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Cited by 32Effects and risks of endocrine disrupting chemicalsSexual Differentiation and DisordersEstrogen and related hormone effects

Developmental Treatment with Ethinyl Estradiol, but Not Bisphenol A, Causes Alterations in Sexually Dimorphic Behaviors in Male and Female Sprague Dawley Rats

Open access
Ferguson, Sherry A., Law, Charles Delbert, Kissling, Grace E.·2014·Toxicological Sciences
DOI 10.1093/toxsci/kfu077PMC PMC4133561
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Abstract

The developing central nervous system may be particularly sensitive to bisphenol A (BPA)-induced alterations. Here, pregnant Sprague Dawley rats (n = 11-12/group) were gavaged daily with vehicle, 2.5 or 25.0 μg/kg BPA, or 5.0 or 10.0 μg/kg ethinyl estradiol (EE2) on gestational days 6-21. The BPA doses were selected to be below the no-observed-adverse-effect level (NOAEL) of 5 mg/kg/day. On postnatal days 1-21, all offspring/litter were orally treated with the same dose. A naïve control group was not gavaged. Body weight, pubertal age, estrous cyclicity, and adult serum hormone levels were measured. Adolescent play, running wheel activity, flavored solution intake, female sex behavior, and manually elicited lordosis were assessed. No significant differences existed between the vehicle and naïve control groups. Vehicle controls exhibited significant sexual dimorphism for most behaviors, indicating these evaluations were sensitive to sex differences. However, only EE2 treatment caused significant effects. Relative to female controls, EE2-treated females were heavier, exhibited delayed vaginal opening, aberrant estrous cyclicity, increased play behavior, decreased running wheel activity, and increased aggression toward the stimulus male during sexual behavior assessments. Relative to male controls, EE2-treated males were older at testes descent and preputial separation and had lower testosterone levels. These results suggest EE2-induced masculinization/defeminization of females and are consistent with increased volume of the sexually dimorphic nucleus of the preoptic area (SDN-POA) at weaning in female siblings of these subjects (He, Z., Paule, M. G. and Ferguson, S. A. (2012) Low oral doses of bisphenol A increase volume of the sexually dimorphic nucleus of the preoptic area in male, but not female, rats at postnatal day 21. Neurotoxicol. Teratol. 34, 331-337). Although EE2 treatment caused pubertal delays and decreased testosterone levels in males, their behaviors were within the range of control males. Conversely, BPA treatment did not alter any measured endpoint. Similar to our previous reports (Ferguson, S. A., Law, C. D. Jr and Abshire, J. S. (2011) Developmental treatment with bisphenol A or ethinyl estradiol causes few alterations on early preweaning measures. Toxicol. Sci. 124, 149-160; Ferguson, S. A., Law, C. D. and Abshire, J. S. (2012) Developmental treatment with bisphenol A causes few alterations on measures of postweaning activity and learning. Neurotoxicol. Teratol. 34, 598-606), the BPA doses and design used here produced few alterations.

AI snapshot

Written from the paper’s full text.

TL;DR

Developmental treatment with ethinyl estradiol (EE2), but not bisphenol A (BPA), altered sexually dimorphic behaviors and pubertal/reproductive endpoints in Sprague Dawley rats. BPA at 2.5 or 25.0 µg/kg/day produced no significant effects on the measured endpoints, while EE2 masculinized/defeminized females and delayed male puberty with lower testosterone.

Key findings

  • BPA treatment did not alter any measured endpoint; no significant differences were found between vehicle and naïve controls.
  • EE2-treated females were heavier (5.0 EE2 from PND 41, 10.0 EE2 from PND 55), had delayed vaginal opening (vehicle 35.95 PND vs 5.0 EE2 45.43 and 10.0 EE2 42.94), aberrant estrous cyclicity (extended estrus transitions 9.34% and 25.00% vs vehicle 4.72%), increased play pinning, decreased running wheel activity, and increased aggression toward stimulus males (29% of 10.0 EE2 sessions vs 7% of vehicle sessions).
  • EE2-treated males had later testes descent (vehicle 23.13 PND vs 5.0 EE2 27.27 and 10.0 EE2 32.88) and preputial separation (vehicle 45.21 PND vs 5.0 EE2 52.50 and 10.0 EE2 70.45), and lower testosterone levels (5.0 EE2 118.387 and 10.0 EE2 108.779 vs vehicle 339.730 ng/dl), but their behaviors remained within the range of control males.
  • Vehicle controls showed significant sexual dimorphism for most behaviors, supporting sensitivity of the assessments to sex differences.

Methods

  • Pregnant Sprague Dawley rats (n = 11–12/group) were gavaged daily on gestational days 6–21 with vehicle, 2.5 or 25.0 µg/kg BPA, or 5.0 or 10.0 µg/kg EE2; a naïve control was not gavaged. Offspring were orally treated with the same dose on postnatal days 1–21.
  • Endpoints included body weight, pubertal landmarks, estrous cyclicity, adult serum hormones, adolescent play, running wheel activity, flavored solution intake, female sex behavior, and manual lordosis.
  • Litter was the experimental unit; analyses used RM ANOVA, MANOVA, chi-square/Markov transition models, logistic regression, and categorical ANOVA-type models, with pairwise comparisons to vehicle controls.

Limitations

  • The paper states that not all females were examined for abnormal genitalia and the number affected per group is unknown; it also notes an inadvertent error in sodium solution concentration for replicate 1, leading to deletion of 14 subjects’ data, and that 15 female hormone samples were collected in the afternoon and analyzed both with and without those subjects.
  • EE2 group sizes were smaller because some females never experienced vaginal opening, and no clear dose-response was apparent for most EE2 effects except estrous cyclicity and dark-period running wheel activity.
  • Evident from the text: BPA was tested at only two doses below the NOAEL in this design, so the absence of BPA effects cannot rule out effects at other doses, exposure windows, or endpoints.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1093/toxsci/kfu077

Cites

99 of 99 references verified0 citation errors · 0 not checkable

Statistics

29 tests recomputed · 0 with the reported and recomputed p on opposite sides of .05

Claims the paper makes

15 of 77 cited sources checked · 10 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    An earlier study of siblings of the subjects of the current study described differences in the naïve and vehicle control groups.

    “This became potentially relevant because an earlier study of siblings of the subjects of the current study described differences in the naïve and vehicle control groups (Cao et al., 2013).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Cao et al., 2013 source
  • Premisenone given

    In none of the three studies (Porrini et al., 2005; Dessi-Fulgheri et al., 2002; Wolstenholme et al., 2011) was the litter statistically controlled.

    “However, in none of those three studies was the litter statistically controlled.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites Porrini et al., 2005 source
    • cites Dessi-Fulgheri et al., 2002 source
    • cites Wolstenholme et al., 2011 source
  • Premisecitation

    These effects were expected given that estradiol, produced via aromatization of androgens, is essential for masculinization of the male rodent central nervous system.

    “These effects were expected given that estradiol, produced via aromatization of androgens, is essential for masculinization of the male rodent central nervous system (reviewed in Lenz and McCarthy, 2010).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Lenz and McCarthy, 2010 source
  • Interpretationcitation

    The relatively few significant alterations in EE2-treated males suggest that they may be less sensitive to developmental EE2 treatment than females.

    “The relatively few significant alterations in EE 2 -treated males suggests that they may be less sensitive to developmental EE 2 treatment than females, a hypothesis that has been presented by Ryan et al. (Howdeshell et al., 2008;Ryan et al., 2010 and see Figure 11 in Ryan et al., 2010).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites Howdeshell et al., 2008 source
    • cites Ryan et al., 2010 source
  • Premisecitation

    The necessity of normal circulating levels of testosterone or estradiol for sex-typical running wheel activity is evident from the effects of adult or preweaning castration and/or exogenous replacement.

    “For example, the necessity of normal circulating levels of testosterone or estradiol for sex-typical running wheel activity is clearly evident from the effects caused by adult or preweaning castration and/or exogenous replacement (Hagenauer et al., 2011;Ibebunjo et al., 2011;Jackson et al., 2011;Rivera and Eckel, 2005).”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites Hagenauer et al., 2011 source

      The Results (Experiment 1, Mean Activity) report that prepubertal gonadectomy reduced wheel-running activity, especially in females. The Discussion also describes adult gonadectomy and hormone-replacement findings from previous studies. But it presents the role of increasing estrogen in the pubertal change as likely, rather than establishing that normal circulating levels of both hormones are necessary for sex-typical activity. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites Ibebunjo et al., 2011

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Jackson et al., 2011 source
  • Classified as probably not a substantial claim. 50
  • May combine two claims. 43

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Previously, the authors described few BPA effects on pre- and postweaning behaviors that are not generally thought to be sexually dimorphic.

    “Previously, we described few BPA effects on pre-and postweaning behaviors that are not generally thought to be sexually dimorphic (Ferguson et al., 2012(Ferguson et al., , 2011)).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Ferguson et al., 2012 source
    • cites Ferguson et al., 2011 source

      The Discussion reports no BPA effects on the two preweaning behaviors tested, supporting the preweaning part of the claim. It says postweaning behavioral results would be reported separately, so this paper does not support the postweaning part. It also does not characterize the tested behaviors as generally non-sexually dimorphic; its results report a sex difference in slant-board performance. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The running wheel model was previously shown to be sensitive to cerebellar development alterations.

    “This running wheel model was previously shown to be sensitive to cerebellar development alterations (Kim et al., 2010).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Kim et al., 2010 source

  • May combine two claims.

    Findingown data

    EE2-treated rats exhibited a significantly lower proportion of days in diestrus and a higher percentage of extended estrus transitions, similar to that reported for dietary EE2 treatment in rats and DES treatment in mice.

    “Here, EE 2 -treated rats exhibited a significantly lower proportion of days in diestrus and a higher percentage of extended estrus transitions, similar to that reported for dietary EE 2 treatment in rats (Delclos et al., 2009) and PNDs 15-18 DES treatment in mice (Nikaido et al., 2005) as well as similar but chronic EE 2 treatment (Delclos et al., 2014).”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites Delclos et al., 2009 source
    • cites Nikaido et al., 2005
    • cites Delclos et al., 2014 source

      The Results (Vaginal Cytology, Table 4; page unavailable) report that chronically EE2-treated rats had markedly more abnormal cycles, including extended estrus and estrus/diestrus transitions. The paper does not report a significantly lower proportion of days in diestrus, so it supports the similarity in extended cycling but not that specific measure. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    As reviewed by Gorski (1986), even slight differences in the timing of exogenous estrogen treatment can affect vaginal opening.

    “As reviewed by Gorski (1986), even slight differences in the timing of exogenous estrogen treatment (e.g., PNDs 26-30 vs. PNDs 33-37) can affect vaginal opening.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Gorski, 1986 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationcitation

    The lack of significance for BPA treatment here is consistent with previous descriptions from the same cohort.

    “Additionally, the lack of significance for BPA treatment here is consistent with our previous descriptions from the same cohort (Ferguson et al., 2011(Ferguson et al., , 2012)).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Ferguson et al., 2011 source
    • cites Ferguson et al., 2012 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Estrogen receptor alpha in the medial preoptic area seems crucially responsible for estrogen-induced increases in running wheel activity.

    “The estrogen receptor alpha in the medial preoptic area seems crucially responsible for estrogen-induced increases in running wheel activity (Fahrbach et al., 1985;Hertrampf et al., 2008;Ogawa et al., 2003;Spiteri et al., 2012).”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites Fahrbach et al., 1985 source
    • cites Hertrampf et al., 2008 source
    • cites Ogawa et al., 2003 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Neonatal estradiol treatment of female rats at doses approximating male-typical levels can cause increased play behavior, but the increase is more related to wrestling/boxing and pouncing than to pinning.

    “A more recent study, however, has demonstrated that neonatal estradiol treatment of females at doses which approximate male-typical levels can cause increased play behavior, although that increase was more related to wrestling/boxing and pouncing behaviors than to pinning behavior (Olesen et al., 2005).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Olesen et al., 2005 source

  • May combine two claims.

    Interpretationcitation

    Our previous descriptions of significant EE2, but few BPA, effects on pre- and postweaning behaviors add to the growing body of literature indicating lack of effect at these orally administered BPA doses.

    “Our previous descriptions of significant EE 2 , but few BPA, effects on pre-and postweaning behaviors (Ferguson et al., 2011(Ferguson et al., , 2012) ) add to the growing body of literature indicating lack of effect at these orally administered BPA doses.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites Ferguson et al., 2011 source

      The Discussion reports few effects of oral BPA at the tested doses, and the preweaning behavioral results show no significant BPA effects. But this paper does not show significant EE2 effects on preweaning behavior: the Righting reflex section reports no treatment group differed significantly from vehicle, and the Slant board behavior section reports no treatment effect. Postweaning behavioral results were reserved for a separate report. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites Ferguson et al., 2012 source

  • May combine two claims.

    Findingown data

    Analyses of whole brain, frontal cortex, and hippocampal weights of males did not indicate any significant treatment effects, nor did ratios to body weight.

    “Analyses of whole brain, frontal cortex, and hippocampal weights of males did not indicate any significant treatment effects, nor did analyses of ratios of those weights to body weight (data not shown).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    On any given week, 5.0 EE2 females consumed 3-14% more and 10.0 EE2 females consumed -4 to 10% more than same-sex vehicle controls.

    “On any given week, females of the 5.0 EE 2 group consumed 3-14% more than same-sex vehicle controls and females of the 10.0 EE 2 group consumed -4 to 10% more than same-sex vehicle controls.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    The amplitude of running wheel activity generally increased across days in all groups.

    “Analyses of these parameters indicated that the amplitude of activity generally increased across days in all groups.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    During euthanasia procedures, some females appeared to exhibit abnormal genitalia.

    “During euthanasia procedures, it was noted that some females appeared to exhibit abnormal genitalia.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    Regular water intake on day 3 was higher than on days 1 or 2 during saccharin availability.

    “Pairwise comparisons indicated that regular water intake on day 3 was higher than that on days 1 or 2 (both comparisons p < 0.046).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    There were significant main effects of sex and day on saccharin-flavored intake.

    “Analysis of saccharin-flavored intake indicated significant main effects of sex (F(1, 128) = 53.54,p < 0.001) and day (F(2, 256) = 69.46,p < 0.001) (see Fig. 5A).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    There was a significant effect of day on regular water intake during saccharin availability.

    “Analysis of regular water intake during the days that saccharin-flavored solution was available indicated a significant effect of day (F(2, 256) = 4.72, p < 0.010) (data not shown).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    There was a significant main effect of treatment on male body weight, but no pairwise differences between vehicle control and any other treatment group.

    “Analysis of male body weight indicated significant main effects of treatment (F(5, 64) = 5.35, p < 0.001) and PND (F(10, 538) = 3638.48,p < 0.001).However, pairwise comparisons did not indicate any significant differences between the vehicle control group and any other treatment group (see Fig. 1A).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Vehicle control males performed an average 34% more pins than vehicle control females, although the difference was not statistically significant.

    “Although a statistically significant effect was not noted for number of pins (Mann-Whitney U = 44.5, p < 0.118), vehicle control males performed an average 34% more pins than vehicle control females.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    All male groups gained weight with age.

    “The main effect of PND indicated that all male groups gained weight with age.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    Regular water intake during each of the three test days differed, with highest intake on day 3.

    “Intake of regular water during each of the three test days was significantly different with the highest intake on day 3 (all comparisons p < 0.022).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    Intake of sodium-flavored solution on each of the 3 days differed.

    “Intake on each of the 3 days differed as well (for all comparisons p < 0.029).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    There was a significant interaction of treatment with sex on sodium preference, but no significant pairwise comparisons to vehicle control groups.

    “Analysis of sodium preference indicated a significant interaction of treatment with sex (F(5, 114) = 2.31, p < 0.049); however, there were no significant pairwise comparisons to either the male or the female vehicle control groups.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    There were significant effects of treatment with sex and day on sodium-flavored intake.

    “Analysis of sodium-flavored intake indicated significant effects of treatment with sex (F(5, 114) = 2.96, p < 0.016) and day (F(2, 223) = 13.96,p < 0.001) (see Fig. 5B).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    There was a significant effect of day on sodium preference, with preference on day 1 less than day 2 but higher than day 3, and day 2 higher than day 3.

    “In addition, analysis of sodium preference indicated a significant effect of day (F(2, 222) = 47.65,p < 0.001) and pairwise comparisons revealed that preference on day 1 was less than day 2 (p < 0.001) but higher than day 3 (p < 0.001).Preference on day 2 was higher than on day 3 (p < 0.001).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    When all females were included, the proportion with a nonzero lordosis response was 75.0% for naive control, 58.3% for vehicle control, 63.6% for 2.5 BPA, 66.7% for 25.0 BPA, 72.7% for 5.0 EE2, and 83.3% for 10.0 EE2.

    “When all females were included, the number (and percentage) of females in each group with a nonzero response on at least one of the three trials on each of the 5 days was: 9/12 (75.0%) for the naïve control group, 7/12 (58.3%) for the vehicle control group, 7/11 (63.6%) for the 2.5 BPA group, 8/12 (66.7%) for the 25.0 BPA group, 8/11 (72.7%) for the 5.0 EE 2 group, and 10/12 (83.3%) for the 10.0 E”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    There were no significant effects of treatment, test day, or interaction on lordosis postures, hops/darts, male aggression directed at female, female investigation of male anogenital area, or male investigation of female anogenital area.

    “There were no significant effects of treatment, test day, or the interaction in the analyses of lordosis postures, hops/darks, male aggression directed at female, female investigation of male anogenital area, or male investigation of female anogenital area.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Each of the seven EE2 females had at least one session with female aggression directed at the male, whereas only 4 of the 12 vehicle control female sessions exhibited a similar event.

    “Each of the seven EE 2 females had at least one session in which there was female aggression directed at the male whereas only 4 of the 12 vehicle control female sessions exhibited a similar event.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    When EE2 females without vaginal opening were excluded, the proportion with a nonzero lordosis response was 60.0% for 5.0 EE2 and 71.4% for 10.0 EE2.

    “When the 5.0 and 10.0 EE 2 females that never experienced vaginal opening were excluded, the number (and percentage) of females in those two groups with a nonzero response on at least one of the three trials on each of the 5 days was: 3/5 (60.0%) for the 5.0 EE 2 group and 5/7 (71.4%) for the 10.0 EE 2 group.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    There was a significant effect of day on saccharin preference, with preference on days 1 and 2 higher than day 3.

    “Analysis of saccharin preference indicated a significant effect of day (F(2, 251) = 7.82, p < 0.001) and pairwise comparisons revealed that preference on days 1 and 2 was higher than on day 3 (p < 0.001).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    On average, 29% of sessions with 10.0 EE2 females contained at least one event of female aggression directed at the male, whereas only 7% of vehicle control female sessions did.

    “On average, 29% of the sessions with 10.0 EE 2 females contained at least one event of female aggression directed at the male whereas only 7% of the vehicle control female sessions contained a similar event.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingown data

    Intake of saccharin-flavored solution on each of the 3 days differed, with highest intake on the first day.

    “Intake on each of the 3 days differed from each other with the highest intake on the first day (all comparisons p < 0.001).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Pinning frequencies of EE2-treated males during play behavior assessments were somewhat higher than those of vehicle control males, but saccharin- and sodium-flavored solution intake and running wheel activity were well within the range of vehicle control males.

    “Pinning frequencies of EE 2 -treated males during play behavior assessments were somewhat higher than those of vehicle control males, but saccharin-and sodium-flavored solution intake as well as running wheel activity were well within the range of vehicle control males.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingcitation

    We previously reported that developmental treatment with 10-38 g/kg/day of dietary EE2 caused increased sodium-flavored solution intake at adulthood in males and females, a finding not replicated here.

    “We previously reported that developmental treatment with 10-38 g/kg/day of dietary EE 2 caused increased sodium-flavored solution intake at adulthood in males and females (Ferguson et al., 2003), a finding not replicated here.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ferguson et al., 2003

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Findingown data

    The pinning frequency of EE2-treated females was almost twice that of vehicle control females and nearly 40% more than that of vehicle control males.

    “Here, the pinning frequency of EE 2 -treated females was almost twice that of vehicle control females and nearly 40% more than that of vehicle con-trol males, indicating pronounced masculinization.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingnone given

    In the current study, all females were gonadally intact and serum estradiol levels were comparable in all treatment groups.

    “In the current study, not only were all females gonadally intact, but serum estradiol levels were comparable in all treatment groups.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingnone given

    In our previous study, that EE2 treatment also decreased postweaning and adult body weights of both sexes, effects not seen here.

    “However, in our previous study, that EE 2 treatment also decreased postweaning and adult body weights of both sexes, effects not seen here.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Males of both EE2-treated groups experienced significant delays of 4-7 days in testes descent and 7-25 days in preputial separation.

    “Males of both EE 2 -treated groups experienced significant delays of 4-7 days in testes descent and 7-25 days in preputial separation.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingcitation

    EE2 treatment increased play behavior pinning frequencies in female rats, similar to the increase caused by developmental androgen treatment.

    “For example, EE 2 treatment resulted in increased play behavior pinning frequencies by female rats similar to that caused by developmental androgen treatment (Meaney, 1989;Meaney and McEwen, 1986;Tonjes et al., 1987).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites Meaney, 1989

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Meaney and McEwen, 1986 source
    • cites Tonjes et al., 1987

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Findingcitation

    Saccharin-flavored intake was not altered in either sex, similar to findings reported by McGivern and Henschel (1990) for adult male and female rats treated with estradiol benzoate on PNDs 2 and 3.

    “Saccharin-flavored intake was not altered in either sex, a finding similar to that reported by McGivern and Henschel (1990) for adult male and female rats treated with estradiol benzoate on PNDs 2 and 3.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites McGivern and Henschel (1990)

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Findingown data

    There were no significant treatment group or sex differences on the dorsal contacts play behavior endpoint.

    “There were no significant treatment group or sex differences on this play behavior endpoint.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingcitation

    In the endpoints measured here, BPA treatment produced no significant effects, despite significantly increased SDN-POA volume in PND 21 males of the 2.5 and 25.0 BPA groups.

    “In the endpoints measured here, BPA treatment produced no significant effects, despite the significantly increased SDN-POA volume in PND 21 males of the 2.5 and 25.0 BPA groups (He et al., 2012), siblings of the subjects described here.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites He et al., 2012

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S4 (based).

    Study design and conductnone given

    The two doses of BPA (2.5 and 25.0 g/kg/day) were specifically selected to be below the no-observed-adverse-effect level (NOAEL) of 5 mg/kg/day.

    “The two doses of BPA (2.5 and 25.0 g/kg/day) were specifically selected to be below the no-observed-adverse-effect level (NOAEL) of 5 mg/kg/day.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Many developmental neurotoxicity effects of BPA have been reported at BPA doses less than the NOAEL.

  • Classified as probably not a substantial claim.

    Premisecitation

    The two doses of EE2 (5.0 and 10 g/kg/day) were selected based on a previous study of dietary EE2 exposure (Ferguson et al., 2003) in which male and female offspring of dams consuming chow containing 200 ppb EE2 exhibited reduced body weight but few behavioral alterations.

    “The two doses of EE 2 (5.0 and 10 g/kg/day) were selected based on our previous study of dietary EE 2 exposure (Ferguson et al., 2003) in which male and female offspring of dams consuming chow containing 200 ppb EE 2 (≈10-13 and 18-38 g/kg/day during pregnancy and lactation, respectively) exhibited reduced body weight but few behavioral alterations.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Ferguson et al., 2003

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Classified as probably not a substantial claim.

    Premisecitation

    CMC was selected as vehicle rather than an oil because certain oils may possess estrogenic activity.

    “CMC was selected as vehicle, rather than an oil, because certain oils may possess estrogenic activity (Ashby and Lefevre, 1997;Ryan, 2005).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Ashby and Lefevre, 1997

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Ryan, 2005

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The general consensus is that there may be some risks of BPA exposure during development, particularly central nervous system exposure.

    “The general consensus is that there may be some risks of BPA exposure during development, particularly central nervous system exposure (National Toxicology Program, 2008).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites National Toxicology Program, 2008

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductreasoning

    Breeding occurred prior to any treatment; thus, success or failure was not treatment-related.

    “Breeding occurred prior to any treatment; thus, success or failure was not treatment-related.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The authors have previously shown that flavored solution intake (ml intake/g body weight) is sexually dimorphic in adult Sprague Dawley rats.

    “We have previously shown this endpoint (ml intake/g body weight) to be sexually dimorphic in adult Sprague Dawley rats (Ferguson and Boctor, 2010;Ferguson et al., 2007;Flynn et al., 2005).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Ferguson and Boctor, 2010 source
    • cites Ferguson et al., 2007 source
    • cites Flynn et al., 2005 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductreasoning

    This assignment yields approximately equal dam body weight distributions across treatments.

    “This assignment yields approximately equal dam body weight distributions across treatments.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductown data

    Not all females were examined and the number affected in each treatment group is not known.

    “Not all females were examined and the number affected in each treatment group is not known.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Litter was the experimental unit in all analyses.

    “Litter was the experimental unit in all analyses.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The naïve control group was included as a potential control for stress induced by gavage, because prenatal stress can alter later behaviors.

    “The naïve control group was included as a potential control for stress induced by gavage, because prenatal stress can alter later behaviors (reviewed in Buynitsky and Mostofsky, 2009;Maccari and Morley-Fletcher, 2007;Weinstock, 2001).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Buynitsky and Mostofsky, 2009 source
    • cites Maccari and Morley-Fletcher, 2007 source
    • cites Weinstock, 2001 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Females for which a sperm plug was detected were randomly assigned to one of the six treatment groups within their body weight stratum.

    “Females for which a sperm plug was detected were randomly assigned to one of the six treatment groups within their body weight stratum.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductown data

    A single tester blind to treatment status scored all play behavior sessions after training to reliability coefficients of 0.94 or better.

    “A single tester blind to treatment status scored all sessions after training to reliability coefficients of 0.94 or better on the two behaviors.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Female sex behavior videorecordings were scored by a single experienced tester blind to treatment status.

    “Videorecordings were later scored by a single experienced tester blind to treatment status of the female.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Manual lordosis assessments were conducted by a tester blind to treatment status.

    “Each female was palpated three times in succession and all palpations were done by the same tester who was blind to treatment status.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductreasoning

    Hormone levels were statistically analyzed twice: once including all subjects and once without the 15 females for which samples were obtained in the afternoon.

    “Thus, hormone levels were statistically analyzed twice: once including all subjects and once without the 15 females for which samples were obtained in the afternoon.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Several hormones measured in this study are associated with estrous phase.

  • Classified as probably not a substantial claim.

    Premisecitation

    Several hormones measured here exhibit circadian rhythms.

    “Several hormones measured here exhibit circadian rhythms (e.g., Kalra and Kalra, 1977;Mock et al., 1978).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Kalra and Kalra, 1977 source
    • cites Mock et al., 1978 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    Omitting data from the 15 females in which blood was obtained in the afternoon did not change the statistical results.

    “Omitting data from the 15 females in which blood was obtained in the afternoon did not change the statistical results.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Prenatal DES or neonatal estradiol benzoate treatment can produce increased body weight in female rats and mice.

    “Further, prenatal DES or neonatal estradiol benzoate treatment can produce increased body weight in female rats and mice (Donohoe and Stevens, 1983;Nikaido et al., 2004).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Donohoe and Stevens, 1983 source
    • cites Nikaido et al., 2004 source

  • Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing).

    Interpretationreasoning

    The historical literature from our laboratory and the effects in the vehicle control group here provide support for the treatment-related effects described here.

    “The historical literature from our laboratory and the effects in the vehicle control group here provide support for the treatment-related effects described here.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    With the possible exceptions of estrous cyclicity and dark period running wheel activity, there were no apparent indications of a dose-response as the effects seemed equally severe in both EE2-treated groups.

    “With the possible exceptions of estrous cyclicity and dark period running wheel activity, there were no apparent indications of a dose-response as the effects seemed equally severe in both EE 2treated groups.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationcitation

    The delayed vaginal opening described for the EE2-treated females here was not unexpected as similar EE2 or estradiol benzoate treatment also resulted in vaginal opening delays.

    “The delayed vaginal opening described for the EE 2 -treated females here was not unexpected as similar EE 2 (e.g., 0.5 or 5.0 g/kg/day) or estradiol benzoate treatment also resulted in vaginal opening delays (Berretti et al., 2014;Delclos et al., 2012Delclos et al., , 2014;;Sawaki et al., 2003a).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Berretti et al., 2014 source
    • cites Delclos et al., 2012

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Sawaki et al., 2003a

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    There appear to be no reports of BPA effects on long-term activity assessments, such as residential running wheel activity.

    “There appear to be no reports of BPA effects on long-term activity assessments, such as residential running wheel activity.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisenone given

    The methodology used here for assessing play behavior is standard in the rodent literature, but others have used different methods for examining BPA-induced alterations.

    “The methodology used here for assessing play behavior (i.e., a brief separation followed by reuniting a same-sex pair) is standard in the rodent literature, but others have used different methods for examin-”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisenone given

    There do not appear to be published descriptions of the effects of BPA treatment on intake or preference of sodium flavored solutions.

    “There do not appear to be published descriptions of the effects of BPA treatment on intake or preference of sodium flavored solutions.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Multiple endpoints in both sexes were significantly affected by EE2 treatment, although the number of affected endpoints and their severity were higher in females.

    “Multiple endpoints in both sexes were significantly affected by EE 2 treatment, although the number of affected endpoints and their severity were higher in females.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    The EE2 effects here replicate those previously described, but the lack of BPA effects at these doses is not entirely consistent with the literature.

    “Although the EE 2 effects here replicate those previously described (discussed below), the lack of BPA effects at these doses is not entirely consistent with the literature.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    The masculinization/defeminization of EE2-treated females was apparent in a heavier body weight, delayed or complete lack of vaginal opening, aberrant estrous cyclicity, increased play behavior, decreased running wheel activity, and increased male-directed aggression.

    “Here, the masculinization/defeminization of EE 2 -treated females was apparent in a heavier body weight, delayed or complete lack of vaginal opening, aberrant estrous cyclicity, increased play behavior, decreased running wheel activity, and increased maledirected aggression.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductnone given

    Due to an inadvertent error, the sodium solution concentration for animals in replicate 1 was 3.0% instead of 0.3%, and data for those 14 subjects were deleted prior to analyses.

    “Due to an inadvertent error, the sodium solution concentration for animals in replicate 1 was 3.0% and not 0.3% as intended.Therefore, data for regular water and sodium-flavored solution intake for those 14 subjects were deleted prior to analyses.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    The lack of BPA-induced long-term activity alterations reported here awaits replication.

    “Thus, the lack of BPA-induced long-term activity alterations reported here awaits replication.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    In the previous study, postnatal exposure to EE2 occurred via lactation and the offspring were likely exposed to a lower dose postnatally than with the direct treatment in the current study.

    “However, in that previous study, postnatal exposure to EE 2 occurred via lactation and it is likely that the offspring were exposed to a lower dose postnatally relative to the direct treatment in the current study.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Circulating levels that are 54%, but not 13%, of normal endogenous levels can maintain normal wheel running activity levels of male mice held under constant light or dark photoperiods.

    “Circulating levels that are 54%, but not 13%, of normal endogenous levels can maintain normal wheel running activity levels of male mice held under constant light or dark photoperiods (Butler et al., 2012).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Butler et al., 2012 source

  • May combine two claims.

    Premisecitation

    EE2 treatment (at doses >5.0 g/kg/day) from GD 7 through lactational day 18 or an injection of estradiol benzoate at birth decreased lordosis behavior of adult female offspring.

    “Likewise, EE 2 treatment (at doses >5.0 g/kg/day) from GD 7 through lactational day 18 or an injection of estradiol benzoate at birth decreased the lordosis behavior of adult female offspring (Berretti et al., 2014;Ryan et al., 2010).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Berretti et al., 2014 source
    • cites Ryan et al., 2010 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Such malformations in females can also result from a single injection of estradiol or DES on GD 19.

    “Such malformations in females can also result from a single injection of estradiol or DES on GD 19 (Henry et al., 1984).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Henry et al., 1984 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationcitation

    The testosterone levels of the EE2-treated males in the current study may have been sufficient to maintain normal wheel running activity.

    “Although low, perhaps the testosterone levels of the EE 2 -treated males of the current study were sufficient to maintain normal wheel running activity.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Males treated with 10 g/kg/day EE2 exhibited a significant increase in SDN-POA volume at PND 21.

    “In contrast, males treated with 10 g/kg/day EE 2 exhibited a significant increase in SDN-POA volume at PND 21 (He et al., 2012).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites He et al., 2012

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Loss of circulating estrogens (e.g., postovariectomy) drastically reduces running wheel activity in rats and mice, and this can be restored by exogenous estradiol treatment.

    “Nevertheless, it is clear that loss of circulating estrogens (e.g., postovariectomy) drastically reduces running wheel activity in rats and mice, which can be restored by exogenous estradiol treatment (Gray et al., 1989;Morgan andPfaff, 2001, 2002;Rivera and Eckel, 2005;Ruiz de Elvira et al., 1992).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Gray et al., 1989 source
    • cites Morgan andPfaff, 2001, 2002

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Rivera and Eckel, 2005 source

  • May combine two claims.

    Interpretationnone given

    Although males were affected, females treated with EE2 appeared more severely affected.

    “Although males were affected, females treated with EE 2 appeared more severely affected.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    The EE2 doses used here were high enough to cause malformations, and such doses are within the range of those that produce adverse effects in adult women.

    “Further, although the EE 2 doses here were high enough to cause malformations, such doses are within the range of those that produce adverse effects in adult women (see argument presented in Gray et al., 2010).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Gray et al., 2010 source
78 of 78

  • The paper relies on this without support.

    Bisphenol A is a weak estrogen.

    “Because BPA is a weak estrogen and because androgens/estrogens are critical for sexual differentiation of the brain and behavior of rodents, we tested the hypothesis that BPA could influence sexually dimorphic behaviors when administered during the developmental period that is known to be affected by estrogens in the rat.”

  • The paper relies on this without support.

    Androgens and estrogens are critical for sexual differentiation of the brain and behavior of rodents.

    “Because BPA is a weak estrogen and because androgens/estrogens are critical for sexual differentiation of the brain and behavior of rodents, we tested the hypothesis that BPA could influence sexually dimorphic behaviors when administered during the developmental period that is known to be affected by estrogens in the rat.”

  • The paper relies on this without support.

    Many developmental neurotoxicity effects of BPA have been reported at BPA doses less than the NOAEL.

    “Many of the developmental neurotoxicity effects of BPA have been reported at BPA doses less than the NOAEL; thus, it was essential to investigate doses within that range.”

  • The paper relies on this without support.

    Several hormones measured in this study are associated with estrous phase.

    “Further, several hormones measured here are associated with estrous phase.”

  • The authors state this as an assumption.

    Activity is cyclical over a 24-h period.

    “In addition to those typical analyses, running wheel data were modeled using a sigmoidally transformed cosine curve described by Marler et al. (2006) in which it is assumed that activity is cyclical over a 24-h period.”

  • The paper relies on this without support.

    Few studies have examined long-term activity levels, such as residential running wheel activity, after developmental estrogen treatment.

    “Few studies have examined long-term activity levels, such as residential running wheel activity, after developmental estrogen treatment.”
6 of 6

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • In mammals, pharmacological levels of EE2 exert important effects on reproductive physiology and behavior.

    “In mammals, pharmacological levels of EE2 exert important effects on reproductive physiology and behavior [18, 31–34].”

    From Developmental exposure to low levels of ethinylestradiol affects social play in juvenile male rats (2020) · cites this paper as [18]

    The Discussion reports that developmental EE2 treatment altered reproductive measures and behavior in male and female Sprague Dawley rats, and Materials and Methods specifies the administered doses. The study supports the claim for rats under this treatment regimen, but does not establish the effect across mammals generally or explicitly characterize the doses as pharmacological. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

  • Estrogens or estrogenic EDCs are involved in the development of play behavior in rats.

    “However, recent research showed that estrogens or estrogenic EDCs are also involved in the development of play behavior in rats [15–19].”

    From Developmental exposure to low levels of ethinylestradiol affects social play in juvenile male rats (2020) · cites this paper as [18]

Statistics that don’t add up

Each reported p-value recomputed from its test statistic and degrees of freedom.

  • p-value fits an unstated one-tailed test: F(1, 128) = 10.47,p < .001

    Minor

    The paper reports p < .001 here; recomputing from the test statistic and its degrees of freedom gives p = .002. The reported value is what a one-tailed test gives, but the paper does not say near this result that a one-tailed test was used.

    Reported p < .001 · recomputed p = .002

  • p-value fits an unstated one-tailed test: F(1, 128) = 9.54, p < .002

    Minor

    The paper reports p < .002 here; recomputing from the test statistic and its degrees of freedom gives p = .002. The reported value is what a one-tailed test gives, but the paper does not say near this result that a one-tailed test was used.

    Reported p < .002 · recomputed p = .002

2 of 2

Statistics recomputed

Every reported test result, recomputed from its test statistic and degrees of freedom. A result shows as not adding up only once an independent check has confirmed it.

  • Doesn’t add upF(1, 128) = 9.54reported p < .002 · recomputed p = .002

    “F(1, 128) = 9.54, p < .002”

  • Doesn’t add upF(1, 128) = 10.47reported p < .001 · recomputed p = .002

    “F(1, 128) = 10.47,p < .001”

  • ConsistentF(5, 64) = 5.35reported p < .001 · recomputed p < .001

    “F(5, 64) = 5.35, p < .001”

  • ConsistentF(10, 538) = 3638.48reported p < .001 · recomputed p < .001

    “F(10, 538) = 3638.48,p < .001”

  • ConsistentF(50, 640) = 4.65reported p < .001 · recomputed p < .001

    “F(50, 640) = 4.65, p < .001”

  • ConsistentF(50, 622) = 1.58reported p < .008 · recomputed p = .008

    “F(50, 622) = 1.58, p < .008”

  • ConsistentF(50, 489) = 1.44reported p < .03 · recomputed p = .030

    “F(50, 489) = 1.44, p < .030”

  • ConsistentF(55, 696) = 2.74reported p < .001 · recomputed p < .001

    “F(55, 696) = 2.74, p < .001”

  • ConsistentF(55, 563) = 1.5reported p < .015 · recomputed p = .014

    “F(55, 563) = 1.50, p < .015”

  • ConsistentF(5, 64) = 41.16reported p < .001 · recomputed p < .001

    “F(5, 64) = 41.16,p < .001”

  • ConsistentF(5, 63) = 34.03reported p < .001 · recomputed p < .001

    “F(5, 63) = 34.03,p < .001”

  • ConsistentF(5, 56) = 4.88reported p < .001 · recomputed p < .001

    “F(5, 56) = 4.88, p < .001”

  • ConsistentF(1, 127) = 109.09reported p < .001 · recomputed p < .001

    “F(1, 127) = 109.09,p < .001”

  • ConsistentF(5, 127) = 7.37reported p < .001 · recomputed p < .001

    “F(5, 127) = 7.37, p < .001”

  • ConsistentF(5, 127) = 6.59reported p < .001 · recomputed p < .001

    “F(5, 127) = 6.59, p < .001”

  • ConsistentF(5, 128) = 7.83reported p < .001 · recomputed p < .001

    “F(5, 128) = 7.83, p < .001”

  • ConsistentF(5, 128) = 6.75reported p < .001 · recomputed p < .001

    “F(5, 128) = 6.75, p < .001”

  • ConsistentF(1, 128) = 126.9reported p < .001 · recomputed p < .001

    “F(1, 128) = 126.90,p < .001”

  • ConsistentF(5, 128) = 5.3reported p < .001 · recomputed p < .001

    “F(5, 128) = 5.30, p < .001”

  • ConsistentF(2, 256) = 4.72reported p < .01 · recomputed p = .010

    “F(2, 256) = 4.72, p < .010”

  • ConsistentF(1, 128) = 53.54reported p < .001 · recomputed p < .001

    “F(1, 128) = 53.54,p < .001”

  • ConsistentF(2, 256) = 69.46reported p < .001 · recomputed p < .001

    “F(2, 256) = 69.46,p < .001”

  • ConsistentF(2, 251) = 7.82reported p < .001 · recomputed p < .001

    “F(2, 251) = 7.82, p < .001”

  • ConsistentF(5, 114) = 3.38reported p < .008 · recomputed p = .007

    “F(5, 114) = 3.38, p < .008”

  • ConsistentF(2, 226) = 16.73reported p < .001 · recomputed p < .001

    “F(2, 226) = 16.73,p < .001”

  • ConsistentF(5, 114) = 2.96reported p < .016 · recomputed p = .015

    “F(5, 114) = 2.96, p < .016”

  • ConsistentF(2, 223) = 13.96reported p < .001 · recomputed p < .001

    “F(2, 223) = 13.96,p < .001”

  • ConsistentF(5, 114) = 2.31reported p < .049 · recomputed p = .049

    “F(5, 114) = 2.31, p < .049”

  • ConsistentF(2, 222) = 47.65reported p < .001 · recomputed p < .001

    “F(2, 222) = 47.65,p < .001”

29 of 29

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Two different dark-cycle start and end times are specified

    Minor

    The running-wheel methods say its lighting schedule matched the housing room's 6 a.m.-6 p.m. schedule, implying darkness from 6 p.m. to 6 a.m. The running-wheel analysis instead places darkness approximately half an hour later at both ends.

1 of 1

Cites

Flagged first, then by citations

Research works printed but not linked: 2

  1. 2012, Effects of Bisphenol A (BPA) and ethinyl estradiol (EE2) in Sprague-Dawley rats exposed orally from GD 6 through PND 90. The Toxicologist (published by Society of Toxicology). Abstract #1719, 371
  2. 2005, Developmental exposure to environmental extrogens alters adult behavior in female rodents. Zoology. Dissertation, 213

Cited by

Sorted by citations

Related

Authors
Ferguson, S. A., Law, C. D., & Kissling, G. E.
Affiliations
  • National Center for Toxicological Research (US)
  • National Institutes of Health (US)
ORCID
Ferguson, Sherry A.
Venue
Toxicological Sciences
Publisher
Oxford University Press (OUP)
Published
2014-08-01
Volume
140
Issue
2
Pages
374-392
ISSN
  • 1096-0929 (ISSN-L)
  • 1096-6080
DOI
10.1093/toxsci/kfu077
PMC
PMC4133561
OpenAlex
W2120150972
Semantic Scholar
6704d493f3d9e19b40eef5d1ef2c2580388aafae
Cited by
32
References
98
Citations by year
2023: 4 · 2021: 2 · 2020: 4 · 2019: 2 · 2018: 4 · 2017: 2 · 2016: 2 · 2015: 9 · 2014: 3
FWCI
3.29
Type
journal article
Language
en
Open access
Licence
public-domain
Locations
  • Toxicological Sciences (publishedVersion, public-domain)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, public-domain)
Funders
  • National Institutes of Health
  • National Institute of Environmental Health Sciences (Z01 ES45003)
Topics
  • Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Effects and risks of endocrine disrupting chemicals
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Sexual Differentiation and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
Keywords
ethinyl estradiol, bisphenol A, developmental neurotoxicity, sexually dimorphic behavior, prenatal exposure, puberty, testosterone, estrous cyclicity, lordosis, aggression, sexually dimorphic nucleus of the preoptic area, masculinization, defeminization, endocrine disruptors, Sprague-Dawley rats, delayed puberty, testicular descent
MeSH terms
Bisphenol A Compounds, Animals, Behavior, Animal, Benzhydryl Compounds, Body Weight, Drinking Behavior, Ethinyl Estradiol, Feeding Behavior, Female, Humans, Male, Organ Size, Phenols, Sex Characteristics, Sexual Maturation, Rats, Sprague-Dawley, Endocrine Disruptors

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-12.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-12.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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