Gene co-expression patterns in Atlantic salmon adipose tissue provide a molecular link among seasonal changes, energy balance and age at maturity.

Sexual maturation in many fishes requires a major physiological change that involves a rapid transition between energy storage and usage. In Atlantic salmon, this transition for the initiation of maturation is tightly controlled by seasonality and requires a high-energy status. Lipid metabolism is at the heart of this transition since lipids are the main energy storing molecules. The balance between lipogenesis (lipid accumulation) and lipolysis (lipid use) determines energy status transitions. A genomic region containing a transcription co-factor of the Hippo pathway, vgll3, is the main determinant of maturation timing in Atlantic salmon. Interestingly, vgll3 acts as an inhibitor of adipogenesis in mice and its genotypes are potentially associated with seasonal heterochrony in lipid storage and usage in juvenile Atlantic salmon. Here, we explored changes in expression of more than 300 genes directly involved in the processes of adipogenesis, lipogenesis and lipolysis, as well as the Hippo pathway in the adipose tissue of immature and mature Atlantic salmon with distinct vgll3 genotypes. We found molecular evidence consistent with a scenario in which immature males with different vgll3 genotypes exhibit contrasting seasonal dynamics in their lipid profiles. We also identified components of the Hippo signalling pathway as potential major drivers of vgll3 genotype-specific differences in adipose tissue gene expression. This study demonstrates the importance of adipose gene expression patterns for directly linking environmental changes with energy balance and age at maturity through genetic factors bridging lipid metabolism, seasonality and sexual maturation.

A cretaceous frog with eggs from northwestern China provides fossil evidence for sexual maturity preceding skeletal maturity in anurans.

Mesozoic fossils of frogs are rare in the palaeontological record, particularly those exhibiting soft tissues that offer limited insights into early life-history characteristics. Here we report on a skeletally immature frog from the Lower Cretaceous of northwest China, with egg masses in the body and eggs in the oviduct, indicative of a gravid female. CT reconstruction of the specimen allows referral to Gansubatrachus qilianensis and we assign it as a paratype complementing the diagnosis of the type species. The new fossil, which might represent a younger individual than the holotype of Gansubatrachus, shows that sexual maturation occurred before full adulthood in this frog and provides evidence of death linked to mating behaviour. We also discuss other potential sources of variation and life-history traits of Gansubatrachus. The new finding represents the oldest Early Cretaceous frog preserving in situ eggs and provides a glimpse into ancient anuran development during Mesozoic times.

Validity and reliability of parent assessments of pubertal maturation among adolescent girls in Isfahan, Iran.

OBJECTIVES: The current paper presents the steps considered for validation of a questionnaire for assessment of sexual maturity among Iranian adolescent girls. METHODS: This cross-sectional study was performed in 2022 in Isfahan, Iran. Based on the Growth and Development Questionnaire that included both the Pubertal Development Scale (PDS) and Sexual Maturation Scale (SMS), two Persian questionnaires were prepared. The face validity, content validity, criterion validity, and reliability of the questionnaire were assessed. We compared agreement of two parent-reported measures of puberty, SMS and PDS, with clinical Tanner stages (TSs) as the gold standard. Percent agreement, Cohen's kappa coefficient, and Kendall's τ b were used to assess the agreement between maternal assessments with clinical TS. The intraclass correlation coefficient (ICC) and the Cronbach's α coefficient were also calculated to evaluate the reliability of the questionnaire. RESULTS: A total of 150 students aged 6-17 years with mean (SD) age of 10 (2.04) completed this study. The percentages of agreement for the mother-reported SMS in relation to clinical TS for breast stage and pubic hair stage were 60 % and 53.8 %, respectively. The percentages of agreement of the mother-reported PDS in relation to clinical TS for breast stage and pubic hair stage were 55.8 and 66 %, respectively. The weighted kappa coefficients showed moderate agreement, with weighted kappa ranging from 0.52 to 0.61. The mother-reported SMS and PDS showed high reliability. The Cronbach's alpha of the PDS and the SMS was 0.88 and 0.83, respectively. The ICC of the mother-reported SMS and the mother-reported PDS was 0.95 (0.92-0.98) and 0.97 (0.94-0.98), respectively. CONCLUSIONS: This study indicated that a maternal assessment of sexual maturity using the PDS or SMS can reliably estimate pubertal development in adolescent girls in an Iranian population.

Characteristics of Puberty in a Population-Based Sample of Danish Adolescents.

PURPOSE: To describe the duration, timing, tempo, and synchronicity of puberty, as well as the correlation between timing and tempo of puberty. METHODS: Overall, 15,819 of 22,439 invited children participated in the Puberty Cohort within the Danish National Birth Cohort. Participants completed a web-based questionnaire every 6 months through maturation with questions on current pubertal status. Girls reported current Tanner stage of breast and pubic hair development, and timing of menarche. Boys reported current Tanner stage of genital and pubic hair development, timing of first ejaculation, and vocal changes. While accounting for this interval-censored puberty information, we estimated the duration of puberty. Then, we used a nonlinear mixed effect growth model to estimate timing, tempo, synchronicity of puberty, and correlation between timing and tempo of puberty. RESULTS: In girls, the average duration of breast development was longer, whereas the average tempo was slower than pubic hair development. The average timing of breast development was earlier than the average timing of pubic hair development. The majority of girls had asynchronous puberty. In boys, the average duration was longer and average tempo slower for genital than pubic hair development. The average timing of genital and pubic hair development were comparable; hence, the majority had synchronous pubertal development. Adolescents who had earlier timing also tended to have a faster tempo. DISCUSSION: Being one of the largest puberty cohorts worldwide, these unique contemporary data can help physicians, parents, and children to understand and anticipate expected progression through pubertal development.

Transcriptome-wide m6A methylation profiling of Wuhua yellow-feathered chicken ovary revealed regulatory pathways underlying sexual maturation and low egg-laying performance.

RNA N6-melthyladenosine (m6A) can play an important role in regulation of various biological processes. Chicken ovary development is closely related to egg laying performance, which is a process primarily controlled by complex gene regulations. In this study, transcriptome-wide m6A methylation of the Wuhua yellow-feathered chicken ovaries before and after sexual maturation was profiled to identify the potential molecular mechanisms underlying chicken ovary development. The results indicated that m6A levels of mRNAs were altered dramatically during sexual maturity. A total of 1,476 differential m6A peaks were found between these two stages with 662 significantly upregulated methylation peaks and 814 downregulated methylation peaks after sexual maturation. A positive correlation was observed between the m6A peaks and gene expression levels, indicating that m6A may play an important role in regulation of chicken ovary development. Functional enrichment analysis indicated that apoptosis related pathways could be the key molecular regulatory pathway underlying the poor reproductive performance of Wuhua yellow-feathered chicken. Overall, the various pathways and corresponding candidate genes identified here could be useful to facilitate molecular design breeding for improving egg production performance in Chinese local chicken breed, and it might also contribute to the genetic resource protection of valuable avian species.

Larval plastic responses to warming and desiccation delay gonadal maturation in postmetamorphic spadefoot toads.

Developmental plasticity allows organisms to adjust life-history traits to varying environmental conditions, which can have concomitant effects across life stages. Many amphibians are suitable model systems to study plasticity because their larvae can adjust growth and differentiation under fluctuating environments. It is unknown, however, whether somatic and gonadal differentiation are equally affected by environmentally induced plasticity or whether their decoupling alters gonadal maturation postmetamorphosis, which may affect fitness. We tested if developmental acceleration in response to warming and desiccation risk results in shifts in gonadal maturation during metamorphosis and postmetamorphic growth in western spadefoot toads (Pelobates cultripes). We found additive effects of increased temperature and desiccation risk on development and growth at metamorphosis, which largely constrained gonadal maturation in metamorphic and postmetamorphic individuals of both sexes. Furthermore, the conditions experienced by larvae incurred sex-specific carryover effects on the gonadal maturation of juveniles 5 months after metamorphosis. In females, high temperature delayed ovarian maturation regardless of the water level. In males, exposure to high temperature and high water levels slightly delayed the testes' maturation. These results highlight the relevance of larval plasticity in the gonadal maturation of species undergoing metamorphosis, which may have implications for population demographics and the evolution of life histories.

Early-life exposure to acephate inhibits sexual development and induces testicular and ovarian toxicity in mice.

Acephate is an organophosphate insecticide that exerts its toxicity by acting on the nervous system of insects. In addition to its action on the mammalian nervous system, acephate can also induce endocrine disruption of the reproductive system in mammals. However, the effects of acephate on sexual maturation and ovary development remain unclear. This study evaluated whether early-life exposure to acephate negatively impacts the male and female sexual maturation process and mature reproductive tissues. C57BL/6N mice were exposed to acephate (0, 0.3, 300 ppm) in drinking water from embryonic day 11.5 to ablactation, when the pups were four weeks old. Both sexes in the high-dose group experienced an early postnatal growth deficit, while females in the low-dose group continued to gain weight until 10 weeks of age. Exposure to acephate altered the anogenital index in females. Furthermore, preputial separation and vaginal opening were delayed in the high-dose group. At maturity, the weight of the seminal vesicles was decreased in the high-dose group. All treated groups exhibited increased vacuolation, accumulation of residual bodies, and degeneration in the testes. Furthermore, follicle regression was observed, and the healthy follicle number at each developmental stage was decreased in all treated groups. These results are probably due to the inhibition of the regulation by the hypothalamic-pituitary-gonadal axis and direct toxicity to reproductive organs. In conclusion, our study demonstrates that early-life exposure to acephate in mice may disrupt normal postnatal development, postpone puberty onset, and adversely affect reproductive functions during the reproductive period in both sexes.

Vasa, Piwi, and Pl10 Expression during Sexual Maturation and Asexual Reproduction in the Annelid Pristina longiseta.

Naidids are tiny, transparent freshwater oligochaetes, which are well known for their ability to propagate asexually. Despite the fact that sexually mature individuals and cocoons with embryos are sometimes found in nature, in long-period laboratory cultures, worms reproduce agametically only. In this paper, we showed, for the first time, the expression of Vasa, Piwi, and Pl10 homologs in mature Pristina longiseta worms with well-developed reproductive system structures and germ cells. Although the animals have been propagated asexually by paratomic fission for over 20 years in our lab, some individuals become sexualized under standard conditions for our laboratory culture and demonstrate various stages of maturation. The fully matured animals developed a complete set of sexual apparatus including spermatheca, atrium, seminal vesicles, and ovisac. They also had a clitellum and were able to form cocoons. The cues for the initiation of sexual maturation are still unknown for P. longiseta; nevertheless, our data suggest that the laboratory strain of P. longiseta maintains the ability to become fully sexually mature and to establish germline products even after a long period of agametic reproduction. On the other hand, many of the sexualized worms formed a fission zone and continued to reproduce asexually. Thus, in this species, the processes of asexual reproduction and sexual maturation do not preclude each other, and Vasa, Piwi, and Pl10 homologs are expressed in both somatic and germline tissue including the posterior growth zone, fission zone, nervous system, germline cells, and gametes.

Mini review shows that a testicular volume of 3 mL was the most reliable clinical sign of pubertal onset in males.

AIM: We aimed to evaluate aspects of pubertal development to identify the most reliable clinical sign of pubertal onset in males. METHODS: We performed a mini review of the literature. RESULTS: In 1951 Reynolds and Wines categorised pubic hair growth and genital development in five stages by visual inspection. Today the Tanner scale is used to assess the five stages of pubertal development, The second genital stage, characterised by enlargement of the scrotum defines pubertal onset in males. Testicular volume may be evaluated by using a calliper or by ultrasound scan. The Prader orchidometer, described in 1966, offers a method for evaluating testicular growth by palpation. Pubertal onset is commonly defined as testicular volume >3 or ≥4 mL. The development of sensitive laboratory methods has enabled studies analysing hormonal activity in the hypothalamus-pituitary-gonadal axis. We review the relationships between physical and hormonal signs of puberty. We also discuss the results of studies assessing different aspects of pubertal development with a focus on identifying the most reliable clinical sign of pubertal onset in males. CONCLUSION: A substantial amount of evidence supports testicular volume of 3 mL as the most reliable clinical sign of male pubertal onset.

Triazine Herbicide and NPK Fertilizer Exposure: Accumulation of Heavy Metals and Rare Earth Elements, Effects on Cuticle Melanization, and Immunocompetence in the Model Species Tenebrio molitor.

The increasing use of agrochemicals, including fertilizers and herbicides, has led to worrying metal contamination of soils and waters and raises serious questions about the effects of their transfer to different levels of the trophic web. Accumulation and biomagnification of essential (K, Na, Mg, Zn, Ca), nonessential (Sr, Hg, Rb, Ba, Se, Cd, Cr, Pb, As), and rare earth elements (REEs) were investigated in newly emerged adults of Tenebrio molitor exposed to field-admitted concentrations of a metribuzin-based herbicide and an NPK blend fertilizer. Chemical analyses were performed using inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) supported by unsupervised pattern recognition techniques. Physiological parameters such as cuticle melanization, cellular (circulating hemocytes), and humoral (phenoloxidase enzyme activity) immune responses and mass loss were tested as exposure markers in both sexes. The results showed that NPK fertilizer application is the main cause of REE accumulation in beetles over time, besides toxic elements (Sr, Hg, Cr, Rb, Ba, Ni, Al, V, U) also present in the herbicide-treated beetles. The biomagnification of Cu and Zn suggested a high potential for food web transfer in agroecosystems. Gender differences in element concentrations suggested that males and females differ in element uptake and excretion. Differences in phenotypic traits show that exposure affects metabolic pathways involving sequestration and detoxification during the transition phase from immature-to-mature beetles, triggering a redistribution of resources between sexual maturation and immune responses. Our findings highlight the importance of setting limits for metals and REEs in herbicides and fertilizers to avoid adverse effects on species that provide ecosystem services and contribute to soil health in agroecosystems.

miR-101-5p overexpression suppresses the proliferation of goat spermatogonial stem cells by targeting EZH2.

MicroRNAs (miRNAs), as post-transcriptional gene mediators, regulate the biological characteristics of spermatogonial stem cells (SSCs), including proliferation, differentiation and apoptosis. However, the potential roles and mechanisms by which miR-101-5p affected the biological characters of goat SSCs have not been fully elucidated. Herein, we reported that miR-101-5p overexpression decreased cell viability (P < 0.01), arrested cell cycle in the G1 phase (P < 0.05), and aggravated apoptosis of goat SSCs (P < 0.01) compared with negative control (NC), as determined by CCK-8 assay and flow cytometry analysis. Additionally, PCNA protein expression was attenuated by miR-101-5p overexpression (P < 0.05). Notably, the expression of SSCs specific genes Oct4 (P < 0.05), PLZF (P < 0.01) and DAZL (P < 0.01) were decreased in miR-101-5p overexpressed SSCs. Furthermore, the dual luciferase reporter assay showed that, when co-transfected with miR-101-5p mimics, the relative luciferase activity of EZH2 wide-type (WT) was inhibited (P < 0.05) compared with the transfection of EZH2 mutant (MUT). EZH2 expression was negatively correlated with miR-101-5p expression in goat SSCs. Collectively, our data implicates that miR-101-5p overexpression aggravates cell apoptosis, and suppresses cell proliferation of goat SSCs via targeting EZH2, which may impair spermatogenesis.

Diet impacts the reproductive system's maturation in the male moth Agrotis ipsilon (Noctuidae, Lepidoptera).

In male moth Agrotis ipsilon, sexual maturation occurs between the third and the fifth day of adult life and is characterized by the development of the reproductive organs such as testes and accessory sex glands. Since sexual maturation requires considerable energy investment, we hypothesized that diet would be an essential regulatory factor in this developmental process. Indeed, the links between the male diet and reproductive physiology have not been described as in females. To test the previous hypothesis, we offered male moths diets corresponding to different flower nectars found in nature, and measured morphological and functional changes in the testes and accessory sex glands. In comparison to a diet composed of sucrose only, males fed with a diet composed of diverse sugars, including glucose, supplemented with sodium led to an earlier increase in the length and the protein content of accessory sex glands, as well as a reduction of the testicular volume accompanied by an acceleration of the sperm bundle transfer from the testes to the duplex. These results show that these specific diets accelerate the maturation of the reproductive system in male moth Agrotis ipsilon.

Effects of adult-diet modifications on sexual maturation rate, body weight, and quantity of sperm transfer by male Mexican fruit fly (Diptera: Tephritidae).

Mass-reared sterile Mexican fruit flies (Anastrepha ludens [Loew]) are released along the US-Mexico border to control fruit fly damage and spread. It is more cost effective for the mass rearing program if males mature at a younger age because they can be held for a shorter time at the facility before release. In this study, adult male Mexican fruit fly diets were tested to determine if the current diet used at the mass rearing facility promotes younger mating and optimal sperm transfer compared to other diet formulations. Hydrolyzed yeast presentation methods were examined: an agar-free dry mixture of yeast and sugar (Y+S), yeast embedded in the diet during the agar boiling process (current method used) and sifted dry on the ager gel surface. Methoprene, a juvenile hormone analog was also tested as a supplement to agar gel diets with and without yeast. A greater proportion of males fed Y+S mated 1 day earlier than males fed other diets. Male mating age and diet did not significantly affect the quantity of sperm transferred; however, a non-significantly higher percentage of males fed diets with embedded yeast transferred enough sperm to fill all 3 spermathecae. The results suggest that the current diet used is optimal for mass-rearing the flies and that yeast presentation method has a significant effect on A. ludens male mating age but not on the quantity of sperm transferred to females.

Impact of growth trajectory on sexual maturation in layer chickens.

Recent studies showed that apart from photostimulation, metabolic triggers may independently activate sexual maturation and egg production in chickens. However, the origin, mode of action, and specific target(s) of this metabolic control remain unknown. Beyond body weight (BW), we hypothesize that body composition (BC) and associated specific metabolic signals are involved. Thus, this study was conducted to determine the BW and BC thresholds triggering spontaneous sexual maturation in layer pullets under different growth trajectories. Day-old Lohman LSL lite and Lohman brown lite chicks (n = 210 each) raised in brooding cages under ad libitum (AL) feeding until 8 weeks of age were randomly allocated into individual cages and assigned to one of 3 experimental growth profiles; AL, breeder's target (T), restricted 20% below target (R), (n = 70 birds/profile/strain). Birds had free access to water throughout the trial. All hens were maintained on 10 h of light (10 lux) throughout the rest of the study. Blood and tissue samples were collected throughout the study to measure plasma estradiol (E2) concentrations and organ weights, respectively. Furthermore, carcasses were subjected to Dual-energy X-ray absorptiometry (DEXA) analyses. All analyses were completed with SAS using the MIXED procedure. Results show that R treatment slowed (p < 0.001) growth, delayed age at first egg (FE) and egg production (p < 0.001) and resulted in lower BW at FE (p < 0.001), lower ovary weight and number of follicles (p < 0.001) compared to AL in both strains, whereas, the strain significantly impacted body weight (p < 0.0001), ovary weight (p < 0.001), BW at FE (p < 0.001), age at FE (p < 0.001), egg production (p < 0.0001), E2 (p < 0.0001) and body composition (p < 0.05). For DEXA, AL feeding (p < 0.001) increased fat deposition compared to R. Furthermore, there was a positive correlation between plasma E2 and bone mineral content (p < 0.01) and bone mineral density (p < 0.01). In conclusion, feed allocation impacted growth and BC in a strain dependent manner which resulted in differing age at sexual maturation and egg production. Furthermore, a body fat threshold between 10% to 15% appears to be required for the occurrence of spontaneously sexual maturation in laying hens.

Exposure to non-persistent pesticides and sexual maturation of Spanish adolescent males.

BACKGROUND: Several non-persistent pesticides are endocrine disrupting chemicals and may impact on sexual maturation. OBJECTIVE: To examine the association between urinary biomarkers of non-persistent pesticides and sexual maturation in adolescent males in the Environment and Childhood (INMA) Project. METHODS: The metabolites of several pesticides were measured in spot urine samples collected from 201 boys aged 14-17 years, including: 3,5,6-trichloro-2-pyridinol (TCPy), metabolite of chlorpyrifos; 2-isopropyl-4-methyl-6-hydroxypyrimidine (IMPy), metabolite of diazinon; malathion diacid (MDA), metabolite of malathion; diethyl thiophosphate (DETP) and diethyl dithiophosphate, non-specific metabolites of organophosphates; 3-phenoxybenzoic acid (3-PBA) and dimethyl cyclopropane carboxylic acid, metabolites of pyrethroids; 1-naphthol (1-NPL), metabolite of carbaryl; and ethylene thiourea (ETU), metabolite of dithiocarbamate fungicides. Sexual maturation was assessed using Tanner stages, self-reported Pubertal Development Scale, and testicular volume (TV). Multivariate logistic regression was employed to examine associations between urinary pesticide metabolites and the odds of being in Tanner stage 5 of genital development (G5) or pubic hair growth (PH5); stage ≥4 of overall pubertal development, gonadarche, and adrenarche; or having mature TV (≥25 mL). RESULTS: DETP concentrations>75th percentile (P75) were associated with lower odds of being in stage G5 (OR = 0.27; 95% CI = 0.10-0.70), detectable TCPy with lower odds of gonadal stage≥4 (OR = 0.50; 95% CI = 0.26-0.96), and intermediate detectable MDA concentrations (

Current Pubertal Development in Chinese Children and the Impact of Overnutrition, Lifestyle, and Perinatal Factors.

CONTEXT: Age of pubertal onset has been decreasing in many countries but there have been no data on pubertal development in Chinese children over the last decade. OBJECTIVE: The primary objective of the study was to evaluate the current status of sexual maturation in Chinese children and adolescents. Secondary objectives were to examine socioeconomic, lifestyle, and auxological associations with pubertal onset. METHODS: In this national, cross-sectional, community-based health survey, a multistage, stratified cluster random sampling method was used to select a nationally representative sample, consisting of 231 575 children and adolescents (123 232 boys and 108 343 girls) between 2017 and 2019. Growth parameters and pubertal staging were assessed by physical examination. RESULTS: Compared to 10 years previously, the median age of Tanner 2 breast development and menarche were similar at 9.65 years and 12.39 years respectively. However, male puberty occurred earlier with a median age of testicular volume ≥4 mL of 10.65 years. Pubertal onset did occur earlier at the extremes, with 3.3% of the girls with breast development at 6.5-6.99 years old, increasing to 5.8% by 7.5-7.99 years old. Early pubertal onset was also noted in boys, with a testicular volume ≥ 4 mL noted in 1.5% at 7.5-7.99 years, increasing to 3.5% at 8.5-8.99 years old. Obesity and overweight increased risk of developing earlier puberty relative to normal weight in both boys and girls. CONCLUSION: Over the past decade, pubertal development is occurring earlier in Chinese children. While the cause is multifactorial, overweight and obesity are associated with earlier puberty onset. The currently used normative pubertal data of precocious puberty may not be applicable to diagnose precocious puberty.

Untargeted Metabolomics Revealed Potential Biomarkers of Small Yellow Follicles of Chickens during Sexual Maturation.

Sexual maturation provides economically important traits in poultry production. Research on the initiation mechanism of sexual maturity is of great significance for breeding high-yield laying hens. However, the underlying mechanisms are not fully clear. Here, one hundred and fifty Chahua No. 2 laying hens (the CH2 group, which has precocious puberty) and one hundred and fifty Wu Liang Shan black-bone laying hens (the WLS group, a late-maturing chicken breed) with similar weights and ages were randomly selected. ELISA was used to determine the secretion levels of luteinizing hormone (LH), estradiol (E2), and progesterone (P4) in 150-day-old serum and small yellow follicle (SYF) tissues. A histology examination, immunohistochemistry, and quantitative real-time PCR (qPCR) were used to explore the molecular mechanism of how some genes related to oxidative stress affect sexual maturation. The results showed that the secretion levels of LH, E2, and P4 in the CH2 group serum and SYF were higher than those in the WLS group. The results of the real-time PCR of all genes showed that the expression levels of cytochrome P450 family 11 subfamily A member 1, steroidogenic acute regulatory protein, follicle-stimulating hormone receptor, and cytochrome P450 family 19 subfamily A member 1 in the CH2 group were significantly higher than those in the WLS groups (p < 0.001). Untargeted metabolomics combined with multivariate statistical analysis was used to identify biomarkers of SYF tissues in the CH2 and WLS groups. A trajectory analysis of the principal component analysis (PCA) results showed that the samples within the group were clustered and that the samples were dispersed between the CH2 and the WLS groups, indicating that the results of the measured data were reliable and could be used for further research. Further analysis showed that a total of 319 metabolites in small yellow follicles of the CH2 and WLS groups were identified, among which 54 downregulated differential metabolites were identified. These 54 metabolites were found as potential CH2 biomarkers compared with WLS at 150 days, and the different expressions of L-arginine, L-prolinamide, (R)-4-hydroxymandelate, glutathione, and homovanillic acid were more significant. Twenty metabolic pathways were found when significantly differential metabolites were queried in the KEGG database. According to the impact values of the metabolic pathways, eighteen differential metabolites belonged to the mTOR signaling pathway, glutathione metabolism, ABC transporters, the cell ferroptosis pathway, and D-arginine and D-ornithine metabolism. Interestingly, we identified that the cell ferroptosis pathway played an important role in chicken follicle selection for the first time. The histology and immunohistochemistry of SYF showed that the number of granulosa cells increased in the CH2 groups and the expression levels of glutathione peroxidase 4, tumor protein p53, ribosomal protein S6 kinase, and sterol regulatory element binding protein 1 in the granulosa cell layer were upregulated in the CH2 group at the time of sexual maturation. Furthermore, we also speculated that the antioxidant system may play an indispensable role in regulating sexual maturity in chickens. Overall, our findings suggest differentially expressed metabolites and metabolic pathways between CH2 and WLS chickens, providing new insights into the initiation mechanism of sexual maturation.

Reproductive condition of the black-lip pearl oyster Pinctada margaritifera during the lunar phase.

This study analyzed the relationship between the lunar phase and the reproductive cycle of Pinctada margaritifera inhabiting Weno Island, Chuuk Lagoon, Micronesia. We measured indicators of maturity (gonadosomatic index [GSI] and sexual maturation-related genes) and investigated changes in the gonadal maturity stages (GMS) of P. margaritifera over lunar cycle. GSI was higher around the full moon. GMS of P. margaritifera were classified as the early gametogenesis stage, ripe and spawning stage, and spent and degenerating stage. A large percentage of oysters was observed in the ripe and spawning stage at the first quarter moon in female and the full moon in male as well as in the spent and degenerating stages at the third quarter moon in both sexes. In addition, the expression of doublesex- and mab-3-related transcription factor 2 (DMRT2) in the male P. margaritifera black-lip pearl oyster was the highest during the full and third quarter moon phases, whereas no difference in expression was observed with the lunar phase in females. In contrast, the expression of vitellogenin (VTG) was the highest in female P. margaritifera during the first and third quarters. No difference in expression was observed according to the lunar phase in males. The results suggest that the lunar phase directly affects the expression of sexually mature gonads in P. margaritifera black-lip pearl oyster.

Cut-off points in the waist:height ratio for the prediction of insulin resistance, second stage of sexual maturation in Brazilian adolescents.

The objective of this study was verify the best waist:height ratio (WHtR) cut-off points for the prediction of insulin resistance (IR) according to stage of sexual maturation in Brazilian adolescents. Data were analysed from 37 759 adolescents between 12 and 17 years of age, of both sexes, participating in the ERICA study on cardiovascular risk factors in adolescents ('Estudo de Riscos Cardiovasculares em Adolescentes - ERICA'). The following variables were included sex, age and sexual maturity according to the Tanner stage, waist circumference, height, WHtR, fasting insulin and fasting blood sugar so as to obtain the homeostasis model assessment-estimated insulin resistance (HOMA-IR). We constructed an Receiver Operating characteristic (ROC) curve for each group in order to determine the best WHtR cut-off points for the sample stratified by sex and stratified by sex and sexual maturation stage. The female sex corresponded to 60% of the sample, with a mean age of 14·7 years (sd ± 0·08 years) and 13·1 % had IR. The prediction of IR through the WHtR indicator was better for boys, where all the areas under the ROC curve were superior to 0·70. When stratifying by sex alone, the best WHtR cut-off points found were equal to 0·45 for girls and 0·44 for boys. When stratifying by Tanner maturation stages, the cut-off points were reduced with advancing sexual maturation in the male group, while the opposite occurred in the female group. Among Brazilian adolescents, WHtR is a good predictor of IR evaluated by HOMA-IR, but different cut-off points should be adopted according to sex and sexual maturation stage.