bluuebunny/crossref_metadata_2025 huggingface dataset:
SAT-308 Effect of Silibinin on ACTH Synthesis and Secretion in Human Adenomatous Corticotropes in Vitro
Authors: Cassarino Maria Francesca , Sesta Antonella , Libera Laura , Bardelli Donatella , Losa Marco , Lasio Giovanni , Giraldi Francesca Pecori
Published: May 2020

Silibinin, a milk thistle extract with known hepatoprotective effects, has recently been shown to act upon tumoral corticotropes and revert the Cushingoid phenotype in an allograft mouse model (Riebold et al 2015). Silbinin is known to inhibit HSP90 -a chaperone to the glucocorticoid receptor- thereby restoring sensitivity to glucocorticoid negative feedback in tumoral corticotropes. Aim of the present study was to assess the effect of silibinin on ACTH synthesis and secretion by human corticotrope adenomas in vitro. Methods: Eight human ACTH-secreting pituitary adenomas were collected during surgery and established in culture as per our protocol (Pecori Giraldi et al 2011). Specimens were treated with 10 - 50 µM silibinin for up to 72 hours. ACTH medium levels were measured by Elisa; POMC expression was assessed by RT-PCR (Cassarino et al 2017). Results. Silibinin reduced spontaneous ACTH secretion to a variable extent in individual adenomas: from 32 to 79% of baseline at 4h, and 54 - 85 % of baseline at 48 and 72h. Silibinin was also effective in reinstating or enhancing sensitivity to steroid negative feedback: ACTH decreases during 10–50 µM silibinin incubation ranged from 10 to 63% of dexamethasone-treated wells at 4 hours, 70 -80% at 48 hours and 36 to 80% at 72 hours, indicating long-lasting effect on glucocorticoid sensitivity. Silibinin induced a variable decrease in POMC expression, both as regards expression in control and dexamethasone-treated wells; some specimens exhibited a marked sensitivity to the inhibitory effect, with POMC expression decreasing to less than 50% of control. Conclusions:, this data suggests that silibinin can inhibit ACTH secretion and POMC synthesis and restore sensitivity to negative glucocorticoid feedback. References: Cassarino et al (2017) Endocrine55: 853–860. Pecori Giraldi et al (2011) Journal of Neuroendocrinology23:1214–21. Riebold et al (2015) Nature Medicine21:276–280.
SUN-117 Growth Hormone-Releasing Hormone (GHRH) Deficiency Promotes Inflammation Associated Carcinogenesis
Authors: Recinella, PharmD Lucia , Leone Sheila , Chiavaroli, PharmD Annalisa , Ferrante, PharmD Claudio , Orlando, PharmD Giustino , Salvatori Roberto , Brunetti Luigi
Published: May 2020

The somatotropic axis, in addition to its well-known metabolic and endocrine effects, plays a pivotal role in modulation of inflammation. Additionally, GH-releasing hormone (GHRH) has been involved in the regulation of the growth of various human tumors. In this work, we aimed to investigate the consequences of GHRH and GH deficiency on the development of inflammation-associated colon carcinogenesis in a mouse model of isolated GH deficiency due to generalized ablation of the GHRH gene [GHRH knock out (GHRHKO)]. Homozygous GHRHKO (-/-) male mice and wild type (C57/BL6, +/+) male mice as control group were used.

After azoxymetane (AOM)/dextran sodium sulfate (DSS) treatment GHRHKO-/- mice displayed higher Disease Activity Index (DAI) score, and more marked weight loss compared to +/+ animals. Additionally, -/- mice showed a significant increase in total colonic tumors, in particular of large size and predominantly localized in distal colon. In colonic tissue of AOM/DSS-treated -/- mice we found the presence of invasive adenocarcinomas, dysplasia and colitis with mucosal ulceration. Conversely, AOM/DSS-treated +/+ mice showed only presence of adenomas, without invasion of sub-mucosa. Treatment with AOM/DSS significantly increased prostaglandin (PG)E2 and 8-iso-PGF2α levels along with cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF)-α, nuclear factor kappa B (NFkB) and inducible nitric oxide synthase (iNOS) gene expression, in colon specimens of both groups. However, the degree of increase of all these parameters was more marked in -/- than +/+ mice.

In conclusion, generalized GHRH ablation increases inflammatory response and colon carcinogenesis in male mice. Whether this results from lack of GH or GHRH remains to be established.
OR06-04 Tissue-Specific Tumorigenesis in Multiple Endocrine Neoplasia Type I
Authors: Gardiner Juliet C
Published: May 2020

While a germline heterozygous mutation in the Multiple Endocrine Neoplasia type 1 (MEN1) gene predisposes tumor formation in specific tissues such as the endocrine pancreas, parathyroid glands and anterior pituitary, this tissue-specific tumorigenesis is not dependent on MEN1 mutations alone. In fact, a homozygous deletion of Men1 in mouse pancreatic exocrine tissue does not result in tumor formation, suggesting a tissue-specific mechanism. Loss of menin activates a menin-interacting protein retinoblastoma-binding-protein 5 (RBBP5). Since RBBP5 transcriptionally regulates DNA methyltransferase 1 (DNMT1), this causes global DNA hypermethylation and subsequent tumorigenesis in MEN1-target endocrine tissues. We hypothesize that while RBBP5 is ubiquitously expressed, it exclusively binds to the DNMT1 promoter in MEN1-target-tissues through its recruitment by tissue-specific factors. Using chromatin immunoprecipitation, we demonstrated that Rbbp5 is bound to the Dnmt1 promoter in MEN1-target-tissues, while not in non-target tissues. Following a high-throughput genome-wide approach, we identified two candidate factors that may recruit Rbbp5 to the Dnmt1 promoter. Immunohistochemistry showed MEN1-target-tissue-specific expression of these target factors. Co-immunoprecipitation revealed MEN1-target-tissue-specific binding of Rbbp5 to the factors. In conclusion, Rbbp5 binds the Dnmt1 promoter in MEN1-target-tissues and we have identified candidates for Rbbp5 recruitment to the Dnmt1 promoter that must be tested further to determine their role in the observed tissue specificity of MEN1-related tumorigenesis.
SUN-349 Characterizing Functional Performance in Adults with Hypophosphatasia
Authors: Dahir Kathryn , Durrough Christina , Hudson Margaret , de Riesthal Michael , Hu Jiun-Ruey , Simmons Jill
Published: May 2020

Introduction:

Limited data exist regarding physical and cognitive functioning of adults with hypophosphatasia (HPP), and there are no guidelines for evaluation by physical therapists (PT), occupational therapists (OT), or speech-language pathologists (SLP). We evaluated physical and cognitive functioning among adults with HPP through comprehensive assessments and patient reported outcome tools.

Methods:

Sixteen participants with HPP (median age 42 (32.5–50.5) yrs, 73% female, 100% Caucasian, 50% on enzyme replacement therapy) completed standardized assessments of mobility, balance, fine motor control, activities of daily living and cognition, as well as self-reported measures of health-related quality of life, fatigue, depression and anxiety.

Results:

Compared with normative data from community dwelling adults, participants traveled less distance on a Six-Minute Walk Test (1,376 ± 431** ft vs 1873±299) and had slower gait on a 10-Meter Walk Test (1.04±0.21 vs 1.39–1.46 m/s). Participants were slower to respond on the Nine Hole Peg Test (20.6±2.4s** for right & 21.7±2.4s** for left hand vs 16.5s to18.5s), and 2 had an abnormally slow reaction time via Dynavision (0.9s* [0.85,0.96], functional speed is <1.15s). 20% scored in the low average/borderline range of performance on the Repeatable Battery for the Assessment of Neuropsychological Status, suggesting potential cognitive impairment. On the Short Form-36, 75% reported limitations in their ability to fulfill life roles due to physical problems (25%±39%ile**), 75% reported below average energy (30%±23ile**), and 100% rated their health as unlikely to improve (32%±15%ile**). Fatigue Severity Scale scores were well above the median for a healthy population (47 [34,60.5]* vs 2.3). Median scores for Depression, Anxiety, and Stress were within the normal range, but moderately severe depression was reported by 4 participants. Participants reported moderate (4), severe (1), or extremely severe (1) anxiety; and 4 reported severe (2) or extremely severe (2) stress.

Conclusions:

Objective functional assessments indicate mild deficits, but participants self-reported significant limitations due to physical dysfunction, indicating that current objective testing may not be sufficient in the HPP population. Impaired reaction time may indicate potential safety concerns with driving or certain occupations, and screening may be indicated. A subgroup of participants was significantly affected by depression, stress, and/ or anxiety. Guidelines and additional assessment tools should be created to further evaluate physical and cognitive functioning among adults with HPP. The use of PT, OT, and SLP specialists can aid in establishing baseline assessment of impairment and developing treatment plans with objective metrics for assessing efficacy of treatment.

*median

**mean
SAT-256 The Ever Confusing Cushing’s Work Up: Is It Real? Is It Pseudo Cushing’s’? Or Could It Be Factitious
Authors: Ghalib Luma , Prevedello Daniel , Mohyeldin Ahmed
Published: May 2020

Case Presentation:

A 50 year old female initially presented with progressive weight gain and mood swings. She had normal 24 h urine cortisol, but an elevated midnight serum cortisol. She underwent transphenoidal surgery for a presumed ACTH-dependent Cushing’s disease. Pathology was not supportive of a pituitary adenoma, showing adenohypophyseal tissue with focal expansion of the acini. The surgery was complicated by hypothyroidism and growth hormone deficiency. She was able to weaned off of the steroids after a few months. She had recurrence of her initial symptoms, she was found to have elevated late evening and morning cortisol levels. She underwent a bilateral adrenelactomy for “recurrence of the cyclical Cushing’s symptoms.” She was started on HC replacement; 10 mg AM and 2.5 mg PM, florinef 0.05 mg daily. She slowly lowered the hydrocortisone dose, and as a result lost 120 lbs.

Three years later she presented with fatigued and gaining weight, by that time she was on Hydrocortisone 3.75 mg AM, 1.25 mg evening, and fludrocortisone 0.1 mg/day. ACTH was 355 (6–48 pg/ml), serum cortisol 10 (8–19 ug/dl) on Hydrocortisone and < 1.0 ug/dl off cortisone. The 24 h urine free cortisol < 1.0 (10–24 ug/34h), and 17 OH- corticosteroids < 4.8(4–14 mg/dl). A possible adrenal remnant was seen on abdominal CT, surgically removed of the lesion showed a lipoma.

She was referred to Neurosurgery for a second pituitary surgery for the concern Cushing’s recurrence. A pituitary MRI revealed a small potential microadenoma. The small dose of hydrocortisone was held for 48 h and an 8 AM test dose: Serum cortisol < 1.20 mcg/dl (3–18), ACTH 1,077 pg/ml (5–72), 24 h urine cortisol < 1.5 mcg/24h (3.5–45), 24 h urine cortisone 10 mcg/24h (17–129), and two midnight salivary cortisol were 128 and 265 ng/dl (< 100 ng/dl). There was a concern raised by the laboratory for a contaminated salivary sample, as the salivary cortisol to cortisone ratio was concerning for contamination with exogenous steroid (1)

Discussion:

Work up for Cushing’s syndrome can be very confusing and frustrating at times for the patient and their physicians. Doing a meticulous work up is necessary to reach an accurate conclusion. Misdiagnosing Cushing’s can lead to a cascade of mistreatment with serious consequences. The case presented highlights the challenges encountered in taking care of such patients. It is necessary to understand the pre-testing probability to reach a precise conclusion. Factitious disorder or sample contamination can be yet another challenge in the differential diagnosis of Cushing’s work up.

(1)

Raff H Measurement of Late-Night, Salivary Cortisol and Cortisone by LC-MS/MS to Assess Preanalytical Sample Contamination with Topical Hydrocortisone. Clinical Chemistry 58:5 (2012)
MON-660 Hypoglycemia Following OGTT Is More Frequent and Pronounced in CF Compared with Controls
Authors: Harris Clea D , Kilberg Marissa J , Sheikh Saba , Stefanovski Darko , Cuchel Marina , Kubrak Christina , Hadjiliadis Denis , Rubenstein Ronald C , Rickels Michael Roehrhoff , Kelly Andrea
Published: May 2020

Glucose homeostasis is often abnormal in people with pancreatic insufficient cystic fibrosis (PI-CF). This dysfunction is viewed on a continuum from “normal” glucose tolerance to cystic fibrosis-related diabetes (CFRD), and may also include postprandial and oral glucose tolerance test (OGTT)-related hypoglycemia. This study aimed to delineate the mechanism(s) underlying OGTT-related hypoglycemia. We compared extended OGTT with frequent blood sampling of glucose and insulin in adolescents and young adults with PI-CF [CF(+)] to historical data from a healthy cohort [CF(-)]. We hypothesized that the subset of CF(+) with hypoglycemia would demonstrate 1-hour glucose ≥ 155 mg/dL and impaired early phase insulin secretion (insulin secretion within first 30 min of OGTT). Hypoglycemia [hypo(+)] was defined as plasma glucose <65 mg/dL and was used to assign subjects to exposure groups. We restricted analyses to 180 minutes given available control data. Glucose and insulin incremental areas under the curve (Glc-AUC; Ins-AUC) for 30-minute intervals were calculated. One-hour glucose, nadir glucose, Glc-AUC0-30, and Ins-AUC0-30 and were compared between CF(+) and CF(-) subjects using Student’s t-test or Wilcoxon Rank Sum depending upon normality. Participants were 60.5% male, age: 25.4±4.8 years, with BMI-Z: 0.06±0.96kg/m2 [no differences for CF(+) vs CF(-)]. FEV1%-predicted for CF(+) was 83±21. 69.6% of CF(+) participants self-reported prior episodes of hypoglycemia, 68.7% of whom reported confirmation via glucometer. Hypoglycemia occurred by 180 minutes [hypo(+)] in 15/23 (65%) CF(+) and 5/15 (33.3%) CF(-) subjects (p=0.028). For hypo(+), nadir glucose occurred on average at 180 minutes for both CF(+) and (-). Hypo(+) CF(+) had higher mean 1-hour glucose (197±49mg/dL vs 134±66mg/dL, p=0.035), lower mean glucose nadir (48±7mg/dL vs 61±4mg/dL, p<0.01), and lower early-phase insulin secretion (Ins-AUC0-30: 263±168 versus 650±275 µU/mL, p<0.01) than hypo(+) CF(-). There was no difference in Glc-AUC0-30 for hypo(+) CF(+) vs CF(-). Hypoglycemia is frequent in CF, and is associated with early glucose dysregulation (elevated 1-hour glucose) and compromised early-phase insulin secretion compared to controls with presumed non-pathologic reactive hypoglycemia. The mechanism of hypoglycemia in CF appears to be different than that seen in healthy individuals, and its association with progression to CFRD warrants further evaluation.
SAT-430 Quality of Life After Thyroidectomy for Patients with Hashimoto’s Disease and Persistent Symptoms
Authors: Thatipamala Priyanka , Noel Julia , Orloff Lisa
Published: May 2020

Objective: To determine whether thyroidectomy improves quality of life in patients with Hashimoto’s thyroiditis with persistent symptoms despite biochemical euthyroidism.

Study Design: A retrospective cohort study of patients undergoing thyroidectomy for Hashimoto’s thyroiditis.

Setting: Tertiary referral center.

Subjects and Methods: Included patients underwent thyroidectomy for Hashimoto’s thyroiditis at our institution between 2014 and 2018. The following variables were collected: age, race, body mass index (BMI), pre-operative symptoms, pre-operative thyroid peroxidase antibody (TPO Ab), thyroglobulin antibody (Tg Ab) thyroid stimulating hormone (TSH), free T4, specimen weight, and presence of thyroiditis or malignancy on the pathology report. The primary outcome was general health score on the Short Form-36 Health Survey (SF-36) ranging from 3–35 months postoperatively. Secondary outcomes included an additional questionnaire specifically addressing disease management after surgery as well as any adverse outcomes.

Results: 19 patients were included in the study. 18 of the 19 patients were female with a mean age of 48 years (SD 13.6 years). The majority of patients were Caucasian. There was no significant difference between the general health score of the Hashimoto’s thyroiditis patients post-operatively compared to a healthy control population. (66.9 vs. 74.1; 95% CI [-16.9 + 2.5], p = 0.16). There were also no significant differences between groups within the 7 SF-36 subscores. Elevation in pre-operative TPO Ab correlated with lower reported post-operative energy levels (r = -0.63, p = 0.016) and emotional well-being (r = -.55, p = 0.041). 87.5% (14/16) of respondents reported to be moderately or extremely happy with their decision to proceed with surgery.

Conclusions: Quality of life in patients with Hashimoto’s thyroiditis who undergo thyroidectomy is equivalent to the general population, and the majority of patients are satisfied with the decision to have undergone surgery. Thyroidectomy is a consideration for patients with Hashimoto’s disease and persistent symptoms despite optimization on medical therapy.
SAT-353 Familial Hyperparathyroidism - Due to a Rare Genetic Mutation
Authors: Taiwo Adeyinka , Dillon Joseph Stephen , Stinson Jennifer
Published: May 2020

Introduction: Hyperparathyroidism occurs most commonly in middle age patients, predominantly in women. It can be caused by parathyroid adenoma, hyperplasia or parathyroid carcinoma. Genetic predisposition can be found in about 10% of primary hyperparathyroidism due to certain gene mutations. This case emphasizes the importance of taking a detailed family history when patients present with hyperparathyroidism at a young age, so that familial hyperparathyroidism, if present, can be detected and relatives screened.

Clinical case: A 26 y.o. male presented with symptoms of fatigue and polydipsia for several years.

He was noted to have a serum calcium of 12.4 mg/dL(8.5–10.5), with parathyroid hormone of

213 pg/ml (15–65). He denied any history of kidney stones, fractures and no palpable neck masses. The patient’s family history was significant for his paternal half-sister who had parathyroidectomy for hyperparathyroidism at 20yrs old and paternal grandmother died of parathyroid cancer in her 50s. The patient’s father died of pancreatic cancer at 41yrs old. A neck ultrasound revealed a mass posterior to the left inferior thyroid. A Sestamibi parathyroid scan revealed a parathyroid adenoma at the posteroinferior aspect of the left hemithyroid. Labs for free metanephrines and normetanephrine, prolactin and gastrin levels were all normal.

Due to his young age and the possibility of having familial hyperparathyroidism, he underwent bilateral neck exploration and parathyroidectomy, with removal of his left inferior, right superior, left superior parathyroid glands and left upper thymus. Surgical pathology revealed, hypercellular parathyroid tissue. Post operatively, his calcium and vitamin D remained within the normal range.

Genetic studies revealed a mutation in the parafibromin gene - CDC73 (also called HRPT2), a tumor suppressor gene, which is on chromosome 1q25. The patient currently has 6 children ranging from age 5 months to 6 years. He was advised to have his children tested any time from age 7 years for the gene mutation.

The patient has remained stable 4yrs post operatively, with normal calcium and PTH levels. He does not have any history of jaw tumor. He never had an ultrasound kidney done. He is being monitored with yearly lab tests.

Conclusion: CDC73 gene mutation-associated disorders are inherited as an autosomal dominant fashion, with variable penetrance. This gene mutation can be found in conditions such as hyperparathyroidism jaw tumor, familial hyperparathyroidism and parathyroid cancer.

Reference:

1.

CDC73-Related Disorders: Clinical Manifestations and Case Detection in Primary Hyperparathyroidism. J Clin Endocrinol Metab. 2017 Dec 1;102(12):4534–4540.
SAT-461 NIFTP: A Painstaking Diagnosis Through the Pathologist’s Eyes
Authors: Chittimoju Sanjita B , Drake Frederick T , Lee Stephanie L
Published: May 2020

Non-invasive encapsulated follicular variant of papillary thyroid cancer (EFVPTC) was recently reclassified as non-invasive follicular thyroid neoplasm with papillary-like nuclear features (NIFT-P).1 In 2018, revised and stricter criteria were proposed for a lesion to qualify as NIFT-P including no well-formed papilla or BRAF-V600E mutation.2 We are presenting an interesting case to highlight the importance of scrutinizing pathology slides to diagnose NIFTP with these more strict criteria.

35-year-old female from Puerto-Rico was diagnosed with Graves’ disease. After 2 years of methimazole treatment, total thyroidectomy was planned for definitive treatment of Graves’ disease. During the work up, she was noted to have a cystic nodule in isthmus, a 1.1 cm hypoechoic nodule in left mid-lobe and a 1.1 cm isoechoic rounded mass in left level III neck, which was initially thought to be a lateral aberrant thyroid remnant. Her thyroid uptake scan was consistent with a multinodular goiter with no uptake in the extrathyroidal mass. The mass was biopsied and showed Atypia of Undetermined Significance (AUS) Bethesda III with washout positive for thyroglobulin (Tg). Total thyroidectomy with bilateral central and left lateral neck dissection was performed. The pathology showed an intrathyroidal 1.2 cm EFVPTC with predominant follicular features and <1% papillae, without tumor capsular invasion. The initial diagnosis was NIFT-P with a background of chronic thyroiditis. However, on pathology, the level III neck mass was a 2 cm metastatic node with classical PTC. ThyroSeq mutational analysis of tissue blocks for both the thyroid nodule and lymph node were positive for NCOA4-RET (RET-PTC3) gene fusion, a BRAF-V600E-like mutation found in classical PTC. On review of her pathology, the thyroid lesion was noted to have more than one papilla, though <1% papillae and was >30% solid, hence not qualifying as NIFT-P and her histological diagnosis was changed to EFVPTC. She was staged as AJCC 8th edition stage 1 with intermediate ATA risk for which she received adjuvant therapy of 101 mCi 131I.

Although classification into NIFT-P has been shown to reduce overtreatment of low risk encapsulated PTC, pathology slides should be closely scrutinized to ensure fulfillment of all criteria in order for a lesion to qualify as NIFT-P. This will minimize failure to recognize PTCs, that would warrant closer follow up and surveillance for recurrence.

1. Rossi, Esther D, et al. Noninvasive Follicular Thyroid Neoplasm with Papillary-Like Nuclear Features: Update and Diagnostic Considerations—a Review. Endocrine Pathology 30.2 (2019)

2. Nikiforov, Yuri E et al. Change in Diagnostic Criteria for Noninvasive Follicular Thyroid Neoplasm With Papillary-like Nuclear Features. JAMA oncology vol. 4,8 (2018)
MON-067 A Perplexing Case of Hyponatremia and Abdominal Pain
Authors: Yeliosof Olga , Balachandar Sadana
Published: May 2020

Previously healthy 20-year-old female presented with diffuse lower abdominal pain, cramping in nature, multiple episodes of emesis as well as urinary frequency. On day 2 of symptoms, she was treated for a urinary tract infection with antibiotics, as well as NSAIDs and opiates for pain relief. Her serum sodium was 133 mmol/L at this time. On day 3 of symptoms, a CT scan of the abdomen was performed however did not reveal pathology. Her serum sodium was 129 mmol/L at this time. She presented to our ED, on day 5 of symptoms, where serum sodium was down to 122 mmol/L. Despite IV fluids, her sodium continued to decrease to a nadir of 117 mmol/L. Further testing, including a serum osmolality of 242 mOsm/kg, urine osmolality of 540 mOsm/kg, and urine sodium of 207 mmol/L, was consistent with a diagnosis of SIADH. Given persistence of abdominal symptoms along with SIADH further imaging studies, including US abdomen, CT brain and Chest XR, were ordered and returned unremarkable. The constellation of SIADH along with persistent abdominal pain, with negative imaging, lead to consideration of acute intermittent porphyria as a diagnosis. Random urine porphobilinogen was found to be elevated to 147.2 mcmol/L (≤ 2.4) leading to the presumptive diagnosis of acute intermittent porphyria presenting as a neurovisceral attack. Biochemical and genetic testing is being pursued to confirm her diagnosis.

Acute intermittent porphyria is an autosomal dominant hematologic disorder characterized by deficiency in porphobilinogen deaminase, an enzyme in the heme synthesis cascade. Acute attacks are caused by accumulation of porphyrin resulting in autonomic and peripheral neuropathy which can present as abdominal pain, urinary retention, polyneuropathy, dark urine and psychiatric disturbance. Hyponatremia is present in 25–60% of cases which is caused by SIADH or sometimes renal and gastrointestinal sodium loss. Triggers for acute attacks include medications, starvation, infections, hormonal changes and alcohol. Treatment includes avoidance of triggers, IV dextrose and high carbohydrate diet. In severe attacks, IV hemin is used.

Our patient’s urinary tract infection likely triggered her acute symptoms, which was further exacerbated by treatment with NSAIDS and opiates. She developed SIADH which improved with hypertonic saline and fluid restriction.

This case illustrates the need to consider acute intermittent porphyria in the differential diagnosis of SIADH presenting with abdominal pain of unknown etiology.
OR29-07 A Novel Estimate of Creatinine Excretion to Determine Adequacy of 24-Hr Urine Collection
Authors: Kondapalli Ananya V , Li Xilong , Huet Beverly Adams , Maalouf Naim
Published: May 2020

24-hour urine collections are used to assess excretion of various analytes. Although concomitant measurement of creatinine excretion adjusted for body weight (BW) is utilized to determine adequacy of collection, no gold standard exists for determining completeness of a collection. Use of current reference ranges for daily creatinine excretion/BW of 15–20 mg creatinine/kg BW/day in women and 20–25 mg creatinine/kg/d in men established prior to the rising prevalence of obesity has resulted in a large proportion of contemporary individuals appearing to have “incomplete” urine collections.

Our objective was to evaluate the range of creatinine excretion in accurately collected urine specimen from adults with a wide spread of age and BW, and to generate an equation that accurately predicts 24-hour urine creatinine excretion rate from readily available clinical parameters to aid in assessing adequacy of a 24-hour urine collection.

We analyzed data from participants who completed two consecutive 24-hour urine collections while consuming a fixed metabolic diet during inpatient research admissions. Results from participants with 24-hour urine creatinine excretion rate differing by >10% between the two consecutive collections were excluded. In the initial 115 pairs of inpatient 24-hour urine collections (50 female, 65 male) participants, creatinine excretion/BW fell outside the currently accepted reference ranges in >50% of collections. The proportion below the reference range increased with higher BMI. In this derivation dataset, linear regression models were then constructed to predict 24-hr urine creatinine excretion from race, sex, age, weight and height. Reliable prediction of observed 24-hr urine creatinine excretion was confirmed in a validation dataset that included 50 pairs of 24-hour urine samples similarly collected in an inpatient research setting.

This new prediction model performed significantly better than the currently used reference ranges in a large outpatient dataset including 1,399 pairs of 24-hour urine collections. In women, actual creatinine excretion fell within the 95% prediction intervals for our derived equation in 90% of cases using the new interval vs 46% using the current reference range. The corresponding values were 90% and 33% in men. In both genders, the superiority of the new prediction over the current reference range was more pronounced at higher BMI.

We therefore propose revision of currently used criteria to define adequacy of 24-hour urine collection to account for the impact of obesity. The proposed equation incorporates readily available demographic parameters to predict urine creatinine excretion. These findings have wide implications on patient care and research studies, and future studies should test this equation in different settings, diets, and populations.
SUN-369 Different Association of Dietary Fat Intake with Femoral Neck Strength According to Gender: A Nationwide Population-Based Study
Authors: Kim Beom-Jun , Kim Hyeonmok
Published: May 2020

Purpose: Despite the general belief that higher fat intake may be harmful for bone health, its impact on bone strength has not been thoroughly studied. Methods: We conducted a population-based cross-sectional study derived from the Korea National Health and Nutrition Examination Surveys, including 2,590 participants. Composite indices of femoral neck strength, such as the compression strength index (CSI), bending strength index (BSI), and impact strength index (ISI), were generated by combining bone mineral density, body mass, and height with the femoral axis length and width. Results: Dietary fat intake (%) was inversely related to CSI and ISI in men, but not in women. Men in the highest three fat intake quintiles had lower CSI, BSI, and/or ISI than those in the lowest quintile (P=0.003–0.024). In women, compared with participants in the third fat intake quintile, those in the other four quintiles had lower CSI, BSI, and/or ISI (P=0.004–0.049). When the participants were allocated to three groups according to the dietary reference intake of fat in Koreans [low (<15%), moderate (15–30%), or high (≥30%)], men with a moderate or high fat intake had significantly lower ISIs than those with a low fat intake (P=0.046 and 0.039, respectively). By contrast, compared with women consuming a moderate amount of fat, those with a high intake had lower CSI, BSI, and ISI (P = 0.025–0.047). Conclusion: These findings suggest that higher fat intake in men and low or high fat intake in women may contribute to deteriorations in bone strength.
SAT-603 Growth Hormone-Releasing Hormone (GHRH) Antagonists Stimulate Feeding in Mice
Authors: Leone Sheila , Recinella, PharmD Lucia , Chiavaroli, PharmD Annalisa , Orlando, PharmD Giustino , Ferrante, PharmD Claudio , Gesmundo Iacopo , Granata Riccarda , Cai Renzhi , Sha Wei , Schally Andrew V , Brunetti Luigi
Published: May 2020

Growth hormone-releasing hormone (GHRH) is a hypothalamic neuropeptide which stimulates the synthesis and secretion of growth hormone (GH) in pituitary gland. GHRH was also found to modulate food intake in mammals. MIA-690 is a synthetic GHRH antagonist of the Miami (MIA) series with potent antitumor effects. To date, its role in hypothalamic feeding modulation has not been evaluated. In the present study, we aimed to investigate the effects of chronic MIA-690 administration on feeding behavior, locomotor activity and hypothalamic dopamine (DA), norepinephrine (NE), serotonin (5-hydroxytriptamine, 5-HT), orexigenic peptides [agouti-related peptide (AgRP) and neuropeptide Y (NPY)] and anorexigenic peptides [cocaine and amphetamine-regulated transcript (CART) and proopiomelanocortin (POMC)] activity.

Adult C57/BL6 mice were treated daily for 4 weeks by subcutaneous administration of (5 µg) MIA-690 or vehicle solution. Food intake and body weight were recorded every 4 days throughout the study. Immediately after the last injection, locomotor activity in the home cage was recorded, and thereafter animals were sacrificed. Visceral, subcutaneous and brown fat depots were quickly excised and weighed. Hypothalamus was also dissected for evaluating gene expression of AgRP, NPY, CART and POMC by real-time reverse transcription polymerase chain reaction. In addition, hypothalamic DA, NE and 5-HT levels were measured by high performance liquid chromatography (HPLC) coupled to electrochemical detection.

Our findings show that administration of MIA-690 increased food intake and body weight, without affecting locomotor activity. No difference was observed in visceral, subcutaneous and brown fat mass in animals treated with MIA-690 or vehicle. As for neuromodulatory effects, a significant increase of AgRP gene expression and NE levels, along with a reduction of 5-HT levels were found after MIA-690 treatment. On the other hand, we did not observe any alteration in NPY, POMC and CART gene expression, as well as DA levels, following MIA-690 administration.

In conclusion, chronic peripheral administration of MIA-690 could play an orexigenic role paralleled by increased body weight. The stimulation of feeding could be mediated, at least in part, by increased AgRP gene expression and NE levels and decreased 5-HT levels, in the hypothalamus.
SAT-092 Positive Effect of an Early Start of Growth Hormone Therapy on Auxology and Metabolic Parameters in Children with Prader Willi Syndrome
Authors: Gohlke Bettina , Magill Lucy , Laemmer Constante , Woelfle Joachim
Published: May 2020

Introduction: Prader-Willi-Syndrom (PWS) is a rare multisystem genetic disorder characterized by hypothalamic-pituitary dysfunction. Various characteristic clinical features and several endocrinological problems such as hypogonadism, hypothyroidism, Growth hormone (GH) and adrenal deficiency have been described. Since GH-Therapy (GHT) was approved the physical benefits of the treatment have been established in many studies. It is now recommended to start treatment as soon as possible. The aim of this study is to shed light on possible differences in height, carbohydrate and lipid metabolism between children with PWS in whom GHT was initiated either during or after their first year of life. Patients and Methods: This retrospective longitudinal study included 62 children (31 males) with genetically confirmed PWS in whom fasting morning blood samples and auxological parameters were obtained before start of therapy and semi-annually thereafter. The early treatment cohort A consisted of 21 (11 males) infants who were recruited at 0.3–0.99 yrs (mean 0.72 yrs) for GHT. The later treatment cohort B entailed 41 individuals (20 males) in whom GHT was initiated at 1.02–2.54 yrs (mean 1.42 yr). Results: Auxology: Mean length/height-SDSPWS differed significantly throughout the entire observation period between the groups: 1 yr: A: 0.37 (±0.83) vs B: 0.05 (±0.56); 5 yrs: A: 0.81 (±0.67) vs. B: 0.54 (±0.64); p=0.012). No significant differences were found in BMI, lean body mass or percent body fat. Endocrinological Parameters: Mean IGF-I SDS in group A did not differ significantly from group B and mean IGF-I SDS were mostly below 0 SDS (within normal range) in both groups.Lipid Metabolism: Low-density lipoprotein (LDL) was statistically significantly lower in Group A than in Group B during the entire course of the study (LDL: 1 yr: A: 79 (±20) mg/dl vs. B: 90 (±19) mg/dl; 5 yrs: A: 91(±18) mg/dl vs. 104 (±26) mg/dl; p=0.024).Carbohydrate Metabolism: Significant differences in mean fasting insulin levels and HOMA-IR between the two groups were found (fasting insulin p=0.012; HOMA-IR p=0.006). Significant differences in HbA1c and blood glucose levels were also determined between the two groups (HbA1c: p<0.001; blood glucose: p=0.022). Conclusion:Our analysis shows that early GHT had a statistically significant favorable effect on height-SDS, LDL, HOMA-IR and fasting insulin. The two groups did not significantly differ in BMI-SDS, body composition or IGF-I SDS.
SUN-687 MODY-14: A Rare Cause of Inherited Diabetes
Authors: Epp Riley , Arnaout Amel , Doyle Mary-Anne
Published: May 2020

Case: A 29-year-old physically active, lean man presented with a three-year history of poorly controlled diabetes diagnosed during a flu-like illness and no history diabetic ketoacidosis. Despite treatment with various oral hypoglycemic medications his glycated hemoglobin was consistently above target (never falling below 9.3%) and correlated with home blood glucose readings. His mother had gestational diabetes with all three of her pregnancies. His maternal grandmother was also known to have diabetes. Pancreatic islet antibodies were negative. C-peptide levels were low. Monogenic diabetes (MODY) was suspected. Discussion: Genetic testing revealed a mutation in the Adaptor Protein, Phosphotyrosine Interaction, PH domain, and leucine zipper containing 1 (APPL1) gene. APPL1 is involved in insulin secretion and insulin signaling via the Akt pathway [1,2,3]. Other mutations of APPL1 causing MODY-14 have been previously described [4]. In our case, the substitution of glutamate for aspartate at the 265 position occurs within the unique fourth alpha helix of the BAR domain of APPL1 [5]. Generally, BAR domains contribute to APPL1 dimerization and plasma membrane association [5]. Substitution of glutamate could disrupt protein folding, impairing dimerization and phosphorylation of Akt, thereby decreased insulin secretion as seen in other mutations of the BAR domain of APPL1 [4]. Although this remains a hypothesis, loss-of-function of APPL1 would explain the lack of insulin secretion without evidence of pancreatic autoimmunity. If the hypothesis is correct, it would be a novel mutation attributable to this MODY subtype. 1. Cheng KKY, Lam KSL, Wu D, et al. APPL1 potentiates insulin secretion in pancreatic β cells by enhancing protein kinase Akt-dependent expression of SNARE proteins in mice. Proc Natl Acad Sci U S A. 2012;109(23):8919–8924. doi:10.1073/pnas.1202435109 2. Ryu J, Galan AK, Xin X, et al. APPL1 potentiates insulin sensitivity by facilitating the binding of IRS1/2 to the insulin receptor. Cell Rep. 2014;7(4):1227–1238. doi:10.1016/j.celrep.2014.04.006 3. Saito T, Jones CC, Huang S, Czech MP, Pilch PF. The interaction of Akt with APPL1 is required for insulin-stimulated Glut4 translocation. J Biol Chem. 2007;282(44):32280–32287. doi:10.1074/jbc.M704150200 4. Prudente S, Jungtrakoon P, Marucci A, et al. Loss-of-Function Mutations in APPL1 in Familial Diabetes Mellitus. Am J Hum Genet. 2015;97(1):177–185. doi:10.1016/j.ajhg.2015.05.011 5. Li J, Mao X, Dong LQ, Liu F, Tong L. Crystal Structures of the BAR-PH and PTB Domains of Human APPL1. Structure. 2007;15(5):525–533. doi:10.1016/j.str.2007.03.011
SUN-213 The Role of Filamin A (FLNA) in the Regulation of Insulin-Like Growth Factor System in Adrenocortical Carcinomas
Authors: Peverelli Erika , Catalano Rosa , Giardino Elena , Treppiedi Donatella , Mangili Federica , Morelli Valentina , Mannelli Massimo , Spada Anna , Arosio Maura , Mantovani Giovanna
Published: May 2020

Adrenocortical carcinomas (ACCs) are rare endocrine tumors with poor prognosis. They overexpress insulin-like growth factor 2 (IGF2), that drives a proliferative autocrine loop by binding to IGF1R and IR, with molecular dynamics still poorly identified. Although promising, IGF1R/IR-targeted therapies have demonstrated a limited efficacy in clinical trials in ACC patients. The cytoskeleton actin-binding protein filamin A (FLNA) was shown to impair IR and IGF1R signalling in melanoma and neural progenitor cells, respectively. The aims of this study were to test in ACC cells: 1) FLNA involvement in regulating IGF1R and IR expression and signalling; 2) FLNA role in modulating responsiveness to IGF1R and IGFR/IR inhibitors; 3) FLNA expression in ACCs and correlation with IGF system. In ACC cells we found by immunoprecipitation that both IGF1R and IR interacted with FLNA in basal condition, with an increased or decreased FLNA recruitment to IGF1R and to IR, respectively, after IGF2 stimulation. Genetic silencing of FLNA in ACC cell lines H295R and SW13 induced a significant increase of IGF1R expression (1.4- and 2.3-fold, respectively) and a reduction of IR (-85.5±9.1%, p<0.001 and -32±19.1%, respectively, p<0.05), with a downstream effect of increased cell proliferation (130±13.4%, p<0.01 in H295R and 144.3±23.3%, p<0.01 in SW13 cells) accompanied by an enhanced ERK phosphorylation. Accordingly, in ACC primary cultured cells FLNA silencing increased IGF1R levels (2.9-fold) and enhanced IGF2 effects on ERK phosphorylation by 2.2-fold. In addition, FLNA knockdown potentiated the antiproliferative effects of IGF1R/IR inhibitor Linsitinib and IGF1R specific inhibitor NVP-ADW742 in H295R cells and SW13. This key role of FLNA was even more evident in A7/M2 melanoma cell model, since IGF2 and Linsitinib exerted the expected effects on ERK phosphorylation in M2 cells, lacking FLNA, but not in FLNA-expressing counterpart (A7 cells). Finally, western blot analysis showed significantly lower, although variable, FLNA expression in ACCs (n=10) than in adrenocortical adenomas (ACAs) (n=10) (FLNA/GAPDH ratio 0.37±0.38 and 0.90±0.63, respectively, p<0.05). Interestingly, FLNA/IGF1R ratio inversely correlated with ERK phosphorylation status in ACCs (p<0.05) but not in ACA. In conclusion, we demonstrated that low levels of FLNA enhance both IGF2 proliferative effects and IGF1R/IR inhibitors efficacy in ACC cells, suggesting FLNA as a new factor possibly influencing tumor clinical behavior and the response to the therapy with IGF1R/IR-targeted drugs.
SAT-396 A Non-Surgical Animal Model of Hypoparathyroidism for Testing PTH Analogues
Authors: Ramezani-Pour Narjes , Esfahani Sayyed Hamid Zarkesh , Pradhananga Sarbendra , Newell-Price John D C , Ross Richard John M , Wilkinson Ian R
Published: May 2020

BACKGROUND:

In vivo animal models for testing the pharmacokinetics and bioactivity of PTH and its analogues require parathyroidectomy by surgery (1, 2). As the parathyroid glands of rodents are very small the surgery often includes thyroidectomy, making this animal model time-limited, single use, complex, and expensive. We have developed a non-surgical rodent model of hypoparathyroidism using the Type II calcimimetic compound, Cinacalcet-HCl, to suppress PTH and thereby serum calcium levels.

Methods:

Normal male Wistar rats were gavaged with 30 mg/kg Cinacalcet-HCl (or vehicle only). To test the effect of PTH 1–34, animals were dosed immediately after Cinacalcet-HCl gavage with either a single subcutaneous injection of PTH at 20 nmol/kg or given as same dose repeated every hour for 6 hrs or vehicle only. Serum samples were analysed for ionised calcium (iCa) using an EasyLyte, fully automated electrolyte analyser (Medica Corporation) and phosphate using a Phosphorus Detection Assay Kit (Pars Azmun, IRAN) and an Hitachi 917 Clinical Chemistry Analyser.

Results:

Rats gavaged with 30 mg/kg Cinacalcet-HCl produced a significant reduction in iCa levels between 2-24hrs returning to baseline at 48-72hrs post dose with the nadir at 8 hours (ANOVA P < 0.0001). This equated to a 25% reduction in iCa at 8 hrs: mean±SD, iCa 1.19 ± 0.09 mmol/L at predose and 0.891 ± 0.04 mmol/L at 8 hours (t-test P < 0.0001). For phosphate there was an initial lowering within the first 2 hrs in all test groups but then a rise such that phosphate was at higher levels than control from 8–24 hrs (ANOVA, ns), returning to baseline at 48 hrs. PTH at 20 nmol/kg given as a single sc dose abrogated the Cinacalcet-HCl induced fall in iCa for up to 2 hrs (AUC±SD (mmol/L).hr, 0.076 ±0.047 versus 0.168±0.0874, t-test P=0.0289).

Conclusions:

We have shown that the administration of Cinacalcet-HCl provides a robust and reproducible lowering of calcium which is line with current published data (3). These studies demonstrate that the use of Cinacalcet-HCl in normal rats produces a hypocalcemic state that can be abrogated by the addition of PTH. This non-surgical animal model of hypoparathyroidism will be of value in testing the pharmacodynamics of PTH analogues.

References

1. Shimizu M, et al. J Bone Miner Res. 2016;31(7):1405–12.

2. Jung SY, et al. PLoS One. 2016;11(10):e0163911-e.

3. Nemeth EF, et al. 2004;308(2):627–35.
SAT-160 Evaluation of Life Style and Anthropometric Parameters in Patients with Nonfunctional Adrenal Incidentalomas
Authors: Dagdemir Arzu Nevin , Akalin Aysen
Published: May 2020

Introduction

Adrenal lesions that are found incidentally during routine imaging of people who do not have any complaints or physical findings related to adrenal gland are called adrenal incidentaloma (1,2). Incidentalomas appear more frequently with the increasing use of imaging techniques and increasing age. It is unclear why most of these masses emerged. It is controversial whether nonfunctional adrenal incidentalomas (NFAI) increase the risk of cardiovascular disease or metabolic syndrome or are more common in people with these diseases. In this study, we aimed to investigate whether lifestyle and body fat have an impact on the occurrence of NFAI.

Materials and Methods

100 patients with NFAI were included in the study. Control group consisted of 50 healthy and similar age groups. Physical activities of these groups (with the International Physical Activity Questionnaire Short Form), smoking were questioned and anthropometric measurements were made (Height, body weight, body mass index (BMI), neck, hip, waist circumference and body fat mass, fat percentage, total body water, fet free mass with bioimpedance method). Laboratory tests were examined from the patient file.

Results

Female dominance was observed in patients with NFAI. BMI, waist circumference, hip circumference, neck circumference, total body fat percentage and mass and smoking were found to be higher in the patient group compared to the healthy group and a statistically significant difference was found. When a subgroup of patients with similar age and BMI among the patients and the control group were constituted (25-29,99 kg / m2) waist circumference and total fat mass were again significantly higher in the patient group compared to controls. In addition, there was a significant positive correlation between mass size and waist circumference, BMI, neck circumference, cortisol after 1 mg dexamethasone suppression test and a significant negative correlation with ACTH.

Conclusion

The data obtained showed that body adiposity and smoking were higher in patients with NFAI. Also, ıt was shown that although the patients were regarded as nonfunctional, suppressibility of the cortisol decreases as the mass size of the incidentaloma increase.

Resources

1. Turkey Endocrinology and Metabolic Diseases Society of Adrenal and Gonadal Working Group. Adrenal and Gonadal Diseases Guide. 2019, pages 77–84.

2. Fassnacht M, Arlt W, Bancos I, Dralle H, Newell-Price J, Sahdev A, et al. Management of adrenal incidentalomas: adrenal tumors. 2016; 175 (2): G1-G34.