Not within spitting distance: Salivary immunoassays of estradiol have subpar validity for predicting cycle phase.

Salivary steroid immunoassays are widely used in psychoneuroendocrinological studies of menstrual cycle phase, puberty, and menopause. Though manufacturers advertise their assays as suitable, they have not been rigorously validated for these purposes. We collated data from eight menstrual cycle studies across > 1200 female participants and > 9500 time points. Seven studies collected saliva and one collected serum. All assayed estradiol and progesterone and had an independent measure of cycle phase (LH-surge, menstrual onset). In serum, cycle phase measures strongly predicted steroid concentrations. In saliva, cycle phase poorly predicted estradiol values, which showed an upward bias compared to expectations from serum. For salivary progesterone, predictability from cycle phase was mixed, low for enzyme-linked assays and moderate for tandem mass spectrometry. Imputing the population-average serum steroid changes from cycle phase may yield more valid values of hormonal changes for an independent person than directly assessing their hormone levels using salivary immunoassays.

Antenatal Classes in the Context of Prenatal Anxiety and Depression during the COVID-19 Pandemic.

Perinatal maternal anxiety and depression negatively affect intrauterine fetal development, birth outcome, breastfeeding initiation, duration, and milk composition. Antenatal classes potentially reduce the anxiety of pregnant women and may thus contribute to healthy infant development. The study investigates the relationship between participation in online or in-person antenatal classes and levels of anxiety and depression in Polish women during the COVID-19 pandemic. The study group included 1774 adult, non-smoking pregnant women. We compared the state anxiety (STAI-State) and depression levels (EPDS) in women who (i) attended antenatal classes in-person, (ii) attended online classes, and (iii) did not attend any of them. The statistical analyses included a GLM model and trend analysis, while controlling for maternal trait anxiety, age, pregnancy complications, trimester of pregnancy, previous pregnancies, and COVID-19 infections. We observed statistically significant differences in the level of anxiety (and depression). Women who did attend antenatal classes in person had the lowest levels of anxiety and depression. Considering the importance of maternal mental well-being on fetal development, birth outcome, and breastfeeding, in-person participation in antenatal classes should be recommended to pregnant women.

Importance of Daily Sex Hormone Measurements Within the Menstrual Cycle for Fertility Estimates in Cyclical Shifts Studies.

Recent discussions have highlighted the importance of fertility measurements for the study of peri-ovulatory shifts in women's mating psychology and mating-related behaviors. Participants in such studies typically attend at least two test sessions, one of which is, at least in theory, scheduled to occur during the high-fertility, peri-ovulatory phase of the menstrual cycle. A crucial part of this debate is whether luteinizing hormone (LH) tests alone are sufficient to accurately assign test sessions to the peri-ovulatory phase. This article adds to this ongoing debate by presenting analyses of a detailed database of daily estradiol levels and LH tests for 102 menstrual cycles. Based on more than 4,000 hormonal measurements, it is clear that individual differences in length of the cycle, length of the luteal phase and, perhaps most importantly, the discrepancy between the timing of the LH surge and the drop in estradiol that follows it are pronounced. Less than 40% of analyzed cycles followed the textbook pattern commonly assumed to occur in fertility-based research, in which the LH surge is assumed to occur not more than 48 hr before the estradiol drop. These results suggest that LH tests alone are not sufficient to assign test sessions to the peri-ovulatory phase and that analyses of sex hormones are essential to identify whether the participant was tested during the peri-ovulatory phase.

Average ovarian hormone levels, rather than daily values and their fluctuations, are related to facial preferences among women.

Hormones are of crucial importance for human behavior. Cyclical changes of ovarian hormones throughout women's menstrual cycle are suggested to underlie fluctuation in masculinity preference for both faces and bodies. In this study we tested this hypothesis based on daily measurements of estradiol and progesterone throughout menstrual cycle, and multiple measurements of women's preference towards masculinity of faces and bodies of men. We expected that due to a large variation among daily hormonal levels we would not observe a direct effect of daily hormone levels, but rather that average levels of ovarian hormones throughout the cycle (a reliable marker of a probability of conception) would better predict women's preferences. We found a negative relationship between average progesterone levels and facial masculinity preference, but only among women who were in long-term relationships. There was no relationship between facial masculinity preference and either of the estradiol or progesterone daily levels. Similarly, only average levels of hormones were significantly related to facial symmetry preference. For women who were in relationships estradiol was positively related to symmetry preference, while for single women this relationship was opposite. For body masculinity preference there were no significant relationships with neither averaged nor daily hormonal levels. Taken together, our results further suggest that overall cycle levels of ovarian hormones (averaged for a cycle) are better predictors of facial masculinity and symmetry preference than daily levels assessed during preferences' tests. Importantly, including information about relationship status in the investigations of hormonal bases of preferences is crucial.

Progesterone and women's anxiety across the menstrual cycle.

Animal models and a few human investigations suggest progesterone may be associated with anxiety. Progesterone naturally fluctuates across the menstrual cycle, offering an opportunity to understand how within-person increases in progesterone and average progesterone levels across the cycle correspond to women's anxiety. Across two longitudinal studies, we simultaneously modeled the between- and within-person associations between progesterone and anxiety using multilevel modeling. In Study 1, 100 Polish women provided saliva samples and reported their anxiety at three phases of the menstrual cycle: follicular, peri-ovulatory, and luteal. A significant between-person effect emerged, revealing that women with higher average progesterone levels across their cycles reported higher levels of anxiety than women with lower progesterone cycles. This effect held controlling for estradiol. In Study 2, 61 American women provided saliva samples and reported their attachment anxiety during laboratory sessions during the same three cycle phases. A significant between-person and within-person association emerged: women with higher average progesterone levels reported higher levels of attachment anxiety, and as women's progesterone levels increased across their cycles, so too did their attachment anxiety. These effects held controlling for cortisol. In sum, both studies provide support for a link between menstrual cycle progesterone levels and subjective anxiety.

Digit ratio (2D:4D) and circulating testosterone, oestradiol, and progesterone levels across the menstrual cycle.

BACKGROUND: Digit ratio (2D:4D) is used by researchers as an indicator of prenatal sex hormone exposure. Two previous studies have examined associations between 2D:4D and circulating sex steroid concentrations across the menstrual cycle in adult females. One reported that digit ratio correlated positively with oestradiol levels, whereas the other found no such effect; neither observed significant associations with progesterone. AIMS: To examine associations between 2D:4D, as well as asymmetry (i.e. right minus left 2D:4D), and circulating sex steroids across the menstrual cycle. STUDY DESIGN: Correlational. SUBJECTS: 32 naturally cycling adult females from rural southern Poland. OUTCOME MEASURES: Salivary oestradiol, progesterone, testosterone, and testosterone to oestradiol ratio (T:O) measured during the follicular, peri-ovulatory, and luteal phases. Average levels across the cycle were also examined. RESULTS AND CONCLUSIONS: Asymmetry in digit ratio correlated positively with oestradiol at each phase, as well as with average levels across the cycle. Each association, other than that relating to average levels, remained statistically significant after a range of covariates had been controlled for. No other significant correlations were observed between digit ratio variables and circulating hormone levels. Our results might suggest that low exposure to androgens and/or high exposure to oestrogens during gestation is a predictor of high oestradiol levels in naturally cycling females of reproductive age. However, considering that it was asymmetry in digit ratio, and not either right or left 2D:4D, that was a significant predictor, it is also possible that these effects reflect more general associations between bilateral asymmetry and circulating oestradiol levels.

La donna è mobile? Lack of cyclical shifts in facial symmetry, and facial and body masculinity preferences-A hormone based study.

Although under investigation for more than two decades, a common agreement on the occurrence of cyclical shifts in women's masculinity and symmetry preferences is still missing. Such shifts are considered to be an important feature of sexual selection as they supposedly direct women's attention towards cues for "good genes" (e.g. masculinity and symmetry) during times when probability of conception is the highest. Multiple studies have, however, failed to find these shifts. We attempt to address this lack of agreement analysing a sample of 110 healthy women, using intra-participant design and repeated measurements of oestradiol and LH during the cycle. To ensure the reliable detection of increased conception probability, both LH- based ovulation tests and multiple oestradiol measurements were used. We found no significant differences between women's preferences during different cycle phases for either body or facial masculinity, or for facial symmetry. Differences remained non-significant after controlling for participants' sexual openness, relationship status, and self-judged attractiveness. We suggest that putative cyclical shifts in preferences for cues for good genes are either very small (impossible to be tracked even with a relatively large sample) or they are far more complex than previously assumed, and further studies accounting for more confounding variables should be undertaken.

Women's preferences for men's beards show no relation to their ovarian cycle phase and sex hormone levels.

According to the ovulatory shift hypothesis, women's mate preferences for male morphology indicative of competitive ability, social dominance, and/or underlying health are strongest at the peri-ovulatory phase of the menstrual cycle. However, recent meta-analyses are divided on the robustness of such effects and the validity of the often-used indirect estimates of fertility and ovulation has been called into question in methodological studies. In the current study, we test whether women's preferences for men's beardedness, a cue of male sexual maturity, androgenic development and social dominance, are stronger at the peri-ovulatory phase of the menstrual cycle compared to during the early follicular or the luteal phase. We also tested whether levels of estradiol, progesterone, and the estradiol to progesterone ratio at each phase were associated with facial hair preferences. Fifty-two heterosexual women completed a two-alternative forced choice preference test for clean-shaven and bearded male faces during the follicular, peri-ovulatory (validated by the surge in luteinizing hormone or the drop in estradiol levels) and luteal phases. Participants also provided for one entire menstrual cycle daily saliva samples for subsequent assaying of estradiol and progesterone. Results showed an overall preference for bearded over clean-shaven faces at each phase of the menstrual cycle. However, preferences for facial hair were not significantly different over the phases of menstrual cycle and were not significantly associated with levels of reproductive hormones. We conclude that women's preferences for men's beardedness may not be related to changes in their likelihood of conception.

Lack of support for relation between woman's masculinity preference, estradiol level and mating context.

It has been proposed that women's preferences for male facial sexual dimorphism are positively correlated with conception probability and differ between short- and long-term mating contexts. In this study, we tested this assumption by analyzing relationships between estradiol levels to the women's preferences of male faces that were manipulated to vary in masculinity. Estradiol was measured in daily saliva samples throughout the entire menstrual cycle collected by Polish women with regular menstrual cycles. In our analyses, we included the three most commonly used definitions of the fertile window in the literature. After computing the overall masculinity preference of each participant and measuring hormone levels, we found that i) the timing of ovulation varied greatly among women (between -11 and -17days from the onset of the next menses, counting backwards), ii) there was no relationship between daily, measured during the day of the test (N=83) or average for the cycle (N=115) estradiol levels and masculinity preferences, iii) there were no differences in masculinity preferences between women in low- and high-conception probability phases of the cycle, and iv) there were no differences in masculinity preferences between short- and long-term mating contexts. Our results do not support the idea that women's preferences for a potential sexual partner's facial masculinity fluctuate throughout the cycle.