Prepubertal Stage Research Articles

The TCONS_00046732/chi-miR-135b-5p/PRLR regulatory axis promotes endometrial epithelial cells growth through the PI3K-Akt signaling pathway

Non-coding RNAs are considered key regulatory factors in biological and reproductive physiological processes in mammals. However, the molecular functions of long noncoding RNAs (lncRNAs) in regulating dynamic uterine development and reproductive capacity during sexual maturation remain unclear. This study analyzed the expression characteristics and molecular functions of lncRNAs in uterine tissues from 20 goats at four specific time points during sexual maturation: day 1 after birth (D1), 2 months (M2), 4 months (M4), and 6 months (M6), finding that stage-specific DE lncRNAs may regulate cell proliferation, apoptosis, hormone secretion, metabolism, and immune response through multiple action modes. Within the lncRNA-miRNA-mRNA network, a novel lncRNA, TCONS_00046732, associated with uterine development, exhibited significantly higher expression during sexual maturity compared to the prepubertal stage, correlating positively with PRLR and negatively with chi-miR-135b-5p. FISH and IF analyses revealed significant co-localization and distinct expression patterns of TCONS_00046732, chi-miR-135b-5p, and PRLR in the endometrial epithelium. Further experiments confirmed that TCONS_00046732 competitively binds to chi-miR-135b-5p to upregulate PRLR, thereby activating the PI3K-Akt signaling pathway, promoting primary endometrial epithelial cell proliferation, G1-to-S phase transition, and inhibiting apoptosis. These findings enhance our understanding of uterine molecular regulation and provide key insights into the molecular basis of goat sexual development.

Pattern of physical growth and pubertal changes in adolescent girls with Systemic Lupus Erythematosus

ABSTRACT Aims: To study the pattern of physical growth and pubertal changes in adolescent girls with Systemic Lupus Erythematosus (SLE). Methods and Material: Weight, height, BMI, waist and hip circumference among 50 adolescent SLE girls (aged 8–17 years) were cross-sectionally studied using standardized techniques and instruments. Breast development stage, presence or absence of pubic and axillary hair and age of attainment of menarche were also noted. Results: With the advancement of age, weight and height of SLE girls increased but they were lighter and shorter compared to normal Indian peers. 18.4% of study girls were short-statured while only one participant was underweight. As per mid-parental height, 34.3% were predicted to have shorter final heights. BMI demonstrated an inconsistent trend with 12.3% and 9.2% being obese and overweight, respectively. Interestingly, 10.7% of SLE girls were at risk of metabolic syndrome (waist circumference >70th centile). About 50% of study girls had attained menarche (mean age: 13.04 ± 1.38 years). Appearance of pubic and axillary hair occurred around 1 year later than attainment of menarche. 33.80% of study subjects were in prepubertal stage of breast development and rest 66.20% were in advanced stages of breast development. SLE girls who had younger age at diagnosis and longer duration of therapy had significant delay in breast development and attainment of menarche. Conclusions: SLE girls show delayed growth and pubertal attainments compared to their normal Indian and Western counterparts. The data presented will provide an understanding of the auxological dynamics and pattern of pubertal changes among adolescent girls with SLE living in north-western India.

Three-dimensional assessment on digital cast of spontaneous upper first molar distorotation after Ni-ti leaf springs expander and rapid maxillary expander: A two-centre randomized controlled trial.

The aim of this randomized controlled trial (RCT) was to evaluate the spontaneous distorotation of upper first permanent molars and the transverse dentoalveolar changes on digital casts in growing patients following maxillary expansion treatment using either the Leaf Expander® or the rapid maxillary expander (RME), both anchored to the deciduous second molar. This study was a two-arm, parallel-assignment, RCT with a dual-centre design conducted at two teaching hospitals in Italy. Inclusion criteria included maxillary transverse deficiency, prepubertal development stage (cervical vertebra maturation stage [CVMS] 1-2) and early mixed dentition with fully erupted upper first permanent molars. Exclusion criteria were systemic diseases or syndromes, CVMS 3-6, agenesis of upper second premolars, unavailability of the second deciduous molar for anchorage and Class III malocclusion. Patients were randomly assigned to the Leaf Expander® or RME group using a computer-generated randomization list created by a central randomization centre. Randomization was conducted immediately before the start of treatment. The intervention involved treatment with either the Leaf Expander® or the RME. Both devices were anchored to the second deciduous molars. Following randomization, patients were further categorized based on the presence of no crossbite, unilateral crossbite or bilateral crossbite. The primary outcome measure was the distorotation of the upper first molar (U6). Secondary outcomes included measurements of interdental linear dimensions, specifically upper inter-canine width (53-63), upper inter-molar width (MV16-MV26) and upper inter-deciduous second molar width (55-65). The examiner analysing the digital casts was blinded to the treatment groups to prevent detection bias and ensure objective assessment. However, due to the nature of the intervention, blinding was not feasible for the patients and clinicians involved in administering the treatment. A total of 150 patients were enrolled and randomly assigned to two groups: 75 to the Leaf Expander® group and 75 to the RME group. Recruitment started in November 2021 and was completed in November 2022. At the time of analysis, the trial was complete with no ongoing follow-ups. ANOVA tests revealed no significant differences between the three subgroups (no-cross, unilateral-cross and bilateral-cross) within both the Leaf Expander® and RME groups at T0. The Leaf Expander® demonstrated significantly greater distorotation in the unilateral crossbite subgroup compared to the RME (p = .014). In terms of total molar distorotation, the Leaf Expander® appliance showed a significantly greater effect (12.66°) compared with conventional RME (7.83°). Linear regression analysis demonstrated a significant correlation between the extent of expansion and the degree of molar rotation. Maxillary expansion resulted in significant spontaneous molar distorotation when the appliance was bonded to the second deciduous molars. The Leaf Expander® exhibited significantly greater molar distorotation compared with conventional RME. The degree of molar distorotation was correlated with the extent of expansion obtained on the second deciduous molar. The trial was registered at ClinicalTrials.gov (ID: NCT05135962).

Multiomics and eXplainable artificial intelligence for decision support in insulin resistance early diagnosis: A pediatric population-based longitudinal study

Pediatric obesity can drastically heighten the risk of cardiometabolic alterations later in life, with insulin resistance standing as the cornerstone linking adiposity to the increased cardiovascular risk. Puberty has been pointed out as a critical stage after which obesity-associated insulin resistance is more difficult to revert. Timely prediction of insulin resistance in pediatric obesity is therefore vital for mitigating the risk of its associated comorbidities. The construction of effective and robust predictive systems for a complex health outcome like insulin resistance during the early stages of life demands the adoption of longitudinal designs for more causal inferences, and the integration of factors of varying nature involved in its onset. In this work, we propose an eXplainable Artificial Intelligence-based decision support pipeline for early diagnosis of insulin resistance in a longitudinal cohort of 90 children. For that, we leverage multi-omics (genomics and epigenomics) and clinical data from the pre-pubertal stage. Different data layers combinations, pre-processing techniques (missing values, feature selection, class imbalance, etc.), algorithms, training procedures were considered following good practices for Machine Learning. SHapley Additive exPlanations were provided for specialists to understand both the decision-making mechanisms of the system and the impact of the features on each automatic decision, an essential issue in high-risk areas such as this one where system decisions may affect people’s lives. The system showed a relevant predictive ability (AUC and G-mean of 0.92). A deep exploration, both at the global and the local level, revealed promising biomarkers of insulin resistance in our population, highlighting classical markers, such as Body Mass Index z-score or leptin/adiponectin ratio, and novel ones such as methylation patterns of relevant genes, such as HDAC4, PTPRN2, MATN2, RASGRF1 and EBF1. Our findings highlight the importance of integrating multi-omics data and following eXplainable Artificial Intelligence trends when building decision support systems.

Estimating mandibular growth stage based on cervical vertebral maturation in lateral cephalometric radiographs using artificial intelligence

IntroductionDetermining the right time for orthodontic treatment is one of the most important factors affecting the treatment plan and its outcome. The aim of this study is to estimate the mandibular growth stage based on cervical vertebral maturation (CVM) in lateral cephalometric radiographs using artificial intelligence. Unlike previous studies, which use conventional CVM stage naming, our proposed method directly correlates cervical vertebrae with mandibular growth slope.Methods and materialsTo conduct this study, first, information of people achieved in American Association of Orthodontics Foundation (AAOF) growth centers was assessed and after considering the entry and exit criteria, a total of 200 people, 108 women and 92 men, were included in the study. Then, the length of the mandible in the lateral cephalometric radiographs that were taken serially from the patients was calculated. The corresponding graphs were labeled based on the growth rate of the mandible in 3 stages; before the growth peak of puberty (pre-pubertal), during the growth peak of puberty (pubertal) and after the growth peak of puberty (post-pubertal). A total of 663 images were selected for evaluation using artificial intelligence. These images were evaluated with different deep learning-based artificial intelligence models considering the diagnostic measures of sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV). We also employed weighted kappa statistics.ResultsIn the diagnosis of pre-pubertal stage, the convolutional neural network (CNN) designed for this study has the higher sensitivity and NPV (0.84, 0.91 respectively) compared to ResNet-18 model. The ResNet-18 model had better performance in other diagnostic measures of the pre-pubertal stage and all measures in the pubertal and post-pubertal stages. The highest overall diagnostic accuracy was also obtained using ResNet-18 model with the amount of 87.5% compared to 81% in designed CNN.ConclusionThe artificial intelligence model trained in this study can receive images of cervical vertebrae and predict mandibular growth status by classifying it into one of three groups; before the growth spurt (pre-pubertal), during the growth spurt (pubertal), and after the growth spurt (post-pubertal). The highest accuracy is in post-pubertal stage with the designed networks.

Pre-Weaning Overfeeding Promotes Prepubertal Obesity and Evokes Sexual Dysfunction in Young Adult Rats

Obesity in early life can contribute to several health issues in adulthood, including diabetes, high blood pressure, and high cholesterol, and these conditions are associated with sexual dysfunction. Here, we hypothesized that pre-weaning overfeeding can have long lasting effects on body fat distribution and erectile function that will be observed in young adult males. On postnatal day (PND 1), Wistar rats were separated into two groups: normal litter (NL) with 10 pups per dam, or small litter (SL) with 3 pups per dam. Male rats (n = 8-10 per group) were then assessed at two different time points: during the prepubertal stage (PND 40) and in adulthood (PND 120). Erectile function was assessed only after sexual maturation (i.e., PND 120). Body weight, retroperitoneal and perigonadal adipose tissue (fat pad weight/100g of the body) were evaluated. Following euthanasia at PND 120, the corpus cavernosum (CC) was removed to perform functional studies using a strip myograph. Contractile and relaxation responses were evaluated by electrical field stimulation (EFS) and concentration-response curves (CRC) to phenylephrine [Phe, 1nM-30μM, millinewton (mN)] or acetylcholine (Ach 1nM-30μM, % relaxation) were performed. At the prepubertal stage, SL rats had higher body weight (NL 149±3 vs. SL 186±5, g, p<0.05), and increased retroperitoneal (NL 0.17±0.01 vs. SL 0.32±0.03, g/100g, p<0.05) and perigonadal (NL 0.38± 0.02 vs. SL 0.45±0.03, g/100g, p<0.05) adipose tissue. At PND 120, SL group presented increased body weight (NL 534.2.7±13 vs. SL 631.6±10, g, p<0.05), retroperitoneal (NL 1.31±0.11 vs. SL 1.82±0.14, g/100g, p<0.05) and perigonadal (NL 1.74±0.10 vs. SL 2.18±0.15, g/100g, p<0.05) adipose tissue when compared with NL group. The CC of SL rats at PND 120 had increased contraction induced by EFS for all frequencies (1 Hz: NL 0.5 ± 0.2 mN vs. SL 0.9 ± 0.2 mN; 4 Hz: NL 1.2 ± 0.3 mN vs. SL 1.8 ± 0.2 mN; 16 Hz: NL 1.9 ± 0.4 mN vs. SL 3 ± 0.4 mN), whereas the relaxation was decreased in SL (1 Hz: NL 0.77 ± 0.18% vs. SL 0.27 ± 0.09%; 4 Hz: NL 2.48 ± 0.26% vs. SL 1.36 ± 0.41%; 16 Hz: NL 3.06 ± 0.28% vs. SL 2.39 ± 0.53%). CRC showed increased contractile response to Phe (Emax NL 2.3±0.3 vs. SL 3.8± 0.6, mN, p=0.05) and reduced potency of ACh (pEC50 NL 6.4±0.6 vs. SL 4.9±0.7, mN, p=0.1) in the CC of SL. Incubation with indomethacin (10μM) abolished the differences in the contractile response induced by EFS or Phe between SL and NL, suggesting a heightened inflammatory response. Moreover, the CC of SL rats presented increased expression of Phospho-p44/42 MAPK (NL 0.9±0.25 vs. SL 1.3±0.11, p=0.2). Finally, proteome profiler cytokine array of serum samples from SL animals showed increased circulating levels of cytokines and chemokines associated with obesity, aging, and vascular dysfunction. Together, these data suggest that early prepubertal obesity has long-lasting negative implications on sexual function in young adulthood. National Institutes of Health (NIH); American Heart Association (AHA); Fulbright Program. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Impact of Mid and Late-gestation Toll-like Receptor 7 Stimulation on Development and Anxiety-like Behavior in Offspring

Maternal immune activation (MIA) can occur during pregnancy due to exposure to infectious diseases or other inflammatory conditions. MIA is a risk factor for various psychiatric disorders, such as major depressive disorder, as well as cardiac disorders in the offspring. Resiquimod (R-848), a toll-like receptor (TLR) 7 agonist, induces systemic inflammatory responses and mimics responses to viral infections. Previous studies have shown that MIA with a TLR 4 agonist lipopolysaccharide or the viral mimic, TLR 3 agonist polyinosinic:polycytidylic acid produces neuropsychiatric effects in the offspring. The current project investigates the degree to which mid and late-gestation MIA with R-848 impacts offspring development and anxiety-like behavior. Pregnant rat dams received vehicle (phosphate-buffered saline) or R-848 (1 mg/kg, s.c.) injections on gestation day (GD) 14 (mid-gestation) or GD18 (late gestation). Maternal body weight as well as offspring body weight, developmental milestones, and anxiety-like behavior were assessed. Although all dams gained weight throughout pregnancy, the R-848 dams from both injection days displayed pronounced weight fluctuations post-injection, with a trend toward attenuated weight gain in dams injected in late gestation. On postnatal day 21 (PND21), the R-848 dams injected on GD14 had significantly lower body weights (p=0.007) compared to the vehicle dams, whereas the body weights of the R-848 dams injected on GD18 did not differ significantly from the vehicles. Litter size, male-to-female ratio, and birth weight were also not affected by R-848. Semi-weekly offspring weights were not statistically different between any experimental groups by sex during any measured week in the mid-gestation-exposed cohort, but both male and female R-848-exposed offspring in the late-gestation exposure cohort displayed significantly decreased body weights at week 7 (males, p=0.0282 and females, p=0.0009). Eye opening, an important developmental milestone, was not different between the vehicle and R-848 pups regardless of the timing of exposure. Open field testing in the mid-gestation-exposed adult offspring revealed no significant locomotion differences or differences in time spent in the center zone. Open field testing in the late-gestation-exposed adult offspring also revealed no locomotion differences, however, the R-848-exposed female offspring spent markedly less time in the center zone than all other groups. These findings suggest that late gestation R-848 exposure results in attenuated maternal weight gain as well as increased anxiety-like behaviors in female offspring. Future exploration of pre-pubertal stages and assessments of autonomic regulation in offspring exposed to R-848 in late gestation will provide further insight into the long-term consequences of immune activation during pregnancy. NIH U54 MH118919. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Body odours as putative chemosignals in the father-child relationship: New insights on paternal olfactory kin recognition and preference from infancy to adolescence

Children's body odours are effective chemical cues in the parent-child relationship. Mothers can recognize the odour of their child and prefer this odour over that of unfamiliar children. This effect is mediated by genetic similarity and developmental stage and is therefore suited to promote parental care at pre-pubertal stage, while facilitating incest avoidance at (post-)pubertal stage. The present study tested whether similar mechanisms apply to fathers. Therefore n = 56 fathers evaluated body odour samples of their own and of unfamiliar children in varying genetic and developmental stages. Genetic status was determined by human leucocyte antigen (HLA) profiling, developmental status by standardized assessment of pubertal status and steroid hormone concentration (estradiol, testosterone). Similar to mothers, fathers identified their own child's body odour above chance and preferred that odour. The paternal preference did not relate to HLA similarity but decreased with increasing age of the child. The decline was associated with higher pubertal stages in daughters only, which supports the hypothesis of odour-mediated incest prevention in opposite-sex parent-child dyads.

The Relationship between Birth Weight and the Risk of Overweight and Obesity among Chinese Children and Adolescents Aged 7-17 Years.

Obesity is a major public health issue in children and adolescents. Our study aimed to examine the impacts of birth weight on overweight and obesity among Chinese children and adolescents. Using data from the China National Nutrition and Health Surveillance of Children and Lactating Mothers in 2016-2017, we included 10,041 participants aged 7-17 years. According to birth weight, participants were categorized into six groups, and the birth weight category of 3000 to 3499 g was chosen as the reference group, containing the largest number of children. Logistic regression analyses were used to investigate the association of birth weight with the risk of being obese at 7 to 17 years of age in multivariable-adjusted models. A restricted cubic spline was utilized to show the odds ratios (ORs) of obesity at different birth weight levels. The adjusted ORs for overweight were 0.98 (95%CI 0.63, 1.53), 1.02 (95%CI 0.84, 1.25), 1.34 (95%CI 1.16, 1.55), 1.72 (95%CI 1.35, 2.18), and 1.17 (95%CI 0.71, 1.96) in several birth weight groups, compared with group C (3000-3499 g). The adjusted ORs for obesity were 0.82 (95%CI 0.48, 1.40), 0.77 (95%CI 0.60, 0.98), 1.33 (95%CI 1.13, 1.57), 1.97 (95%CI 1.53, 2.53), and 2.01 (95%CI 1.27, 3.19). Furthermore, children in the post-pubertal stage had a slightly higher risk of overweight and obesity than those in the pre-pubertal and pubertal stage. Moreover, these associations were stronger among boys. The lower part of normal birth weight range is associated with a lower risk of overweight and obesity in children and adolescents. However, higher levels of birth weight increase risk.

(054) Erectile Dysfunction in Young Adulthood as a Consequence of Early Childhood Obesity

Abstract Introduction Childhood obesity is a worrisome health concern for children and teenagers, as it can contribute to several health issues in adulthood, including diabetes, high blood pressure, and high cholesterol. These conditions have been associated with sexual dysfunction. However, the precise impact of early childhood obesity on erectile function in adult males remains uncertain. Objective We hypothesized that overfeeding during early childhood can have lasting effects on body fat distribution, blood pressure, and erectile function in young adult males. Methods On postnatal day (PND 1), Wistar rats were separated into two groups: normal litter (NL) with 10 pups per dam, or small litter (SL) with 3 pups per dam. Male rats (n = 8-10 per group) were then assessed at two different time points: during the prepubertal stage (PND 40) and in adulthood (PND 120). Erectile function was assessed only after sexual maturation (i.e., on PND 120). Body weight, retroperitoneal and perigonadal adipose tissue (fat pad weight/100g of the body) were evaluated. The mean blood pressure (MBP) was recorded in conscious rats using the indirect tail-cuff method. Following euthanasia at PND 120, the corpus cavernosum was removed to perform functional studies using a strip myograph. Contractile and relaxation responses were evaluated by electrical field stimulation (EFS) and concentration-response curves (CRC) to phenylephrine [Phe, 1nM-30μM, millinewton (mN)] or acetylcholine (Ach 1nM-30μM, % relaxation) were performed. Results At the prepubertal stage, SL rats had higher body weight (NL 149±3 vs. SL 186±5, g, p<0.05), and increased retroperitoneal (NL 0.17±0.01 vs. SL 0.32±0.03, g, p<0.05) and perigonadal (NL 0.38± 0.02 vs. SL 0.45±0.03, g, p<0.05) adipose tissue. At PND 120, SL group presented increased body weight (NL 534±13vs. SL 631.6±10, g/100g, p<0.05), retroperitoneal (NL 1.31±0.11vs. SL 1.82±0.14, g/100g, p<0.05) and perigonadal (NL1.74±0.10vs. SL 2.18±0.15, g/100g, p<0.05) adipose tissue when compared with NL group. Moreover, SL rats presented increased MBP (NL 114.9±0.58 vs SL 120.5±1.2, mmHg). The corpus cavernosum of SL rats at PND 120 had increased contraction induced by EFS for all frequencies (1 Hz: NL 0.5 ± 0.2 mN vs SL 0.9 ± 0.2 mN; 4 Hz: NL 1.2 ± 0.3 mN vs SL 1.8 ± 0.2 mN; 16 Hz: NL 1.9 ± 0.4 mN vs SL 3 ± 0.4 mN), whereas the relaxation was decreased (1 Hz: NL 0.77 ± 0.18% vs SL 0.27 ± 0.09%; 4 Hz: NL 2.48 ± 0.26% vs SL 1.36 ± 0.41%; 16 Hz: NL 3.06 ± 0.28% vs SL 2.39 ± 0.53%). CRC showed increased contractile response to Phe (Emax NL 2.3±0.3 vs. SL 3.8± 0.6, mN, p=0.05) and reduced potency of ACh (pEC50 NL 6.4±0.6 vs SL 4.9±0.7, mN, p=0.1) in the corpus cavernosum of SL. Incubation with indomethacin (10uM) abolished the differences in the contractile response induced by EFS or Phe between SL and NL, suggesting a heightened inflammatory response. Conclusions These data suggest that early childhood obesity has long-lasting negative implications on sexual function in adulthood. Disclosure No.

Biometric and ultrasonographic evaluation on mammary gland of goat

Gross and ultrasonographic studies were conducted on the mammary gland of goat (n=18 for each study) during pre-pubertal, lactating and non-lactating stages. Mammary gland was located into the inguinal region and divided into two halves by mammary groove which was prominent during lactation stage. Each halve had separate glandular body and conical teat divisible into tip, mid and base regions. The mammary gland contained numerous lactiferous ducts which opened into the gland sinus that continued distally as teat sinus and teat canal. The gland sinus and teat sinus were separated from each other by annular folds, which were more prominent in lactating animals. At the junction of teat sinus and teat canal, small mucosal elevation was observed as Furstenberg’s rosette. Teat wall was lined by skin, teat internal mucosal layer and teat sinus from outside to inside. Ultrasonographic observations showed that the lactating mammary gland contained an echogenic structure of variable size indicating gland sinus, lactiferous ducts, blood vessels, glandular parenchyma, while the homogenous echogenic component depicting more connective tissue stroma in non-lactating and pre-pubertal goat. The teat wall was made up of innermost hyperechogenic layer (teat mucosa), middle hypoechogenic (fibromuscular) layer and mild hyperechogenic teat skin. Teat sinus was anechogenic, however teat canal was demarcated by hyperechogenic streak. Biometrical and ultrasonographic observations were recorded on teat canal length, maximum diameter of teat, maximum teat wall thickness. Present study showed that there was no significant difference in biometric and ultrasonographic measurements of goat teat. Therefore, it may be concluded that the gross and ultrasonographic observations may be used to diagnose any abnormality in the udder during clinical examination.

Prepubertal ve Pubertal Hastalarda Hashimoto Tiroiditi’nin Dağılımı, Klinik Özellikleri ve Laboratuvar Bulguları

Aims:Hashimoto's thyroiditis (HT) is the most common cause of acquired hypothyroidism in childhood. This disease, which develops on an autoimmune basis, often appears in adolescence. However, in recent years, However, in recent years, cases diagnosed with this disease in prepubertal and infant period have been reported. This is considered to regarding early exposure to environmental factors that disrupt the endocrine system and trigger autoimmunity. We aimed to evaluate the distribution, clinical features, and laboratory findings of our patients diagnosed with Hashimoto's thyroiditis in our clinic in prepubertal and pubertal periods. Material and Method:In the study, the cases diagnosed with Hashimoto's Thyroiditis in Ankara City Hospital Pediatric Endocrinology 5th outpatient clinic between August 2019 and May 2022 were divided into two groups prepubertal (Tanner stage 1) and pubertal (Tanner stage 2-3-4-5). The two groups were compared retrospectively in terms of clinical, laboratory, and ultrasound findings. Results: The mean±SDS of 134 patients diagnosed with HT was 12.4±3.4 years. While 25% (n:33) of the cases were prepubertal, 75% (n:101) were pubertal. The cases' female/male ratio (F/M) was 4.1/1, F/M: 2.3/1 in the prepubertal period, and F/M: 5.3/1 in the pubertal period. The euthyroid phase was detected most frequently in all cases at diagnosis. While the rate of euthyroid and subclinical hypothyroidism was the same in the prepubertal group, it was most detected in these two phases. In the pubertal group, it was mostly detected in the euthyroid phase. When we compared the prepubertal and pubertal groups, it was seen that subclinical hypothyroidism and hypothyroidism were most common in the prepubertal group. In contrast, subclinical hyperthyroidism and hyperthyroidism were more common in the pubertal group. Conclusion:In our study, HT is more prevalent in boys at the prepubertal stage than in the pubertal stage. While the patients in the prepubertal period applied in the hypothyroid phase, it was remarkable that the pubertal group had hyperthyroidism in their admissions.

Analysis of OCT4 and PGP9.5 gene expression in prenatal and postnatal buffalo (Bubalus bubalis) testes

This study aimed to investigate and characterize the spermatogonial stem cells (SSCs) in buffaloes at different stages of development, including prenatal, neonatal, prepubertal, and adult testes. We sought a comprehensive understanding of these cells through a combination of histological, immunohistochemical, and ultrastructural analyses. Specifically, we examined changes in the expression of two potential SSC markers, OCT4 and PGP9.5, using immunohistochemistry. Additionally, we conducted a real-time quantitative polymerase chain reaction (RT-qPCR) to assess the relative gene expression of OCT4 and PGP9.5. The relative expression of the OCT4 gene was down-regulated in the adult testes compared to its expression during prepubertal and neonatal life. The relative expression of the PGP9.5 gene was up-regulated in the neonatal testes and down-regulated in the prepubertal and adult testes. The spermatogonia were round, oval-to-ellipsoidal cells lying over the basement membrane (BM) with a round-to-oval nucleus. Based on the immunoexpression of the putative SSC markers, OCT4 and PGP9.5, we concluded that the proportion of stem cells was highest during the neonatal stage, followed by the prepubertal and prenatal stages. This finding sheds light on the dynamics of spermatogonial stem cells in buffalo testes at different developmental stages, providing valuable insights into these cells’ regulation and potential applications.

Effect of SEC III protocol on upper airway dimensions in growing class III patients: a retrospective study

BackgroundThe primary objective of the study was to evaluate the effects of SEC III (Splints, Class III Elastics, and Chin cup) protocol on the upper airway dimensions using lateral cephalometric radiographs. The secondary objectives were to evaluate the skeletal and dental effects of the SEC III protocol using lateral cephalometric radiographs.MethodsThe pre- and post-treatment lateral cephalometric radiographs of 24 patients treated using the SEC III protocol were used to address the aim of the study. Children in the pre-pubertal (CS-1 or CS-2) or circumpubertal (CS-3 or CS-4) skeletal maturation stage and having class III dentoskeletal malocclusion were included in the study. Patients with a history of previous orthodontic treatment, maxillofacial surgery or trauma, tonsillectomy, adenoidectomy, or craniofacial malformations were excluded. The pre-treatment and post-treatment lateral cephalometric radiographs were traced, then airway measurements, skeletal measurements, and dental measurements were performed. The results were analysed using paired samples t-test or Wilcoxon signed rank test according to the data normality.ResultsData of 6 males and 18 females were analysed (Mean age = 11.21 ± 1.02 years). Duration of active treatment was 5.75 ± 1.03 months. Treatment using SEC III protocol resulted in a significant increase in ANB angle (2.92 ± 1.50 degrees, p < 0.001) and Wits appraisal (3.31 ± 1.99 mm) (p < 0.001). The increase in the mandibular plane angle (0.75 ± 1.42 degrees, p = 0.02) and the maxillary length (2.29 ± 2.69 mm, p < 0.001) was statistically significant. Contrarily, the mandibular length did not change significantly (p = 0.10). The maxillary incisors were significantly proclined (4.38 ± 4.28 degrees; p < 0.001), while the mandibular incisors were significantly retroclined (-5.79 ± 6.21 degrees; p < 0.001) following treatment. The change in the nasopharyngeal airway and the retropalatal airway was not statistically significant. The middle and inferior pharyngeal space (retroglossal airway) significantly decreased by 1.33 ± 1.97 mm (p = 0.003) and 1.96 ± 2.48 mm (p = 0.001), respectively.ConclusionsEarly class III correction using SEC III protocol reduced the retroglossal airway dimensions but did not affect the nasopharyngeal and retropalatal airway dimensions. Correction of the class III dentoskeletal relationship was obtained through both skeletal and dental changes.

Quantification of overnight urinary gonadotropin excretion predicts imminent puberty in girls: a semi-longitudinal study

PurposeWe explored the alternative of using overnight fold change in gonadotropin levels by comparing the last-night-voided (LNV) and first-morning-voided (FMV) urine concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) as a conceptual analogy to the invasive gonadotropin-releasing hormone (GnRH) stimulation test setting.MethodsWe investigated the nocturnal changes in the immunoreactivity levels of urinary gonadotropins between early and late prepubertal stages as well as between early and late pubertal stages in FMV and LNV urine samples from 30 girls, of whom those who were prepubertal were further investigated through follow-up visits within the 1-year period from the start of the study.ResultsROC analysis revealed that the FMV total U-LH and FMV U-FSH concentrations at or above 0.3 IU/L and 2.5 IU/L, respectively, were excellent predictors of forthcoming onset of puberty within 1 year (100% sensitivity, 100% specificity, AUC: 1.00, and n = 10, for both). FMV total U-LH concentration at or above 0.8 IU/L represented the cut-off for clinical signs of puberty. FMV/LNV total U-LH and FMV/LNV U-FSH ratios at or below 4.11 and 1.38, respectively, were also good predictors of the onset of clinical puberty within 1 year. An overnight increase (FMV/LNV ratio) in total U-LH concentrations and in the U-LH/U-FSH ratio at or below 1.2-fold in pubertal girls was associated with the postmenarcheal pubertal stage.ConclusionFMV total U-LH and U-FSH above 0.3 IU/L and 2.5 IU/L, respectively, can be used as cut-off values to predict the manifestation of the clinical signs of puberty within 1 year. FMV total U-LH concentrations 0.3–0.8 IU/L and 0.6 IU/L may represent the range and the threshold, respectively, that reflect the loosening of the central brake on the GnRH pulse generator. An overnight increase of 20% or less in total U-LH concentrations and in the U-LH/U-FSH ratio in an early pubertal girl may serve as an indicator of imminent menarche, a presumed timing of which can be unraveled by future longitudinal studies.

Protective Effect of L-Carnitine against Deltamethrin Induced Male Reproductive Toxicity in Adult Rats Exposed at their Prepubertal Stage

The present study aims to investigate the protective role of L-Carnitine (LC) against Deltamethrin-induced testicular toxicity in male rats exposed during their early developmental period. DTM was administered at a dose of 6 mg/kg body weight to the prepubertal rat’s Post-Natal Day (PND) 23 rats through oral route and exposure was continued until they reach PND 90. After completion of the exposure period, male rats were assessed for reproductive endpoints. Deltamethrin exposure caused a significant reduction in testosterone production and decreased the sperm parameters such as sperm count, sperm motility, sperm viability and sperm membrane integrity. Deltamethrin intoxication also reduced the reproductive organs’ weight and testicular steroidogenic enzymes (3β-hydroxysteriod dehydrogenase and 17β-hydroxysteriod dehydrogenase) levels. Further DTM exposure induced the oxidative stress as evidenced by the significant reduction in the superoxide dismutase, catalase, and glutathione levels with significant elevation in the malondialdehyde. DTM also induced the significant sperm DNA damage. In addition, DTM exposure significantly declined the testosterone levels over the control group, indicating the impaired steroidogenesis. In addition to the compromised steroidogenesis DTM exposure also deteriorated the testicular architecture according to the histological observations. On the other hand, therapy with LC (100 mg/kg body weight) improved the biomass of reproductive organs, the features of the sperm, testicular steroidogenesis and testicular antioxidant enzymes levels. Concluding that DTM might disturb the testicular antioxidant level that eventually impaired the reproductive health of rats. However, LC supplementation mitigated the reproductive toxicity through its antioxidant property.

Association of HDL-Cholesterol, hypertension and left ventricular hypertrophy in youths with overweight or obesity

Background and aimTo evaluate the relationship between HDL-Cholesterol (HDL-C), hypertension, and left ventricular hypertrophy (LVH) in a large sample of Caucasian youths with overweight/obesity (OW/OB). Methods and resultsA cross-sectional multicenter study was performed in 1469 youths (age 6–16 years) with OW/OB observed in the period 2016–2020. An additional independent sample of 244 youths with an echocardiographic evaluation, observed in a single center was analyzed. The sample was divided in six quantiles (Q) of HDL-C: Q1: >56, Q2: ≤56 > 51, Q3: ≤51 > 45, Q4: ≤45 > 41, Q5: ≤41 > 39, Q6: <39 mg/dL. The nadir of the relationship was identified in youths in the first quantile. Among HDL-Cholesterol quantiles the distribution of hypertension was non-linear with a percentage of 25.0%, 40.1%, 33.6%, 31.3%, 35.2% and 39.7% in the six quantiles, respectively. The percentage of LVH was 21.8%, 43.6%, 48.8%, 35.5%, 38.5% and 52.0% in the six quantiles, respectively. The highest odds [95%Cl] of hypertension were 2.05 (1.33–3.16) (P < 0.01) in Q2, 1.67 (1.10–2.55) (P < 0.05) in Q3 and 1.59 (1.05–2.41) (P < 0.05) in Q6 vs Q1. The odds of LVH were 3.86 (1.15–10.24) (P < 0.05) in Q2, 4.16 (1.58–10.91) (P < 0.05) in Q3 and 3.60 (1.44–9.02) (P < 0.05) in Q6 vs Q1, independently by centers, age, sex, prepubertal stage, and body mass index. ConclusionContrary to the common belief, the present study shows that high levels of HDL-C may be not considered a negative predictor of hypertension and LVH, two risk factors for future CV disease.

Single-cell RNA sequencing technology in human spermatogenesis: Progresses and perspectives.

Spermatogenesis, a key part of the spermiation process, is regulated by a combination of key cells, such as primordial germ cells, spermatogonial stem cells, and somatic cells, such as Sertoli cells. Abnormal spermatogenesis can lead to azoospermia, testicular tumors, and other diseases related to male infertility. The application of single-cell RNA sequencing (scRNA-seq) technology in male reproduction is gradually increasing with its unique insight into deep mining and analysis. The data cover different periods of neonatal, prepubertal, pubertal, and adult stages. Different types of male infertility diseases including obstructive and non-obstructive azoospermia (NOA), Klinefelter Syndrome (KS), Sertoli Cell Only Syndrome (SCOS), and testicular tumors are also covered. We briefly review the principles and application of scRNA-seq and summarize the research results and application directions in spermatogenesis in different periods and pathological states. Moreover, we discuss the challenges of applying this technology in male reproduction and the prospects of combining it with other technologies.

Epigenetic regulation of functional candidate genes for milk production traits in dairy sheep subjected to protein restriction in the prepubertal stage

BackgroundAs the prepubertal stage is a crucial point for the proper development of the mammary gland and milk production, this study aims to evaluate how protein restriction at this stage can affect methylation marks in milk somatic cells. Here, 28 Assaf ewes were subjected to 42.3% nutritional protein restriction (14 animals, NPR) or fed standard diets (14 animals, C) during the prepubertal stage. During the second lactation, the milk somatic cells of these ewes were sampled, and the extracted DNA was subjected to whole-genome bisulfite sequencing.ResultsA total of 1154 differentially methylated regions (DMRs) were identified between the NPR and C groups. Indeed, the results of functional enrichment analyses of the genes harboring these DMRs suggested their relevant effects on the development of the mammary gland and lipid metabolism in sheep. The additional analysis of the correlations of the mean methylation levels within these DMRs with fat, protein, and dry extract percentages in the milk and milk somatic cell counts suggested associations between several DMRs and milk production traits. However, there were no phenotypic differences in these traits between the NPR and C groups.ConclusionIn light of the above, the results obtained in the current study might suggest potential candidate genes for the regulation of milk production traits in the sheep mammary gland. Further studies focusing on elucidating the genetic mechanisms affected by the identified DMRs may help to better understand the biological mechanisms modified in the mammary gland of dairy sheep as a response to nutritional challenges and their potential effects on milk production.

Srd5a1 is Differentially Regulated and Methylated During Prepubertal Development in the Ovary and Hypothalamus.

5α-reductase-1 catalyzes production of various steroids, including neurosteroids. We reported previously that expression of its encoding gene, Srd5a1, drops in murine ovaries and hypothalamic preoptic area (POA) after early-life immune stress, seemingly contributing to delayed puberty and ovarian follicle depletion, and in the ovaries the first intron was more methylated at two CpGs. Here, we hypothesized that this CpG-containing locus comprises a methylation-sensitive transcriptional enhancer for Srd5a1. We found that ovarian Srd5a1 mRNA increased 8-fold and methylation of the same two CpGs decreased up to 75% between postnatal days 10 and 30. Estradiol (E2) levels rise during this prepubertal stage, and exposure of ovarian cells to E2 increased Srd5a1 expression. Chromatin immunoprecipitation in an ovarian cell line confirmed ESR1 binding to this differentially methylated genomic region and enrichment of the enhancer modification, H3K4me1. Targeting dCas9-DNMT3 to this locus increased CpG2 methylation 2.5-fold and abolished the Srd5a1 response to E2. In the POA, Srd5a1 mRNA levels decreased 70% between postnatal days 7 and 10 and then remained constant without correlation to CpG methylation levels. Srd5a1 mRNA levels did not respond to E2 in hypothalamic GT1-7 cells, even after dCas9-TET1 reduced CpG1 methylation by 50%. The neonatal drop in POA Srd5a1 expression occurs at a time of increasing glucocorticoids, and treatment of GT1-7 cells with dexamethasone reduced Srd5a1 mRNA levels; chromatin immunoprecipitation confirmed glucocorticoid receptor binding at the enhancer. Our findings on the tissue-specific regulation of Srd5a1 and its methylation-sensitive control by E2 in the ovaries illuminate epigenetic mechanisms underlying reproductive phenotypic variation that impact life-long health.