Potential Predictor of Epigenetic Age Acceleration in Men: 2D:4D Finger Pattern.

OBJECTIVES: Second to fourth digit ratio is widely known indicator of prenatal sex hormones proportion. Higher prenatal androgenization results in longer fourth finger and lower 2D:4D index. The aim of this study was to determine whether the 2D:4D digit ratio is associated with DNA methylation (DNAm) age dependently on sex. MATERIAL AND METHODS: The study included 182 adults (106 females and 76 males) with a mean age of 51.5 ± 13 years. The investigation consisted of three main parts: a survey, anthropometric dimensions measurements (fingers length) and methylome analysis using collected blood samples. Genome-wide methylation was analyzed using EPIC microarray technology. Epigenetic age and epigenetic age acceleration were calculated using several widely applied algorithms. RESULTS: Males with the female left hand pattern had more accelerated epigenetic age than those with the male pattern as calculated with PhenoAge and DNAmTL clocks. CONCLUSIONS: Finger female pattern 2D:4D above or equal to 1 in males is associated with epigenetic age acceleration, indicating that prenatal exposure to estrogens in males may be related to aging process in the later ontogenesis.

Cortisol concentration affects fat and muscle mass among Polish children aged 6-13 years.

BACKGROUND: Cortisol is a steroid hormone acting as a stress hormone, which is crucial in regulating homeostasis. Previous studies have linked cortisol concentration to body mass and body composition. METHODS: The investigations were carried out in 2016-2017. A total of 176 children aged 6-13 years in primary schools in central Poland were investigated. Three types of measurements were performed: anthropometric (body weight and height, waist and hip circumferences), body composition (fat mass FM (%), muscle mass - MM (%), body cellular mass - BCM (%), total body water - TBW (%)), and cortisol concentration using saliva of the investigated individuals. Information about standard of living, type of feeding after birth, parental education and maternal trauma during pregnancy was obtained with questionnaires. RESULTS: The results of regression models after removing the environmental factors (parental education, standard of living, type of feeding after birth, and maternal trauma during pregnancy) indicate a statistically significant association between the cortisol concentration and fat mass and muscle mass. The cortisol concentration was negatively associated with FM (%) (Beta=-0.171; p = 0.026), explaining 2.32 % of the fat mass variability and positively associated with MM (%) (Beta = 0.192; p = 0.012) explaining 3.09 % of the muscle mass variability. CONCLUSIONS: Cortisol concentration affects fat and muscle mass among Polish children. TRIAL REGISTRATION: The Ethical Commission at the University of Lodz (nr 19/KBBN-UL/II/2016).

Prenatal oestrogen-testosterone balance as a risk factor of migraine in adults.

BACKGROUND: Migraine is a common neurological disease with extremely debilitating, but fully reversible symptoms. Women suffer from migraine more often than men. It was assumed that fluctuation of oestrogen level during menstrual cycle is one of many factors responsible for more frequent migraine attacks. The second-to-fourth digit ratio (2D:4D) is considered as an indicator of prenatal sex steroids. Balance of prenatal androgens (testosterone) and oestrogen has been studied in numerous diseases that are affected by hormones. However, the relationship between migraine and the sex steroids balance in prenatal development is still unexplained. The aim of this paper is to provide an evidence of relationship between prenatal oestrogen and testosterone exposure following 2D:4D digit ratio, and migraine prevalence in adults. METHODS: We examined a group of 151 adults (33 males, 118 females) with migraine and a control group of 111 adults (45 males, 66 females). 2D:4D digit ratio of both hands was measured using sliding Vernier calliper. RESULTS: Significant differences were found in the right hand. Female migraineurs had lower value of 2D:4D ratio than the control group and the right 2D:4D was lower than left 2D:4D (Δ2D:4D), suggesting prenatal testosterone dominance. The opposite relationship was observed in males. Male migraineurs had higher value of 2D:4D ratio and Δ2D:4D was greater than the control group, suggesting prenatal oestrogen dominance. CONCLUSIONS: Our results suggest that depending on sex, different proportion of prenatal sex steroids might be a risk factor of migraine in adults. Women with migraine were presumably exposed in prenatal life to higher testosterone levels relative to oestrogen, while men with migraine were probably exposed in prenatal life to higher levels of oestrogen relative to testosterone.

Association of saliva 25(OH)D concentration with body composition and proportion among pre-pubertal and pubertal Polish children.

OBJECTIVES: Due to increasing problems with obesity and vitamin D deficiency among children, studies that tackle both problems together are needed. METHODS: Data were collected from 182 randomly selected children aged 6-13 years in primary schools in central Poland. Measures included anthropometric dimensions, body composition, questionnaires completed by participants' parents, and saliva samples. The level of 25(OH)D was assessed from the saliva samples using an enzyme-linked immunosorbent assay kit. The children were divided into two groups: pre-pubertal (girls below 10 years and boys below 11 years) and pubertal individuals (girls above 10 years and boys above 11 years). RESULTS: The 25(OH)D concentrations were higher in late spring (June) among pre-pubertal children than in the autumn (November-December) among pubertal children. The level of 25(OH)D was positively correlated with body cell mass (BCM,%) among all children (pubertal: R = 0.20, P = .044; pre-pubertal: R = 0.23, P = .041) and inversely associated with waist-to-hip ratio (WHR) among pubertal children of both sexes (R = -0.25; P = .031). The stepwise regression analysis revealed that investigation in spring (June) and breastfeeding was associated with increased muscle mass (MM, %) (beta = 0.253, P = .003 and beta = 0.225, P = .005, respectively) and total body water (TBW, %) (beta = 0.276, P = .004 and beta = 0.246, P = .011, respectively) and was associated with decreased body mass index (BMI; beta = -0.222, P = .024 and beta = -0.269, P = .009, respectively) and fat mass (%) (beta = -0.288, P = .003 and beta = -0.266, P = .005, respectively). CONCLUSIONS: Season of salivary sampling and breastfeeding status were more strongly associated with body components, BMI and WHR, than 25(OH)D concentrations.

Do skin and hair pigmentation in prepubertal and early pubertal stages correlate with 2D:4D?

OBJECTIVES: In most human populations women are less pigmented than men. However, before puberty, boys tend to have lighter skin than girls. Previous research has shown that skin pigmentation in European women is negatively correlated with digit ratio (2D:4D). Therefore, it has been hypothesized that skin pigmentation in females is partially controlled by prenatal hormones. The aim of this study was to examine the correlation between 2D:4D and pigmentation in girls and boys in the prepubertal or early pubertal stages of ontogenesis. METHODS: The study group consisted of 598 Polish children (275 boys), aged 6.5-10.5 years. All underwent spectrophotometric measurements of skin and hair melanin index (SMI and HMI) and had the second and the fourth finger on both hands measured directly. RESULTS: SMI (in girls) and HMI (in girls and boys) were positively correlated with age but 2D:4D was not. We regressed SMI and HMI on age and used the residuals as a measure of age-independent pigmentation. In girls, positive correlations were found between resSMI and 2D:4D for the right hand, but there was no correlation with left 2D:4D. In boys, resSMI did not correlate with 2D:4D. The resHMI was not related to 2D:4D in girls or boys. CONCLUSION: Skin pigmentation in prepubertal or early pubertal girls is related to 2D:4D such that high 2D:4D is associated with higher pigmentation. This relationship may be mediated by the individual rate of sexual maturation.

FURTHER EVIDENCE OF AN ASSOCIATION BETWEEN LOW SECOND-TO-FOURTH DIGIT RATIO (2D:4D) AND SELECTION FOR THE UNIFORMED SERVICES: A STUDY AMONG POLICE PERSONNEL IN WROCLAW, POLAND.

Males and females differ in their preference for occupations and sporting activities, and differ also in risk-taking attitudes. In addition to other explanations, prenatal hormone exposure has been implicated in these gender-associated differences. The ratio of the relative lengths of the second-to-fourth digits (2D:4D) is a proxy indicator of prenatal exposure to testosterone relative to oestrogen. The 2D:4D ratio has been found to be associated with choice of occupation, particularly among females. This study investigated whether 2D:4D differed between police officers and a control group of civilians in Wroclaw, Poland. Participants were 147 male and 55 female police officers and 91 male and 75 female civilian controls. The police officers had to undergo rigorous physical ability tests during recruitment and their job bore relatively higher risk, whereas the controls had a normal civilian lifestyle. Height, weight, hand grip strength and lengths of the second and fourth digits were measured. Analyses of variance and covariance were employed to assess the significance of difference in digit ratio between groups (police officers and civilians) allowing for interaction with sex. The policewomen, compared with the female controls, were taller and had stronger hand grip strength, but had lower 2D:4D in the right hand and average 2D:4D of both hands. However, male and female police officers slightly differed only in the right hand digit ratio but not in the left hand ratio or the average for the two hands. However, the control group showed significant sex differences in all digit ratios with higher (feminine) mean values in females. The study provides further evidence that prenatal testosterone exposure, as reflected in the 2D:4D ratio, might have an association with choice of occupation, particularly among females.

Skin color parameters and Fitzpatrick phototypes in estimating the risk of skin cancer: A case-control study in the Polish population.

BACKGROUND: Light skin pigmentation is a known risk factor for skin cancer. OBJECTIVE: Skin color parameters and Fitzpatrick phototypes were evaluated in terms of their usefulness in predicting the risk of skin cancer. METHODS: A case-control study involved 133 individuals with skin cancer (100 with basal cell carcinoma, 21 with squamous cell carcinoma, 12 with melanoma) and 156 healthy individuals. All of them had skin phototype determined and spectrophotometric skin color measurements were done on the inner surfaces of their arms and on the buttock. Using those data, prediction models were built and subjected to 17-fold stratified cross-validation. RESULTS: A model, based on skin phototypes, was characterized by area under the receiver operating characteristic curve = 0.576 and exhibited a lower predictive power than the models, which were mostly based on spectrophotometric variables describing pigmentation levels. The best predictors of skin cancer were R coordinate of RGB color space (area under the receiver operating characteristic curve 0.687) and melanin index (area under the receiver operating characteristic curve 0.683) for skin on the buttock. LIMITATIONS: A small number of patients were studied. Models were not externally validated. CONCLUSIONS: Skin color parameters are more accurate predictors of skin cancer occurrence than skin phototypes. Spectrophotometry is a quick, easy, and affordable method offering relatively good predictive power.

Relationship between pre-natal factors, the perinatal environment, motor development in the first year of life and the timing of first deciduous tooth emergence.

BACKGROUND: The emergence of deciduous teeth, despite being genetically determined, shows significant correlation with the pre-natal environment, maternal factors, method of infant feeding and also family socioeconomic status. However, reported results are often contradictory and rarely concern healthy, full-term children. AIM: The objective of this study was to evaluate the influence of pre-natal and maternal factors as well as the method of infant feeding on the timing of first deciduous tooth emergence in healthy, full-term infants and to examine the relationship between the psychomotor development rate and the age at first tooth. SUBJECTS AND METHODS: The database contained 480 records for healthy, term-born children (272 boys and 208 girls born at 37-42 weeks of gestation) aged 9-54 months. Multiple regression analysis and multi-factor analysis of variance were used to identify significant explanatory variables for the age at first tooth. RESULTS: The onset of deciduous tooth emergence is negatively correlated with birth weight and maternal smoking during pregnancy and positively correlated with breastfeeding and the age at which the child begins to sit up unaided. These factors have an additive effect on the age at first tooth. CONCLUSION: An earlier onset of tooth emergence in children exposed to maternal smoking during pregnancy seems to provide further evidence for disturbed foetal development in a smoke-induced hypoxic environment.

Digit ratio and the femoral circumflex artery perforator: A sex-dependent association between the vascular system of the thigh and prenatal sex hormones?

BACKGROUND: The anterolateral thigh flap (ALTF) is a type of free flap which is commonly used in reconstructive surgery procedures and is based on the perforator from femoral circumflex artery. Some authors found that the location of the perforator differs between sexes. OBJECTIVE: To verify the influence of prenatal sex hormones (as measured by 2D:4D) on the morphology of the perforator of the descending branch of the femoral circumflex artery. METHODS: 88 participants (43 women) of a mean age of 28.5 years (SD 8.1 years) had the following measurements performed: height and weight; right and left-hand 2nd and 4th fingers' lengths; and distance from anterior superior iliac spine to the perforator from descending branch of lateral femoral circumflex artery related to the thigh length (right and left) (AP ratio). RESULTS: Right 2D:4D and Dr-l (right minus left 2D:4D) were dimorphic, similarly to AP ratio (right and left), which in women was significantly higher than in men. A significant correlation was found between right and left 2D:4D and location of the perforator of the left thigh in men. The higher right and left 2D:4D (more feminine) the higher the left AP ratio (more feminine). CONCLUSION: Our findings suggest that the location of perforator of femoral circumflex artery may be influenced by prenatal sex steroids such that low prenatal testosterone and high prenatal oestrogen are correlated to high AP (lower branching of the artery).

Do Intraoperative Platelet-Rich Plasma Injections Influence the Final Appearance of Vertical Scars after Breast Reduction? Spectrophotometric Analysis.

Background: Platelet-rich plasma (PRP) has been shown to support wound healing and tissue regeneration due to its high concentration of growth factors and cytokines. This study aims to investigate the effect of intraoperative PRP injections on the final appearance of vertical scars after breast reduction, as well as to identify potential predictors of a scar's aesthetic assessment using spectrophotometric parameters. Methods: In this prospective, randomized trial, 82 scars from 41 women who underwent bilateral breast reduction with an inverted T pattern were analyzed. PRP or a placebo substance (0.9% sodium chloride solution) was injected intraoperatively into the edges of vertical wounds. Spectrophotometric measurements of scar pigmentation were performed 3 and 6 months after surgery; additionally, two independent observers evaluated the aesthetic appearance of scars based on photographs. Results: The results showed that the use of intraoperative PRP injections did not significantly influence the final appearance of vertical scars after breast reduction. Conclusions: We indicated spectrophotometric variables (b) in the early stages of wound healing (after 3 months) that can be predictors of the final scar's aesthetic outcome. This can be helpful in detecting scars that may need additional interventions to optimize the healing process.

Mental Health, and Eating Disorders in Patients After Roux-en-Y Gastric Bypass Surgery (RYGB).

<b>Introduction:</b> Obesity, as one of the main health problems worldwide, is associated with an increased risk of developing mental and eating disorders and negative eating habits. Bariatric surgery allows for rapid weight loss and alleviates the symptoms of concomitant diseases in obese patients.<b>Aim:</b> Pre- and postoperative estimation of mental disorders and eating behaviors in patients after Roux-en-Y Gastric Bypass (RYGB).<b>Material and methods:</b> Analysis of data from up to 5 years of follow-up including clinical examination and questionnaires.<b>Results:</b> Following parameters decreased after RYGB: anxiety and hyperactivity from 32.81% to 21.88%, mood disorders - 31.25% to 20.31%, substance abuse - 40.63% to 28.13%, emotional eating - 76.56% to 29.69%, binge eating - 50% to 6.25%, night eating - 87.5% to 20.31%. Postoperative rates of: negative eating habits, daily intake of calories and sweetened beverages, flatulence, constipation, and abdominal pain decreased, while the rate of food intolerance and emesis increased.<b>Conclusions:</b> In our patients, the occurrence of: mental and eating disorders, negative eating habits, daily calories, sweetened beverages, coffee intake decreased after weight loss (as a result of RYGB), but water, vegetables and fruit consumption increased. Lower rate of flatulence, constipation, and abdominal pain, but higher of food intolerance and emesis were also confirmed after RYGB.

Analysis of epigenetic clocks links yoga, sleep, education, reduced meat intake, coffee, and a SOCS2 gene variant to slower epigenetic aging.

DNA methylation (DNAm) clocks hold promise for measuring biological age, useful for guiding clinical interventions and forensic identification. This study compared the commonly used DNAm clocks, using DNA methylation and SNP data generated from nearly 1000 human blood or buccal swab samples. We evaluated different preprocessing methods for age estimation, investigated the association of epigenetic age acceleration (EAA) with various lifestyle and sociodemographic factors, and undertook a series of novel genome-wide association analyses for different EAA measures to find associated genetic variants. Our results highlighted the Skin&Blood clock with ssNoob normalization as the most accurate predictor of chronological age. We provided novel evidence for an association between the practice of yoga and a reduction in the pace of aging (DunedinPACE). Increased sleep and physical activity were associated with lower mortality risk score (MRS) in our dataset. University degree, vegetable consumption, and coffee intake were associated with reduced levels of epigenetic aging, whereas smoking, higher BMI, meat consumption, and manual occupation correlated well with faster epigenetic aging, with FitAge, GrimAge, and DunedinPACE clocks showing the most robust associations. In addition, we found a novel association signal for SOCS2 rs73218878 (p = 2.87 × 10-8) and accelerated GrimAge. Our study emphasizes the importance of an optimized DNAm analysis workflow for accurate estimation of epigenetic age, which may influence downstream analyses. The results support the influence of genetic background on EAA. The associated SOCS2 is a member of the suppressor of cytokine signaling family known for its role in human longevity. The reported association between various risk factors and EAA has practical implications for the development of health programs to improve quality of life and reduce premature mortality associated with age-related diseases.

Correlations between antimicrobial peptides and spectrophotometric skin color parameters in patients with basal cell carcinoma.

BACKGROUND: Antimicrobial peptides (AMPs) are active molecules in the human innate immune system, that participate in host defense and regulate the inflammation process. Previous reports have confirmed that antimicrobial peptides play a critical role in carcinogenesis. OBJECTIVE: The present study aimed to evaluate the correlations between plasma concentrations of AMPs and spectrophotometric parameters of skin color in patients with basal cell carcinoma and compare the results with those of healthy controls. METHODS: The plasma concentrations of cathelicidin and beta-defensin-2 in 100 patients (50 with skin cancer and 50 healthy control subjects) were measured, and skin color parameters were tested using a DermaSpectrophotometer. RESULTS: In patients with basal cell carcinoma, the concentrations of cathelicidin and beta-defensin-2 were significantly higher than those in healthy controls. In healthy control patients, when erythema increases, the levels of cathelicidin and beta-defensin-2 also increase. The direction of the relationship is opposite in people with basal cell carcinoma-the concentration of antimicrobial peptides decreases and the level of erythema increases. CONCLUSION: A significantly higher level of plasma concentrations of cathelicidin and HBD-2 are correspondent to the presence of basal cell carcinoma. Skin cancer modifies the relationship between intensity of skin erythema and the levels of cathelicidin and HBD-2. This can be related to inadequate immunological response in patients with skin cancers. New direction of research may be pioneered in searching for cytokine or mast cells disorders.

Variability of anterolateral thigh flap perforator locations - clinical implications.

<b><br>Introduction:</b> The anterolateral thigh flap is an example of a free flap widely applied for reconstruction of tissues within various areas of the body.</br> <b><br>Aim:</b> The aim of the study was to determine the most common locations of the ALT flap perforators in men and women as well as to clarify any potential differences in the location of the vessel in male vs. female subjects.</br> <b><br>Material and method:</b> In years 2021-2022, a study was conducted at the Plastic, Reconstructive, and Aesthetic Surgery Clinic in a group of 90 volunteers (45 women and 45 men, a total of 180 lower extremities) to determine the location of the anterolateral thigh flap perforator against the flap surface using a portable Doppler probe.</br> <b><br>Results:</b> Variable locations were revealed for the anterolateral thigh flap perforator within the study. In women, the perforator is located more distally from the anterior superior iliac spine, most frequently within the inferolateral quadrant, while being located closer to the anterior superior iliac spine, and most frequently within the superolateral quadrant, in men. No perforator was found in 3.3% of the examined limbs. No statistically significant differences were observed between the course of the perforators within the left and right lower limbs in individual patients.</br> <b><br>Conclusions:</b> The location of the ALT flap perforator is subject to sexual dimorphism, which facilitates the investigation area being narrowed down to the well-defined locations of its most frequent occurrence when planning anterolateral thigh flap reconstructive procedures.</br>.

Selected factors affecting the rate of reduction of body weight components during the first six months after bariatric surgery: A cohort study.

<b><br>Aim:</b> To determine whether the rate of slimming following bariatric surgery depends on the sex, type of bariatric surgery, time elapsed since surgery or body weight components.</br> <b><br>Materials and methods:</b> The material is comprised of the results of three series of anthropometric measurements in 91 obese patients (before bariatric surgery, about 3 months after bariatric surgery and about 6 months afterwards). The inclusion criteria were patients of Polish origin admitted to the hospital from July 1, 2017 to January 31, 2019 for surgical treatment of obesity and written consent for the surgery and participation in the study. The measurements included body weight components assessed by bioelectrical impedance analysis.</br> <b><br>Results:</b> The reduction of all features was greater after SG than GB (p<0.0001), greater in the first quartile than in the second quartile after surgery (p<0.0001), and higher for fat mass than for other body weight components (p<0.0001). The sex of the patients, type of bariatric surgery, time elapsed since the operation, and type of body weight component constituted interactive modifiers of the rate of reduction.</br> <b><br>Conclusion:</b> After bariatric surgery, the reduction of fat mass was quicker in men than in women. SG and RYGB lead to a greater reduction of fat mass than GB. Among all three analyzed procedures, only for SG did the rate of body weight component reduction not decrease in the second quartile after surgery. This finding should be taken into account when creating an algorithm for treating a patient after bariatric surgery</br>.

The impact of exposure to tobacco smoking and maternal trauma in fetal life on risk of migraine.

Introduction: Prenatal period is the key time in human development. Many prenatal factors are well-known and increase the risk of developing diseases' after birth. Few studies indicated the link between the prenatal period and the prevalence of migraine in childhood and adolescence so far. We decided to broaden current knowledge and investigate whether the prenatal factors influence the prevalence of migraine in adulthood. The objective of this study is to provide evidence of relationship between in utero environment and risk of migraine. Methods: In total 266 females (136 in the migraine group, 130 in the control group) and 80 males (35 in the migraine group, 45 in the control group), aged 18-65 participated in the study. The quality of prenatal environment was characterized on the basis of mother's and father's education, tobacco smoke exposure, alcohol consumption, and traumatic event during pregnancy, which are considered as prenatal factors and affect on fetal development. Results: Migraine occurrence in adulthood was significantly associated with maternal tobacco smoking during pregnancy (OR 3.42, 95% CI 1.54-7.61, p = 0.036) and traumatic event during pregnancy (OR 2.27, 95% CI 1.24-4.13, p = 0.020). Discussion: Our study suggests that the fetal programming effect of tobacco smoking exposure and maternal trauma is not limited to prenatal life and is suggested as having a role in adulthood. Our findings support evidence that migraine adulthood can be partly influenced by early life conditions.

Digit ratio (2D:4D), laryngeal cancer and vocal fold leukoplakia.

BACKGROUND: To date, there are no studies that have analyzed the possible influence of exposure to prenatal sex hormones on the risk of laryngeal cancer (LC) and premalignant laryngeal lesion-vocal fold leukoplakia (VFL). Digit ratio (2D:4D) is suggested to be a proxy of prenatal sex hormone exposure. OBJECTIVE: To examine 2D:4D in patients with LC and clarify if it could add to the verified risk factors in estimating the overall risk of LC. METHODS: 511 subjects participated in the study. The study group included 269 patients: with LC (N = 114, 64 men) and VFL (N = 155, 116 men). Controls included 242 healthy individuals (66.40 ± 4.50 years (106 men)). RESULTS: Predictive models estimating the risk of VFL and LC in women, based solely on predictors like smoking and alcohol consumption had a lower area under the ROC curve (AUC) than the model with left 2D:4D. AUC for the model estimating the likelihood of VFL increased from 0.83 to 0.85, and for LC from 0.76 to 0.79. CONCLUSIONS: Low left 2D:4D may be associated with an increased risk of developing leukoplakia and laryngeal cancer in women. In the case of laryngeal cancer, left 2D:4D may serve as additional variable (to other known risk factors, such as smoking and/or alcohol consumption), which can improve cancer risk prediction.

Introduction of a multiplex amplicon sequencing assay to quantify DNA methylation in target cytosine markers underlying four selected epigenetic clocks.

BACKGROUND: DNA methylation analysis has proven to be a powerful tool for age assessment. However, the implementation of epigenetic age prediction in diagnostics or routine forensic casework requires appropriate laboratory methods. In this study, we aimed to compare the performance of large-scale DNA methylation analysis protocols that show promise in terms of accuracy, throughput, multiplexing capacity, and high sensitivity. RESULTS: The protocols were designed to target a predefined panel of 161 genomic CG/CA sites from four known estimators of epigenetic age-related parameters, optimized and validated using artificially methylated controls or blood samples. We successfully targeted 96% of these loci using two enrichment protocols: Ion AmpliSeq™, an amplicon-based method integrated with Ion Torrent S5, and SureSelectXT Methyl-Seq, a hybridization-based method followed by MiSeq FGx sequencing. Both protocols demonstrated high accuracy and robustness. Although hybridization assays have greater multiplexing capabilities, the best overall performance was observed for the amplicon-based protocol with the lowest variability in DNA methylation at 25 ng of starting DNA, mean observed marker coverage of ~ 6.7 k reads, and accuracy of methylation quantification with a mean absolute difference between observed and expected methylation beta value of 0.054. The Ion AmpliSeq method correlated strongly with genome-scale EPIC microarray data (R = 0.91) and showed superiority in terms of methylation measurement accuracy. Method-to-method bias was accounted for by the use of linear transformation, which provided a highly accurate prediction of calendar age with a mean absolute error of less than 5 years for the VISAGE and Hannum age clocks used. The pace of aging (PoAm) and the mortality risk score (MRS) estimators included in our panel represent next-generation clocks, were found to have low to moderate correlations with the VISAGE and Hannum models (R < 0.75), and thus may capture different aspects of epigenetic aging. CONCLUSIONS: We propose a laboratory tool that allows the quantification of DNA methylation in cytosines underlying four different clocks, thus providing broad information on epigenetic aging while maintaining a reasonable number of CpG markers, opening the way to a wide range of applications in forensics, medicine, and healthcare.

DNAmFitAge: biological age indicator incorporating physical fitness.

Physical fitness is a well-known correlate of health and the aging process and DNA methylation (DNAm) data can capture aging via epigenetic clocks. However, current epigenetic clocks did not yet use measures of mobility, strength, lung, or endurance fitness in their construction. We develop blood-based DNAm biomarkers for fitness parameters gait speed (walking speed), maximum handgrip strength, forced expiratory volume in one second (FEV1), and maximal oxygen uptake (VO2max) which have modest correlation with fitness parameters in five large-scale validation datasets (average r between 0.16-0.48). We then use these DNAm fitness parameter biomarkers with DNAmGrimAge, a DNAm mortality risk estimate, to construct DNAmFitAge, a new biological age indicator that incorporates physical fitness. DNAmFitAge is associated with low-intermediate physical activity levels across validation datasets (p = 6.4E-13), and younger/fitter DNAmFitAge corresponds to stronger DNAm fitness parameters in both males and females. DNAmFitAge is lower (p = 0.046) and DNAmVO2max is higher (p = 0.023) in male body builders compared to controls. Physically fit people have a younger DNAmFitAge and experience better age-related outcomes: lower mortality risk (p = 7.2E-51), coronary heart disease risk (p = 2.6E-8), and increased disease-free status (p = 1.1E-7). These new DNAm biomarkers provide researchers a new method to incorporate physical fitness into epigenetic clocks.

Biometric characteristics of the pelvis in female-to-male transsexuals.

The objective of the study was to evaluate the metric features of pelvises of 24 female-to-male (FtM) transsexuals as compared to control groups of 24 healthy males and 24 healthy females. The participants had their pelvises X-rayed with the same X-ray apparatus and in the same position. Seventeen measurements were taken on the basis of X-ray pictures of FtM transsexuals' pelvises and both comparison groups. Additionally, their body height was compared. The results showed that FtM transsexuals having female body height represent an intermediate size of three pelvic features and male values of five variables. In order to develop a model based on metric variables of the pelvis that would best discriminate the FtM transsexuals, the control females, and the control males, a discriminant analysis was applied. The model included four variables out of 17 metric features: the height of the pubic symphysis, the greatest pelvic breadth, the interischial distance, and the acetabular diameter. The model was found to be the best in discriminating males from females and FtM transsexuals, but considerably less effective in discriminating transsexuals from the two control groups. The results demonstrate that a number of FtM transsexuals' pelvic measurements reveal "masculinization," which confirms current results demonstrating a shift in the somatometric traits of transsexual females towards male traits. A discriminant analysis based only on pelvic metric features shows some differences between the size of the pelvis and chromosomal sex in FtM transsexuals, which might indicate a biological basis for gender identity disorder.