Higher paracetamol levels are associated with elevated glucocorticoid concentrations in hair: findings from a large cohort of young adults.

Paracetamol is one of the most commonly used over-the-counter medications. Experimental studies suggest a possible stress-suppressing effect of paracetamol in humans facing experimental stress-inducing paradigms. However, no study has investigated whether paracetamol and steroid hormones covary over longer time frames and under real-life conditions. This study addresses this gap by investigating associations between steroid hormones (cortisol, cortisone, and testosterone) and paracetamol concentrations measured in human hair, indexing a timeframe of approximately three months. The data came from a large community sample of young adults (N = 1002). Hair data were assayed using liquid chromatography-tandem mass spectrometry. Multiple regression models tested associations between paracetamol and  steroid hormones, while adjusting for a wide range of potential confounders, such as sex, stressful live events, psychoactive substance use, hair colour, and body mass index. Almost one in four young adults from the community had detectable paracetamol in their hair (23%). Higher paracetamol hair concentrations were robustly associated with more cortisol (ß = 0.13, ηp = 0.016, p < 0.001) and cortisone (ß = 0.16, ηp = 0.025, p < 0.001) in hair. Paracetamol and testosterone hair concentrations were not associated. Paracetamol use intensity positively correlated with corticosteroid functioning across several months. However, a potential corticosteroid-inducing effect of chronic paracetamol use has yet to be tested in future experimental designs.

Substance use in sexual minority youth: prevalence in an urban cohort.

BACKGROUND: Little comparative data on substance use (SU) between sexual minority youth (SMY) and heterosexual youth (HET) is available. This study compares the prevalence of SU in an urban cohort between SMY and HET and evaluates demographic and psychosocial predictors of SU. METHODS: Data came from a prospective-longitudinal cohort study in an urban setting (N = 1297). SU and psychosocial variables such as internalizing symptoms, self-control, sensation-seeking, bullying-victimization, subjective stress, leisure activities, and peer influences were assessed with self-reports at age 17 and 20. SU was stratified by sex and sexual attraction, and the groups were compared using regression models, with demographic and psychosocial variables included as covariates. RESULTS: SMY- and HET-youth displayed differences in a number of psychosocial variables. Overall, SMY- and HET-youth differed in their 12-months prevalence of SU: At age 17, SMY-females had significantly higher rates of SU than HET-females for cannabis (aOR = 2.14, p = 0.04), ecstasy/MDMA (aOR = 4.29, p = 0.01), and hallucinogens (aOR = 5.59, p = 0.02). At age 20, SMY-females had significantly higher rates of SU than HET-females for tobacco (aOR = 2.06, p = 0.03), cannabis (aOR = 2.24, p = 0.004), ecstasy/MDMA (aOR = 3.93, p < 0.001), stimulants (aOR = 3.45, p = 0.002), and hallucinogens (aOR = 6.65, p < 0.001). SMY-males reported significantly lower rates for tobacco and cannabis than HET-males at age 17. At age 20, they reported significantly higher rates for the use of ecstasy/MDMA (aOR = 2.30, p = 0.04) and hallucinogens (aOR = 2.43, p = 0.03). CONCLUSIONS: Given that psychosocial variables were significant covariates of SMY-status and SU, our results underline the importance of accounting for these when explaining differences in SU between adolescents. While differentiation by sex is established in most studies, such standardized comparisons are lacking with regards to sexual identities. But knowledge about SU of SMY is critical for designing effective interventions. This is especially true for SMY-females: Thus, SU in SMY-females early in life needs to be explored more thoroughly and addressed with adequate prevention measures.

Associations of psychoactive substances and steroid hormones in hair: Findings relevant to stress research from a large cohort of young adults.

OBJECTIVE: Epidemiological studies increasingly use hair samples to assess people's cumulative exposure to steroid hormones, but how the use of different psychoactive substances may affect steroid hormone levels in hair is, so far, largely unknown. The current study addresses this gap by establishing the substance exposure correlates of cortisol, cortisone, and testosterone in hair, while also accounting for a number of relevant covariates. METHOD: Data came from a large urban community-sample of young adults with a high prevalence of substance use (N = 1002, mean age=20.6 years, 50.2% female), who provided 3 cm of hair samples. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantified cortisol, cortisone, and testosterone, as well as delta-9-tetrahydrocannabinol (THC), 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy"), cocaine, several opioids, and their respective metabolites. Multiple linear regression models with covariates were used to predict steroid hormone levels from substance exposure in a four-step approach: In the full sample, low and high substance hair concentrations (median split) were first tested against no use for each substance individually (step 1) and for all substances together (step 2). Then, within the participants with any substance in hair only, the continuous hair concentration of each substance in pg/mg (step 3) and finally of all substances together, were regressed (step 4). RESULTS: Low, high, and continuous levels of THC in hair were robustly associated with higher levels of cortisol (sig. in step 1 low THC: ß = 0.29, p = .021; high THC: ß = 0.42, p = .001; step 2: low THC: ß = 0.27, p = 0.036, and high THC: ß = 0.40, p = .004, and step 4: ß = 0.12, p = .041). Participants with high MDMA levels had higher levels of cortisone without adjusting for other substances (step 1: ß = 0.34, p = .026), but this effect was not significant in the other models. While high THC levels were associated with lower levels of testosterone in step 2 (ß = -0.35, p = .018), MDMA concentration was positively related to testosterone concentration with and without adjusting for other substances (step 3: ß = 0.24, p = .041; step 4: ß = 0.17, 95%, p = .015) in male participants. CONCLUSION: The use of psychoactive substances, especially of cannabis and ecstasy, should be considered in studies investigating steroid hormones in hair.

The worst and the best: new insights into risk and resilience in young adults from the COVID-19 pandemic.

Historic declines in young people's mental health began to emerge before the COVID-19 pandemic. In the face of this youth mental health crisis, the pandemic constituted a naturalistic stressor paradigm that came with the potential to uncover new knowledge for the science of risk and resilience. Surprisingly, approximately 19-35% of people reported better well-being in the first few months of the COVID-19 pandemic than before. Therefore, in May and September 2020, we asked N=517 young adults from a cohort study to describe the best and the worst aspects of their pandemic lives (N=1,462 descriptions). Inductive thematic analysis revealed that the best aspects included the deceleration of life and a greater abundance of free time, which was used for hobbies, healthy activities, strengthening relationships, and for personal growth and building resilience skills. Positive aspects also included a reduction in educational pressures and work load and temporary relief from climate change concerns. The worst aspects included disruptions and changes to daily life; social distancing and restrictions of freedoms; negative emotions that arose in the pandemic situation, including uncertainty about the future; and the growing polarization of society. Science that aims to reverse the youth mental health crisis must pay increased attention to sources of young people's distress that are not commonly measured (e.g., their educational, work, and time pressures; their fears and uncertainties about their personal, society's, and the global future), and also to previously untapped sources of well-being - including those that young people identified for themselves while facing the COVID-19 pandemic. Supplementary Information: The online version contains supplementary material available at 10.1007/s42844-023-00096-y.

Associations of different hormonal contraceptive methods with hair concentrations of cortisol, cortisone, and testosterone in young women.

Hair concentrations of cortisol, cortisone, and testosterone are non-invasive measures of cumulative steroid hormone levels. Use of contraceptives co-varies with levels of cortisol and cortisone in women's hair. It is unclear, however, how different contraceptive methods (i.e., that differ in their steroid hormone composition) affect corticosteroid and testosterone hair levels. The current study examines associations of contraceptives with hair steroid hormone concentrations in females from the community (N = 464, M = 20.6 years old, age range = 19-22). Self-reported contraceptives were first categorized as combined estrogen-progestin or progestin-only, and then analyzed individually in follow-up analyses. Multiple regressions adjusting for body mass index (BMI) and hair characteristics revealed that levels of hair cortisol, cortisone, and testosterone were significantly lower in women who used combined estrogen-progestin methods than in women who did not use hormonal contraception (ßcortisol(log) = -0.29; ßcortisone(log) = -0.28; ßtestosterone(log) = -0.36), showing moderate to large effect sizes (d = 0.64, d = 0.71, and d = 0.81, respectively). Concentrations of hair cortisol were lower in women who used progestin-only contraceptives (ß = -0.49) compared to no contraceptive use, with a large effect size (d = 1.67). Follow-up analyses revealed that the association of the three steroid hormones with estrogen-progestin methods was strongest for the combined oral "micro-pill." Future studies of hair steroid hormones should take into account the specific type of contraceptive used, as this may affect study results.

Constitutive Notch Signaling Causes Abnormal Development of the Oviducts, Abnormal Angiogenesis, and Cyst Formation in Mouse Female Reproductive Tract.

The Notch signaling pathway is critical for the differentiation of many tissues and organs in the embryo. To study the consequences of Notch1 gain-of-function signaling on female reproductive tract development, we used a cre-loxP strategy and Amhr2-cre transgene to generate mice with conditionally activated Notch1 (Rosa(Notch1)). The Amhr2-cre transgene is expressed in the mesenchyme of developing female reproductive tract and in granulosa cells in the ovary. Double transgenic Amhr2-cre, Rosa(Notch1) females were infertile, whereas control Rosa(Notch1) mice had normal fertility. All female reproductive organs in mutants showed hemorrhaging of blood vessels progressing with age. The mutant oviducts did not develop coiling, and were instead looped around the ovary. There were multiple blockages in the lumen along the oviduct length, creating a barrier for sperm or oocyte passage. Mutant females demonstrated inflamed uteri with increased vascularization and an influx of inflammatory cells. Additionally, older females developed ovarian, oviductal, and uterine cysts. The significant change in gene expression was detected in the mutant oviduct expression of Wnt4, essential for female reproductive tract development. Similar oviductal phenotypes have been detected previously in mice with activated Smo and in beta-catenin, Wnt4, Wnt7a, and Dicer conditional knockouts, indicating a common regulatory pathway disrupted by these genetic abnormalities.

Genetic ablation of androgen receptor signaling in fetal Leydig cell lineage affects Leydig cell functions in adult testis.

It is commonly accepted that androgen-producing fetal Leydig cells (FLC) are substituted by adult Leydig cells (ALC) during perinatal testis development. The mechanisms influencing this process are unclear. We used mice with a retinoid acid receptor 2 promoter-Cre recombinase transgene (Rarb-cre) expressed in embryonic FLC precursors, but not in postnatal testis, and a dual fluorescent Cre recombinase reporter to label FLC and ALC in vivo. All FLC in newborn testis had the recombinant, whereas the majority of LC in adult testis had the nonrecombinant reporter. Primary LC cultures from adult testis had either recombinant (20%) or nonrecombinant (80%) cells, demonstrating that the FLC survive in adult testis and their ontogeny is distinct from ALC. Conditional inactivation of androgen receptor (AR) allele using the Rarb-cre transgene resulted in a 50% increase of AR-negative LC in adult testis. The mutant males became infertile with age, with all LC in older testis showing signs of incomplete differentiation, such as a large number of big lipid droplets, an increase of finger-like protrusions, and a misexpression of steroidogenic or FLC- and ALC-specific genes. We propose that the antiandrogenic exposure during early development may similarly result in an increase of FLC in adult testis, leading to abnormal LC differentiation.

The fate of spermatogonial stem cells in the cryptorchid testes of RXFP2 deficient mice.

The environmental niche of the spermatogonial stem cell pool is critical to ensure the continued generation of the germ cell population. To study the consequences of an aberrant testicular environment in cryptorchidism we used a mouse model with a deletion of Rxfp2 gene resulting in a high intra-abdominal testicular position. Mutant males were infertile with the gross morphology of the cryptorchid testis progressively deteriorating with age. Few spermatogonia were identifiable in 12 month old cryptorchid testes. Gene expression analysis showed no difference between mutant and control testes at postnatal day 10. In three month old males a decrease in expression of spermatogonial stem cell (SSC) markers Id4, Nanos2, and Ret was shown. The direct counting of ID4+ cells supported a significant decrease of SSCs. In contrast, the expression of Plzf, a marker for undifferentiated and differentiating spermatogonia was not reduced, and the number of PLZF+ cells in the cryptorchid testis was higher in three month old testes, but equal to control in six month old mutants. The PLZF+ cells did not show a higher rate of apoptosis in cryptorchid testis. The expression of the Sertoli cell FGF2 gene required for SSC maintenance was significantly reduced in mutant testis. Based on these findings we propose that the deregulation of somatic and germ cell genes in the cryptorchid testis, directs the SSCs towards the differentiation pathway. This leads to a depletion of the SSC pool and an increase in the number of PLZF+ spermatogonial cells, which too, eventually decreases with the exhaustion of the stem cell pool. Such a dynamic suggests that an early correction of cryptorchidism is critical for the retention of the SSC pool.

Testicular cancer and cryptorchidism.

The failure of testicular descent or cryptorchidism is the most common defect in newborn boys. The descent of the testes during development is controlled by insulin-like 3 peptide and steroid hormones produced in testicular Leydig cells, as well as by various genetic and developmental factors. While in some cases the association with genetic abnormalities and environmental causes has been shown, the etiology of cryptorchidism remains uncertain. Cryptorchidism is an established risk factor for infertility and testicular germ cell tumors (TGCT). Experimental animal models suggest a causative role for an abnormal testicular position on the disruption of spermatogenesis however the link between cryptorchidism and TGCT is less clear. The most common type of TGCT in cryptorchid testes is seminoma, believed to be derived from pluripotent prenatal germ cells. Recent studies have shown that seminoma cells and their precursor carcinoma in situ cells express a number of spermatogonial stem cell (SSC) markers suggesting that TGCTs might originate from adult stem cells. We review here the data on changes in the SSC somatic cell niche observed in cryptorchid testes of mouse models and in human patients. We propose that the misregulation of growth factors' expression may alter the balance between SSC self-renewal and differentiation and shift stem cells toward neoplastic transformation.

Spermatogenesis in cryptorchidism.

Cryptorchidism or undescended testis is the most frequent congenital abnormality in newborn boys. The process of testicular descent to the scrotum is controlled by hormones produced in Leydig cells, insulin-like3, and androgens. Variation in genetic and environmental factors might affect testicular descent. A failure of spermatogenesis and germ cell apoptosis resulting in infertility as well as an increased risk of neoplastic transformation of germ cell are the direct consequences of cryptorchidism in adulthood.

Two novel mouse genes mapped to chromosome Yp are expressed specifically in spermatids.

The male-specific region of the Y chromosome is evolutionarily predisposed to accumulate genes important for spermatogenesis. Recent work in this laboratory identified two novel Y-linked transcripts that were upregulated in the testis in response to deletions on the chromosome arm Yq. This article reports the further characterisation of these two transcripts and their comparison to related X and autosomal genes. Both map to chromosome arm Yp, outside the Sxr ( b ) deletion interval, both are present in at least two copies on the Y, and both are expressed specifically in spermatids. Given the testicular phenotype of mice with deletions on the Y chromosome, both genes are therefore likely to function in spermatid differentiation. AK006152 is a novel mouse-specific gene with a single potential open reading frame, and it is unusual in that there appears to be no X-linked relative. H2al2y is a novel histone in the H2A superfamily and has multiple X-linked relatives and a single autosomal relative in mouse. The presence of a single X-linked copy in rat suggests that H2al amplification is mouse-specific, with the alternative explanation being an earlier amplification followed by gene loss. A phylogenetic analysis of H2al genes together with other H2A genes indicates that H2al is most closely related to the mammalian-specific H2A.Bbd family of histones. Interestingly, K (a)/K (s) analysis indicates that the X and Y members of the H2al family may be under positive selection in mouse, while the autosomal copy is under purifying selection and presumably retains the ancestral function.

Bidirectional transcription of a novel chimeric gene mapping to mouse chromosome Yq.

BACKGROUND: The male-specific region of the mouse Y chromosome long arm (MSYq) contains three known highly multi-copy X-Y homologous gene families, Ssty1/2, Sly and Asty. Deletions on MSYq lead to teratozoospermia and subfertility or infertility, with a sex ratio skew in the offspring of subfertile MSYqdel males RESULTS: We report the highly unusual genomic structure of a novel MSYq locus, Orly, and a diverse set of spermatid-specific transcripts arising from copies of this locus. Orly is composed of partial copies of Ssty1, Asty and Sly arranged in sequence. The Ssty1- and Sly-derived segments are in antisense orientation relative to each other, leading to bi-directional transcription of Orly. Genome search and phylogenetic tree analysis is used to determine the order of events in mouse Yq evolution. We find that Orly is the most recent gene to arise on Yq, and that subsequently there was massive expansion in copy number of all Yq-linked genes. CONCLUSION: Orly has an unprecedented chimeric structure, and generates both "forward" (Orly) and "reverse" (Orlyos) transcripts arising from the promoters at each end of the locus. The region of overlap of known Orly and Orlyos transcripts is homologous to Sly intron 2. We propose that Orly may be involved in an intragenomic conflict between mouse X and Y chromosomes, and that this process underlies the massive expansion in copy number of the genes on MSYq and their X homologues.

Deletions on mouse Yq lead to upregulation of multiple X- and Y-linked transcripts in spermatids.

Deletions on the mouse Y-chromosome long arm (MSYq) lead to teratozoospermia and in severe cases to infertility. We find that the downstream transcriptional changes in the testis resulting from the loss of MSYq-encoded transcripts involve upregulation of multiple X- and Y-linked spermatid-expressed genes, but not related autosomal genes. Therefore, this indicates that in normal males, there is a specific repression of X and Y (gonosomal) transcription in post-meiotic cells, which depends on MSYq-encoded transcripts. Together with the known sex ratio skew in favour of females in the offspring of fertile MSYqdel males, this strongly suggests the existence of an intragenomic conflict between X- and Y-linked genes. Two potential antagonists in this conflict are the X-linked multicopy gene Xmr and its multicopy MSYq-linked relative Sly, which are upregulated and downregulated, respectively, in the testes of MSYqdel males. Xmr is also expressed during meiotic sex chromosome inactivation (MSCI), indicating a link between the MSCI and the MSYq-dependent gonosomal repression in spermatids. We therefore propose that this repression and MSCI itself are evolutionary adaptations to maintain a normal sex ratio in the face of X/Y antagonism.