Point Absorber Limits to Future Gravitational-Wave Detectors.

High-quality optical resonant cavities require low optical loss, typically on the scale of parts per million. However, unintended micron-scale contaminants on the resonator mirrors that absorb the light circulating in the cavity can deform the surface thermoelastically and thus increase losses by scattering light out of the resonant mode. The point absorber effect is a limiting factor in some high-power cavity experiments, for example, the Advanced LIGO gravitational-wave detector. In this Letter, we present a general approach to the point absorber effect from first principles and simulate its contribution to the increased scattering. The achievable circulating power in current and future gravitational-wave detectors is calculated statistically given different point absorber configurations. Our formulation is further confirmed experimentally in comparison with the scattered power in the arm cavity of Advanced LIGO measured by in situ photodiodes. The understanding presented here provides an important tool in the global effort to design future gravitational-wave detectors that support high optical power and thus reduce quantum noise.

An integrated "4-phase" approach for setting endocrine disruption screening priorities--phase I and II predictions of estrogen receptor binding affinity.

Recent legislation mandates the US Environmental Protection Agency (EPA) to develop a screening and testing program for potential endocrine disrupting chemicals (EDCs), of which xenoestrogens figure prominently. Under the legislation, a large number of chemicals will undergo various in vitro and in vivo assays for their potential estrogenicity, as well as other hormonal activities. There is a crucial need for priority setting before this strategy can be effectively implemented. Here we report an integrated computational approach to priority setting using estrogen receptor (ER) binding as an example. This approach rationally integrates different predictive computational models into a "Four-Phase" scheme so that it can effectively identify potential estrogenic EDCs based on their predicted ER relative binding affinity (RBA). The system has been validated using an in-house ER binding assay dataset for 232 chemicals that was designed to have both broad structural diversity and a wide range of binding affinities. When applied to 58,000 chemicals identified by Walker et al. as candidates for endocrine disruption screening, some 9100 chemicals were predicted to bind to ER. Of these, only 3600 were expected to bind to ER at RBA values up to 100,000-fold less than that of 17beta-estradiol. The method ruled out 83% of the chemicals as non-binders with a very low rate of false negatives. We believe that the same integrated scheme will be equally applicable to endpoints of other endocrine disrupting mechanisms, e.g. androgen receptor binding.

Threshold analysis of selected dose-response data for endocrine active chemicals.

Using a biologically relevant mathematical model, the Michaelis-Menten equation, we examined published data from endocrine active chemicals for evidence of no-threshold dose-response curves. Data were fit to a modified Michaelis-Menten equation which accounted for total background response. Subsequently, the data sets were analyzed using non-linear regression in order to estimate the four parameters of interest (non-hormone controlled background (Bnh), maximum response (Rmax), endogenous hormone level (D0), and the dose at which a half-maximal response was observed (ED50)) and to determine the fit to the fully modified Michaelis-Menten equation. Subsequently, response data were adjusted to account for Bnh and then normalized to Rmax, while dose data were adjusted to account for D0 and then normalized to the ED50. This data set was combined into a single, composite data set and fit to the fully modified Michaelis-Menten equation. We examined 31 data sets (24 endpoints) from studies on 9 different chemical/hormone treatments. Twenty-six of the data sets fit the modified Michaelis-Menten equation with high multiple correlation coefficients (r>0.90). The normalized data demonstrated a good fit to the modified Michaelis-Menten equation. These results indicate that a variety of biological responses fit the modified Michaelis-Menten equation, which does not have a threshold dose term.

QSAR models using a large diverse set of estrogens.

Endocrine disruptors (EDs) have a variety of adverse effects in humans and animals. About 58,000 chemicals, most having little safety data, must be tested in a group of tiered assays. As assays will take years, it is important to develop rapid methods to help in priority setting. For application to large data sets, we have developed an integrated system that contains sequential four phases to predict the ability of chemicals to bind to the estrogen receptor (ER), a prevalent mechanism for estrogenic EDs. Here we report the results of evaluating two types of QSAR models for inclusion in phase III to quantitatively predict chemical binding to the ER. Our data set for the relative binding affinities (RBAs) to the ER consists of 130 chemicals covering a wide range of structural diversity and a 6 orders of magnitude spread of RBAs. CoMFA and HQSAR models were constructed and compared for performance. The CoMFA model had a r2 = 0.91 and a q2LOO = 0.66. HQSAR showed reduced performance compared to CoMFA with r2 = 0.76 and q2LOO = 0.59. A number of parameters were examined to improve the CoMFA model. Of these, a phenol indicator increased the q2LOO to 0.71. When up to 50% of the chemicals were left out in the leave-N-out cross-validation, the q2 remained significant. Finally, the models were tested by using two test sets; the q2pred for these were 0.71 and 0.62, a significant result which demonstrates the utility of the CoMFA model for predicting the RBAs of chemicals not included in the training set. If used in conjunction with phases I and II, which reduced the size of the data set dramatically by eliminating most inactive chemicals, the current CoMFA model (phase III) can be used to predict the RBA of chemicals with sufficient accuracy and to provide quantitative information for priority setting.

The estrogen receptor relative binding affinities of 188 natural and xenochemicals: structural diversity of ligands.

We have utilized a validated (standardized) estrogen receptor (ER) competitive-binding assay to determine the ER affinity for a large, structurally diverse group of chemicals. Uteri from ovariectomized Sprague-Dawley rats were the ER source for the competitive-binding assay. Initially, test chemicals were screened at high concentrations to determine whether a chemical competed with [3H]-estradiol for the ER. Test chemicals that exhibited affinity for the ER in the first tier were subsequently assayed using a wide range of concentrations to characterize the binding curve and to determine each chemical's IC50 and relative binding affinity (RBA) values. Overall, we assayed 188 chemicals, covering a 1 x 10(6)-fold range of RBAs from several different chemical or use categories, including steroidal estrogens, synthetic estrogens, antiestrogens, other miscellaneous steroids, alkylphenols, diphenyl derivatives, organochlorines, pesticides, alkylhydroxybenzoate preservatives (parabens), phthalates, benzophenone compounds, and a number of other miscellaneous chemicals. Of the 188 chemicals tested, 100 bound to the ER while 88 were non-binders. Included in the 100 chemicals that bound to the ER were 4-benzyloxyphenol, 2,4-dihydroxybenzophenone, and 2,2'-methylenebis(4-chlorophenol), compounds that have not been shown previously to bind the ER. It was also evident that certain structural features, such as an overall ring structure, were important for ER binding. The current study provides the most structurally diverse ER RBA data set with the widest range of RBA values published to date.

High environmental temperature and humidity decrease oocyte quality in Bos taurus but not in Bos indicus cows.

Two experiments were conducted to assess the effects of environmental temperature and humidity on the quality and developmental capabilities of bovine oocytes. In Experiment 1, Bos taurus (Holstein and crossbred Angus) cows were subjected to 5 weekly sessions of ultrasound-guided follicle aspiration from February 16 through March 23 (cool season) and 5 sessions from May 22 through June 20 (hot season). In Experiment 2, Bos taurus (Holstein) and Bos indicus (Brahman) cows were superstimulated (Super-Ov) during the months of August (hot season) or January (cool season), and each cow was subjected to a single oocyte aspiration session. In each experiment, oocytes were classified as normal or abnormal based on ooplasm morphology and cumulus cell layers. In Experiment 1, oocytes classified as normal were in vitro matured and fertilized (IVM/IVF), and the resulting embryos cultured for 8 d. All oocytes recovered from superstimulated cows in Experiment 2 were matured and fertilized in vitro and the subsequent embryos cultured for 8 d, regardless of their morphological appearance. In Experiment 1, Bos taurus cows produced a higher (P = 0.02) percentage of normal oocytes during the cool season (75.9 +/- 8.0) than during the hot season (41.0 +/- 9.5). The percentage of fertilized oocytes developing to the 2-cell (82.4), 8-cell (65.4) and morula (46.6) stages were also greater (P < or = 0.06) during the cool season than the hot season (45.0, 21.2, 6.0 for 2-cell, 8-cell and morula stages, respectively). In Experiment 2, Bos taurus cows (Holstein) had a lower (P = 0.01) percentage of normal oocytes in the hot season (24.5 vs 80.0) and a lower (P < or = 0.003) percentage of fertilized oocytes developing to the 8-cell, morula and blastocyst stages. No difference (P > or = 0.57) in the percentage of normal oocytes or in embryo development was detected between seasons in Bos indicus (Brahman) cows. In conclusion, high environmental temperature and humidity resulted in a marked decline in the quality of oocytes retrieved from Bos taurus cows and markedly decreased their in vitro developmental capabilities. In contrast, a high percentage of oocytes retrieved from Bos indicus cows exhibited normal morphology and yielded a high proportion of blastocysts, regardless of season.

Roles of leukotrienes in bovine corpus luteum regression: an in vivo microdialysis study.

A microdialysis system (MDS) was surgically implanted into the corpora lutea (CL) of 12 normally cycling Holstein heifers. Heifers were either allowed to undergo spontaneous luteolysis (Spontaneous, n = 6) or received an intramuscular injection of prostaglandin F2 alpha (PGF2 alpha) on Day 12 of the estrous cycle (Induced, n = 6). The MDS was implanted on Day 11 in the induced heifers and on Day 17 in Spontaneous heifers. CL were perfused with Ringer's solution at a flow rate of 3 ml/hr beginning immediately after surgery. Dialysate samples were collected hourly for 3-4 days. Samples were assayed for progesterone (P4), oxytocin (OT), PGF, and leukotrienes B (LTB) and C (LTC). Dialysate OT was undetected in all but one Spontaneous and one induced heifer. Lipoxygenase products of arachidonic acid (AA) metabolism (LTB and LTC) in the dialysate were found to be closely associated with luteal regression. In Spontaneous heifers, the mean interval from the first hormone peak to the onset of P4 decline was similar for PGF, LTB, and LTC, with the first peak occurring at 12.8 +/- 8.1, 22.0 +/- 6.1, and 11.0 +/- 8.9 hr before the onset of P4 decline, respectively. The peak LTC value was greater (P < 0.05) than peak LTB or PGF. The 12-hr sampling interval with the highest LTC peak frequency was highly correlated (r = 1.0; P < 0.01) with the onset of P4 decline, but the highest LTB and PGF peak frequencies were not associated with the onset of P4 decline. Indeed, the mean numbers of PGF and LTB hormone peaks were higher (P < 0.05) after the onset of P4 decline than before. Administration of PGF2 alpha on Day 12 of the estrous cycle stimulated a decline in P4 secretion and an increase in the secretion of PGF, LTB, and LTC from the CL. In induced animals, the peak level of PGF was greater (P < 0.05) than peak LTB. These results suggest that the AA metabolites LTB and, especially, LTC play important roles during normal regression of the bovine CL.

Uterine response to progesterone in prepubertal gilts.

During early pregnancy, progesterone stimulates the secretion of proteins and other molecules that support the developing conceptus. Some gilts are able to support conceptus development as early as 110 days of age. The objective of this study was to evaluate the onset of responsiveness of the prepubertal uterus to progesterone. Thirty gilts were assigned to receive 2.2 mg progesterone kg-1 body mass per day or corn oil daily for 14 days starting at 6, 46, 76, 106, and 136 days of age. Hysterectomies were performed the day after the last treatment of progesterone, and the uterine horns were weighed and flushed with sterile saline (0.85% NaCl). Recovered flushings were analysed for total luminal protein, retinol binding protein, uteroferrin, prostaglandin E and prostaglandin F. An interaction between age and progesterone occurred for uterine wet mass (P < 0.001). Progesterone did not affect the uterine mass of gilts that underwent hysterectomy at 20 days of age, but did increase the uterine mass (P < 0.05) in other age groups. Progesterone increased (P < 0.01) the amount of total luminal protein in all but the youngest gilts. An increase in the amounts of retinol binding protein and uteroferrin (P < 0.001) by progesterone was first observed in 90-day-old gilts. Prostaglandins exhibited a different age-related pattern. The amount of prostaglandin E was increased (P < 0.001) by progesterone treatment in gilts aged 90-150 days, with a greater (P < 0.05) response at 120 days than at 90 days old. The response at 150 days old decreased (P < 0.05) to that observed at day 90. The response of prostaglandin F to progesterone followed a similar age-related pattern. Therefore, uterine responsiveness to progesterone develops between 20 and 90 days after birth, and uterine mass responds earlier than the secretory responses measured in our study.

Characterization and proteolytic activity of a cathepsin L-like polypeptide in endometrium and uterine flushings of cycling, pregnant and steroid-treated ovariectomized gilts.

Cathepsin L has been proposed to be involved with the endothelial-chorial type of placentation in the cat. Little information concerning the presence and secretion of cathepsin L is available for a species with noninvasive epitheliochorial placentation such as the pig. Cathepsin L activity in uterine flushings and endometrium from gilts during different days of the oestrous cycle and early pregnancy was analysed through specific substrate metabolism and Western blot analyses with antiserum against cat endometrial cathepsin L. This antiserum was utilized to determine the cellular localization of the enzyme within porcine endometrium. Cathepsin L activity within uterine flushings was elevated on Day 15 of the oestrous cycle and early pregnancy, with activity declining on Day 18. Cat cathepsin L antiserum cross-reacted with a group of 46, 40 and 38 kDa uterine proteins and detected a product within the surface and glandular epithelium of the endometrium. The appearance of the 40 kDa protein was first detected on Day 10 of the oestrous cycle with the 38 kDa proteins appearing on Day 15 and 18 of pregnancy. The 40 and 38 kDa uterine proteins appear to be steroid regulated as 12 days of progesterone administration is necessary to detect the proteins and cathepsin L activity.

Effect of unilateral ovariectomy, gonadotropin stimulation and immunization against a synthetic peptide of the inhibin alpha-subunit on follicular development and oocyte recovery in cattle.

Nulliparous Holstein cows were randomly distributed among 4 treatment groups to test the effects of treatments, including unilateral ovariectomy, anti-inhibin immunization and gonadotropin stimulation on ovarian follicle population and oocyte recovery. The Control treatment consisted of intact cows (I-Control). Unilaterally ovariectomized cows were included in the 3 remaining treatments consisting of ovariectomy alone (U-Control), cows immunized against a synthetic peptide of the alpha(c)-subunit of bovine inhibin (alpha(c)I; U-IH), and cows stimulated with FSH (Super-Ov; 75 units/female/week) and also immunized with alpha(c)I as in the previous treatment (U-IH/FSH). Oocytes were collected by transvaginal ultrasound-guided aspiration on a weekly basis from cows in each treatment for 5 consecutive weeks. Intact Control cows had a greater (P<0.05) number of follicles > or = 3 mm per female (4.7) than the U-Control and U-IH cows (2.6 and 2.9, respectively), and had a similar number of follicles as the U-IH/FSH treatment group (3.5). The numbers of follicles aspirated (2.7 to 3.6) and oocytes recovered/cow (1.6 to 2.6) were similar for cows in the I-Control, U-IH and U-IH/FSH treatment groups. Cows in the U-Control treatment group had a lower (P<0.05) number of aspirated follicles (2.0) and recovered oocytes (1.1) than the I-Control cows. Cows in the U-IH/FSH and U-IH treatments had follicles with larger (P<0.01) diameters (8.7 and 8.2 mm, respectively) than cows in the I-Control (6.6 mm) and U-Control (5.7 mm) treatments. In conclusion, unilateral ovariectomy did not result in compensatory increase of follicle number or size in the intact ovary; cows in the U-IH/FSH treatment group had a greater number of follicles aspirated than the U-Control cows. In addition, the anti-alpha(c)I immunization may have played a role in increasing the number and diameter of the follicles. None of the treatments evaluated in this study improved oocyte retrieval over that of the intact, nontreated cows.

Peri-oestrous hormone profiles, embryonic survival and variation in embryonic development in gilts and primiparous sows.

The primary objective of this study was to determine whether embryo survival in gilts and primiparous sows is related to variations in the peri-oestrous profiles of oestradiol, progesterone and LH. A secondary objective of the present work was to compare embryo development and certain endocrine characteristics in gilts and primiparous sows. Sows (n = 6) and gilts (n = 6) were catheterized in the jugular vein on the day after weaning or on day 14 of the oestrous cycle, respectively. Additional females (one gilt and seven sows) were examined only for characteristics of embryonic development. Embryos were recovered on day 11.5-11.75 of gestation, and size and volume of individual embryos were recorded. Minimal differences were observed between sows and gilts for endocrine and embryo data. Embryo recovery was 71.38 +/- 4.77% based on the number of corpora lutea. However, endocrine differences were noted for pigs with high embryo survival (> 71% recovery) compared with those with low survival. Peak oestradiol concentration occurred closer (P < 0.05) to the onset of oestrus in pigs with high embryo survival than in pigs with low embryo survival (3.3 +/- 4.6 h after oestrus versus 13.0 +/- 5.5 h before oestrus) and peak LH concentration occurred later (P < 0.05) after the onset of oestrus for pigs with high embryo survival. Peak oestradiol concentration tended (P = 0.07) to be higher in pigs with low embryo survival (35.21 +/- 2.56 pg ml-1) compared with pigs with high embryo survival (28.17 +/- 2.14 pg ml-1).(ABSTRACT TRUNCATED AT 250 WORDS)

Electronic animal identification for controlling feed delivery and detecting estrus in gilts and sows in outside pens.

The objective of the present study was to evaluate the feasibility of delivering feed and detecting estrous behavior by computer-controlled equipment in a nonconfinement environment. In Exp. 1, gilts were assigned to treatment when detected pregnant by ultrasound at 30 to 35 d after artificial insemination. They were assigned to be fed individually in stalls once/day (0830) with a scoop (controls, n = 20) or with an electronic sow feeding station (ESF, n = 20). The ESF gilts received their feed in 98.6-g aliquots at 80-s intervals as they visited the feeding station. Control vs ESF gilts did not differ (P > .8) for backfat (2.2 vs 2.1 cm) or weight (170 vs 172 kg) before farrowing, total and live pigs/litter (9.3 and 8.7 vs 9.1 and 8.8), or litter birth weight (12.7 vs 12.1). In Exp. 2, proceptive behavior, as measured by visits to a boar's pen, were recorded electronically, and observed estrus was evaluated in two groups of sows during their first (n = 11) and second and third (n = 19) estrous cycles and in one group of gilts (n = 14). A partition prevented visual and physical contact between the boar and the visiting females except where the electronic estrus detection (EED) station was installed. Feed delivery software was used to monitor boar visitation even though no feed delivery equipment was present at the boar pen.(ABSTRACT TRUNCATED AT 250 WORDS)

Uterine cellular changes in 2',5'-oligoadenylate synthetase during the bovine estrous cycle and early pregnancy.

The signal for maternal recognition of pregnancy in cattle is bovine trophoblast protein-1 (bTP-1), a Type I trophoblast interferon. One of the many functions of interferons is the induction of the 2',5'-oligoadenylate (2-5[A]) system, which is involved in cell division and selective degradation of mRNA. The present study focused on the cellular changes of 2-5(A) synthetase in bovine endometrium during the estrous cycle and early pregnancy. Ability of recombinant bTP-1 to stimulate activity of the enzyme in uterine cells throughout the estrous cycle was also evaluated in vitro. Surface epithelium, glandular epithelium, and stroma were enzymatically separated in cows on Day 5, 10, 15, or 18 postestrus and on Day 15 or 18 of pregnancy. Cell samples were lysed and frozen for determination of the endogenous cellular content of 2-5(A) synthetase. Additional cells obtained during the estrous cycle were cultured and treated with increasing doses of recombinant bovine interferon alpha (rbIFN-alpha) or recombinant bTP-1 (rbTP-1). During the estrous cycle, 2-5(A) synthetase was greatest on Day 5 and declined approximately 10-fold by Day 15 (p < 0.01). Cellular content of 2-5(A) synthetase was similar among all three endometrial cell types. In the gravid horn of pregnant animals, presence of a conceptus significantly increased (p < 0.01) 2-5(A) synthetase in all endometrial cell types compared to levels on Days 15 and 18 of the estrous cycle. On Day 18, levels of 2-5(A) synthetase were 30-fold greater (p < 0.01) in epithelium (surface and glandular) from pregnant cows compared to that from cyclic cows.(ABSTRACT TRUNCATED AT 250 WORDS)

Studies of uterine secretions and products of primary cultures of endometrial cells in pigs.

The uterus plays a central role in the reproductive biology of mammals. Adaptation of the uterus from an oviparous to a viviparous nature required changes that involved production of a uterine environment that could support the development of the embryo and fetus. Production of a suitable environment includes the synthesis and secretion of products by the uterine endometrium. However, the uterine endometrium is not a single homogeneous unit, but rather consists of several cell populations. Recent accomplishments in cell culture techniques provide a means for examining the contributions and secretory control of different endometrial cell populations. Furthermore, it is possible to recombine specific cell types to study their interaction. It is clear that the luminal epithelium, glandular epithelium and endometrial stroma produce different secretory products. Some secretions (for example uteroferrin) are secreted by only one cell type; others (for example prostaglandins, PGs) are secreted by all types of cell. There is much to be learned about the functions and regulations of endometrial secretions and there are important aspects of the role of the endometrium in pregnancy that present concepts do not address. For example, there is no explanation for the required synchrony between the embryo and uterus before day 10 and the implications of control of the uterine environment by progesterone from day 4 to day 10 are not understood. Almost all of the uterine secretory proteins are produced after day 10. In this review, we consider the protein and prostaglandin products from the different cell populations of the pig endometrium and propose a model to explain the integration of multiple sources of PGs and multiple regulators of PG secretion. Our purpose is to facilitate a more complete understanding of the individual uterine cell populations and a better understanding of how these cell types interact to function as a complete unit.

Effects of three methods of selection for litter size in mice on pre-implantation embryonic development.

Characteristics of preimplantation embryonic development to Day 3.5 of gestation were evaluated in lines of mice after 21 generations of selection for litter size or components of litter size. Selection criteria were direct selection for number born (LS), selection on an index of ovulation rate and the proportion of ova shed that resulted in fully formed pups (IX), selection for number born in unilaterally ovariectomized females as an indication of uterine capacity (UT), and an unselected control (LC). Comparison of the average distributions of embryonic stage of development on the left side of the uterus showed that selection (average effect of LS, IX, and UT vs. LC) tended to advance (p = 0.07) the average stage of embryonic development at Day 3.5 and shift the distribution (p = 0.10) by increasing the frequency of expanded blastocysts and decreasing the frequency of pre-morula embryos. A similar shift in the distribution on the right side of the uterus was not statistically significant. Selection decreased (p = 0.06) variability in developmental stage among embryos within the right uterine horn. These selection criteria evaluated in the mouse appear to have changed the frequencies of genes that affect some determinants of average stage of embryonic development and uniformity of development within a uterine horn at Day 3.5 of gestation.

Porcine conceptus proteins: antiviral activity and effect on 2',5'-oligoadenylate synthetase.

Interferon-like proteins synthesized by conceptuses of domestic ruminants inhibit luteolysis during early pregnancy. Although pig conceptuses secrete trophoblast interferons during the period of CL maintenance, estrogen is involved with maintenance of the CL. The principal purposes of this work were to confirm production of trophoblast interferons by porcine conceptuses and to compare the effect of trophoblast interferons on endometrium of pigs and cattle. When measured using Madin-Darby bovine kidney (MDBK) cells challenged with vesicular stomatitis virus, antiviral activity in uterine flushings from cyclic gilts was not detectable throughout the estrous cycle; however, in pregnant gilts, antiviral activity increased from undetectable amounts to 4-11 x 10(3) U on Days 14, 16, and 18. Porcine embryos in culture produced 1,100 U/embryo/ml/24 h. Porcine conceptus secretory proteins induced 2',5'-oligo(A) synthetase in MDBK cells and in endometrial explants of cows but had no measurable effect on 2',5'-oligo(A) synthetase activity of endometrial explants of pigs. Similarly, endometrial 2',5'-oligo(A) synthetase of pregnant pigs was unaffected in vivo during the period of maximal synthesis of conceptus secretory proteins. Porcine conceptus secretory proteins produced no detectable increase in serum antiviral activity or 2',5'-oligo(A) synthetase activity of blood mononuclear leukocytes in utero-ovarian venous blood. These results suggest that conceptus interferons of pigs play different roles in the establishment of pregnancy compared to their roles in ruminants.

Effect of asynchronous transfer and oestrogen administration on survival and development of porcine embryos.

Results indicate that recovery of embryos on Days 11 and 13 of pregnancy was reduced for Day 5 embryos transferred to recipients on Day 6 of their oestrous cycle and was greatly reduced when embryos were transferred to recipients on Day 7 of the cycle (P less than 0.01). Administration of oestradiol-17 beta on Day 11 of the recipient's cycle did not appear to affect embryo development on Day 13. Day 6 embryos transferred to recipients on Day 8 of the oestrous cycle deteriorated rapidly within 24 h of transfer; there was no recovery of embryos from the uterus after 36 h. Treatment of pregnant gilts with 1 mg oestradiol-17 beta (i.v.) on Day 10.5 resulted in total embryonic loss by Day 23, but pregnancy rates of gilts treated with oestradiol-17 beta on Day 12 were similar to those of vehicle-treated gilts (60.6 vs. 71.4%).

Endometrial surface and secretory alterations associated with embryonic mortality in gilts administered estradiol valerate on days 9 and 10 of gestation.

Administration of estrogen to gilts on Days 9 and 10 of pregnancy results in total embryonic loss by Day 18. The present study examined changes in the uterine endometrial surface and secretion during conceptus attachment in control and estrogen-treated (Days 9 and 10) pregnant gilts. Gilts were unilaterally hysterectomized on either Days 12 and 14 or Days 16 and 18 of gestation. Uterine horns were flushed with saline and conceptuses were evaluated. Intact conceptuses were recovered from all control gilts, whereas estrogen-treated gilts contained normal intact conceptuses only on Day 12 of gestation. Antiviral activity, which reflects conceptus viability, was reduced (p less than 0.01) in uterine flushings after Day 14 in estrogen-treated gilts. Culture of endometrial explants with [3H]glucosamine revealed several glycoproteins that are synthesized during the period of conceptus attachment; however, no difference in glycoprotein synthesis between treatment groups was detected by analysis with two-dimensional PAGE and fluorography. Analyses of the uterine epithelium by scanning and transmission electron microscopy demonstrated that estrogen administration caused an alteration in the uterine surface, a thinning of the uterine epithelial glycocalyx, and a reduction of cationic ferritin binding to the microvilli of the uterine epithelium. Results indicate that conceptus mortality after early administration of estrogen is associated with alterations in the uterine endometrial surface during the period of conceptus attachment in the pig.

Identification and characterization of bovine oviductal glycoproteins synthesized at estrus.

Published reports indicate that in several mammalian species the oviduct synthesizes and secretes specific glycoproteins which are components of the luminal fluids at the time of ovulation and fertilization. The present study characterized the secretory glycoproteins synthesized by the bovine oviduct at estrus. Oviducts obtained from four crossbred cows in estrus were flushed with saline, and segments of the ampullary and isthmic regions were fixed for immunocytochemical analyses. The remainder of the tissue was subjected to explant culture for 24 h in medium containing either 3H-leucine or 3H-glucosamine. Analysis of culture media by one- and two-dimensional SDS-PAGE followed by fluorography indicated that both the ampullary and isthmic regions synthesized a major class of Mr 97,000 glycoproteins with isoelectric points ranging from 5.5 to 8.1. A polyclonal antibody was generated to the glycoproteins after their isolation by gel filtration followed by electrophoretic separation. Western blot analysis of oviductal culture media indicated that the antisera cross-reacted with a doublet at Mr 97,000 and to a lesser extent with two additional bands at Mr greater than 200,000. Immunoreactive antigens were not identified in serum or in culture media of ovary, uterus, and nonreproductive tract tissues. The Mr 97,000 glycoproteins were present in oviductal flushings obtained from cows in estrus. They were also detected to a lesser degree in oviductal flushings obtained from cows at Days 5, 10, 15, and 18 of the estrous cycle, with the least amount of immunoreactivity being observed in Day 10 samples.(ABSTRACT TRUNCATED AT 250 WORDS)