Gestational and Early Postnatal Exposure to an Environmentally Relevant Mixture of Brominated Flame Retardants: General Toxicity and Skeletal Variations.

Brominated flame retardants (BFRs) are stable environmental contaminants known to exert endocrine-disrupting effects. Developmental exposure to polybrominated diphenyl ethers (PBDEs) is correlated with impaired thyroid hormone signaling, as well as estrogenic and anti-androgenic effects. As previous studies have focused on a single congener or technical mixture, the purpose of the current study was to examine the effects of gestational and early postnatal exposure to an environmentally relevant mixture of BFRs designed to reflect house dust levels of PBDEs and hexabromocyclododecane on postnatal developmental outcomes. Pregnant Sprague-Dawley rats were exposed to the PBDE mixture from preconception to weaning (PND 21) through the diet containing 0, 0.75, 250, and 750 mg mixture/kg diet. BFR exposure induced transient reductions in body weight at PND 35 in male and from PND 30-45 in female offspring (250 and 750 mg/kg). Liver weights (PND 21) and xenobiotic metabolizing enzyme activities (PND 21 and 46) were increased in both male and female offspring exposed to 250 and 750 mg/kg diets. Furthermore, serum T4 levels were reduced at PND 21 in both,male and female offspring (250 and 750 mg/kg). At PND 21, Serum alkaline phosphatase (ALP) was decreased in males exposed to 750 mg/kg dietat, and females exposed to 250 and 750 mg/kg diets. At PND 46 ALP was significantly elevated in males (250 and 750 mg/kg). Variations in the cervical vertebrae and phalanges were observed in pups at PND 4 (250 and 750 mg/kg). Therefore, BFR exposure during gestation through to weaning alters developmental programming in the offspring. The persistence of BFRs in the environment remains a cause for concern with regards to developmental toxicity.

A multiple lines of evidence approach for the ecological risk assessment of an accidental bitumen release from a steam assisted gravity drainage (SAGD) well in the Athabasca oil sands region.

To assess the ecological impacts of two independent accidental bitumen releases from two steam assisted gravity drainage (SAGD) wells in the Athabasca oil sands region, a multiple lines of evidence (LOE) approach was developed. Following the release in 2010, action was taken to minimize environmental impact, including the selective removal of the most highly impacted vegetation and the use of oil socks to minimize possible runoff. An ecological risk assessment (ERA) was then conducted based on reported concentrations of bitumen related contaminants in soil, vegetation, and water. Results of biological assessments conducted at the site were also included in the risk characterization. Overall, the conclusion of the ERA was that the likelihood of long-term adverse health effects to ecological receptors in the area was negligible. To provide evidence for this conclusion, a small mammal sampling plan targeting Southern red-back voles (Myodes gapperi) was carried out at two sites and two relevant reference areas. Voles were readily collected at all locations and no statistically significant differences in morphometric measurements (i.e., body mass, length, foot length, and adjusted liver weight) were found between animals collected from impact zones of varying levels of coverage. Additionally, no trends corresponding with bitumen coverage were observed with respect to metal body burden in voles for metals that were previously identified in the source bitumen. Hepatic ethoxyresorufin-O-deethylase (EROD) activity was statistically significantly elevated in voles collected from the high impact zones of sites compared to those collected from the reference areas, a finding that is indicative of continued exposure to contaminants. However, this increase in EROD was not correlated with any observable adverse population-wide biological outcomes. Therefore the biological sampling program supported the conclusion of the initial ERA and supported the hypothesis of no significant long-term population-wide ecological impact of the accidental bitumen releases.

Exposure of Female Rats to an Environmentally Relevant Mixture of Brominated Flame Retardants Targets the Ovary, Affecting Folliculogenesis and Steroidogenesis.

Brominated flame retardants (BFRs) are incorporated into various consumer products to prevent flame propagation. These compounds leach into the domestic environment, resulting in chronic exposure and contamination. Pregnancy failure is associated with high levels of BFRs in human follicular fluid, raising serious questions regarding their impact on female reproductive health. The goal of this study is to elucidate the effects of an environmentally relevant BFR mixture on female rat ovarian functions (i.e., folliculogenesis and steroidogenesis). A BFR dietary mixture formulated to mimic the relative BFR congener levels in North American house dust was administered to adult female Sprague-Dawley rats from 2 to 3 wk before mating until Gestational Day 20; these diets were designed to deliver nominal doses of 0, 0.06, 20, or 60 mg/kg/day of the BFR mixture. Exposure to BFRs triggered an approximately 50% increase in the numbers of preantral and antral follicles and an enlargement of the antral follicles in the ovaries of the dams. A significant reduction in the expression of catalase, an antioxidant enzyme, and downregulation of the expression of insulin-like factor 3 (Insl3) and 17alpha-hydroxylase (Cyp17a1) were observed in the ovary. In addition, BFR exposure affected steroidogenesis; we observed a significant decrease in circulating 17-hydroxypregnenolone and an increase in testosterone concentrations in BFR-exposed dams. Thus, BFRs target ovarian function in the rat, adversely affecting both folliculogenesis and steroidogenesis.

Progesterone and HMOX-1 promote fetal growth by CD8+ T cell modulation.

Intrauterine growth restriction (IUGR) affects up to 10% of pregnancies in Western societies. IUGR is a strong predictor of reduced short-term neonatal survival and impairs long-term health in children. Placental insufficiency is often associated with IUGR; however, the molecular mechanisms involved in the pathogenesis of placental insufficiency and IUGR are largely unknown. Here, we developed a mouse model of fetal-growth restriction and placental insufficiency that is induced by a midgestational stress challenge. Compared with control animals, pregnant dams subjected to gestational stress exhibited reduced progesterone levels and placental heme oxygenase 1 (Hmox1) expression and increased methylation at distinct regions of the placental Hmox1 promoter. These stress-triggered changes were accompanied by an altered CD8+ T cell response, as evidenced by a reduction of tolerogenic CD8+CD122+ T cells and an increase of cytotoxic CD8+ T cells. Using progesterone receptor- or Hmox1-deficient mice, we identified progesterone as an upstream modulator of placental Hmox1 expression. Supplementation of progesterone or depletion of CD8+ T cells revealed that progesterone suppresses CD8+ T cell cytotoxicity, whereas the generation of CD8+CD122+ T cells is supported by Hmox1 and ameliorates fetal-growth restriction in Hmox1 deficiency. These observations in mice could promote the identification of pregnancies at risk for IUGR and the generation of clinical interventional strategies.

Health-based audible noise guidelines account for infrasound and low-frequency noise produced by wind turbines.

Setbacks for wind turbines have been established in many jurisdictions to address potential health concerns associated with audible noise. However, in recent years, it has been suggested that infrasound (IS) and low-frequency noise (LFN) could be responsible for the onset of adverse health effects self-reported by some individuals living in proximity to wind turbines, even when audible noise limits are met. The purpose of this paper was to investigate whether current audible noise-based guidelines for wind turbines account for the protection of human health, given the levels of IS and LFN typically produced by wind turbines. New field measurements of indoor IS and outdoor LFN at locations between 400 and 900 m from the nearest turbine, which were previously underrepresented in the scientific literature, are reported and put into context with existing published works. Our analysis showed that indoor IS levels were below auditory threshold levels while LFN levels at distances >500 m were similar to background LFN levels. A clear contribution to LFN due to wind turbine operation (i.e., measured with turbines on in comparison to with turbines off) was noted at a distance of 480 m. However, this corresponded to an increase in overall audible sound measures as reported in dB(A), supporting the hypothesis that controlling audible sound produced by normally operating wind turbines will also control for LFN. Overall, the available data from this and other studies suggest that health-based audible noise wind turbine siting guidelines provide an effective means to evaluate, monitor, and protect potential receptors from audible noise as well as IS and LFN.

The bioaccessibility of soil-based mercury as determined by physiological based extraction tests and human biomonitoring in children.

Environmental contaminants associated with soil particles are generally less bioavailable than contaminants associated with other exposure media where chemicals are often found in more soluble forms. In vitro methods, such as Physiological Based Extraction Tests (PBET), can provide estimates of bioaccessibility for soil-based contaminants. The results of these tests can be used to predict exposure to contaminants from soil ingestion pathways within human health risk assessment (HHRA). In the current investigation, an HHRA was conducted to examine the risks associated with elevated concentrations of mercury in soils in the northern Canadian smelter community of Flin Flon, Manitoba. A PBET was completed for residential soils and indicated mean bioaccessibilities of 1.2% and 3.0% for total mercury using gastric phase and gastric+intestinal phase methodologies, respectively. However, as many regulators only allow for the consideration of in vitro results for lead and arsenic in the HHRA process, in vitro bioaccessibility results for mercury were not utilized in the current HHRA. Based on the need to assume 100% bioaccessibility for inorganic mercury in soil, results from the HHRA indicated the need for further assessment of exposure and risk. A biomonitoring study was undertaken for children between 2 and 15 years of age in the community to examine urinary inorganic mercury concentrations. Overall, 375 children provided valid urine samples for analysis. Approximately 50% of urine samples had concentrations of urinary inorganic mercury below the limit of detection (0.1 µg/L), with an average creatinine adjusted concentration of 0.11 µg/g. Despite high variability in mercury soil concentrations within sub-communities, soil concentrations did not appear to influence urinary mercury concentrations. The results of the current investigation indicate that mercury bioaccessibility in residential soils in the Flin Flon area was likely limited and that HHRA estimates would have been better approximated through inclusion of the in vitro study results.

Wind turbines and human health.

The association between wind turbines and health effects is highly debated. Some argue that reported health effects are related to wind turbine operation [electromagnetic fields (EMF), shadow flicker, audible noise, low-frequency noise, infrasound]. Others suggest that when turbines are sited correctly, effects are more likely attributable to a number of subjective variables that result in an annoyed/stressed state. In this review, we provide a bibliographic-like summary and analysis of the science around this issue specifically in terms of noise (including audible, low-frequency noise, and infrasound), EMF, and shadow flicker. Now there are roughly 60 scientific peer-reviewed articles on this issue. The available scientific evidence suggests that EMF, shadow flicker, low-frequency noise, and infrasound from wind turbines are not likely to affect human health; some studies have found that audible noise from wind turbines can be annoying to some. Annoyance may be associated with some self-reported health effects (e.g., sleep disturbance) especially at sound pressure levels >40 dB(A). Because environmental noise above certain levels is a recognized factor in a number of health issues, siting restrictions have been implemented in many jurisdictions to limit noise exposure. These setbacks should help alleviate annoyance from noise. Subjective variables (attitudes and expectations) are also linked to annoyance and have the potential to facilitate other health complaints via the nocebo effect. Therefore, it is possible that a segment of the population may remain annoyed (or report other health impacts) even when noise limits are enforced. Based on the findings and scientific merit of the available studies, the weight of evidence suggests that when sited properly, wind turbines are not related to adverse health. Stemming from this review, we provide a number of recommended best practices for wind turbine development in the context of human health.

Exposure to an environmentally relevant mixture of brominated flame retardants affects fetal development in Sprague-Dawley rats.

Brominated flame retardants are incorporated into a wide variety of consumer products and are known to enter into the surrounding environment, leading to human exposure. There is accumulating evidence that these compounds have adverse effects on reproduction and development in humans and animal models. Animal studies have generally characterized the outcome of exposure to a single technical mixture or congener. Here, we determined the impact of exposure of rats prior to mating and during gestation to a mixture representative of congener levels found in North American household dust. Adult female Sprague-Dawley rats were fed a diet containing 0, 0.75, 250 or 750mg/kg of a mixture of flame retardants (polybrominated diphenyl ethers, hexabromocyclododecane) from two weeks prior to mating to gestation day 20. This formulation delivered nominal doses of 0, 0.06, 20 and 60mg/kg body weight/day. The lowest dose approximates high human exposures based on house dust levels and the dust ingestion rates of toddlers. Litter size and resorption sites were counted and fetal development evaluated. No effects on maternal health, litter size, fetal viability, weights, crown rump lengths or sex ratios were detected. The proportion of litters with fetuses with anomalies of the digits (soft tissue syndactyly or malposition of the distal phalanges) was increased significantly in the low (0.06mg/kg/day) dose group. Skeletal analysis revealed a decreased ossification of the sixth sternebra at all exposure levels. Thus, exposure to an environmentally relevant mixture of brominated flame retardants results in developmental abnormalities in the absence of apparent maternal toxicity. The relevance of these findings for predicting human risk is yet to be determined.

Perturbation of male sexual behavior in mice (Mus musculus) within a discrete range of perinatal bisphenol-A doses in the context of a high- or low-phytoestrogen diet.

Differentiation of masculine and feminine behavior in mammals depends on perinatal sex steroids. As bisphenol-A (BPA) can be estrogenic and anti-androgenic, we examined impacts of perinatal exposure upon adult sexual behavior and morphology of male mice. In Experiment 1, dams were fed either a high- or low-phytoestrogen diet and received daily oral doses of 0, 0.175, 1.75, or 17.5µg BPA from gestation day 10 through post-partum day 9. Male offspring from the high-phytoestrogen plus 17.5µg BPA condition showed reduced mass of vesicular-coagulating but not other male glands, and showed increased latency to insemination when paired with females. In Experiment 2, these procedures were replicated but with all animals fed the high-phytoestrogen diet and perinatal BPA doses of 0, 17.5, 175, or 1750µg/day. Adult masses of testes and male-accessory glands and levels of urinary steroids were not significantly affected. When males each encountered a sexually receptive female, there were fewer intromissions among those given 17.5 or 175µg and fewer ejaculations among those given 17.5µg, but the 1750µg dose had no effect. Perinatal BPA dosages thus influenced male sexual behavior non-monotonically, with impairment evident in a discrete dose range among males on a high-phytoestrogen diet.

Effects of chronic exposure to an environmentally relevant mixture of brominated flame retardants on the reproductive and thyroid system in adult male rats.

Brominated flame retardants (BFRs) are incorporated into a wide variety of consumer products, are readily released into home and work environments, and are present in house dust. Studies using animal models have revealed that exposure to polybrominated diphenyl ethers (PBDEs) may impair adult male reproductive function and thyroid hormone physiology. Such studies have generally characterized the outcome of acute or chronic exposure to a single BFR technical mixture or congener but not the impact of environmentally relevant BFR mixtures. We tested whether exposure to the BFRs found in house dust would have an adverse impact on the adult male rat reproductive system and thyroid function. Adult male Sprague Dawley rats were exposed to a complex BFR mixture composed of three commercial brominated diphenyl ethers (52.1% DE-71, 0.4% DE-79, and 44.2% decaBDE-209) and hexabromocyclododecane (3.3%), formulated to mimic the relative congener levels in house dust. BFRs were delivered in the diet at target doses of 0, 0.02, 0.2, 2, or 20 mg/kg/day for 70 days. Compared with controls, males exposed to the highest dose of BFRs displayed a significant increase in the weights of the kidneys and liver, which was accompanied by induction of CYP1A and CYP2B P450 hepatic drug-metabolizing enzymes. BFR exposure did not affect reproductive organ weights, serum testosterone levels, testicular function, or sperm DNA integrity. The highest dose caused thyroid toxicity as indicated by decreased serum thyroxine (T4) and hypertrophy of the thyroid gland epithelium. At lower doses, the thickness of the thyroid gland epithelium was reduced, but no changes in hormone levels (T4 and thyroid-stimulating hormone) were observed. Thus, exposure to BFRs affected liver and thyroid physiology but not male reproductive parameters.

Rounds reports: Early experiences of using printed summaries of electronic medical records in a large teaching medical hospital.

This article describes the rationale, processes, technology, and results of creating of a paper-based rounds report that is now used by our entire institution for efficient inpatient work rounds and checkout rounds that are routinely done in virtually every hospital, both academic and private, in the US. The results of a survey of clinicians suggests that printed rounds reports have markedly improved rounding efficiency, saved substantial amounts of physician time, standardized checkout processes, and improved patient safety.

A case of progressive multifocal leukoencephalopathy in a patient treated with infliximab.

We describe a 72-year-old white man with erosive rheumatoid arthritis in whom subacute neurologic and psychiatric symptoms developed after 3 years of treatment with infliximab, prednisone, and methotrexate. White matter demyelination was seen on magnetic resonance imaging of the brain, and progressive multifocal leukoencephalopathy (PML) was ultimately confirmed by brain biopsy. The patient was treated with supportive therapy and discontinuation of disease-modifying antirheumatic drugs, resulting in stabilization of the disease process. The patient survived, but neurologic and cognitive deficits persisted. The distribution and pathology of this patient's disease are unique from almost all reported incidents of oral methotrexate-associated leukoencephalopathy. The pathogenesis of disease may be linked to a T cell-mediated process that is potentially impacted by infliximab. This case provides the first reported evidence that PML can be seen in association with infliximab therapy.

Bisphenol-A exposure during the period of blastocyst implantation alters uterine morphology and perturbs measures of estrogen and progesterone receptor expression in mice.

Bisphenol-A (BPA) has estrogenic properties both in vitro and in vivo. We investigated its impacts upon uterine morphology and estrogen and progesterone receptors after injection on gestational days 1-4 in doses known to disrupt pregnancy. Blastocyst implantation was significantly reduced by doses of 6.75 and 10.125 mg/animal. Uterine luminal area expanded substantially in response to increasing doses of BPA. Luminal epithelial cell height increased following exposure to 10.125 mg/animal, whereas there were no differences in the number of corpora lutea among conditions. The proportion of cells staining positively for estrogen receptors was affected non-monotonically, showing highest levels at 3.375 mg/animal and lowest levels at 10.125 mg/animal. Similarly progesterone receptor expression measured through western blots related non-monotonically to dose, being highest at 3.375 mg/animal and diminishing with increasing dose. These results suggest that BPA exposure during early gestation acts at the uterus to disrupt intrauterine implantation, consistent with an estrogenic effect.

Excretion and binding of tritium-labelled oestradiol in mice (Mus musculus): implications for the Bruce effect.

Male mouse urine contains 17beta-oestradiol (E(2)) and other steroids. Given that males actively direct urine at proximate females and intrauterine implantation of blastocysts is vulnerable to minute amounts of exogenous oestrogens, males' capacity to disrupt early pregnancy could be mediated by steroids in their urine. When male mice were implanted with osmotic pumps containing tritium-labelled E(2) ((3)H-E(2)) or injected i.p. with (3)H-E(2), radioactivity was reliably detected in their urine. Following intranasal administration of (3)H-E(2) to inseminated females, radioactivity was detected in diverse tissue samples, with there being significantly more in reproductive tissues than in brain tissues. When urine was taken from males injected with (3)H-E(2), and then intranasally administered to inseminated females, radioactivity was detected in the uterus, olfactory bulbs, and mesencephalon and diencephalon (MC+DC). When inseminated and ovariectomised females were perfused at the point of killing to remove blood from tissues, more radioactivity was detected in the uterus than in muscle, olfactory bulbs, MC+DC and cerebral cortex. Pre-treatment with unlabelled E(2) significantly reduced the uptake of (3)H-E(2) in the uterus. Taken with evidence that males deliver their urine to the nasal area of females, these results indicate that male urinary E(2) arrives in tissues, including the uterus, where it could lead to the disruption of blastocyst implantation.

Preputialectomised and intact adult male mice exhibit an elevated urinary ratio of oestradiol to creatinine in the presence of developing females, whilst promoting uterine and ovarian growth of these females.

Exposure to novel adult males and their urine can hasten the onset of sexual maturity in female mice. Some evidence implicates chemosignals from males' preputial glands, while other evidence suggests that male urinary steroids, especially 17beta-oestradiol, contribute to this effect. The present experiment was designed to determine whether preputial gland removal would influence the capacity of males to accelerate female sexual development, and to measure male urinary oestradiol and testosterone in the presence or absence of these glands. Juvenile females aged 28 days were housed for two weeks in isolation or underneath two outbred males that had undergone preputialectomy or sham surgery. Urine samples were collected non-invasively from males that were isolated or exposed to females, then assayed for oestradiol, testosterone and creatinine. Combined uterine and ovarian mass from females sacrificed at 43 days of age was increased by exposure to males, regardless of whether or not these males had been preputialectomised. Male urinary creatinine was reduced by exposure to developing females. Creatinine-adjusted oestradiol and testosterone were significantly greater in female-exposed than in isolated males, in both preputialectomised and intact males. These data suggest that the preputials are not necessary for the capacity of males to hasten female uterine and ovarian growth. As exogenous oestrogens can promote uterine growth and other parameters of female reproductive maturation, oestradiol in males' urine may contribute to earlier sexual maturity in male-exposed females.

The realities of implementation of Clinical Context Object Workgroup (CCOW) standards for integration of vendor disparate clinical software in a large medical center.

CCOW standards have been touted to currently be the best way to integrate disparate clinical applications by passing user identification and passwords as well as patient context between these applications at the desktop. However, nothing has been published in academic journals on the actual realities of implementation of CCOW. In this report, we describe the implementation of CCOW for our three main clinical applications and compare this with the simultaneous development and implementation of a portal session manager for the same purpose. We found the portal session manager much easier to develop and implement than CCOW. The resulting functionality was almost equivalent as judged by our clinical end users who compared both solutions. We now have the portal session manager functional across the institution and have stopped any further work on CCOW.

Exposure to developing females induces polyuria, polydipsia, and altered urinary levels of creatinine, 17beta-estradiol, and testosterone in adult male mice (Mus musculus).

Novel male mice can accelerate reproductive maturation in proximal developing females, an effect mediated by the chemistry of the males' urine. Exogenous estrogens can similarly accelerate female sexual development. In Experiment 1, adult male mice were housed across wire grid from either empty compartments or those containing post-weanling females. Proximity of females caused males to urinate more, progressively over days of exposure, with most urination directed towards females' compartments. Male urine collected after 5 days in these conditions was analyzed by enzyme immunoassay for 17beta-estradiol, testosterone, and creatinine. Urinary creatinine of isolated males significantly exceeded that of female-exposed males. Unadjusted urinary steroids also trended toward higher levels in isolates, but creatinine-adjusted estradiol and testosterone of female-exposed males significantly exceeded that of isolated males. In Experiment 2, measurement of water consumption indicated significantly greater drinking by female-exposed as opposed to isolated males. In Experiment 3, males were housed in isolation or beside post-weanling intact (sham-operated) females, ovariectomized females, or intact (sham-operated) males. Male water consumption was elevated in all conditions involving social contact. Urinary creatinine was significantly lower in female-exposed males compared to isolated controls, while unadjusted testosterone was significantly lower in males in all social conditions. Again, creatinine-adjusted estradiol in female-exposed males significantly exceeded that of isolates. These data indicate that adult males drink and urinate more, have more dilute urine, and have a higher ratio of estradiol to creatinine when they are near developing females. These dynamics increase females' exposure to urinary steroids and other urinary constituents that can hasten sexual maturity.

Impact of acute bisphenol-A exposure upon intrauterine implantation of fertilized ova and urinary levels of progesterone and 17beta-estradiol.

Bisphenol-A (BPA), a monomer used in production of polycarbonate plastics and epoxy resins, has established estrogenic properties. We assessed the impact of acute and repeated subcutaneous BPA administration upon intrauterine implantation of fertilized ova and urinary levels of 17beta-estradiol and progesterone in inseminated female mice. In Experiment 1, females received varied doses of BPA on days 1-4 of gestation. Daily doses of 6.75 and 10.125mg/animal significantly reduced the number of implantation sites. Urinary progesterone was significantly reduced by the higher dose, but no other dose had an effect on progesterone levels and no dose altered estradiol levels. In Experiment 2, inseminated females received a single dose of BPA on days 0, 1, or 2 of gestation. A single dose of 10.125mg reduced the number of implantation sites when given on day 0 or day 1, and 6.75mg on day 1 also produced fewer implantation sites, but there was no such effect of any dose when administered on day 2. These data show a lower threshold for BPA-induced pregnancy disruption than previously reported, also indicating effects of just one exposure. They confirm that this disruption is due to the actions of BPA upon implantation sites, and show that higher doses can influence systemic progesterone levels.

The onset of puberty in female mice as reflected in urinary steroids and uterine/ovarian mass: interactions of exposure to males, phyto-oestrogen content of diet, and ano-genital distance.

Development of puberty in female mice was examined in relationship with the ano-genital distance index (AGDI), phyto-oestrogen content of diet and exposure to males post weaning. Throughout gestation and post-natal development, females were exposed to a regular diet or a nutritionally similar diet deficient in phyto-oestrogens. After segregation at weaning on the basis of short or long AGDI, an indirect measure of in utero androgen exposure, females were housed alone or underneath two outbred adult males for 2 weeks. Female urinary samples were collected non-invasively throughout this exposure, then assayed for oestradiol, progesterone and creatinine. Females were then killed and uterine and ovarian mass was determined. Urinary oestradiol was substantially reduced in females raised on the phyto-oestrogen-free diet. Oestradiol levels were more dynamic over days in urine of male-exposed females, especially among those on the regular diet. Urinary progesterone was not strongly influenced by diet. Progesterone was more dynamic in urine of male-exposed females, and was generally elevated compared with levels in isolated females, the size of this effect dependent on AGDI, diet and whether the measure was adjusted for creatinine. Urinary creatinine was elevated by the phyto-oestrogen-free diet and reduced by male exposure, tending to decline over days in females exposed to males. Male exposure increased uterine and ovarian mass and was influenced by AGDI in interaction with diet and male exposure.

Influence of oral and subcutaneous bisphenol-A on intrauterine implantation of fertilized ova in inseminated female mice.

Intrauterine implantation of fertilized ova in inseminated females is sensitive to minute levels of natural estrogens. Bisphenol-A (BPA), a widely used chemical in the production of polycarbonate plastics and epoxy resins, can be estrogenic. Here we administered BPA during the period of implantation to determine levels of exposure required to terminate pregnancy in mice. Varied doses were given through either injection or ingestion. Subcutaneous injections during days 1-4 of gestation significantly reduced litter size at 3.375 mg/day and substantially reduced the proportion of females that were parturient at 10.125 mg/day. Uterine implantation sites were also significantly reduced in females sacrificed at day 6 after receiving 10.125 mg/day. Exposure to lower doses was without significant effect. When inseminated females' diets were supplemented on days 1-5 with peanut butter contaminated by 0.11-9.0% BPA, litter size and percent parturient were not affected. However, when the animals' diet was exclusively comprised of a mixture of BPA, peanut butter, and powdered chow during days 1-4, an average daily intake of 68.84 mg BPA terminated all pregnancies. No significant effects at lower doses of BPA were seen in number of births or other measures through either mode of administration.