bluuebunny/crossref_metadata_2025_split huggingface dataset:
SAT001 Membrane Progesterone Receptors Induce Glycogenolysis In Uterine Epithelial Cells
Authors: Berg Malia , Dean Matthew 
Published: October 2023

Abstract
Disclosure: M. Berg: None. M. Dean: None.
In humans and cattle, 40-60% of pregnancies fail, with most losses occurring prior to or during implantation. Throughout this time, the developing embryo needs glucose to survive. The demand for glucose increases dramatically at the morula stage as the embryo enters the uterus. Yet, how glucose is regulated to match the changing needs of the embryo is unclear. Glycogen is composed of thousands of glucose molecules and is present in the uterine epithelium of many species. Our lab has shown that the glycogen content of the bovine uterine epithelium was lower during the luteal phase than at estrus. Therefore, our objective was to elucidate the role of progesterone in glycogenolysis of uterine epithelial cells. Bovine uterine epithelial (BUTE) cells were treated with insulin-like growth factor 1 (IGF1) to stimulate glycogenesis, followed by treatment for 48 hours. Progesterone decreased glycogen levels in BUTE cells by 99% (P=0.0002). RU486 did not block progesterone's effect, indicating that the effect of progesterone was not mediated by nuclear progesterone receptors (nPR). Thus, we hypothesized that the effect of progesterone is mediated by membrane progesterone receptors (mPRs). RT-PCR confirmed that BUTE cells expressed all five mPRs (α, β, γ, δ, and ε). Like progesterone, a specific mPR agonist (Org OD 02-0) decreased glycogen levels in BUTE cells by 99% (P&lt;0.0001). Indicating a potential for mPRs to regulate glycogen in vivo, immunohistochemistry showed that the bovine uterine epithelium expressed high levels of mPRα. These results validate that progesterone is acting through mPRs to stimulate glycogen breakdown in BUTE cells. Once progesterone binds to the mPR, two signaling pathways can be activated, cAMP and AMPK. Neither progesterone nor Org OD 02-0 changed intracellular cAMP concentrations. In agreement, the adenylyl cyclase activator forskolin increased cAMP concentration but did not decrease glycogen. BUTE cells treated with progesterone had increased pAMPK levels at 24 hours compared to the control (P=0.0001). BUTE cells treated with an AMPK activator (D942) had a decrease in glycogen (P=0.0051). Supporting these results in vivo, pAMPK levels in the uterine epithelium were high in the bovine uterine epithelium during the luteal phase when glycogen levels were low. In a human model, progesterone and Org OD 02-0 decreased glycogen levels to a similar extent in Ishikawa cells that express mPRs but not nPRs (P=0.0027). In conclusion, progesterone acting through the mPR and AMPK to stimulate glycogenolysis may play a crucial role in providing glucose to endometrial tissue or the growing embryo.
Presentation: Saturday, June 17, 2023
SAT012 In Vivo Mapping Of Subcellular Proteomes In Mice
Authors: Kim Kwang-eun , Park Isaac , Kim Jeesoo , Kim Jong-Seo , Rhee Hyun-Woo , Myoung Suh Jae 
Published: October 2023

Abstract
Disclosure: K. Kim: None. I. Park: None. J. Kim: None. J. Kim: None. H. Rhee: None. J. Suh: None.
To facilitate the understanding of metabolic changes associated with aging and disease processes we have developed new in vivo tools that enable tissue-specific profiling of subcellular proteomes. First we describe a method to profile in vivo mitochondrial proteomes utilizing transgenic mice expressing mitoAPEX, a peroxidase-based proximity labeling enzyme containing a mitochondrial matrix targeting sequence. Upon label activating conditions, mitoAPEX rapidly (&lt;1 min) catalyzes production of biotin radicals which biotinylate proteins within a 20 nm radius. Mass analysis of biotinylated proteomes from proximity labeled mitoAPEX mouse tissues confirmed specific and efficient labeling of the mitochondrial proteome and revealed tissue-specific patterns of the matrix proteome. The labeled muscle proteomes from young and old mitoAPEX mice revealed significant changes in the quantity and composition of protein species. Of these, RTN4IP1, was shown to be downregulated in muscle tissue of old mice. However, in contrast to previous reports, our analysis of RTN4IP1 shows RTN4IP1 is localized to the mitochondrial matrix and not the outer membrane. In addition to the mitoAPEX mice, we generated another in vivo proximity labeling tool iSLET (in situ Secretory protein Labeling via ER-anchored TurboID) which labels secretory pathway proteins, via proximity labeling activity by ER lumen targeted TurboID, an engineered biotin ligase. We expressed iSLET in the mouse liver and demonstrate efficient labeling of the liver secreted proteome which could be tracked and identified within circulating blood plasma. We expect mitoAPEX and iSLET mice will facilitate our understanding of mitochondrial function and interorgan communication networks in aging and associated disease processes.
Presentation: Saturday, June 17, 2023
OR22-01 Sex Differences In Enterohepatic Circulation Mediated By Hepatic FXR And SHP Depletions In Mice
Authors: Reichardt Francois , Galuke Christopher A , Anakk Sayeepriyadarshini 
Published: October 2023

Abstract
Disclosure: F. Reichardt: None. C. Galuke: None. S. Anakk: None.
Bile acids are transported via the portal vein to the liver after their absorption through the distal intestine mucosa. In hepatocytes, they activate the nuclear receptors, Farnesoid-X-receptor (FXR) and Small Heterodimer Partner (SHP), which exert negative feedback on bile acids synthesis. The aim of this project is to decipher how hepatic FXR/SHP deletion impact BA composition, enterohepatic circulation, and its cross talk with the gut microbiota. Enterohepatic circulation was investigated by measuring total bile acids in intestinal, hepatic tissues and fecal tissues and in portal and systemic blood, using 3-5 months-old fed female and male wildtype (WT) and LDKO (liver specific FXR/SHP depletions) mice. We also examined if intestinal dysmotility mediated the alteration in bile acids absorption by following methylene blue dye progression in overnight fasted WT and LDKO mice. Portal and systemic total bile acids concentrations were higher in LDKO mice compared to WT, despite of no significant change in hepatic and intestinal levels. Sex differences were seen only in LDKO mice with higher systemic and portal concentrations in females compared to males. Intestinal transit time and fecal total bile acids were equivalent between all mice. Intriguingly, we observed that portal bile acids levels correlated to the liver weight to body weight ratio and to cecum weight to body weight ratio, suggesting that bile acids alterations within the enterohepatic loop could be associated to cecal dysbiosis. These preliminary findings highlight altered BA recirculation in LDKO mice compared to WT, but also between females and males LDKO. We hypothesize that these sex-specific modifications are associated with intestinal dysbiosis impacting bile acid compositions, and to test this we will be analyzing the influence of sex, and BA composition on the microbiome. Further investigations using fasted or HFD-fed LDKO and WT mice will help us to better understand the role of hepatic FXR and SHP in bile acids circulation between intestine and liver. Acknowledgement: University of Illinois Urbana-Champaign, American Cancer Society and Cancer Center at Illinois Seed grant
Presentation: Saturday, June 17, 2023
OR22-02 Sex Differences In 11-Oxyandrogen Correlations With Adipose Tissue Compartments
Authors: Turcu Adina F , Lee Chaelin , Joseph Auchus Richard 
Published: October 2023

Abstract
Disclosure: A.F. Turcu: None. C. Lee: None. R.J. Auchus: None.
Background: Adrenal-derived 11-oxygenated C19 steroids (11-oxyandrogens) are mediators of adrenarche and several disorders of androgen excess. 11β-hydroxyandrostenedione (11OHA4), a major adrenal product, is converted to the bioactive androgen 11-ketotestosterone (11KT) via 11β-hydroxysteroid dehydrogenase type 2 (HSD11B2) and aldo-keto reductase 1C3 (AKR1C3). The adipose tissue expresses both AKR1C3 and the type 1 isoenzyme HSD11B1, which participate in modulating peripheral androgen production. The impact of adipose tissue compartments on circulating 11-oxyandrogens in men and women is not well understood. Setting and Participants: A subset of participants in the population-based, multiethnic cohort of the Dallas Heart Study, phase 2 were included. Measurements: All participants underwent subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) measurements by abdominal magnetic resonance imaging (MRI). Testosterone (T), androstenedione (A4), and four 11-oxyandrogens were quantified simultaneously in peripheral blood using liquid chromatography-tandem mass spectrometry. Associations between adipose tissue compartments and androgens were assessed by Pearson correlation tests, stratified by sex. Results: Of the 498 participants included, 282 (56.6%) were women, 49% were Black, 32% White, 18% Hispanic. The mean age of study participants was 54.3 +/- 10.7 years, and mean BMI was 30.9 +/- 6.9 kg/m2. There was no difference in age and race distribution between men and women. 11OHA4, 11β-hydroxytestosterone (11OHT), and 11KT correlated positively with BMI in women (R2 = 0.15-0.18, p&lt;0.01) but not in men. Conversely, T correlated negatively with BMI only in men (R2 = -0.26, p&lt;.0001). 11OHA4 and 11OHT correlated directly with both SAT and VAT in women (R2 = 0.14-0.18, p&lt;0.05), and a similar correlation was observed between 11OHT and VAT in men. Conversely, T and A4 displayed an inverse correlation with SAT and/or VAT in both sexes. 11KT and 11-ketoandrostenedione (11KA4) displayed no association with adipose tissue compartments. Conclusions: 11OHA4 and 11OHT, but not their respective 11-keto metabolites correlate with all adipose tissue compartments in women, while T correlates inversely with adiposity in men, particularly with SAT. Sexual dimorphism in adipose tissue and impact on androgen modulation might contribute to sex differences in cardiometabolic morbidity.
Presentation: Saturday, June 17, 2023
OR33-03 Circadian Analysis Of Nuclear Receptor Protein Expression In Mouse Liver Reveals Peak COUP-TFII, EAR2, LXRa, LXRb And TR4 Expression At The End Of The Mouse Active Cycle ZT0
Authors: Fadi Saikali Michael , Xu Li Jia , Cummins Carolyn L 
Published: October 2023

Abstract
Disclosure: M.F. Saikali: None. J. Li: None. C.L. Cummins: None.
Nuclear Receptors (NRs) are a family of ligand-mediated transcription factors that control the expression of genes involved in a wide range of physiological processes. The ability to study NRs quantitatively at the protein level has been hindered by the poor quality of available antibodies, as well as the lack of available high throughput methods to study more than a few receptors at a time. To address this need, we have developed a mass spectrometry based targeted proteomic assay to quantify the absolute amounts of NR protein in mouse livers over a circadian period. C57Bl6 mice were sacrificed at 12 weeks every 4 hours from zeitgeber time ZT0 to ZT20. Their livers were processed using a transcription factor extraction protocol1. For each sample, 100 µg nuclear protein was processed using the NR assay described previously2. Briefly, the lysate is digested following the Multi-Enzyme Digestion Filter Aided Sample Preparation protocol. The sample is then spiked with 50 fmol of our internal standard mix. This mix is composed of 66 carefully selected peptides that account for 49 NRs, 11 histone peptides, and 6 circadian clock proteins. The resulting digest that contains the internal standards is desalted on C18 tips, followed by separation on an EASY-Spray C18 column (75 um x 50 cm, 3Å), and analyzed on a Thermo QExactive HF mass spectrometer in parallel reaction monitoring mode. We used this assay to quantify the changes in NR protein expression in mouse livers for 9 receptors; RORα, CAR, COUP-TFII, EAR2, GR, LXRα, LXRβ, PPARα, and TR4. RORα showed peak nuclear expression at ZT0, with a peak-to-trough change of 2.2-fold. COUP-TFII, EAR2, LXRα, LXRβ, and TR4 also showed peak expression at ZT0 (the end of the active phase) with changes of 3.3, 2.4, 1.7, 2.0, and 1.6-fold respectively. GR showed peak nuclear expression at ZT12 with a 1.8-fold change. CAR showed 2.3-fold peak expression at ZT16. COUP-TFII, EAR2, and TR4 are considered orphan receptors with no known endogenous ligands. These data will be used to uncover endogenous tissue-specific ligands by analyzing extracts isolated at the zeitgeber time that the receptor is maximally expressed. Sources of Research Support: This project has been supported by the J. P. Bickell Foundation and the Canadian Institutes of Health Research New Frontiers Research Fund and International Development Research Fund. 1.Gillespie, M. A. et al. Absolute Quantification of Transcription Factors Reveals Principles of Gene Regulation in Erythropoiesis. Mol Cell (2020) doi:10.1016/j.molcel.2020.03.031.2.Saikali, M. F. & Cummins, C. L. Quantifying the Protein Levels of All Nuclear Hormone Receptors by Mass Spectrometry. J Endocr Soc5, (2021).
Presentation: Sunday, June 18, 2023
THU659 Plasmapheresis As A Therapy For Amiodarone-induced Thyrotoxicosis: A Case Report And Literature Review
Authors: Loughner Chelsea L , Lynn Bononi Patricia 
Published: October 2023

Abstract
Disclosure: C.L. Loughner: None. P.L. Bononi: None.
Introduction: Amiodarone-induced thyrotoxicosis (AIT) is a common cause of hyperthyroidism. Treatment consists of stabilizing the thyroid followed by thyroidectomy1. Plasmapheresis has been suggested as a temporary measure when illness severity precludes standard therapy. Here, we review current literature and provide a case of AIT treated with plasmapheresis. Clinical Case: A 71-year-old male was seen for progressive subclinical hyperthyroidism, diagnosed several years prior. Three years ago, he started taking amiodarone for paroxysmal ventricular tachycardia. One month prior to admission, he was admitted for decompensated heart failure and had TSH 0.02 mcU/mL (n 0.4-4.4 mcU/mL), free T4 4.48 ng/dL (n 0.7-1.9 ng/dL), and free T3 2.58 pg/mL (n 2.0-4.4 pg/mL). Amiodarone was halved after being diagnosed with AIT. Methimazole and prednisone were started. He was readmitted one month later. TSH was undetectable at admission and free T4 was 4.04 ng/dL. Amiodarone was discontinued. His hepatic enzymes were elevated, necessitating the discontinuation of methimazole. His heart failure precluded thyroidectomy. Due to the limitation of treatment options, the patient was given three rounds of plasmapheresis to reduce thyroid hormone levels. His free T4 dropped to 2.81 ng/dL. The patient underwent total thyroidectomy once stable. His free T4 was 1.73 after and was started on levothyroxine. Clinical Lesson: When patients are unable to have standard treatment of AIT, other options must be considered. While there are some case reports3,4, there are currently no RCTs for plasmapheresis for AIT. In the case of this patient with cardiac complications from AIT and concurrent hepatic impairment, plasmapheresis provided stability of thyroid hormones until surgical correction could be achieved.
References: 1. Tizianel I, Sabbadin C, Censi S, et al. Therapeutic Plasma Exchange for the Treatment of Hyperthyroidism: Approach to the Patient with Thyrotoxicosis or Antithyroid-Drugs Induced Agranulocytosis. J Pers Med. 2023;13(3):517. Published 2023 Mar 13. doi:10.3390/jpm13030517. 2. Padmanabhan A, Connelly-Smith L, Aqui N, et al. Guidelines on the Use of Therapeutic Apheresis in Clinical Practice - Evidence-Based Approach from the Writing Committee of the American Society for Apheresis: The Eighth Special Issue. J Clin Apher. 2019;34(3):171-354. doi:10.1002/jca.217053. Miller A, Silver KD. Thyroid Storm with Multiorgan Failure Treated with Plasmapheresis. Case Rep Endocrinol. 2019;2019:247584. 3. Published 2019 Oct 9. doi:10.1155/2019/24758434. Zhu L, Zainudin SB, Kaushik M, Khor LY, Chng CL. Plasma exchange in the treatment of thyroid storm secondary to type II amiodarone-induced thyrotoxicosis. Endocrinol Diabetes Metab Case Rep. 2016;2016:160039. doi:10.1530/EDM-16-0039.
Presentation: Thursday, June 15, 2023
THU660 Pericardial Effusion Secondary To Uncontrolled Hypothyroidism
Authors: Deniz Cain Gozde , Goldsmith Daniel , Ruel Ewa 
Published: October 2023

Abstract
Disclosure: G.D. Cain: None. D. Goldsmith: None. E. Ruel: None.
Introduction: Hypothyroidism is an endocrine disorder that can effect multiple organ systems. Pericardial effusion is a rare but significant complication of hypothyroidism. Early diagnosis and management of the pericardial effusion is important to prevent progression to cardiac tamponade and hemodynamic instability. We present a case of pericardial effusion in a 62-year-old female with hypothyroidism and medication non-compliance. Case Presentation: A 62-year-old female with past medical history of hypothyroidism and hypertension presented to the emergency department due to altered mental status. Patient's family reported that she had a progressive decline in her speech for more than a month, until she became completely aphasic on day of presentation, which prompted them to call EMS. Patient had diminished exercise capacity, dyspnea on exertion, bilateral pedal edema over the past couple of months. Family also reported that the patient had been depressed and negligent of her health following the passing of her husband three years ago. She was not taking her medications including levothyroxine. In the ED, patient was hypertensive, blood pressure was 190/100. CT head stroke protocol did not show stroke. Chest x-ray showed massive cardiac enlargement. 2D-Echo confirmed a large pericardial effusion without cardiac tamponade. Thyroid function panel showed high TSH of 77, free T4 was undetectable. Pericardial effusion was most likely secondary to hypothyroidism. Myxedema coma was suspected due to patient's progressive mental deterioration. Patient also had myxedema of bilateral lower extremities. IV levothyroxine and hydrocortisone were started with improvement in her symptoms. A pericardial window was performed by the thoracic surgeon for her large pericardial effusion. Patient had large amounts of drainage from JP pericardial drain for more than two weeks following the surgery. JP drain was removed after the amount of drainage decreased to less than 100 ml a day. Cultures and cytology for malignancy were sent and all came back negative. Patient’s symptoms improved with treatment and the she was discharged on levothyroxine 137 mcg daily. Patient was counseled to take her medication regularly. Follow-up appointments were arranged with primary care provider and endocrinology. Discussion: Pericardial effusion is a rare complication of hypothyroidism, more likely to be seen in severe, untreated hypothyroidism such as in this case. The mechanism of pericardial effusion in hypothyroidism is through increased capillary permeability in heart and decreased lymphatic drainage of albumin, resulting in accumulation of fluid in the pericardial space. The effusion develops over time and patients can have large amounts of volume at the time of diagnosis without tamponade physiology. Early diagnosis is important to avoid tamponade and hemodynamic instability.
Presentation: Thursday, June 15, 2023
THU667 Severe Hypothyroidism With Paradoxically Elevated Direct Free T4 Due To Anti-thyroxine Antibodies
Authors: Magvanjav Oyunbileg , John McPhaul Michael , Choucair Ibrahim , Holt Elizabeth 
Published: October 2023

Abstract
Disclosure: O. Magvanjav: None. M.J. McPhaul: None. I. Choucair: None. E. Holt: None.
Introduction Laboratory interference may cause abnormal thyroid function test (TFT) results, leading to misdiagnosis and inappropriate patient care. There are several major types of interference. Here, we describe a hypothyroid patient with an elevated thyroid stimulating hormone (TSH) level with a paradoxically elevated direct free thyroxine (FT4) level caused by interference from anti-thyroxine autoantibodies (THAAB). Clinical Case Our patient is a 38-year-old man who was found to have markedly elevated TSH and FT4 levels during an outpatient workup for Lyme disease. The patient was overall well-appearing but reported some symptoms suggestive of hypothyroidism including excessive fatigue, non-pitting edema, and dry skin. Physical exam was notable elevated blood pressure (systolic blood pressure of 150 mmHg and diastolic blood pressure of 95 mmHg), normal temperature, normal heart rate (72 bpm), regular heart rhythm, and normal body mass index. He had non-tender thyromegaly without nodules, non-pitting edema of bilateral hands and legs, decreased leg hair over bilateral shins, and coarse, dry skin of his hands. Initial TSH level was &gt;150 mIU/ml, with a paradoxically elevated direct FT4 of 10 ng/dl (normal range 0.80-1.70 ng/dL). The FT4 level by equilibrium dialysis was &lt;0.2 ng/dl. Outside laboratory testing to eliminate the effect of heterophile antibodies using two different assays again returned with elevated FT4 levels. Screening for macro TSH confirmed the markedly elevated TSH level. Further investigations at a second outside laboratory revealed that he had THAAB. A repeat FT4 level by dialysis confirmed the markedly low FT4 levels. He was empirically started on low dose levothyroxine and after increasing the levothyroxine dose, he showed improvement in clinical symptoms and the expected decline in TSH. Conclusions Here we present a rare case of THAAB that caused laboratory interference during thyroid function testing, leading to falsely elevated thyroid hormone levels on immunoassays. This case shows the importance of suspecting interference when there is a discrepancy between clinical symptoms and laboratory results or between results obtained from different testing methods. Our patient likely had a genetic predilection for autoimmunity, and Lyme disease is a known trigger for various autoimmune conditions. Our case also emphasizes the importance of taking a systematic approach to investigate the cause of the laboratory interference. This will help prevent misdiagnosis of thyroid disorders that may lead to additional unnecessary testing and treatment.
Presentation: Thursday, June 15, 2023