bluuebunny/crossref_metadata_2025_split huggingface dataset:
RNA-Binding Protein Musashi-2 Inhibits Aldosterone Production
Authors: Bartholomay Kathryn , Baldwin Amber , Mukherjee Neelanjan 
Published: May 2021

Abstract
The adrenal cortex is the site of steroid hormone synthesis. These hormones control important physiological processes like metabolism, blood pressure and volume, and sexual characteristic development. While the signaling pathways, transcription factors, and steroidogenic enzymes are well-characterized, surprisingly little is known about the contribution RNA-binding proteins (RBPs). RBPs exert post-transcriptional control by interacting with specific elements within target mRNAs. Here we focus on the RBP, Mushashi-2 (MSI2), which binds to UAG sequences in the 3’UTR of its target transcripts. MSI2 is required for development of steroidogenic tissues which is consistent with its higher mRNA levels in human ovaries and testis. MSI2 also exhibits high expression levels in human adrenal tissue and the immortalized human adrenocortical cell line (H295R). Based on the compelling MSI2 expression pattern, we set out to determine the role of MSI2 on aldosterone production. Depletion of MSI2 using siRNA led to significantly lower aldosterone levels in H295R cells stimulated with AngII. We also employed an orthogonal loss-of-function approach by co-treating cells with AngII and increasing concentrations of Ro-08-2750 (Ro), a direct and selective inhibitor of MSI2-RNA interactions. Ro inhibited aldosterone production in a dose-dependent manner at 1 µM with almost complete inhibition at 5 µM. The molecular mechanism by which MSI2 regulates target RNA translation and/or decay is unknown. Moreover, whether MSI2 acts as a repressor or activator appears to be context dependent. Our goal is to determine the precise molecular mechanism by which MSI2 promotes aldosterone production. Specifically, we will identify MSI2 targets, temporally resolved consequences of MSI2 inhibition, and protein interaction partners. This work will impact our understanding of fundamental principles of RBP-mediated regulation, as well as novel regulatory mechanisms underlying human steroid hormone synthesis. Indeed, Ro (or further optimized compounds) may represent new therapeutic avenues for adrenal disease.
Spiral Steroids During Pregnancy - It’s All About Potassium
Authors: Chasalow Fred I 
Published: May 2021

Abstract
This presentation has two parts. The 1st section describes processes during pregnancy with unknown, underlying biochemistry. The 2nd section shows the role of spiral steroids (SS) in these processes. 1] Fetal nutrition is provided through the placenta. Plasma electrolytes are 145 mM Na+ and 3-5 mM K+. Fetal K+ requirements reach a maximum during the 3rd trimester and must be pumped into cells against the plasma- intracellular gradient. 2] During the 3rd trimester, aldosterone is present in fetal plasma but the signal for endothelial sodium channel (ENaC) synthesis is blocked, leading to fetal Na+ wasting. The mechanism is unknown. 3] After parturition, infants are fed by nursing. Human milk contains 100 mM K+ and 10 mM Na+. Newborn infants are Na+ wasting, despite normal levels of aldosterone. Na+ wasting ends during the 2nd week post-partum without change in serum aldosterone levels. Infant physiology changes from K+ saving to Na+ saving by an unknown mechanism.4] Pre-eclampsia syndrome (proteinuria and hypertension during the 2nd half of pregnancy) affects 3-5 % of pregnant women. These symptoms usually resolve after parturition. 5] Women who have had pre-eclampsia have long-term, excess risk of cardiac and renal disorders.
In 2018, we discovered Ionotropin, a SS. SS are phosphocholine esters of steroids with a lactone E-ring, similar to that of spironolactone. SS compounds function as aldosterone antagonists and regulate the NaK-ATPase. SS are involved in each of the 5 steps.1] SS are present in high levels in cord serum and were probably present in fetal plasma throughout gestation. SS stimulate the NaK-ATPase to pump K+ into cells against the gradient, just as does spironolactone.2] SS interfere with aldosterone signaling, just as does amiloride. This leads to increased fetal Na+ wasting, which becomes amniotic fluid.3] SS disappear from the infant circulation during the 1st week after parturition and decrease to adult levels during the 2nd week post-partum. Simultaneously, Na+ wasting ends and growth resumes.4] Women with pre-eclampsia have excess precursors for SS. These would be converted to SS in the fetal-placental unit and, during the 3rd trimester, diffuse into the maternal circulation and could cause pre-eclampsia.5] Ouabain, a plant toxin with a lactone E ring, causes renal and cardiac disorders in rat models. In women with pre-eclampsia, persistent excess SS may cause long-term damage.
During gestation, the fetus requires K+ for growth. Our theory is, if (when) a fetus has inadequate K+ (hypokalemia), [a] the mother is signaled to produce SS precursors, [b] the feto-placental unit converts the precursors to SS, [c] fetal SS increase K+ transfer into tissues, and [d] excess SS transfuse back into the maternal circulation and damage maternal organs. We propose that pre-eclampsia is a side effect of fetal efforts to increase supply of K+. There are many possible origins of fetal hypokalemia.
Time of Day Regulates Renal Mineralocorticoid Receptor Transcriptional Control of Electrolyte Balance
Authors: Kanki Monica , Morgan James , Fuller Peter J , Young Morag Jennifer 
Published: May 2021

Abstract
The mineralocorticoid receptor (MR) has an established role in blood pressure control and cardiovascular homeostasis via many actions in the heart and kidney. We recently identified a role for the MR in controlling the circadian clock in cardiac cells and demonstrated that time-of-day impacts MR activation in the heart. While time dependent behaviours such as upright posture and fluid intake control aldosterone release via the renin-angiotensin-aldosterone system (RAAS), we hypothesise that the circadian clock controls aldosterone signalling by modifying MR transcriptional outcomes. Two established MR target genes and core circadian clock genes are period 1 (Per1) and period 2 (Per2). We have previously shown that a bolus dose of aldosterone (i.p.) induced cardiac expression of Per1 and Per2 in wildtype mice treated at 8AM (start of rest period) but not when administered at 8PM (start of active period). Whether MR activation in the kidney is similarly dependent on time of day and aligns with MR actions in the heart remains to be assessed. We also sought to determine if the MR directly regulates the molecular clock in a ligand-dependent manner. In contrast to the heart, renal Per1 and Per2 expression was not upregulated at four hours following aldosterone administration at either 8AM or 8PM. Interestingly, aldosterone administered at 8AM, but not at 8PM, significantly down-regulated expression of the circadian clock gene, nuclear receptor reverse strand c-ERBA (ReverbA); ReverbA was not regulated in the heart following aldosterone treatment. Investigation of renal-MR target genes involved in sodium ion transfer, showed a down-regulation of epithelial sodium channel 1 alpha (ENaC-α), by aldosterone administered only at 8AM. Conversely, aldosterone administered at 8AM induced gene expression of FKBP Prolyl Isomerase 5 (Fkbp5), an immunophilin that is important in nuclear trafficking of the MR. These data provide new insights for tissue sensitivity to MR activation in the heart and the kidney, which may play differing roles in contributing to regulating circadian patterns of several cardiovascular and renal parameters. Moreover, our data suggest that in addition to the accepted entrainment role of the glucocorticoid receptor (GR), the MR may also play a role in the entrainment of the circadian clock in peripheral tissues. Ongoing mechanistic studies will determine whether the MR can directly control the periodicity of the renal cellular clock, as it does in the heart. Key outcomes will be assessed in population databases to determine the clinical relevance of these findings. We propose that understanding of time-of-day dependent vulnerability to MR signalling in the heart versus the kidney may offer the rationale for the development of novel temporal or tissue specific-MR modulators in the management of cardiovascular disease.
Vitamin D and Lumisterol Hydroxyderivatives Can Act on Liver X Receptors (LXRs)
Authors: Slominski Andrzej , Kim Tae-Kang , Qayyum Shariq , Song Yuwei , Janjetovic Zorica , Oak Allen , Slominski Radomir , Raman Chander , Stefan Joanna , Mier-Aguilar Carlos , Atigadda Venkatram , Crossman David , Golub Andriy , Bilokin Yaroslav , Tang Edith , Tuckey Robert C , Jetten Anton M , Song Yuhua 
Published: May 2021

Abstract
New pathways of vitamin D3 (D3) activation initiated by CYP11A1 and involving other CYPs have been discovered. At least 15 hydroxyderivatives, including 20(OH)D3 as the major product, are generated by these pathways (1,2) with some being present in human serum, epidermis, and pig adrenals. CYP11A1 can also metabolize 7-dehydrocholesterol to produce 7-dehydropregnenolone, which can be further modified by steroidogenic enzymes generating Δ7-steroids (1,2). Lastly, CYP11A1 and CYP27A1 act on lumisterol (L3) producing at least 9 biologically active derivatives (1,2). Thus, new pathways generating a large number of biologically active secosteroids and lumisterol-derivatives have now been described. These compounds interact with the vitamin D receptor (VDR), retinoic acid receptors (RORs) α and γ, and the aryl hydrocarbon receptor (AhR)(1). These findings challenge dogmas that lumisterol is biologically inactive and that 1,25(OH)2D3 is the only active form of D3 exerting its effects exclusively through interaction with the VDR. In view of the above and since liver X receptors (LXRs) can be activated by oxysterols, we investigated the interactions of novel products of L3 and D3 metabolism with LXRs. Molecular docking, using crystal structures of the ligand binding domains (LBDs) of LXRα and β, revealed high docking scores for L3 and D3 hydroxymetabolites, like those of the natural ligands, predicting good receptor binding. RNA sequencing of murine dermal fibroblasts stimulated with D3-hydroxyderivatives revealed LXR as the second major nuclear receptor signaling pathway for several D3-hydroxyderivatives, including 1,25(OH)2D3. The involvement of LXRs was validated by the induction of several genes downstream of LXR. Furthermore, L3 and D3-hydroxyderivatives activated an LXR-response element (LXRE)-driven reporter in CHO cells and human keratinocytes. For keratinocytes, enhanced expression of LXR target genes was also observed supporting the involvement of LXR. Importantly, L3 and D3 derivatives showed high affinity binding to the LBD of the LXRα and β in LanthaScreen TR-FRET LXRα and β coactivator assays. The majority of metabolites functioned as LXRα/β agonists; however, 1,20,25(OH)3D3, 1,25(OH)2D3, 1,20(OH)2D3 and 25(OH)D3 acted as inverse agonists of LXRα, but as agonists of LXRβ. Molecular dynamics simulations performed for selected compounds, including 1,25(OH)2D3, 1,20(OH)2D3, 25(OH)D3, 20(OH)D3, 20(OH)L3 and 20,22(OH)2L3, showed overlapping and different interactions with LXRs. Identification of D3 and L3 derivatives as ligands for LXRs changes the accepted paradigms on their biological role and mechanism of action. 1. Cell Biochem Biophys. 2020;78(2):165-180. 2. J Steroid Biochem Mol Biol. 2019;186:4-21.
Daily Versus Weekly Levothyroxine Tablet Replacement in Adults With Hypothyroidism: A Meta-Analysis
Authors: Chang Chiu Harold Henrison , Larrazabal Ramon Bagaporo , Uy Angelique Bea Cariaga , Jimeno Cecilia A 
Published: May 2021

Abstract
Background and Objectives: Hypothyroidism is a common hormone deficiency with a prevalence ranging from 4-5% worldwide. It is a very treatable condition with treatment in the form of thyroid hormone replacement, with an overall excellent prognosis if patients are compliant to regular treatment. Daily levothyroxine (LT4) is the treatment of choice and standard of care, sufficient to restore the thyroid stimulating hormone (TSH) to the normal range. For many patients, daily and lifelong therapy is required, and compliance/adherence then becomes a major issue. In such cases, weekly replacement may be a suitable alternative in terms of improving patient compliance. In this study, we aimed to determine the efficacy and safety of weekly versus daily levothyroxine replacement in patients with hypothyroidism.
Methods: Electronic databases were searched, supplemented with manual searches. Two reviewers independently screened the abstracts, reviewed full-text papers, independently critically appraised the quality of included studies and abstracted the data. A meta-analysis was performed using the random-effects model on randomized controlled trials (RCTs) that reported standard doses of daily versus weekly levothyroxine administration in the treatment of hypothyroidism. The primary outcome is the difference in serum TSH levels between daily versus weekly levothyroxine administration, while secondary outcomes included clinical symptoms and adverse events using the hypothyroidism symptom scale.
Results: The study included two randomized trials (N = 109) in the primary analysis. The difference in TSH levels was 1.78 mIU/mL higher (95% CI: 1.28, 2.28; P &lt; 0.00001) at 6 weeks and 1.22 mIU/mL higher (95% CI: 0.76,1.67; P &lt; 0.00001) at 12 weeks for the weekly replacement regimen, respectively. There was no significant heterogeneity noted between the two groups. There was no significant difference in terms hyperthyroid symptoms and adverse events measured by the hypothyroid symptom scales and echocardiographic parameters, respectively, before and after LT4 within each group.
Conclusions: Our results showed that weekly LT4 administration has less suppression of TSH levels, while still remaining within the reference range of normal. It may be an alternative for patients especially in setting of noncompliance. However, more randomized trials with larger sample sizes and a longer duration of follow-up are needed to firmly establish the definite role of weekly LT4.
The Pitfalls of Using Research Grade Immunoassays for Thyroid Health Measurements
Authors: Boggs Ashley S P , Galligan Thomas M , Kemp Jennifer V , Ravinder Singh J , Grebe Stefan Karl Gunther 
Published: May 2021

Abstract
Introduction: Immunoassay technology is subject to matrix interferences that can produce inaccurate results and incorrect conclusions when using samples not previously validated. While many commercially available research grade (RG) immunoassay kits are available, caution should be applied when using RG kits for thyroid health assessments, particularly on samples from pregnant individuals whose blood chemistry is unique to non-pregnant individuals. Question: Do RG immunoassay kits reliably provide precise and accurate measurements of thyroid health biomarkers in serum Standard Reference Materials (SRMs) from pregnant and non-pregnant donors? Methods: T4, T3, rT3, Tg and TSH measurements were conducted on SRMs 971, Hormones in Frozen Human Serum, and SRM 1949, Frozen Prenatal Serum, using RG immunoassay kits. When available, performance was assessed against validated FDA approved immunoassays or mass spectrometric (MS) methods. Results: RG kits were variable, inaccurate, or imprecise for four of the six biomarkers assessed. RG kit total thyroid hormone measurements overall performed comparably to MS methods, except rT3 measurements, which were twofold greater than mass spectrometric measurements (971M RG mean = 0.51 ng/mL, MS mean = 0.20 ng/mL, p &lt; 0.0001; 971F RG = 0.48 ng/mL, MS = 0.18 ng/mL, p &lt; 0.0001) and had CVs over 30 %. RG kit Tg measurements varied sometimes by as much as tenfold (971M means of 6.50 ng/mL up to 63.3 ng/mL, p &lt; 0.0001; 971F means of 0.350 ng/mL up to 14.5 mg/mL, p &lt; 0.0001). TSH values differed by RG kit manufacturer (971M means of 1.27 µIU/mL up to 1.82 µIU/mL, p &lt; 0.0001; 971F means of 1.36 µIU/mL up to 2.27 µIU/mL, p &lt; 0.0001) and by dilution scheme using the same manufacturer with one case indicating a diagnosis of hypothyroid versus normal TSH levels (1949 non-pregnant undiluted mean = 2325 pg/mL, half dilution mean = 1631 pg/mL, p &lt; 0.0001). Conclusions: RG immunoassays are often used for research projects because they do not require expensive equipment and are simple to conducted. However, we demonstrate here that not all kits are accurate for all patient samples. By utilizing a matrix matched SRM with well-defined quantities of thyroid health biomarkers, one can assess method accuracy, making measurements from different methods comparable. Thereby, data can be harmonized to contribute reliable data on thyroid biomarkers to advance the field of thyroid health.
The Efficacy of Teprotumumab for Chronic, Progressive Thyroid Eye Disease Greater Than 9 Months
Authors: Sears Connie Martin , Kang Julia Y , Cockerham Kimberly , Kossler Andrea L 
Published: May 2021

Abstract
Purpose: To evaluate the efficacy of teprotumumab for the treatment of patients with chronic, progressive Thyroid Eye Disease (TED) who do not meet the inclusion and exclusion criteria used in prior clinical trials. Methods: This is a prospective case series of patients with chronic, progressive TED, defined as ocular symptoms of more than 9 months prior to treatment initiation with teprotumumab, an insulin-like growth factor I receptor inhibitor (10 mg/kg for the first infusion then 20 mg/kg for subsequent infusions, every three weeks for a total 8 infusions). This study included patients with a Clinical Activity Score (CAS) greater than or equal to 4. Data collected included patients’ age, sex, race, baseline thyroid status, thyroid stimulating immunoglobulin, CAS, smoking status, best corrected visual acuity, intraocular pressure, Schirmer’s, pupil exam, extraocular motility, Gorman diplopia score, and exophthalmometer measurements. Pre-infusion and post-infusion external photos in all 9 gazes, orbital imaging (CT or MRI), Humphrey visual fields and optical coherence tomography were obtained when possible.
Results: Ten patients with chronic TED were treated with teprotumumab either as a primary treatment or following other interventions including oral prednisone, intravenous methylprednisolone, orbital radiation, orbital decompression and/or eyelid surgery. The age range was 55 to 79, with 5 female patients and 5 male patients. Four of the patients were African American and 6 of the patients were White/Non-Hispanic. The number of years the patients had been diagnosed with Graves’ disease ranged from 4 to 33 years, with ocular involvement spanning 4 to 15 years. Patients had CT scans of the orbits to document muscle size prior to initiation, and external photos were taken at each visit. All patients improved subjectively after the first infusion and objectively by the fourth infusion. We will report the long-term follow-up after 8 infusions at ENDO2021.
Discussion: Teprotumumab is a fully human monoclonal IgG1 antibody that blocks the IGF-1 receptor, disrupting the cell-to-cell signaling of the immunologic cascade and is independent of the duration or severity of TED. At this time, FDA approval for teprotumumab is for the treatment of TED without any specific restrictions. However, our teprotumumab-treated patients included those who would have been excluded in the phase 2 and 3 clinical trials due to: (1) poor thyroid control, (2) previous orbital surgery, (3) previous therapeutic interventions including steroids and radiation, (4) visual loss due to compressive optic neuropathy or exposure keratopathy and (5) chronic TED greater than 9 months. In our series, we found that teprotumumab was as effective in a wider population of chronic TED patients than included in the clinical trials.
The Immune Microenvironment of Hashimoto’s Thyroiditis Regulates the Glycosylation Modification of IgG
Authors: Cao Yedi , Song Zhijing , Gong Yan , Zhao Keli , Zhao Xue , Huang Youyuan , Qu Chenxue , Li Yan , Gao Ying , Zhang Junqing , Guo Xiaohui 
Published: May 2021

Abstract
Objectives: Elevation of anti-thyroglobulin antibodies that are primarily IgG isotype is a hallmark of Hashimoto’s thyroiditis (HT). As for IgG,it bears two conserved repertoire of N-linked glycans attached to its crystallizable fragment (Fc) at the 297 asparagine residue (Asn297). In our previous study, we found that serum TgAb IgG from HT patients exhibits higher glycosylation levels than those observed from healthy controls. Previous studies confirmed that imbalance of Th1/Th2 and Th17/Treg leading to altered immune microenvironment with elevation of certain cytokines was found in the thyroid tissue of HT, including IFN-γ, TNF-α, IL-21, IL-17A, IL-6, BAFF, APRIL. Thus, the aim of our study was to investigate the influence of the elevated cytokines on the differentiation process of B cells and the glycosylation levels of IgG. Methods: We formed a two-phase culture system in vitro to promote B cells to differentiate to antibody-secreting cells (ASCs). In the process of cell culture, B cells were co-cultured with cytokines as followed: IFN-γ, TNF-α, IL-21, IL-17A, IL-6, BAFF and APRIL. Flow cytometry was performed to identify the percentage of plasmablasts (CD38+CD27high) and plasma cells (CD20-CD138+). ELISA was used to measure the yield of IgG in culture supernatants. The glycosylation levels of secreted IgG under different stimulation conditions were detected by lectin microarray. Results: We found that IL-21, TNF-α and BAFF can significantly promote the differentiation of B cells into ASCs in vitro culture system, and augment the production of IgG to over 4-fold. In addition, cytokines affected the glycosylation modification profile of IgG diversely: 1) IL-21, IL-17A, TNF-α, BAFF significantly increased the glycosylation level of sialic acid of total IgG; 2) IFN-γ significantly increased the level of galactose; 3) IL-21, IL-17A, IFN-γ, BAFF, and APRIL significantly increased the level of mannose; 4) IL-6 significantly decreased the level of sialic acid, galactose and mannose; 5) IL-17A, IFN-γ, TNF-α, BAFF significantly increased the level of GalNAc that was a component of O-Glycan,which only exists in the hinge region of IgG3 subclass. Conclusions: The abnormally elevated cytokines in microenvironment participated in the regulation of B cell terminal differentiation process and glycosylation level of IgG, thereby involving in the pathogenesis of AITD.