bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of Vitamin D Receptor Pathway Genes in Subcutaneous Adipose Tissue of Obese Individuals
Authors: Ginnard Olivia Z B , Sisley Stephanie 
Published: May 2021

Abstract
Introduction: Vitamin D deficiency is a substantial comorbidity in 50% of pediatric patients and is linked with poorer health outcomes in children. Vitamin D levels are also shown to be inversely related to BMI. Therefore, there are many more children with low vitamin D levels due to the increasing prevalence of pediatric obesity. Pediatric patients with obesity and vitamin D deficiency also have a uniquely increased risk of metabolic syndrome, as compared to their lean peers. Measured levels of vitamin D correlate with other physiological markers of vitamin D effects in lean individuals but not obese individuals. It is possible that vitamin D levels reflect a storage form of vitamin D rather than a true reflection of vitamin D action in the body in this particular population. The aim of this study was to provide foundational knowledge to understand if expression of vitamin D receptor (VDR)-target genes may be used as a reference standard for vitamin D status in the body. Methods: We performed a secondary analysis of samples obtained from 33 obese adolescents that were consented under a past IRB-approved protocol. They were between the ages of 13 to 18 years that underwent bariatric surgery between 2004 and 2019. Data comprised of age, gender, race/ethnicity, and BMI. Samples collected included blood and subcutaneous adipose tissue. The tissue was analyzed via Real Time-PCR to obtain quantitative levels of VDR-target gene expression, which included PPARg, TLR4, THBD, CYP24A1, and VDR. Gene expression levels were normalized to the average of two housekeeping genes, GAPDH and RPLPO. Blood samples provided vitamin D levels (serum 25(OH)D). Results: VDR-target gene expression was significantly correlated between THBD, VDR, and TLR4 (p &lt;0.05), and PPARg with THBD and TLR4 (p &lt;0.05). There was no correlation observed between CYP24A1 gene expression and the other genes that were evaluated (p &gt;0.05). PPARg, THBD, TLR4, CYP24A1, and VDR gene expression levels did not correlate with circulating serum 25(OH)D levels (p &gt;0.05). Conclusion: These preliminary findings suggest that VDR-target gene expression correlates with each other but not with circulating serum 25(OH)D levels. This discrepancy supports that 25(OH)D levels do not indicate levels of vitamin D action and may not be an appropriate indicator of vitamin D deficiency in the obese population. Also, the observed CYP24A1 gene expression was limited in subcutaneous adipose tissue yet expression was seen in multiple other VDR-target genes. This emphasizes the tissue-specific nature of gene regulation of vitamin D. Further work should investigate VDR-target gene expression levels across multiple tissues of obese individuals to determine if markers of vitamin D action in one tissue are reflective of action across the body. This study may provide the first step in determining a new and more accurate biomarker for vitamin D deficiency and treatment in obesity.
Exploring the Potential Role of DLK1 in Pubertal Initiation
Authors: Jee Youn Hee , Delaney Angela , Jennings Melissa , Yanovski Jack Adam , Baron Jeffrey 
Published: May 2021

Abstract
The mechanisms that orchestrate the initiation of puberty are not well understood. DLK1 encodes a transmembrane protein that interacts with NOTCH1 receptor to negatively regulate NOTCH signaling. Loss-of-function mutations in DLK1 cause central precocious puberty, suggesting that DLK1 normally inhibits the reproductive axis centrally. The soluble form of DLK1, which is generated by proteolytic cleavage of the DLK1 extracellular domain, is measurable in human serum. We hypothesized that serum soluble DLK1 concentrations decline with age and that the decline, either in circulating levels or in tissue expression, contributes to the physiological mechanisms triggering pubertal initiation. Serum DLK1 was measured by immunoassay in 102 healthy subjects (age newborn - 26 yrs, 54 male). DLK1 concentrations did not differ by sex, BMI SDS, height, or status of fasting. DLK1 concentrations declined overall with age (R2=0.04, P&lt;0.001). However, there was not a substantial decline in the peripubertal period (mean± SEM, at Tanner stage 1, 2, 3, 4, 5: 14.8 ± 1.9, 16.4 ± 1.2, 17.0 ± 5, 13.6 ± 3, 9.7 ± 0.9 ng/mL). Serum DLK1, measured in 12 subjects (2 male) with a previous history of idiopathic central precocious puberty, did not differ from healthy controls. We next hypothesized that declining expression of Dlk1 or increasing expression of competing canonical NOTCH ligands in hypothalamus contributes to pubertal onset. The preoptic area (POA) was microdissected from rat brains (age 4 d, 2 w, 6 w, and 8-16 w, n=5 each) and expression was measured by RT-PCR. Dlk1 expression increased with age in both female and male rats (P&lt;0.001). Notch1 expression did not change with age. Expression of two ligands, Jag1 and Dll4, showed a peak at age 6 w, around the time of puberty, but only in males, and none of the other ligands (Jag1, Dll1, and Dll3) showed increasing expression at the age of puberty. In conclusion, we did not find evidence that declining serum soluble DLK1 concentrations in humans, declining DLK1 expression in rat preoptic area, or increasing NOTCH ligand expression in rat preoptic area contribute to pubertal onset.
Evaluation of the Molecular Pathogenesis of Adrenocortical Tumors by Whole-Genome Sequencing
Authors: Neininger Kerstin , May Patrick , Altieri Barbara , Lippert Juliane L , Roomp Kirsten , Dalmazi Guido Di , Canu Letizia , Ceccato Filippo , Riester Anna , Herterich Sabine L , Fassnacht Martin , Schneider Jochen G , Ronchi Cristina L 
Published: May 2021

Abstract
Pathogenesis of autonomous steroid secretion and adrenocortical tumorigenesis remains partially obscure. Our aim was to identify novel genetic alterations in adrenocortical adenomas (ACA) without somatic mutations in known driver genes. Whole-genome sequencing was performed on 26 ACA/blood-derived DNA pairs without driver mutations in PRKACA, GNAS and CTNNB1 genes at previous WES (ENSAT study JCEM 2016). These included 12 cortisol-producing adenomas with Cushing syndrome (CS-CPAs), 7 with mild autonomous cortisol secretion (MACS-CPAs), and 7 endocrine-inactive ACAs (EIAs). Seven adrenocortical carcinomas (ACC) were added to the cohort. We developed a bioinformatics pipeline for a comprehensive genome analysis and to reveal differences in variant distribution. Strelka, VarScan2 and ANNOVAR software and an in-house confidence score were used for variant calling and functional annotation. Combined Annotation-Dependent-Depletion (CADD) values were used to prioritize pathogenic variants. Additional focus relied on variants in pathogenically known pathways (Wnt/β-catenin, cAMP/PKA pathway). NovoBreak algorithm was applied to discover structural variations. Two hypermutated CS-CPA samples were excluded from further analysis. Using different filters, we detected variants in driver genes not observed at WES (one p.S45P in CTNNB1 and one p.R206L in PRKACA in two different CS-CPAs). In total, we report 179,830 variations (179,598 SNVs; 232 indels) throughout all samples, being more abundant in ACC (88,954) compared to ACA (CS-CPAs: 31,821; MACS-CPAs: 35,008; EIAs: 29,963). Most alterations were in intergenic (&gt;50%), followed by intronic and ncRNA intronic regions. A total of 32 predicted pathogenic variants were found in both coding (CADD values ≥ 15) and non-coding (CADD values ≥ 5) regions. We found 3,301 possibly damaging and recurrent variants (intergenic mutations removed) (CS-CPAs: 1,463; MACS-CPAs: 1,549; EIAs: 1,268; ACC: 1,660), mostly accumulated in intronic regions. Some of these were detected in members of the Wnt/β-catenin (CS-CPAs: 6; MACS-CPAs: 2; EIA: 1) and cAMP/PKA (CS-CPAs: 6; MACS-CPAs: 7; EIA: 4) pathways (e.g. ADCY1, ADCY2, GNA13, PDE11A). We also found a slightly higher number of structural variations in EIA (3,620) and ACC (3,486) compared to CS-CPAs (977) and MACS-CPAs (2,119). In conclusion, still unrevealed genetic alterations, especially in intronic regions, may accompany early adrenal tumorigenesis and/or autonomous cortisol secretion.
Anastrozole Improves Near Adult Height in Boys With Compromised Height Potential, as Monotherapy or in Combination With a GnRH Analogue
Authors: Papadimitriou Dimitrios T , Dermitzaki Eleni , Papagianni Maria , Kleanthous Kleanthis , Papadimitriou Anastasios , Mastorakos George 
Published: May 2021

Abstract
Background: Bone maturation depends mainly on locally produced estrogens by aromatization. Third generation aromatase inhibitors (AIs) are being widely used off-label to improve predicted adult height (PAH) in boys as well as in girls, either as monotherapy or in combination with growth hormone and/or puberty inhibition. They induce reverse binding inhibiting the activity of aromatase (a cytochrome P450 enzyme), which catalyzes the conversion of androstenedione and testosterone to estrone and estradiol, respectively. While numerous studies have shown that AIs delay bone maturation and improve PAH, data on near-adult height (NAH) of children treated with AIs are lacking. Aims: To compare results on NAH of boys treated with anastrozole either as monotherapy or in combination with pubertal inhibition (for at least 1yr at onset). Methods: 159 boys with advanced bone age (BA) and PAH &lt;170 cm that received anastrozole 1 mg/day p.o. either as monotherapy (n=76, group A) or as co-therapy with a GnRH analogue for at least 1yr and then as monotherapy (n=83, group B) until bone age of 15-16 yrs were included. Data on boys that reached NAH (BA at least 16 yrs with height velocity &lt;2 cm/yr) were analyzed: group A, n=16 with PAH 167.3 and TH 170.9 and group B, n=10 with PAH 165.5 and TH 171.7 cm. Measurements were made on the same height meter by the same examiner. The choice of therapeutic intervention was made randomly. Groups A and B did not differ in terms of age at intervention onset, TH or PAH. During treatment, they underwent a 6-month follow-up that included clinical examination, BA, and laboratory tests at 8:00 hrs (general blood count, lipid chart, LH, FSH, testosterone, estradiol, estrone, and complete calcium metabolism), with lumbar spine DEXA (Dual Energy X-ray Absorptiometry) and X-ray performed annually. Results: The duration of anastrozole treatment was 3.9 yrs in group A, and 4.6 yrs in group B (where the GnRHa was administered for at least 1 yr) and the median age at intervention onset was 11.04 and 11.8 yrs, respectively. Both groups had a statistically significant gain in NAH with no difference between them: for group A 3.6 cm (+0.53 SD, p=0.002) and for group B 4.8 cm (+0.71 SD, p=0.0007). Thus, distance from TH was finally 0 cm for group A and -1.5 cm (0.19 SD) for group B. According to the definition of NAH, the adult height of the two groups is expected to be about 2% higher. Follow-up showed no side effects on their biochemical or lipid profile, bone density and vertebral architecture. Conclusions: Anastrozole therapy is safe and effective in improving adult height in boys with advanced puberty and poor height prediction, either as monotherapy or in combination with pubertal inhibition.
Characteristics Associated With Diabetes Device Use Among Youth With Type 1 Diabetes
Authors: Chen Charlotte , Tinsley Liane , Volkening Lisa , Anderson Barbara , Laffel Lori M 
Published: May 2021

Abstract
Type 1 diabetes (T1D) is a common illness of childhood, requiring lifelong, daily complex management to prevent acute and chronic complications. Studies have shown that use of insulin pumps and continuous glucose monitors (CGM) offers benefit for glycemic control. However, such device use is not universal in adolescents. We aimed to compare baseline socio-demographic and diabetes characteristics associated with diabetes technology (pump and CGM) uptake and continued use in 13-17 year old teens with T1D. Data were derived from a multicenter clinical trial aimed at optimizing self-care and glycemic control in teens with T1D. Socio-demographic and diabetes data were collected quarterly by parent-youth interview and electronic medical record review prospectively over 18 months. Chi-square and t-tests compared characteristics of device and non-device users (pump vs no pump; CGM vs no CGM). The study sample comprised 301 teens (41% male) with mean±SD age 15.0±1.3 years, T1D duration 6.5±3.7 years, and A1c 8.5±1.1%. Most (65%) used a pump at entry or initiated pump therapy during the study; 35% used injection therapy at entry or stopped pump therapy. In contrast, 27% used a CGM at entry or started a CGM during the study, while 73% never used or stopped using CGM. Device users at entry and those who began use had similar characteristics, as did those who never used and those who discontinued device use. Pump users were more likely to use CGM than non-pump users (36% vs 10%, p&lt;.0001). Neither age, sex, nor T1D duration was related to pump or CGM use. Pump users (vs non-pump users) were less likely to have another medical condition (44% vs 59%, p=.01) and more likely to be non-Hispanic white (83% vs 61%, p=.0001); have family annual household income ≥$150,000 (34% vs 19%, p=.0003), private health insurance (92% vs 74%, p&lt;.0001), a parent with college education or higher (67% vs 46%, p=.0005), and a 2-parent household (88% vs 78%, p=.03). Pump users also had lower z-BMI (0.73±0.80 vs 0.97±0.79, p=.01), performed more frequent daily BG monitoring (4.8±1.8 vs 3.9±2.0, p&lt;.0001), and were less likely to have HbA1c ≥9% at initial and last visits (25% vs 43%, p=.005; 31% vs 49%, p=.01). CGM users (vs non-CGM users) were more likely to be non-Hispanic white (88% vs 70%, p=.009); have family annual household income ≥$150,000 (44% vs 23%, p=.0001), a parent with college education or higher (78% vs 53%, p=.0004), and private health insurance (95% vs 82%, p=.005). CGM users also performed more frequent daily BG monitoring (5.2±1.9 vs 4.2±1.9, p=.0002) and were less likely to have HbA1c ≥9% after 18 months (27% vs 42%, p=.03). In summary, we found distinct socio-demographic and diabetes-specific factors associated with device use in adolescents with T1D. These findings provide an opportunity to address barriers associated with device non-use in order to expand device implementation, especially in underserved adolescents with T1D.
22q13 Duplication in Newborn With Dysmorphic Features: The Role of SOX10 in Disorders of Sex Development
Authors: Hammoud Batoul , Baldauff Natalie Marie Hecht , Yatsenko Svetlana , Witchel Selma Feldman 
Published: May 2021

Abstract
Background: The 46,XX testicular disorder of sex development (DSD), also known as 46,XX male reversal, is a rare form of DSD and clinical phenotype shows complete sex reversal from female to male. The sex-determining region Y (SRY) gene can be identified in most 46,XX testicular DSD patients; however, approximately 20% are SRY-negative. Here we present a 2 week old with discrepant prenatal karyotype and infant phenotype.
Case: This 2-week-old had dysmorphic features (bitemporal narrowing, broad and flat nasal bridge, bilateral epicanthal folds) and multiple congenital anomalies; IUGR, hypertelorism, cleft lip and palate, ASD, small kidneys, sacral dimple. Physical exam revealed palpable inguinal masses and microphallus without hypospadias. Postnatal karyotype showed a 46,XX chromosome complement with duplication of 22q13-qter. He was negative for SRY gene by FISH. Microarray analysis confirmed the duplication encompassing the SOX10 gene. Lab evaluation showed LH 10.81 mIU/ml, FSH 3.21 mIU/ml and total testosterone 241 ng/dl. These values are consistent with activation of the hypothalamic–pituitary–gonadal axis during the neonatal period in males.
Conclusion: Our case along with previous cases supports the existence of a gene on chromosome 22q that can trigger testis determination in the absence of SRY. Potential mechanisms responsible for ovotesticular disorder in the XX (SRY−) individual could involve activation of testis specifying genes in the absence of SRY and/or inadequate expression of pro-ovary/anti-testis genes such as SOX 8 and SOX9. SOX10, a gene closely related to SOX9 and SOX8, and its overexpression has been suggested as a candidate for 46,XX DSD. Over-expression of SOX10 in mice resulted in the XX DSD. The spectrum of sex reversal phenotypes and gonadal asymmetry
seen in the Sox10 transgenic mice closely mirrors the range of gonadal and reproductive tract anomalies seen in cases of partial duplication of human chromosome 22q13. Although SRY-negative 46,XX testicular DSD is a rare condition, an effort to make an accurate diagnosis is important for the provision of proper genetic counseling and for guiding patients in their long-term management.
A Case of Autoantibody Negative Pediatric Diabetes Mellitus With Marked Insulin Resistance Concomitant With COVID-19: A Novel Form of Disease?
Authors: Martinez Alfonso Hoyos , Hicks Kelly Anne , Moorjani Tracy Patel , Bell Jennifer , Lin Yuezhen 
Published: May 2021

Abstract
Background: SARS-CoV-2 uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter human cells. This receptor is avidly expressed in the pancreatic islets, suggesting the virus may target β-cell function. 17% of adults with COVID-19 have evidence of pancreatic injury. There is a direct relation between insulin resistance and COVID-19 severity and mortality with patients with higher insulin resistance presenting with higher inflammatory response. Fulminant type 1 diabetes (fT1D) has abrupt onset of symptoms with insulinopenia without evidence of autoimmunity, usually preceded by viral illness. Clinical case: A 12-year-old Hispanic male presented with a week history of polyuria, polydipsia, headache, and fatigue but no weight loss, fever, cough, anosmia, or diarrhea. Laboratory testing revealed new onset diabetes with DKA with HbA1C 11.3%, blood pH 7.04, glucose 381 mg/dL, C-peptide 0.6 ng/mL, β-hydroxybutyrate 8.2 mmol/L, and a positive nasopharyngeal PCR for SARS-CoV-2 but no elevation of inflammatory markers (CRP, ESR and ferritin). There was evidence of mild pancreatic injury (lipase 179 U/L, n:15-110 U/L), and all autoantibodies for autoimmune diabetes were negative. He was pubertal (Tanner 3), non-obese (BMI Z score -0.3) and without acanthosis nigricans. Past medical and family history were non-contributory. He was treated with IV insulin at 0.1 u/kg/h until DKA resolved within 24 h then transitioned to subcutaneous insulin at 1 u/kg/day. He did not have signs of systemic inflammatory response, need for respiratory support, or glucocorticoids but had persistent hyperglycemia prompting an increase of insulin dosing and resumption of IV insulin. His insulin requirements continued to increase up to 4 u/kg/d with sustained hyperglycemia indicative of an exceptional state of insulin resistance. On Day 6 of admission metformin was initiated, on Day 9 insulin requirements declined and he was discharged on an insulin regimen close to 1.5 u/kg/d. Eventually metformin was discontinued on Day 20 after diagnosis without rebound in insulin requirements. On Day 28 he had improved glycemic control (HbA1c 8.8%) at insulin doses more appropriate for his age and pubertal status (0.9 u/kg/d). Clinical lessons: We report a case of autoantibody negative diabetes with pancreatic injury and marked insulin resistance unrelated to systemic inflammatory response associated with COVID-19. The clinical presentation was reminiscent of fT1D with the exception of exceptional insulin resistance. This is perhaps a novel form of diabetes mellitus not otherwise classified. Although the pathophysiology remains obscure, it is plausible that it is mediated by direct islet injury. Notably, there was a discordance between the severity of illness and insulin resistance suggesting an intrinsic effect of SARS-CoV-2 on glucose hemostasis.
Investigating the Role of Cholesterol and Lipid Trafficking in Mitotane Resistance in Adrenocortical Carcinoma
Authors: Warde Kate , Lim Yi-Jan , Beuschlein Felix , Hantel Constanze , Dennedy Michael Conall 
Published: May 2021

Abstract
Introduction: Adrenocortical Carcinoma (ACC) is a rare aggressive cancer which carries a poor prognosis. Adjuvant mitotane improves survival but is limited by poor response rates and resistance following tumour recurrence. Mitotane’s efficacy has been attributed to intracellular accumulation of toxic free cholesterol (FC) predominantly through inhibition of cholesterol storage through SOAT1. Yet SOAT1 specific inhibitors demonstrate inferior efficacy to mitotane in inducing ACC cell death. We hypothesize that mitotane’s efficacy to induce toxic FC accumulation in ACC cells is also mediated through enhanced breakdown of stored cholesterol within intracellular lipid droplets (LDs). Methodology: ATCC-H295R (mitotane sensitive) and MUC-1 (mitotane resistant) ACC cells were evaluated for neutral lipid content using BODIPY493/503 under baseline and cholesterol loaded conditions using Amnis ImageStream, additionally cells were treated with mitotane (H295R - 20, 40, 50µM; MUC1 - 50, 100, 200µM) for 6hr. Analysis of LDs using CE-BODIPY and FA-BODIPY identified cholesterol ester (CE) and triacylglycerol (TAG)-containing LDs, respectively. Lipid droplet-associated proteins (LDAPs) Perilipin (PLIN) 1–4 and hormone sensitive lipase (HSL) were evaluated using western blotting and PCR. Lipid uptake receptors; SRB1, LDLR, LRP1 and CD36 were measured by flow cytometry. Results: Mitotane treatment, within therapeutic range, decreased staining for LDs significantly in H295R. This was also reflected by decreased expression of LDAPs, PLIN1 and PLIN3. The decrease in H295R LDs was associated with increased activation of HSL (pHSL and LIPE). However, this effect was only evident in MUC-1 at supratherapeutic mitotane (200µM). H295R and MUC-1 demonstrated similar overall LD numbers at baseline and under cholesterol supplementation. Expression of PLIN3 was high in both cell lines, while PLIN1, PLIN2 and PLIN4 demonstrated distinct LD profiles in each. Investigation of LD content showed that H295R preferentially store CEs while MUC-1 store only TAG, irrespective of cholesterol-loading. Mitotane treatment significantly reduces both CE and TAG LDs in H295R and MUC-1. Expression of lipid uptake receptors also demonstrated significant variability between cell lines including SRB1 and LRP1. Conclusion: We highlight that lipolysis through LD breakdown and activation of HSL represents a putative additional mechanism for mitotane induced FC cytotoxicity in ACC. We also demonstrate significant differences in cholesterol handling and LDAPs between mitotane sensitive and mitotane resistant models, in particular, the absence of CE LDs in MUC-1. We therefore propose a mechanism of resistance to mitotane through absent CE storage. Further understanding of cholesterol and lipid handling in ACC offers novel therapeutic exploitation, especially in the setting of mitotane resistant disease.