Background/Objectives: This study reports on findings from the first preconception screening performed in Russia and provides a comprehensive discussion of the significant results and challenges faced during the implementation of the project. Methods: Using a targeted sequencing panel of 33 genes (associated with 29 autosomal recessive and 4 X-linked diseases), we analyzed 165 couples considering pregnancy. The screening design also included analysis of the frequent pathogenic variants in the SMN1, DMD, CFTR, and CYP21A2 genes that may not be detected through the next-generation sequencing approach. The sequential screening protocol, wherein the female partner was tested first, was used. Results: The results revealed that 35.8% of women (n = 59) were carriers of at least one pathogenic or likely pathogenic (P/LP) variant, with 7.9% of women (n = 13) carrying variants in two or more genes. Notably, the analysis identified 5 deletions of exon 7 in the SMN1 gene, 1 deletion of the CYP21A2 gene, and 1 large duplication in the DMD gene in female participants. The most frequently identified pathogenic variants occurred in the CYP21A2, GJB2, SERPINA1, and ATP7B genes. The screening identified six couples (3.6% of the cohort) at high risk of having a child with an autosomal recessive or X-linked genetic disorder. Conclusions: This pilot study confirms the high clinical utility of the gene panel, effectively evaluating reproductive risk in couples without a known family history of monogenic diseases. The findings indicate that the observed frequencies of identified gene variants differ from those theoretically expected, with a notable percentage of identified couples being at relatively high risk. Furthermore, these results highlight the indispensable role of comprehensive genetic counseling both before and after testing to ensure an appropriate preconception testing algorithm and informed reproductive decision-making.

This study was designed to assess the effectiveness of neonatal congenital adrenal hyperplasia (CAH) screening in Guangzhou, China. A total of 818,417 newborns were screened for CAH by measuring 17-hydroxyprogesterone (17-OHP) concentrations. Cut-off values were stratified based on gestational age (GA) and the timing of sample collection. Neonates with initial positive results (17-OHP ≥ cut-off value) were recalled for a second dried blood spot sample to reassess 17-OHP levels. Confirmatory testing involved biochemical analyses, Sanger sequencing, and multiplex ligation-dependent probe amplification of the CYP21A2 gene. From 2018 to 2024, a total of 40 patients with classical 21-hydroxylase deficiency were identified, including 28 cases (70%) of the salt-wasting form and 12 cases (30%) of the simple virilizing form. The overall incidence of CAH was 1 in 20,653 (95% confidence interval: 1:34,928, 1:14,661). No statistically significant differences in prevalence were observed between sexes or between preterm and full-term infants (p > 0.05). 17-OHP concentrations are influenced by GA and the timing of sample collection. The screening efficiency for CAH could be improved by adopting a multitiered cut-off value system adjusted for GA and collection time.

Background21‐hydroxylase deficiency (21‐OHD) represents the most common form of congenital adrenal hyperplasia (CAH) caused by pathogenic variants in the CYP21A2 gene. To date the molecular analysis of the CYP21A2 gene is based on the selective CYP21A2 gene amplification followed by Sanger sequencing. Herein we present the clinical manifestations, hormonal profile, and application of different molecular strategies to accurately investigate the CYP21A2 gene in a newborn presenting with the salt wasting (SW) form of CAH.MethodsThe patient, his parents, and the paternal grandparents underwent CYP21A2 genotyping employing Sanger sequencing and multiplex ligation‐dependent probe amplification (MLPA). Furthermore, the patient and his parents underwent a more extended protocol to determine the location of the pathogenic variants identified.ResultsThe newborn carried three CYP21A2 gene copies (two paternal and one maternal), each of them harboring a pathogenic variant, which is in concordance with the clinical manifestations of the SW form of CAH.ConclusionThe molecular investigation of the proband presenting with the SW form of CAH revealed a complex genotype that could only be determined by employing different molecular strategies. Laboratories should be aware of the possibility of complex genotypes in the CYP21A2 gene and employ different protocols to avoid the possibility of a genetic misdiagnosis.

Mycobacteriumabscessus CYP123 is a steroid hydroxylase with an implication in host infection.

Reproductive toxicity of chronic exposure to cortisone in western mosquitofish (Gambusia affinis).

A Hispanic male in his 20s presented to the hospital with altered mental status and severe hypercalcemia. During his hospitalization, he developed clinical deterioration requiring intubation and ICU transfer. Initial work-up revealed low PTH and PTHrP levels, along with elevated vitamin D 25-OH and 1,25-OH levels. He developed progressive acute kidney injury, and a kidney biopsy later confirmed macrophage activation syndrome (MAS) in a background of type III lupus nephritis. His hypercalcemia resolved following pulse methylprednisolone therapy. Once clinically improved, the patient reported recent use of vitamin D supplementation. After excluding other causes of non-PTH-dependent hypercalcemia with elevated 1,25-OH vitamin D, MAS was determined to be the driver of increased 1-alpha hydroxylase activity, exacerbated by exogenous vitamin D. This is the first reported case of severe hypercalcemia due to MAS from lupus nephritis, compounded by vitamin D supplementation.

Disclosure: S. Muneer: None. M. Munoz: None. J. Iyer: None. D.R. Weber: None. S.M. Willi: None.Background: Hypercalcemia in infants is rare but potentially serious. While often asymptomatic, it may present with failure to thrive, constipation, dehydration, poor feeding, and in severe cases, hypotonia or seizures. Differential diagnosis includes hypervitaminosis, malignancy, endocrinopathies, granulomatous disease, and fat necrosis. Case: A full-term male infant with congenital lymphatic malformation and neonatal chylothorax developed candidal fungemia on day of life (DOL) 30. Before this diagnosis, he had been noted to have hypercalcemia, 11.5mg/dL (ref 7.8-11.3) on DOL 28. His fungemia was treated with amphotericin B on DOL 32-35 and then fluconazole DOL 36-47. During this time his Ca remained elevated and did not improve. Initial evaluation for hypercalcemia commenced on DOL 48 which showed hypercalcemia 12.7mg/dL, normal 25-OH Vitamin D, low 1,25-dihydroxy Vitamin D, appropriately suppressed PTH, and elevated urinary Ca/creatinine. Bone turnover markers, including c-telopeptide and bone-specific alkaline phosphatase, were normal. The patient initially responded to hyperhydration and removal of calcium from feeds, but hypercalcemia recurred. On DOL 63, he required calcitonin at 2 IU/kg BID. Calcitonin was discontinued after 17 days as calcium normalized, and 1,25-dihydroxy Vitamin D returned to normal. Discussion: Infant hypercalcemia is rare, and its association with fungemia is even more unusual. Hypercalcemia secondary to fungemia is suspected to result from extra-renal 1,25OHD2 production by activated macrophages expressing CYP27B1 (1-alpha hydroxylase). Excess 1,25OHD2 causes hypercalcemia via increased intestinal absorption and increased bone resorption of calcium. In this case, hypercalcemia in the setting of elevated 1,25OHD2 and suppressed PTH is consistent with extra-renal production by activated macrophages. Fluconazole is an inhibitor of CYP27B1, and therefore exerts a beneficial effect on hypercalcemia by reducing 1,25OHD2 synthesis, in addition to treating the fungemia. Calcitonin, an inhibitor of osteoclast activity, is effective in the acute management of hypercalcemia driven in whole or in part by excess bone resorption. Steroids are also an effective treatment of hypercalcemia secondary to activated macrophages, however, are typically not an option with a fungal infection because of the risk of reactivating a systemic infection. This case adds to the limited literature on hypercalcemia secondary to systemic infections in infants. It was noted that the patient started to develop hypercalcemia shortly after the diagnosis of the fungemia and failed to improve while receiving fluconazole. Calcitonin proved effective in rapidly reducing serum Ca levels, highlighting its utility in acute management. Further studies are warranted to understand the pathophysiological mechanisms linking infection and hypercalcemia in infants.Presentation: Sunday, July 13, 2025

Disclosure: B. AlHamer: None. M. Al Mukaddam: None.A 43-year-old transgender woman with a history of illicit soft tissue filler (ISTF) injections was admitted for hypoxic respiratory failure. Endocrinology was consulted for severe hypercalcemia. She notably underwent ISTF injections around 11 years prior to presentation to the cheek, jawline and buttocks. She later went on to develop severe diffuse parenchymal lung disease (DPLD); this was felt to likely be secondary to silicone emboli as liver biopsies demonstrated lipogranulomas consistent with silicone emboli. Initial evaluation showed a 1,25-Dihydroxyvitamin D (Calcitriol) mediated hypercalcemia. Serum calcium was 14.6 mg/dL, PTH was <0.6 pmol/L, PTHrp was 4.5 pmol/L (normal 0.0-2.3 pmol/L, likely due to renal impairment with an eGFR of 27ml/min), 25-Hydroxyvitamin D was 6ng/mL and Calcitriol was elevated to 111 pg/mL (normal 9.9-79.3 pg/mL). FDG PET CT showed gluteal and thigh low level uptake associated with heterogeneity and not distinct musculature on CT. Given prior biopsies negative for other granulomatous diseases, she was presumed to have severe hypercalcemia secondary to silicosis. She was treated with isotonic fluids, calcitonin and pamidronate without significant improvement in calcium levels. High dose methylprednisolone was started as treatment for her DPLD with improvement of calcium to 11.9 mg/dL. Ketoconazole was then started at an initial dose of 200mg twice daily. Calcium improved further to 8.1 mg/dL. Calcitriol was later remeasured to be 62.5mg/mL. Over her hospitalization, ketoconazole was variably held, restarted and adjusted in dose. At the time of writing, she has been maintained on ketoconazole 200mg twice daily with stably improved serum calcium measurements. The use of ISTF injections, including with silicone, has been reported in literature. Injections vary in both site and volume. Complications are often delayed and range from infection to formation of granulomas. Granulomas can induce hypercalcemia through macrophage mediated extra-renal 1 alpha hydroxylase activity, increasing conversion of 25-Hydroxyvitamin D into activated Calcitriol. Evaluation of granuloma mediated hypercalcemia reveals a PTH independent process, often but not always with elevated calcitriol levels. Localization of granulomas can be challenging, with FDG-PET CT having been shown to be useful in identifying areas of hypermetabolic granulomatous disease. Hypercalcemia has been seldom reported in the literature in association with cosmetic injections, commonly of silicone. In these cases, treatment often involves the use of bisphosphonates and steroids, with few reports describing the use of ketoconazole. Ketoconazole inhibits the effects of 1 alpha hydroxylase to reduce calcitriol production. This case represents a rare presentation of hypercalcemia related to silicosis, and the utility of ketoconazole in the treatment of granuloma mediated hypercalcemia.Presentation: Sunday, July 13, 2025

Abstract Disclosure: L.B. Farias: None. L.A. Gross: None. R. Nolasco: None. J.S. Rossi: None. F.A. Barbosa: None. C.E. Kater: None. M.D. Walker: None. J.P. Bilezikian: None. M. Lazaretti-Castro: None. Introduction: 21-Hydroxylase Deficiency (21OHD) is an autosomal recessive disorder characterized by variable degrees of cortisol deficiency and androgen excess. It’s the main cause of congenital adrenal hyperplasia, and according to the pathogenic variation affecting the CYP21A2 gene, 21OHD will manifest in two clinical forms: Classic (CL) and non-classic (NC). The former is a more severe and even fatal condition if not correctly treated. In the latter, symptoms of hyperandrogenism may appear during infancy or adulthood, notably in girls. Management of both forms requires glucocorticoids (GC), often at supraphysiologic doses. While GC negatively impacts bone, high androgen levels may be protective. However, data on skeletal involvement in 21OHD are controversial and limited. Objective: This study aims to evaluate bone health of patients with 21OHD on chronic GC therapy, using high-resolution peripheral quantitative computed tomography (HRpQCT) to assess volumetric (v) BMD, microarchitecture, and mechanical properties of the tibia and radius. We hypothesized androgens may mitigate the deleterious effects of GC on bone. Materials and Methods: Cross-sectional study of patients with CL and NC 21OHD. DXA and HRpQCT were performed, and groups were compared to matched controls (CT). Mean±SE is shown. Results: 33 patients (26 CL, 7 NC) and 66 CT (age: CL 33±2, NC 37±4, CT 35±1 years, p=0.56; 73-86% female) were compared. 21OHD patients were shorter (CL 154±0.02; NC 154±0.03, CT 162±0.01 cm, p&amp;lt;0.001), and weight was higher in CL vs NC (73±3 vs 61±5 kg, p&amp;lt;0.04) with no difference to CT (68±2 kg). Compared to other groups, CL patients had higher BMI (CL 30.5±0.9, NC 25.5±1.8, CT 25.7±0.6 kg/m2, p&amp;lt;0.001), and fat indices by DXA: Body Fat +16% vs NC and +21% vs CT, p&amp;lt;0.004; Fat Mass Index +42% vs NC, +41% vs CT, p&amp;lt;0.0004. The GC dose was 44% higher in CL vs NC (p&amp;lt;0.05). Compared to CT, NC had lower BMD at the spine and hip (-10.3% to -19.5%; all p&amp;lt;0.04) with no difference from the CL group. At the radius, 21OHD patients had lower trabecular (Tb) bone volume fraction, TbvBMD, and Tb area, and greater Tb separation (11-13% for CL vs CT and 13-29% for NC vs CT, all p≤0.03; no difference between 21OHD groups). At the tibia, patients had lower Tb bone volume fraction, TbvBMD, Tb area, and total area (9-14% for CL vs CT and 15-20% for NC vs CT, all p≤0.02; no difference between 21OHD groups). Compared to CT, CL and NC had 13% higher cortical thickness (p&amp;lt;0.03), and higher cortical vBMD at the radius (+2% for CL vs CT, +5% for NC vs CT, p&amp;lt;0.02) and tibia (+3% for CL vs CT, +4% for NC vs CT, p=0.002) with no difference between 21OHD groups. Conclusion: Patients with 21OHD, regardless of severity, exhibit trabecular deterioration despite preserved cortical bone. These findings may suggest compartment-specific alterations reflecting opposing effects of chronic GC exposure and androgen excess on the skeleton. Presentation: Saturday, July 12, 2025

Abstract Disclosure: E. Silverstein: None. C. Eliazo: None. A. Diaz: None. Introduction: Classic congenital adrenal hyperplasia (CAH) is a rare condition affecting 1:16,000 newborns and is characterized by elevated testosterone levels from abnormal production of cortisol. Mutations of the CYP21A2 gene are the most common cause of CAH and can present as mild cases: non-classic/late onset CAH, or severe forms, as in classic CAH. Classic CAH can be simple virilizing or salt-wasting types, the latter, a life-threatening condition if not diagnosed soon after birth. In the U.S., screening for CAH is included on newborn screening; however, this evaluation is not often done in developing countries. Lack of early diagnosis and, in some cases surgical intervention, can be life threatening. Case report: We present the case of a 16-year-old girl born in rural Honduras and assigned as male at birth based on her male-appearing genitalia: a normal-sized and formed phallus with bilateral cryptorchidism. During infancy, this child had recurrent hospitalizations due to severe dehydration, hyponatremia, and hypotension. At 4-months of life, she was transferred to Tegucigalpa, the country's capital, and diagnosed with salt-wasting CAH. At that time, the family was asked to change her name and her gender to female, and she was started on hydrocortisone and fludrocortisone. The family moved to the U.S. when the child was 7 months old; and at that time, she was evaluated by endocrinology. A karyotype was reported as 46, XX and molecular analysis confirmed the diagnosis of CAH (Intron-2 deletions: homozygous in CYP21A2). Although legally the child appears as a male with a masculine name, the family raised her as a girl and socially changed her name. Due to non-typical gender behavior, a gender role evaluation was done by psychology at 8 years of age. However, due to learning difficulties, this evaluation was inconclusive; she identified as a girl, which matches her gender expression. Upon physical examination she has a Tanner V phallus with the urethral opening at the tip (Prader V) and bilateral cryptorchidism. She reached menarche at age 11 and has monthly bleeding through the urethra. She has been evaluated by urology and underwent cystourethroscopy and vaginogram, which showed bladder diverticula. She has had multiple urinary tract infections, some with septic shock, requiring PICU admissions. Due to lack of insurance, surgical repair is not possible. She receives prophylactic antibiotics with suboptimal compliance. Conclusion: Early CAH diagnosis is critical to prevent severe medical and social complications. While the timing of genital surgery in children with differences in sex development remains controversial, multidisciplinary evaluation is critical, especially for CAH patients, to minimize severe complications. Many adult patients report preferring genital surgery in early childhood. Unfortunately, lack of insurance coverage and surgical costs hinder access to care in the U.S. Presentation: Monday, July 14, 2025

To investigate the clinical phenotype and genetic diagnosis process of fetuses with 21 hydroxylase deficiency (21-OHD) caused by compound heterozygous variant of the CYP21A2 gene . A fetus who was diagnosed at Taizhou Hospital in Zhejiang Province on December 4, 2020 due to unclear characteristics of external genitalia on ultrasound was selected as the study subject. Chromosome copy number variation sequencing (CNV-seq) and whole exome sequencing (WES) were performed on amniotic fluid samples. Candidate variants were validated by Sanger sequencing and multiplex ligation-dependent probe amplification (MLPA), and short tandem repeat (STR) analysis was used to exclude maternal blood contamination. The pathogenic mechanism of the variants was further explored. The procedure followed by this study was approved by the Medical Ethics Committee of Taizhou Hospital (Ethics No.: K20201009). The MRI examination of the fetal external genitalia showed thickening of labia minora and enlargement of the clitoris. The CNV-seq results of the fetus showed no significant abnormality. The WES results showed that the fetus had a homozygous c.293-13C>G variant in the CYP21A2 gene (NM-000500.9). STR testing excluded maternal blood contamination. Sanger sequencing verified the presence of heterozygous c.293-13C>G variant of the CYP21A2 gene in the fetus and its mother, while its father did not detect this mutation. Further MLPA testing results showed that the fetus and its father had heterozygous deletion (I2G-C locus) mutations in exon 1~7 of the CYP21A2 gene. Based on the "Standards and Guidelines for Interpretation of Sequence Variants" jointly developed by the American College of Medical Genetics and Genomics (ACMG) and the Association for Molecular Pathology (AMP), both variants of the CYP21A2 gene carried by the fetus were predicted to be pathogenic. According to the imaging and genetic testing results of the external genitalia of the fetus, the fetus was prenatally diagnosed as 21-OHD caused by the CYP21A2 gene variant. Follow-up after prenatal diagnosis showed that the couple had opted to terminate the pregnancy at a local hospital at 31+ weeks of gestation, and the clinical phenotype of the abortion fetus was consistent with the imaging and molecular genetic diagnosis. The imaging features of this fetus are suspected to be congenital adrenal hyperplasia (CAH). Combined with WES, Sanger sequencing, and MLPA testing results, the fetus was diagnosed with 21-OHD caused by compound heterozygous variants of the CYP21A2 gene, which provided a basis for prenatal diagnosis.

Congenital adrenal hyperplasia (CAH) is a group of autosomal recessive endocrine disorders caused by impaired steroidogenesis and insufficient cortisol production from the adrenal cortex. The most common form of CAH is 21-hydroxylase deficiency, caused by mutations in the CYP21A2 gene. This enzyme deficiency causes failure to progress through the steroidogenic pathways, consequently there is an increased synthesis of steroid hormone precursors that result in adrenal androgens excess, leading to virilisation in female patients. Over the past two decades, significant progress has been made in optimising corticosteroid replacement strategies, with a focus on reducing androgen excess while minimising glucocorticoid exposure. Modified-release hydrocortisone formulations, such as Efmody® and Plenadren®, as well as continuous subcutaneous infusion therapies, have been developed to help better mimic physiological cortisol rhythms. In addition, corticotropin-releasing hormone-1 (CRH1) receptor antagonists have been investigated and have now been approved as a novel approach to reducing adrenocorticotropic hormone (ACTH)-driven adrenal hyperplasia and androgen excess. More recently, promising novel approaches have been developed to control androgen excess and reduce glucocorticoid exposure. This review provides an overview of current standard-of-care treatments for CAH and highlights recent advancements in therapeutic strategies. By addressing the limitations of traditional glucocorticoid replacement, these innovations have the potential to improve long-term outcomes and quality of life for patients with CAH.

Background and Objectives: Congenital adrenal hyperplasia (CAH) belongs to a group of disorders caused by the deficiency of the steroid 21-hydroxylase enzyme that impairs the production of adrenal steroids. Many at-risk families opt for prenatal diagnosis (PND) to prevent the birth of an affected child in view of increased mortality in the salt-losing severe cases, genital ambiguity in females, and precocious puberty in males. Before prenatal testing, it is imperative to know the pathogenic variant(s) in the affected child before PND is offered to the families. However, in reality, many families either do not have access to molecular testing or the proband is unavailable for testing. In this study, we report the experience at a tertiary care genetics center of PND in 139 families at risk, utilizing multiple genetic and molecular methods for testing. Materials and Methods: Most families recruited in this study had at least one clinically diagnosed child with CAH. Molecular methods like multiplex ligation probe amplification, Sanger sequencing, polyacrylamide gel electrophoresis, and linkage were used for variant identification in the proband, if available, or in the parents. Variant segregation in parents was done in each family. Maternal cell contamination was ruled out in every sample for which prenatal testing was performed. Results: A total of 139 prenatal samples were analyzed, of which 33 (23.7%) fetuses were affected and 106 (76.3%) were unaffected. Of the 106, 64 (60.4%) were heterozygous (carriers) and 42 (39.6%) were homozygous for wild-type variants. Conclusion: With different molecular methodologies performed on chorionic villus or amniotic fluid, 100% cases were correctly reported, indicating the efficacy of multiple methodologies. Linkage was useful in families where molecular workup was not done. Despite the absence of an affected child in the families, prenatal diagnostic testing could be offered after comprehensive carrier screening and determining the carrier status of the couple.

ObjectiveThis study aimed to develop a novel homologous sequence analysis technique using high-throughput sequencing data to enhance CYP21A2 mutation detection. The approach leverages next-generation sequencing to overcome existing limitations and improve 21-hydroxylase deficiency diagnostic accuracy.MethodsFrom April 21, 2022, to February 21, 2023, a total of 100 unrelated participants were enrolled at the Women and Children’s Hospital of Ningbo University, selected based on clinical manifestations and genetic testing results. The study used next-generation sequencing combined with a homologous sequence alignment (HSA) algorithm, which calculated the sequencing read ratios from homologous regions to identify pathogenic or likely pathogenic variants in the CYP21A2 gene. All detected variants were further validated using long-range PCR or multiplex ligation-dependent probe amplification. The accuracy of the HSA algorithm was systematically assessed.ResultsAmong the 100 participants, 84 were identified as carriers of CYP21A2 mutations, while 16 were diagnosed with 21-hydroxylase deficiency. A total of 107 pathogenic mutations were detected using the homologous sequence alignment algorithm, comprising of 99 single nucleotide variants or insertions/deletions, 6 copy number variants, and 8 fusion mutations. Additionally, eight cases of CYP21A2-CYP21A1P gene conversions were identified based on HSA scores and confirmed through long-range PCR or multiplex ligation-dependent probe amplification. The algorithm demonstrated a positive predictive value of 96.26% for identifying mutations in CYP21A2. The most frequently observed mutations included c.955C > T, c.844G > T, c.293–13C > G, c.518T > A, and exon-level deletions.ConclusionIn genetic testing, particularly when addressing misalignment challenges associated with highly homologous genes such as CYP21A2, application of the HSA algorithm enables accurate mutation detection using commonly employed short-read sequencing methods. Through the characterization of homologous sequence features and optimization of the HSA algorithm, accurate mutation detection can be achieved in more homologous gene families (eg, HBA1/HBA2, SMN1/SMN2, GBA/GBAP1).

ABSTRACTThe prevalence of metabolic disease is increased in congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency. However, the underlying molecular mechanisms causing these problems are not fully understood. We aimed to elucidate the metabolic phenotype and conduct a transcriptomic analysis of a 21-hydroxylase-deficient zebrafish model, to unravel the molecular mechanisms underlying the metabolic pathophysiology of CAH. The morphology, anatomy and transcriptomic analysis of whole larvae, adult liver tissue from 18-month-old cyp21a2−/− zebrafish were compared to those of wild-type siblings. Our main phenotypical finding was that adult mutants were larger, with increased fat deposition compared to controls, in-keeping with the transcriptomic analysis showing the dysregulation of several biological processes involved in lipid metabolism. Importantly, we found that ATP synthesis and provision of energy precursors were included among the most significantly suppressed processes in both larvae and adult livers. We conclude that cortisol deficiency in cyp21a2−/− mutants causes growth and body fat abnormalities at adult stages, as well as transcriptomic dysregulation of metabolic processes, energy homeostasis and inflammatory responses in both larvae and adults. These findings reveal how GC deficiency in zebrafish contributes to the development of the metabolic comorbidities that are similar to those observed in patients with CAH.

Exercise-induced mitochondrial protection in skeletal muscle of ovariectomized mice: A myogenic E2 synthesis-independent mechanism.

Neonicotinoid insecticides and triazole fungicides are widely used in agriculture, often in combination with other pesticides, leading to concerns about potential health effects. This study investigated the combined effect of these chemicals using the Comparative Toxicogenomics Database (CTD) to identify common target genes, followed by functional enrichment analysis and gene-gene and protein-protein interaction assessments. In this study, it was determined that pesticides may interfere with biological processes such as steroid hydroxylase activity, oxidoreductase activity, and steroid metabolism, and cause hormonal imbalances and endocrine system disorders. In addition, among the 10 genes identified, CYP3A5 and CYP3A7 gene expression differed significantly between prostate cancer and normal prostate tissues, and this was supported by UALCAN data. In addition, previous studies have confirmed that hsa-miR-27b, one of the prominent miRNAs in this study, and transcription factors (PROX1 and ESR2) are associated with prostate cancer. Similar to our study, previous studies have confirmed that triazole fungicides disrupt testosterone homeostasis and steroidogenesis, while neonicotinoids damage the prostate due to their effects on androgen receptors. These genes, miRNAs, and transcription factors appear to mediate the effects of these pesticides on cancer pathways and suggest a link to prostate cancer. In conclusion, this study demonstrated that concurrent exposure to neonicotinoid insecticides and triazole fungicides may damage the prostate and potentially contribute to the development of prostate neoplasia. These findings emphasise the importance of further in vitro and in vivo validation to establish a definitive causal relationship and provide insight into the toxicological effects of pesticide exposure on prostate health.

The CYP21A2 gene, mapped to the RCCX module in 6p21.3, is responsible for 21-hydroxylase deficiency (21-HD). In this work, we leveraged Oxford Nanopore Technology (ONT) Long Read sequencing (LRS) to analyze samples from an Argentinian cohort of 21-HD. A total of 34 samples were sequenced in 2 amplicons of 8.5 Kb covering the centromeric and telomeric RCCX modules. The number of variants found varied between 3 and 106 and all expected pathogenic variants and new ones were obtained with the LR sequencing workflow developed. We defined with higher accuracy the breakpoints of the rearrangements allowing the reclassification of chimeras and/or converted genes in 18.75% of the samples, some of them with clinical implications. By addressing the study of the telomeric RCCX module/s in depth, we found 19 genetic variants (GVs) for CYP21A1P and 29 GVs for TNXA not previously described in Latin American populations. This study may represent the first application of ONT LRS for clinical evaluation of Latin American subjects, highlighting the importance of LRS as a high-resolution method of diagnosis. It would allow a better understanding of the diversity of the RCCX modules and improve our knowledge of the variation of genetic mechanisms behind the disease.

Pathogenic variation underlying rare diseases in an Arab population: Implications for screening programs

The aim of this study is to investigate the main active substances of Qilin pills by high performance liquid chromatogre-electrostatic field orbitrap mass spectrometry (HPLC-Q-Orbitrap /MS), and explore the mechanism of its action in the treatment of asthenozoospermia by combining network pharmacology and molecular docking. (1) Qilin pills were quantitatively and qualitatively analyzed by HPLC-Q-Orbitrap /MS. (2) The top 100 compounds in Qilin pills were screened by content analysis and SwissADME, and their targets were predicted. The asthenozoospermia targets were searched through the database. And a "protein-protein interaction" (PPI) network was constructed. KEGG and GO analysis was performed using the DAVID database. And a "drug-target-pathway" network was constructed. (3) SailVina was used for molecular docking. (1) A total of 1 275 known components were found and ranked in Qilin pills by HPLC-Q-Orbitrap /MS analysis. (2) The top 100 compounds in Qilin pills predicted a total of 1 053 targets and 184 potential therapeutic targets for asthenozoospermia. KEGG pathway analysis and GO analysis showed that the treatment of asthenozoospermia by Qilin pills may be related to the steroid hormone synthesis pathway, the response to steroid hormones, the chromosomal region of cells and the activity of steroid hydroxylase. The mechanism of Qilin pills in treating asthenozoospermia may be related to regulating the synthesis, metabolism and reaction process of sex hormone in the body. (3) The molecular docking results of its key targets (CYP19A1, ESR1, HSP90AA1, p53, HIF1α and BCL2) showed that the key active ingredients M030, M039, M043, M050, M055 and M073 of Qilin pills had spontaneous binding. It had a binding energy of less than －5 kJ /mol. The material basis of Qilin pills has been explored by this study. And the mechanism of action of Qilin pills in the treatment of asthenozoospermia is highly bound to the expression and response process of steroid hormones, which provides a theoretical basis for the clinical application of Qilin pills.