Hair Steroid Research Articles

Hair steroid before and after COVID-19 in preschoolers: the moderation of family characteristics

BackgroundFrequent or prolonged exposure to stressors may jeopardize young children’s health. The onset of the COVID-19 pandemic, coupled with disruptions in daily routines and social isolation resulting from public health preventive measures, have raised concerns about its potential impact on children’ experienced stress, particularly for young children and vulnerable families. However, whether the pandemic was accompanied by changes in physiological stress remains unknown as perceived stress is not a good proxy of physiological stress. This study examined if preschoolers showed increasing hair steroid concentrations following the onset of the COVID-19 pandemic and whether family characteristics may have exacerbated or buffered these changes. Methods136 preschoolers (2–4 years) provided hair for steroid measurement (cortisol, dehydroepiandrosterone (DHEA), cortisone, cortisol-to-DHEA ratio, cortisol-to-cortisone ratio) in October-November 2019 (T0) and in July-August 2020 (T1). A 2-centimeter hair segment was analyzed, reflecting steroid production over the two months leading up to collection. Family income, conflict resolution and lack of cohesion, as well as parents’ COVID-19 stress were reported by parents. Linear mixed models for repeated measures and Bayes factors were used. ResultsNo significant changes were noted from before to after the onset of the COVID-19 pandemic for most hair steroids. However, a moderating role of family conflict resolution was noted. Children living with parents with a better ability to resolve conflicts had lower levels of DHEA compared to those who had more difficulty managing conflicts. Additionally, lower levels of family cohesion and income were linked to some steroids, especially DHEA, suggesting that these factors may relate to children’s physiological stress. Finally, boys had higher DHEA levels than girls. ConclusionOur findings suggest that stress biomarkers were comparable from before to during the COVID-19 pandemic. This observation holds true despite the pandemic being perceived by many as a novel, unpredictable, and potentially threatening event. Findings further suggest that family characteristics are associated with hair steroid, especially DHEA, which deserves further investigation.

Analysing and quantifying chronic stress-associated endogenous steroids in hair samples.

In previous studies, various steroids have been associated with stress and have therefore been quantified to investigate stress-related questions. Since the main stress-related steroid cortisol follows a circadian rhythm, often hair is analysed to quantify this steroid. Further, hair analysis gives the unique possibility of long-time monitoring by analysing a certain segment of hair, since hair grows on average 1 cm per month. Hair is a difficult matrix due to the complex sample preparation with many steps including washing and grinding, followed by various extraction steps. Additionally, steroids are endogenous and are therefore present in the hair matrix. Hence, no analyte free matrix is available, which is needed for the quantification via external calibrators. To overcome this problem, the so-called surrogate methods can be used, for which a 13 C3 labelled or deuterated reference compound of the steroid of interest is used for quantification. In the present study, a surrogate method was developed and fully validated for the quantitative analysis of seven steroids in human hair. Validation experiments showed that the method is further suitable for semi-quantitative analysis of estradiol. However, it is not suitable for the analysis of androsterone and DHEAS. The method was successfully used to analyse steroids in a comprehensive study of 360 adolescent hair samples, enabling research into stress markers.

Postpartum hair cortisol, dehydroepiandrosterone sulfate and their ratio in beef cows: Exploring association with parity and conception outcome

Hair steroid measurement has received increasing attention for monitoring hypothalamic-pituitary-adrenal axis function, as it offers the advantages of being noninvasive, fast, and able to indicate steroid concentrations over long periods. The objects of the study were to evaluate cortisol (C) and dehydroepiandrosterone sulfate (DHEA-S) hair concentrations and their ratio (C/DHEA-S) in beef cows from calving to 100 days (d) postpartum (pp) and to assess possible differences related to parity (primiparous vs multiparous) and conception outcome (pregnant vs not pregnant). Hair samples were collected from 6 primiparous and 5 multiparous pregnant beef cows by clipping the coat at calving (T0) and every 20 d for 5 times (T1-T5), collecting only the regrown hair. Starting from the 6th-week pp, cows were submitted to artificial insemination at spontaneous estrus; by 100 d pp, 7 cows were pregnant and 4 were not pregnant. Statistical analysis showed higher hair C concentrations in the 11 cows at calving (T0) compared to all the subsequent samplings except for T1, and higher C concentrations at T1 compared to T3, T4, and T5. These results indicate that hair C concentrations in beef cows are affected by sampling time, with a decrease from calving, as reported in other matrices. When exploring changes within parity groups, no differences were found in the multiparous among sampling times, while hair C concentrations at T0 and T1 tended to be higher than at T2 (0.01 ≤ p < 0.05) and were higher (p < 0.01) than in all the subsequent samplings (T3, T4 and T5) within the primiparous group. Higher hair C concentrations were found at T0 and T1 in the primiparous compared to multiparous (p < 0.01), suggesting that primiparous cows undergo a greater stress level before and around parturition compared to multiparous, probably due to the novelty of the calving experience. No differences were detected in C hair concentrations according to conception outcome (pregnant versus not pregnant) in each sampling time. Hair DHEA-S concentrations were neither affected by time nor by parity or conception outcome. Differences in the C/DHEA-S ratio were found at T1, with higher C/DHEA-S in the multiparous compared to primiparous cows (p < 0.001), and a tendency for higher ratio in the not pregnant compared to the pregnant (0.01 ≤ p < 0.05). These results support the choice of hair as a valuable biological matrix when investigating long-time periods such as postpartum in cows and suggest an enhanced immunoprotective effect of DHEA-S in the postpartum of primiparous cows, and in cows that get pregnant within 100 d postpartum.

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

ObjectiveEpidemiological 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. MethodData 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). ResultsLow, high, and continuous levels of THC in hair were robustly associated with higher levels of cortisol (sig. in step 1 low THC: β = 0.29, p = .021; high THC: β = 0.42, p = .001; step 2: low THC: β = 0.27, p = 0.036, and high THC: β = 0.40, p = .004, and step 4: β = 0.12, p = .041). Participants with high MDMA levels had higher levels of cortisone without adjusting for other substances (step 1: β = 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 (β = −0.35, p = .018), MDMA concentration was positively related to testosterone concentration with and without adjusting for other substances (step 3: β = 0.24, p = .041; step 4: β = 0.17, 95%, p = .015) in male participants. ConclusionThe use of psychoactive substances, especially of cannabis and ecstasy, should be considered in studies investigating steroid hormones in hair.

Psychological prediction of stress-related hair steroid hormone levels in young men: a person-centered approach

The aim was to: (1) identify individual profiles in young men regarding personality and cognitive appraisal style employing easy-to-use instruments, and (2) to explore how such profiles relate to biological stress indicators. The sample consisted of 173 male Lithuanian conscripts. An assessment was made after one month into their basic military training. Levels of cortisol, cortisone and testosterone were established through analysis of hair samples. Psychological assessments included the Big Five model of personality as well as cognitive appraisal style and perceived stress during the previous month of service. Four unique personality profiles were identified in a cluster analysis that differed significantly as theoretically expected on the Perceived Stress Scale. Statistically significant between-profile differences were found on cortisol and cortisone but not on testosterone. The personality profile with the highest scores on all three biological markers was characterized by high scores on Extraversion and Openness and low scores on Agreeableness. Second, three distinct cognitive appraisal style profiles emerged. They were related as theoretically expected to perceived stress, but they were unrelated to the hair steroid hormone concentration levels. Third, the combination of the most psychologically vulnerable personality profile, which included low scores on Emotional stability, and the most stressed cognitive appraisal style profile, yielded the clearest result and showed that it was possible to detect individuals with significantly higher stress-related hair steroid hormone levels using psychological instruments. Practical potential implications include identification of individuals who are most psychologically vulnerable and in need of close monitoring.

Incorporation of Testicular Ultrasonography and Hair Steroid Concentrations in Bull Breeding Soundness Evaluation.

Testicular ultrasonography and steroid concentrations (cortisol, dehydroepiandrosterone sulfate (DHEA-S), cortisol/DHEA-S ratio, testosterone) in hair were examined for their utility in the bull breeding soundness evaluation (BBSE). Beef and dairy bulls (n = 16; 2.7 ± 0.4 years old; body condition score 3.2 ± 0.1) of five breeds were maintained under the same conditions at an accredited semen collection center. Bulls underwent routine semen collection twice weekly for 12 weeks and semen was processed and cryopreserved. Ultrasonography and hair sampling were undertaken at the last semen collection. Bulls with homogeneous testicular parenchyma (n = 8) had a higher (p < 0.05) percentage of motile sperm post-thawing compared with bulls with heterogeneous parenchyma (n = 8). There were no differences (p > 0.05) in the hair concentrations of cortisol, DHEA-S, and testosterone between bulls with homogeneous and heterogeneous parenchyma. In bulls with homogeneous parenchyma, hair DHEA-S concentration was positively correlated with percentage motile sperm (R2 = 0.76), progressively motile sperm (R2 = 0.70), and motility yield (R2 = 0.71). The findings indicate that the integration of testicular ultrasonography and hair DHEA-S status in the BBSE could provide a more comprehensive assessment of indicative fertility in bulls. Additionally, ultrasonography can be used in the BBSE when the evaluation of semen parameters is not available.

Serum and hair steroid profiles in patients with nonfunctioning pituitary adenoma undergoing surgery: A prospective observational study

Patients who undergo transsphenoidal surgery (TSS) experience perioperative hormonal changes, but there are few studies on the perioperative changes of serum and hair steroid profiles. This study investigated the perioperative changes in steroid metabolic signatures in patients with nonfunctioning pituitary adenoma (NFPA) who underwent transsphenoidal surgery (TSS). A total of 55 participants who underwent TSS for NFPA at a single center between July 2017 and October 2018 were enrolled. Fifteen serum steroids and their metabolic ratios were profiled using gas chromatography-mass spectrometry (GC-MS) before and 1 day, 1 week, and 3 months after TSS. Five steroids from hair samples collected 1 day and 3 months after TSS were also quantitatively compared. Serum cortisol and its A-ring reductive metabolites, as well as 6β-hydroxycortisol, increased dramatically 1 day after TSS and then gradually decreased. Seven serum steroids, including adrenal androgens and mineralocorticoids, and hair cortisone levels were significantly lower in patients with preoperative adrenocorticotropic hormone (ACTH) deficiency (N = 7) than in those without ACTH deficiency (N = 48). Serum levels of dehydroepiandrosterone (DHEA) levels 1 week after TSS predicted ACTH deficiency 3 months after TSS, with 100 % sensitivity and 86 % specificity. A significant positive correlation between the preoperative serum and hair DHEA levels (r = 0.356, P = 0.008) was observed. These findings suggest that the levels of DHEA in both the serum and hair could be an early marker of ACTH deficiency after TSS. In addition, hair cortisone may be a useful preoperative indicator of chronic ACTH deficiency.

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 (βcortisol(log) = −0.29; βcortisone(log) = −0.28; βtestosterone(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 (β = −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.

Application of Felid Hair for Non-Invasive Tracking of Animal Reproductive Status and Adrenal Activity.

Simple SummaryHair can be a useful matrix to examine the hormonal status of an animal, although it is difficult to correlate the results to a specific time point. We measured seasonal changes in cortisol and testosterone levels in four feline species with different breeding strategies. Both hormones showed annual dynamic fluctuations, which coincided with the reproductive traits of the studied species. Seasonal increases in testosterone were observed in three seasonal breeders (lynx, Amur wildcat and domestic cat) but not in caracal, an aseasonal species, which sustained high testosterone levels over the whole year. Sexual differences in testosterone level were found for these species only during the mating season, when testosterone levels were higher in males than in females. Cortisol levels increased during the mating period, and in caracal an increase in cortisol was associated with the transfer of animals to smaller cages.Hair can be a useful matrix to examine the hormonal status of an animal, although it is difficult to correlate the results to a specific time point. The aim of this study was to evaluate seasonal changes in cortisol and testosterone levels in the hair of four feline species (lynx, Lynx lynx, n = 8; Amur wildcat, Prionailurus bengalensis euptilurus, n = 8; caracal, Caracal caracal, n = 6 and domestic cat, Felis catus, n = 17) with different breeding strategies. Animals of both sexes were sampled over the year, once per season (every three months), and the concentrations of hair testosterone and cortisol were measured by EIA. Both hormones showed annual dynamic changes, which coincided with the reproductive seasonality of the studied species. Sexual differences in testosterone level were found only during the mating season (spring for lynx and Amur wildcats, spring–summer for domestic cats), when testosterone levels were higher in males than in females. Cortisol levels were higher in males than in females in domestic cats and Amur wildcat, but also only during the mating season. Seasonal increases in testosterone were observed in three seasonal breeders (lynx, Amur wildcat and domestic cat) but not in caracal, which had high testosterone levels over the whole year. In lynx and Amur wildcat, it decreased sharply in the summer. Cortisol levels increased during the mating period in domestic cat males and lynx of both sexes; in caracal, an increase in cortisol was related to the transfer of animals to smaller winter cages. Measurements of steroids in hair can provide a reliable method for evaluating the reproductive status and the activity of the hypothalamus–pituitary–adrenal axis in several felid species.

Association between hair steroid hormone levels and cardiometabolic risk factors in a group of women participating in the national CVD prevention program: A cross-sectional study

Background and Aims : Cardiovascular disease (CVD) is the major cause of death worldwide. Although women develop CVD later than men, women’s morbidity and mortality from CVD are similar compared with men. Such a high prevalence of CVD cannot be explained completely by conventional risk factors. Recent studies indicate that chronic stress may be an independent CVD risk factor. Thus, we aimed to investigate the relationship between the biomarkers of chronic stress (hair cortisol, cortisone, DHEA levels) and traditional cardiovascular risk factors, as well as Systematic COronary Risk Evaluation (SCORE2) index, in a group of women (50-64 years) participating in the national CVD prevention program.

Validation and measurement of physiological stress and reproductive hormones in wolf hair and claws

AbstractThe use of keratinized tissues (e.g., hair, claws) to investigate physiological effects of environmental and anthropogenic stressors in free‐ranging wildlife populations has increased because these tissues retain steroid hormones during growth and are relatively easy to collect and store in the field. We measured reproductive and stress‐related steroid hormones in wolves (Canis lupus ligoni; n = 31) captured on Prince of Wales Island, Alaska, USA, during 1993–1994 and 2012–2014, representing periods of time when both wolf harvest and densities ranged from high to moderate. We validated enzyme immunoassay kits to measure steroid hormone concentrations in wolf guard hair, undercoat hair, and claw tip samples. Progesterone, testosterone, and cortisol were extracted and measured in the 3 keratinous tissues from wolves of different age class, sex, residency status, and collection periods. Within each tissue type, progesterone and testosterone were positively correlated (guard hair, r = 0.59, P = 0.003; undercoat hair, r = 0.55, P = 0.011; claws, r = 0.62, P ≤ 0.001) and cortisol concentrations were not related to either reproductive hormone. We were able to measure hormone concentrations in archived keratinous tissues collected up to 25 years earlier to assess stress and reproductive activity in historical samples. Our study validates a method for measuring steroid hormones in hair, and for the first time, continuously growing claws in wolves. Measurement of hormone concentrations in keratinous tissues may aid in the assessment of reproductive activity and physiological stress responses in wolf populations over long‐term time periods (i.e., decades) to enhance conservation efforts of an important apex predator.

Measuring steroids in hair opens up possibilities to identify congenital adrenal hyperplasia in developing countries.

Patients with congenital adrenal hyperplasia (CAH) in developing countries have limited access to appropriate laboratory facilities for diagnosis and follow-up. The aim of this study is to evaluate steroid measurement in hair as a diagnostic tool to identify and monitor CAH in these patients. A method was developed to measure steroids in hair, the stability of steroids in hair was assessed, and the concentration range in healthy volunteers was determined. Hair samples of patients, before and after starting therapy, were transported at ambient temperature to The Netherlands for analysis. Twenty-two Indonesian CAH patients and 84 healthy volunteers participated. Cortisol, 17-hydroxyprogesterone (17OHP), androstenedione, and testosterone in hair were measured by liquid chromatography with tandem mass spectrometry. Steroids in hair could be measured and remained stable (<4.9% deviation) for at least 3 weeks at 4°C and 30°C. In each of the untreated patients, hair concentrations of 17OHP (9.43-1135 pmol/g), androstenedione (36.1-432 pmol/g), and testosterone (2.85-69.2 pmol/g) were all above the upper limit of the corresponding range in healthy volunteers; 5.5 pmol/g, 13 pmol/g, and 1.8 pmol/g, respectively. After starting glucocorticoid treatment, the steroid concentrations in the hair of CAH patients decreased significantly for androstenedione (73%) and testosterone (59%) after 6 months. CAH could be confirmed in Indonesian patients based on the concentration of 17OHP, androstenedione, and testosterone in hair, and a treatment effect was observed. These findings open up opportunities to diagnose and/or monitor CAH in developing countries with a simple noninvasive technique.

A quantitative ultra-performance liquid chromatography high-resolution mass spectrometry analysis of steroids from human scalp hair

Hair steroid analysis offers the possibility to evaluate long-term steroid metabolism. It is especially beneficial when studying the steroid milieu in newborns and children, for example it can provide new insights into steroid metabolism over months and is unaffected by diurnal fluctuations in steroid concentrations. This study describes a quantitative and highly selective high-resolution mass spectrometry method for the simultaneous analysis of multiple steroids from human scalp hair. The developed method utilizes parallel reaction monitoring in the analysis of hydroxylamine derivatized steroids from hair samples first washed with isopropanol, extracted with methanol, and further purified with solid-phase extraction and liquid-liquid extraction. The method was proven suitable for the quantitative analysis of endogenous glucocorticoids (cortisol [1.5–364 pg/mg]; cortisone [3.6–355 pg/mg]; corticosterone [0.7–347 pg/mg]; 11-deoxycortisol [0.1–343 pg/mg]), androgens (testosterone [0.1–288 pg/mg]; dehydroepiandrosterone [0.3–288 pg/mg]; androstenedione [0.1–285 pg/mg]; 11β-hydroxyandrostenedione [0.3–304 pg/mg]), progestogens (pregnenolone [0.1–313 pg/mg]; progesterone [0.1–313 pg/mg]; 17α-hydroxypregnenolone [0.3–315 pg/mg]; 17α-hydroxyprogesterone [0.7–330 pg/mg]; 21-hydroxyprogesterone [0.6–320 pg/mg]), and estrogens (estrone [0.1–267 pg/mg]; estradiol [0.5–274 pg/mg]). In addition, 11-ketoandrostenedione [0.6–60 pg/mg]; dihydrotestosterone [0.3–290 pg/mg]; 11-ketotestosterone [0.1–12 pg/mg]; and 5α-androstanedione [0.6–281 pg/mg] could be analyzed semi-quantitatively. Aldosterone [3.5–346 pg/mg]; 11β-hydroxytestosterone [0.3–300 pg/mg]; and 11-ketodihydrotestosterone [0.3–300 pg/mg] were also included in the method, but their concentrations were below the lower limit of quantification in all tested hair samples. The method was applied for the analysis of authentic hair samples from different age groups ranging from newborns to adults, including mothers within 48 h after delivery. The newborn hair samples displayed the widest variety and had also the highest amounts of steroids in comparison to the samples of the other groups.

Higher Levels of Stress-Related Hair Steroid Hormones Are Associated with the Increased SCORE2 Risk Prediction Algorithm in Apparently Healthy Women.

Cardiovascular diseases (CVDs) are the major cause of death worldwide. Although the importance of conventional CVD risk factors, including older age, male gender, hypertension, obesity, dyslipidemia and hyperglycemia, is well-studied, psychosocial stress, which is considered an independent CVD risk factor, requires further investigation. Thus, we aimed to investigate the association between long-term secretion of stress-related steroid hormones, including cortisol, cortisone and dehydroepiandrosterone, and the 10-year fatal and non-fatal CVD risk estimated by the SCORE2 risk prediction algorithm, as well as traditional CVD risk factors in a group of apparently healthy women. A total of 145 women (aged 50–64 years) participating in the national CVD prevention program were enrolled in the study. Sociodemographic, lifestyle, health-related characteristics, stress, anxiety and sleep quality indicators were evaluated using specific questionnaires. Anthropometric and arterial blood pressure measures were assessed by trained personnel, lipid and glucose metabolism biomarkers were measured using routine methods, and hair steroid hormone levels were determined by ultra-high-performance liquid chromatography-tandem mass spectrometry. The results showed that higher levels of hair cortisol and cortisone are associated with increased SCORE2 values. Moreover, significant associations between hair glucocorticoids and individual cardiovascular risk factors, including obesity, hypertension, dyslipidemia and hyperglycemia, were found. These findings indicate that stress-related hair steroid hormones might be valuable biomarkers for CVD prediction and prevention.

Three-Faceted Approach to Perceived Stress: A Longitudinal Study of Stress Hormones, Personality, and Group Cohesion in the Real-Life Setting of Compulsory Basic Military Training

Compulsory basic military training is characterized not only by being challenging but also by being stressful. Assuming a high frequency of perceived stress events as a negative outcome of training, this article provides evidence on how the perceived frequency of stressful situations is affected by three types of factors: (i) biological stress response variables measured by hair steroid hormone levels, (ii) personality traits measured using the Big Five personality test, and (iii) group cohesion measures in military squads. A total of 112 conscripts in 11 squads participated in the research at the beginning (T1), in the middle (T2), and at the end (T3) of compulsory basic military training. Hair steroid hormone levels (cortisol, cortisone, and dehydroepiandrosterone (DHEA)) levels were measured by liquid mass spectrometry; other data were collected using self-report questionnaires. The results of the Poisson regression analysis indicated that hair steroid hormone cortisol had a statistically significantly impact and could increase the perceived frequency of stressful situations by up to 1.317 (e0.275, T2) times. The concentrations of other hormones (cortisone = 1.157, e0.146, T3 and DHEA = 1.020, e0.020, T3) also had a statistically significant effect. Other factors had a decreasing effect on the frequency. Extraversion was significant with an effect of 0.907 (e−0.098, T2) and 0.847 (e−0.166, T3), while task cohesion had an effect of 0.946 (e−0.056) and norm cohesion of 0.954 (e−0.047). The research indicates that the three groups of factors affect the perceived frequency of stressful situations during compulsory basic military training, but their impacts are considerably different.

Stability and validity of steroid hormones in hair and saliva across two ovulatory cycles

Steroid hormones are often assessed via saliva samples, as they are noninvasive and easy to collect. However, hormone levels in saliva can fluctuate from moment-to-moment, are influenced by factors such as momentary emotional states and food intake, and some vary strongly across women's ovulatory cycle. In contrast, hormone levels in hair seem to be more robust against these influences and were previously suggested to be a good alternative to obtain women's baseline hormone levels. In the current study, we investigated whether hormone levels are stable across multiple assays and whether hormone levels from saliva and hair samples correlate. We collected saliva and hair samples from N = 155 naturally cycling women across two ovulatory cycles. All samples were analyzed for progesterone, testosterone and cortisol levels via mass spectrometry (LC-MS/MS). Results showed that both averaged saliva and hair hormone levels were moderately stable across cycles. Hair progesterone levels showed higher stability than the respective levels from saliva. Saliva and hair levels for progesterone and testosterone were moderately correlated, whereas cortisol levels from saliva and hair were only weakly correlated. Results suggest that the type of sample from which baseline hormone levels are assessed and the cycle phase in which saliva samples are collected may have a high impact on the obtained results. Implications for future studies are suggested.

Steroid hormones in hair and fresh wounds reveal sex specific costs of reproductive engagement and reproductive success in wild house mice (Mus musculus domesticus)

Not only males but also females compete over reproduction. In a population of free-living house mice (Mus musculus domesticus), we analyzed how (metabolic) costs of aggressive interactions (reflected in fresh wounds and long-term corticosterone concentrations in hair) are predicted by individual reproductive physiology and reproductive success in males and females. Over eight years, we studied wounds and reproduction of more than 2800 adults under naturally varying environmental conditions and analyzed steroid hormones from more than 1000 hair samples. Hair corticosterone were higher and wounds more frequent in males than females. In males, wound occurrence increased with increasing breeding activity in the population, without affecting hair corticosterone levels. Unexpectedly, individual male reproductive success did not predict wounds, while hair corticosterone increased with increasing levels of hair testosterone and reproductive success. High corticosterone in hair of males might therefore reflect metabolic costs of fighting over reproduction. In females, hair corticosterone was generally lower than in males and high levels did not impede pregnancy. Reproductive investment (reflected in hair progesterone) was dissociated from reproductive success. Occasional wounds in females indicated individuals without recent reproductive success and revealed reproductive competition, presumably driven by instability in the social environment. In both sexes, corticosterone increased with age, but there was no evidence that received overt aggression, as indicated by wounds or elevated corticosterone, suppressed reproductive physiology. Our results diverge from laboratory findings and emphasize the need to also study animals in their natural environment in order to understand the complexity of their behavioral physiology.

Single sample preparation for the simultaneous extraction of drugs, pharmaceuticals, cannabinoids and endogenous steroids in hair.

Recently, we published a multi-analyte method for the simultaneous analysis of 116 drugs and pharmaceuticals including different substance groups like opioids, stimulants, benzodiazepines, z-drugs, antidepressants and neuroleptics based on a single sample workup followed by a single analytical measurement with LC-MS/MS. However, in some cases, additional analysis of further substance groups, such as cannabinoids and endogenous steroids, is required, which are analyzed in our laboratory using separate sample preparation and separate analytical methods. The goal of this study was to use the knowledge from the different sample preparations and combine them into a single sample preparation and extraction workflow for the simultaneous extraction of drugs, pharmaceuticals, cannabinoids, and endogenous steroids to be analyzed with the appropriate analytical methods. A partial validation of selected parameters such as selectivity, linearity, limit of quantification (LOQ), accuracy, precision and robustness for the different analytical methods was carried out and revalidated. In addition, comparative measurements of quality controls and authentic pools were performed and statistically evaluated using the unpaired t-test or the non-parametric Mann-Whitney test. The results using the newly established sample preparation and extraction were in good agreement with the original data. In conclusion, the newly established sample preparation is suitable for the combined extraction of drugs, pharmaceuticals, cannabinoids and endogenous steroids, and gives reliable results for quantification of various substances.

The Association between Endogenous Hair Steroid Hormones and Social Environmental Factors in a Group of Conscripts during Basic Military Training.

This study aimed to analyse the association between endogenous hair steroid hormones as reliable biological indicators of an individual’s stress level and the social environmental factors experienced during military training that are manifested at the beginning of compulsory military service. Hair steroid hormone concentrations—cortisol, cortisone, dehydroepiandrosterone (DHEA), and testosterone—in a group of 185 conscripts were measured using the ultra-high performance liquid chromatography-tandem mass spectrometry method. Six subjective social environmental factors in the military—attitude towards the military and military service, adaptation to the military environment, team, task, and norm cohesion, as well as psychological (un)safety in the group—were evaluated using military-specific research questionnaires. Weak but significant negative correlations were identified between cortisol and adaptation (r = −0.176, p < 0.05), attitude (r = −0.147, p < 0.05) as well as between testosterone and task cohesion (r = −0.230, p < 0.01) levels. Additionally, a multiple forward stepwise regression analysis highlighted that cortisone variation might be partially explained by task cohesion; the DHEA—determined by psychological (un)safety in the group, attitude towards the military and military service, and norm cohesion; and the testosterone—determined by task cohesion and adaptation to the new military environment. The results of this study suggest that subjective measures of social factors can be used to predict hair steroid hormone levels as objective measures of the chronic stress perceived by conscripts during their basic military training.

Analysis of hair steroid hormone concentrations at different parts of the head by liquid chromatography-tandem mass spectrometry

BackgroundSteroid hormones are known to be associated with diseases like androgenetic alopecia (AGA) resulting in hair loss. The lack of a detailed analysis of the local concentration of steroids in different parts of the head underlies the rationale and purpose of this study. MethodsTo evaluate the concentration distributions of steroid hormones in hair in different parts of the head, hair samples of 8 healthy men from 9 point-areas covering the frontal, parietal, and occipital regions were collected. Eight steroid hormones were measured by using the LC-MS/MS and region-wise comparison for different hormones was done using the mean z-score and Tukey’s HSD. ResultsFive of the 8 hormones had a high concentration in the parietal region, with dihydrotestosterone (DHT) showing a peak in the central parietal region (z = 1.59) suggesting a correlation with AGA’s clinical presentation. Whereas, no significant differences were observed for testosterone and cortisol between the parietal and occipital regions. Higher DHT levels at the parietal region were also verified with a small group of AGA patients. ConclusionsThis research expands upon the role of steroid hormones in hair follicle tissue elucidating their relationship with disease, thus contributing to disease management.