported to have an increased risk of spontaneous abortion (SAB). Although previous studies have included only clinically recognized SABs, more than two-thirds of SABs may be clinically unrecognized. To determine whether fab work is associated with SAB, we recruited 152 fab and 251 nonfab workers, who collected urine samples for a 6-month period. Samples were analyzed by immunoradiometric assay for the presence of human chorionic gonadotropin to detect early fetal losses. Approximately 63% of fab and 46% of nonfab pregnancies ended in SAB [adjusted relative risk (RR) = 1.25; 95% confidence interval (CI) = 0.63-1.76]. Similar RR were seen for women who worked in dopant and thin-film processes (adjusted RR = 1.30; 95% CI = 0.51-1.96) or in masking (adjusted RR = 1.30; 95% CI = 0.59-1.84). The four pregnancies among women who worked with ethylene-based glycol ethers ended in SAB.

Abortion, Spontaneous

The use of urinary hormonal assessments in human studies.

The collection and analysis of urine samples provides a practical method for monitoring female reproductive events in non-laboratory and non-clinic populations. Collection of biologic samples permits objective assessment of reproductive health endpoints in epidemiologic studies and for epidemiologic research purposes can provide validation of information provided by the subjects, especially outcomes which are usually concealed and thus unknown to the participant. Urine sampling has several advantages over the collection of blood samples, such as simplicity, non-invasiveness, and cost efficiency. Several studies have shown that endocrine information similar to that obtained in blood samples can be obtained from assays of daily urine samples. The measurement of human chorionic gonadotropin in daily and selected urine samples has been incorporated into several recent epidemiologic studies focusing on early fetal loss, and ovarian and pituitary hormone metabolites have been measured in daily urine samples to evaluate ovarian function in studies focusing on women's reproductive health. As the strategy of urinary monitoring becomes more accepted as a legitimate research tool, laboratory methods are being modified to improve performance, reduce costs and adapted to sophisticated algorithms using multiple hormonal measurements to identify a number of end-points.

Biomarkers

The postpartum llama: fertility after parturition.

Fertility was evaluated at various times during the postpartum period in the llama. Fifty-six parous female llamas chosen at random were bred at 10, 20, and 30 days postpartum with six intact males. Half of the females copulated only once and the other half twice within an interval of 24 h. Ovarian activity was monitored by ultrasonography and analysis of urinary estrone sulfate and pregnanediol glucuronide (PdG). At the time of copulation, all females had developed ovulatory-size follicles. Ovulation was confirmed by the presence of a CL at 8 days after breeding and PdG concentrations > 1 ng/mg creatinine (Cr). Conception was defined as PdG concentrations > 1 ng/mg Cr at 15 days after breeding and the presence of a CL. Pregnancy was defined as the presence of a CL, PdG concentrations > 1 ng/mg Cr and an embryonic vesicle 20 days postbreeding. There were no significant differences in the proportion of females ovulating after breeding at different times postpartum; however, conception and pregnancy were significantly greater in females bred at Day 20 or 30. Ovarian follicle size was significantly larger at 30 days (9.1 mm) than at 10 and 20 days (7.9 and 8.8 mm, respectively) of breeding, with no difference in concentrations of estrone sulfate. The proportion of females conceiving as a result of breeding at 10 days postpartum (6 out of 10) was significantly less than for females bred at 20 (13 out of 15) or 30 days (16 out of 18).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Immunocytochemical localization of nonluteal ovarian relaxin.

Relaxin has been demonstrated to be produced by the corpus luteum of the menstrual cycle and pregnancy and is also produced by the endometrium and decidua, although these nonluteal sources may not contribute to circulating relaxin concentrations. The reports of luteal production of relaxin have failed to consider nonluteal ovarian sources. To look for sources of nonluteal ovarian relaxin, human ovaries were collected from patients who underwent removal of the ovary for a variety of reasons. Tissues were fixed in formalin and embedded in paraffin. Two monoclonal antibodies were used for immunocytochemical staining, one directed against human relaxin and the other against the C peptide of prorelaxin. In addition to the expected staining of corpora lutea, the luteinized theca interna but not granulosa of developing follicles from ovaries with an active corpus luteum of the cycle also stained positive for both relaxin and prorelaxin. Ovaries from term pregnant women with luteinized theca also demonstrated staining for relaxin and prorelaxin. In addition to luteal and thecal cell staining, small clusters of pseudodecidual cells in the periphery of the ovary stained positive for relaxin and prorelaxin. These data indicate that the ovary contains theca interna-derived structural elements in addition to the corpus luteum that produce relaxin when a corpus luteum is active, while granulosa-derived elements do not. This suggests that luteal production of relaxin is from theca-derived elements and may explain instances of independent relaxin and progesterone secretion.

Adult

Ovarian and endocrine patterns associated with reproductive abnormalities in llamas and alpacas.

Ovarian activity was characterized in llamas and alpacas with hypoplastic ovaries, cystic follicles, or ovulatory failure. Ovarian follicular activity was determined by transrectal ultrasonography and urinary estrone sulfate analysis; pituitary response was determined by measurement of plasma concentrations of luteinizing hormone. Llamas and alpacas with hypoplastic ovaries had follicles < or = 6 mm (minimal ovulatory size, 7 mm). Cystic follicles, defined as > 12 mm, were maintained for a mean of 9 days. Follicular activity in the ovary contralateral to the cystic follicle tended to be suppressed while the cystic follicle was present. Ovarian response to copulation in females with cystic follicles varied according to the stage of the cystic follicle. Animals with ovulatory failure did not release adequate luteinizing hormone after copulation, even though they had mature and normal follicles (8 to 12 mm). The cystic follicle syndrome appears to be temporary, whereas the syndromes involving hypoplastic ovaries and ovulatory failure may permanently affect fertility.

Animals

Tamoxifen as an antifertility agent in the long-tailed macaque (Macaca fascicularis).

The potential use of tamoxifen, a nonsteroidal antiestrogen, as an antifertility agent was studied in the long-tailed macaque (Macaca fascicularis). Twenty-six cycling females were bred, then treated with a single oral dose of tamoxifen (5 mg/kg) (N = 13) or vehicle (N = 13) on day 4 post-ovulation. Serum progesterone (P4) and tamoxifen concentrations were evaluated on post-ovulation days 4, 8, 12, 16, and 18. No effects of treatment were observed on P4 concentrations or on the fertility rate--pregnancy was achieved in 4/13 controls (31%) and 6/13 treated females (46%). Analysis for serum tamoxifen concentrations in samples collected during the fertility and a supplemental pharmacokinetic study (N = 3; single oral dose of 10 mg/kg; urine and serum evaluated) failed to reveal any detectable tamoxifen levels. It was concluded that (1) absorption of tamoxifen may be negligible under the described treatment regimens or (2) tamoxifen metabolism/clearance occurs at a rapid rate.

Administration, Oral

Simple extraction and enzyme immunoassays for estrogen and progesterone metabolites in the feces of Macaca fascicularis during non-conceptive and conceptive ovarian cycles.

A simple method for extracting ovarian steroids from feces is presented, together with enzyme immunoassay systems for measuring estrogen and progesterone metabolites. Small amounts of feces were combined in a 1:10 proportion with a modified phosphate buffer, shaken for 24 h, centrifuged, and decanted; the supernatant was directly measured for estrogen and progesterone metabolites by enzyme immunoassays. Serum estradiol and progesterone profiles were compared to urinary and fecal profiles in the same animals to determine the degree to which each reflected the ovarian events detectable in serum. The correlation coefficients for the relationship between serum, urinary, and fecal hormones for individual animal cycles were found to be statistically significant in every case but one, where the relationship between serum estradiol and urinary estrone conjugates was not significant. Urinary and fecal measurements were used to determine whether estrogen and progesterone metabolism and excretion varied within and between animals. Variation in unconjugated estrogen and progesterone metabolites was observed in the follicular phase, the luteal phase, and early pregnancy.

Animals