bluuebunny/crossref_metadata_2025 huggingface dataset:
Endocrine Dysfunction in Covid-19
Authors: Das Liza , Dutta Pinaki , Bhadada Sanjay Kumar , Rastogi Ashu , Walia Rama , Mukherjee Soham , Puri Goverdhan Dutt , Bhansali Anil
Published: May 2021

Introduction: Evidence pertaining to new-onset endocrine dysfunction in patients with COVID-19 is currently limited and extrapolated from prior SARS epidemics. Further, identifying whether the quantum of this dysfunction is associated with the severity of disease in patients with COVID-19 is unknown. We aimed to to comprehensively explore the prevalence, nature and degree of endocrine dysfunction stratified based on disease severity at a dedicated COVID care centre.

Patients and Methods: Consecutive patients enrolled at PGIMER Chandigarh, were stratified on the basis of disease severity as: group I (moderate to severe disease including oxygen saturation <94% on room air or those with comorbidities) and group II (mild disease, with oxygen saturation >94% and without comorbidities). Hypothalamo-pituitary-adrenal, thyroid, gonadal axes and lactotroph function were evaluated. Inflammatory and cell-injury markers were also analysed.

Results: Patients in group I had higher prevalence of hypocortisolism (38.5 vs 6.8%, p=0.012), lower ACTH (16.3 vs 32.1pg/ml, p=0.234) and DHEAS (86.29 vs 117.8µg/dl, p= 0.086) as compared to group II. Low T3 syndrome was a universal finding, irrespective of disease severity. Sick euthyroid syndrome (apart from low T3 syndrome) (80.9 vs 73.1%, p= 0.046) and atypical thyroiditis (low T3, high T4, low or normal TSH) (14.3 vs 2.4%, p= 0.046) were more frequent in group I than group II. Male hypogonadism was also more prevalent in group I (75.6% vs 20.6%, p=0.006) than group II, with higher prevalence of both secondary (56.8 vs 15.3%, p=0.006) and primary (18.8 vs 5.3%, p=0.006) hypogonadism. Hyperprolactinemia was observed in 42.4% patients, without significant difference between both groups.

Conclusion: COVID-19 can involve multiple endocrine organs and axes, with a greater prevalence and degree of endocrine dysfunction in those with more severe disease. Involvement of multiple axes, particularly at hypothalamo-pituitary level suggests the possibility of hypophysitis as an underlying etiology. We also observed less characterised findings like atypical thyroiditis and normal DHEAS despite secondary hypocortisolism. Follow-up surveillance of these patients at periodic intervals and estimation of anti-pituitary antibodies could be considered to elucidate viral cytopathic effect or inflammation as the major underlying mechanism of endocrine dysfunction.
Oxytocin Levels in Response to Pituitary Provocation Tests in Healthy Volunteers
Authors: Sailer Clara Odilia , Winzeler Bettina Felicitas , Urwyler Sandrine Andrea , Schnyder Ingeborg , Refardt Julie , Eckert Anne , Verghese Nimmy , Fassnacht Martin , Chifu Irina Oana , Lawson Elizabeth Austen , Verbalis Joseph G , Fenske Wiebke Kristin , Christ-Crain Mirjam
Published: May 2021

Background: Oxytocin, secreted into the circulation through the posterior pituitary, regulates lactation, weight, and socio-behavioral functioning. Oxytocin deficiency has been suggested in patients with hypopituitarism, however, diagnostic testing for oxytocin deficiency has not been developed. Known stimuli used in the diagnosis of pituitary deficiencies - the hypertonic saline and arginine infusion tests stimulating copeptin levels, and the oral macimorelin test stimulating growth hormone levels - have also been shown to stimulate oxytocin secretion in animal models. We hypothesized that these provocation tests would stimulate plasma oxytocin levels in humans.

Methods: Basal plasma oxytocin levels were measured for all three tests. Stimulated plasma oxytocin was measured once plasma sodium >150 mmol/l for the hypertonic saline and after 45 minutes for the arginine infusion and the oral macimorelin test, expected peak of copeptin and growth hormone levels, respectively. Primary outcome was change between basal and stimulated oxytocin levels using a paired t-test.

Results: Median (IQR) age of all participants was 24 years (22, 28), 51% were female. As expected, copeptin increased in response to hypertonic saline from 4.0 pmol/L [3.3, 6.7] to 34.2 pmol/L [23.2, 45.4] (p-value <0.001) and in response to arginine infusion from 4.6 pmol/L [3.2, 6.2] to 8.3 pmol/L [6.4, 10.8] (p-value <0.001). Growth hormone increased in response to oral macimorelin from 1.6 ng/mL [0.3, 17.2] to 106.0 ng/mL [73.3, 127.2] (p-value <0.001). Oxytocin levels increased in response to hypertonic saline infusion from 0.3 pg/mL [0.3, 0.5] to 0.6 pg/mL [0.4, 0.7] (p-value 0.007), while there was no change in response to arginine infusion (basal 0.4 pg/mL [0.4, 0.6], stimulated 0.4 pg/mL [0.3, 0.6], p-value 0.6), nor to oral macimorelin (basal 38.7 pg/mL [31.1, 66.9], stimulated 34.2 pg/mL [31.2, 48.2], p-value 0.3).

Conclusion: We found that hypertonic saline infusion results in doubling of oxytocin levels. Further research will be important to determine whether this test could be used diagnostically to identify patients with oxytocin deficiency. In contrast to animal data, arginine and macimorelin did not stimulate oxytocin.
Relationship Between Oxytocin Levels and Brain Volume in Anorexia Nervosa Compared to Healthy Controls
Authors: Sailer Clara Odilia , Breithaupt Lauren , Lyall Amanda E , Vessey Brynn , Plessow Franziska , Kahn Danielle L , Slattery Meghan , Holsen Laura , Eddy Kamryn T , Misra Madhusmita , Lawson Elizabeth Austen
Published: May 2021

Background: A disruption in food motivation pathways has been described in females with anorexia nervosa (AN), a psychiatric disorder characterized by food restriction despite low weight. We have shown that in AN and healthy controls (HC), levels of oxytocin (OXT), a hormone involved in lactation, social behavior and weight regulation, decrease after a meal. In HC, we identified a relationship between postprandial change in OXT and subjective appetite, yet this association was absent in females with AN, suggesting a disconnect between OXT and appetite regulation in AN. Prior studies have shown that gray matter volume of the amygdala and hippocampus, areas rich in OXT receptors, correlate with OXT levels in HC. Furthermore, these regions play a central role in food reward and decreased volume has been reported in AN. We hypothesized that the relationship between postprandial change in OXT and amygdala and hippocampal gray matter volume would differ between AN and HC.

Methods: We performed a cross-sectional study of 51 females (23 restrictive AN; 28 HC). We drew blood for OXT levels fasting and 60 min after a standard meal and performed T1-weighted MRI scans of the brain in the fasted state. MRI data was quality controlled and processed with FreeSurfer. Average gray matter brain volumes were extracted from the bilateral amygdala and hippocampus for each subject. Linear regression models were used to determine differences between AN and HC of postprandial percent change in OXT on amygdala and hippocampus gray matter volume.

Results: Median [IQR] age was higher in females with AN (20.6 years [19.3, 21.5]) than HC (18.8 years [IQR 17.6, 20.3], p=0.02), and percentage of ideal body weight was lower in AN (75.5%) than HC (97.4%, p<0.01). Right hippocampus volume, adjusted for age and total intracranial volume, was significantly lower in AN (estimated difference -188 dm3 [95%-CI -360, -17], p=0.04). Percent change in OXT was not different (p=0.5) but there was a trend for a positive interaction effect (p=0.08) for AN and percent change in OXT on right hippocampus volume. Posthoc exploratory analysis indicated a positive correlation in AN (R2=0.41, p=0.02) and no correlation in HC (R2=0.17,p=0.4) between percent change in OXT and right hippocampus volume. There was no significant between group difference in volume nor postprandial change in OXT for the bilateral amygdala or left hippocampus between groups.

Discussion: Our results indicate smaller right hippocampus volume and a trend towards a positive association with postprandial change in OXT in AN compared to HC. Future studies will be important to better define the relationships between OXT secretion and food motivation brain regions and the impact on eating behavior in AN.
Assessment of the Severity of Carpal Tunnel Syndrome in Patients With Acromegaly: A Focus on Clinical, Biochemical and Image Outcomes
Authors: Paredes Vinueza Diego Fernando , Socorro Gómez-Caballero María del , Solis-Pazmino Andrea Paola , Salazar-Vega Jorge Luis
Published: May 2021

This is a descriptive cross-sectional study designed to analyze the prevalence and severity of carpal tunnel syndrome (CTS) in addition to its association with clinical, biochemical, and imaging variables in a cohort of patients with acromegaly. Data collection was performed by entering them in our database and we subsequently analyzed in the SPSS system. Of the 71 patients registered with acromegaly, 68 met the inclusion criteria. Among the results, 97.8% presented CTS evidenced by electroconduction. Of these, 43 (63.1%) showed moderate involvement, followed by 17 severe cases (25%) and 6 mild cases (8.8%). The level of GH in serum, the years of delay in diagnosis, and the size of the adenoma were related to the presence and severity of CTS (p less than 0.05). The multivariate analysis showed that severe grade had a significant association with these variables compared with moderate and mild grade.
Cardiac Remodeling in Patients with Childhood-Onset Craniopharyngioma - Results of HIT-Endo and KRANIOPHARYNGEOM 2000/2007
Authors: Sowithayasakul Panjarat , Buschmann Leona Katharin , Boekhoff Svenja , Muller Hermann Lothar
Published: May 2021

Hypothalamic obesity caused by childhood-onset craniopharyngioma results in long-term cardiovascular morbidity. Knowledge about clinical markers and risk factors is rare. A cross-sectional study on transthoracic echocardiographic parameters was performed to determine the associations with clinical and anthropometric parameters in 36 craniopharyngioma patients recruited in HIT-Endo and KRANIOPHARYNGEOM 2000/2007. BMI correlated with the thickness of interventricular septum in diastole (IVSd) (r=0.604, p<0.001) and left ventricular posterior wall thickness in diastole (LVPWd) (r=0.460, p=0.011). In multivariate analyses on risk factors for cardiac remodeling, sex hormone replacement therapy, BMI and male gender were positively correlated with increased left ventricular internal diameter in diastole (LVIDd), R2=0.596, F=10.323, p<0.001. BMI and insulin resistance were selected as significant independent determinants of IVSd, produced R2=0.655, F=29.441, p<0.001. Due to wide range of disease duration, 17 pediatric and 19 adult patients were analyzed separately. In the adult subgroup (age at study ≥18 years), BMI correlated with IVSd (r=0.707, p=0.003), LVPWd (r=0.592, p=0.020) and LVIDd (r=0.571, p=0.026). In the pediatric subgroup (age at study <18 years), no correlation between cardiac parameters and BMI was observed. Only LVIDd correlated with disease duration (r=0.645, p<0.001). All cardiac functions were within the normal range, indicating no association with functional impairments. We conclude that cardiac remodeling in patients with childhood-onset craniopharyngioma correlated with the degree of hypothalamic obesity, disease duration, sex hormone replacement therapy, male gender and insulin resistance. As echocardiography has limited sensitivity in patients with obesity, further research on more sensitive techniques for cardiac diagnostics in craniopharyngioma patients is warranted.
Cerebral Infarction in Childhood-Onset Craniopharyngioma Patients - Results of KRANIOPHARYNGEOM 2007
Authors: Boekhoff Svenja , Bison Brigitte , Genzel Daniela , Eveslage Maria , Otte Anna , Friedrich Carsten , Flitsch Joerg , Muller Hermann Lothar
Published: May 2021

Purpose: Cerebral infarction (CI) represents a vascular complication following treatment of suprasellar tumors. Risk factors for CI, incidence rate, and long-term prognosis are unknown for patients with childhood-onset craniopharyngioma (CP). Methods: MRI of 242 CP patients, recruited 2007-2019 in KRANIOPHARYNGEOM 2007, were reviewed for CI. Risk factors for CI and outcome after CI were analyzed. Results: Twenty-eight of 242 patients (11%) presented with CI based on reference assessment of MRI. One CI occurred before initial surgery and one case of CI after release of pressure via intracystic catheter. 26 of 28 CI were detected after CP resecting surgical procedures at a median postoperative interval of one day (range: 0.5-53 days). Surgical intraoperative vascular lesions were documented in 7 cases with CI. There was a trend (p=0.069) towards higher initial presurgical tumor volume in CI patients (21.7 cm3, range: 0.01-187.6 cm3) compared with non-CI patients (15.5 cm3, range: 0.01-286.3 cm3). The CI rate was lower in cases operated via transsphenoidal approach (4%) when compared with transcranial approach (13%). CP patient load of neurosurgical centers as a potential measure of surgical expertise was not associated with CI. In 12 irradiated patients, CI occurred before irradiation in all cases. Multivariate analyses showed that hydrocephalus and gross-total resection (GTR) at the time of primary diagnosis/surgery were independent risk factors for CI. Two-years progression-free survival rate was lower (p=0.023) after CI (0.310±0.095) when compared with the subgroup of patients without CI (0.604±0.034). After CI, quality of life (PEDQOL) and functional capacity (FMH) were impaired when compared with patients without CI. Conclusions: CI occurs in about 11% of CP cases mainly after surgery. Degree of resection and increased intracranial pressure are risk factors, which should be considered in the planning of surgical procedures for prevention of CI.
Effects and Side Effects of Pasireotide Treatment in Cushing’s Disease: Experience of a Single Tertiary Center
Authors: Ozkaya Hande Mefkure , Sahin Serdar , Kerimova Güler , Durcan Emre , Sulu Cem , Kadioglu Pinar
Published: May 2021

Context: Cushing’s disease (CD), when uncontrolled despite surgery, requires medical treatment. In this vein, treatments with long-term effectiveness and safety profiles are needed. We aimed to evaluate the effects and side effects of pasireotide treatment in Cushing’s disease in the real world. Methods: Patients who were followed up for Cushing’s disease and treated with pasireotide between 2001-2019 at Cerrahpaşa Medical Faculty Endocrinology, Metabolism and Diabetes Department were evaluated. Pasireotide efficacy and side effects were evaluated. Results: Thirty-four patients were included in the study. The mean duration of treatment was 24.59 (SD ± 15.21) months. Urinary free cortisol (UFC) decreased in 40.59 % of the patients and normalized in 33.3 %. A total of %71.4 of the patients who were in remission at the 3rd month of the treatment preserved their remission status at last follow-up. A significant difference was found between pre-treatment and final UFC (p = 0.001), but the absence of remission at 3 months did not predict final remission (p = 0.274). ACTH decreased by 9.16 % (SD ± 46.67). A significant difference was found between ACTH levels at 3rd, 6 months, and last control (p = 0.014, 0.017, and 0.017, respectively). Serum cortisol decreased by 13.89 %. A significant difference was found between pre-treatment and 3rd month, pre-treatment and final cortisol (p = 0.034 and 0.013, respectively). The body weight decreased by an average of 5.5 [IQR (0.0-11.5)] kg. Cholecystectomy was performed in 3 patients (9%). The percentage of patients requiring antidiabetic medication increased from 33.3% at diagnosis to 81.8% at the last follow-up visit. A significant difference was found between pre-treatment and 3rd month fasting plasma glucose levels ​​(p = 0.008). HbA1c increased by 17 %. There was a positive correlation between UFC and 3rd month HbA1c. All patients with hyperglycemia were controlled with effective antidiabetic therapy. Conclusion: Pasireotide treatment in CD is effective and safe, remission occurs in the first months of treatment and continues for a long time. Although hyperglycemia is the most common side effect, it is successfully controlled with antidiabetic medication.
Genome-Wide DNA Methylation Differences in Patients With Non-Functioning Pituitary Adenomas With or Without Postsurgical Intervention
Authors: Hallén Tobias , Johannsson Gudmundur , Dahlén Rahil , Andersson-Assarsson Johanna C , Glad Camilla A , Örndal Charlotte , Engvall Angelica , Carén Helena , Skoglund Thomas , Olsson Daniel S
Published: May 2021

Background: More than half of all surgically treated patients with non-functioning pituitary adenomas (NFPA) experience tumor progression during their follow-up, with some of these adenomas exhibiting locally aggressive behavior. Tumor progression is strongly associated with excess morbidity and mortality. Tumor control is, therefore, of vital importance for the patient outcome. There is no reliable method to predict tumor behavior after the initial surgical treatment. The aim of this study was therefore to study associations between DNA methylation patterns and tumor progression in patients with NFPA. Methods: In a case-controlled trial, 43 patients operated for NFPA with immunohistochemical characteristics of gonadotroph subtype were selected. Cases included patients who needed reintervention due to tumor progression (Reintervention group, n = 26), and controls had residual tumor without tumor progression during at least five years of follow-up (Radiologically stable group, n = 17). Genome-wide methylation analysis of the tumor tissue from each patient was performed on the Infinium MethylationEPIC BeadChip platform. Results: Principal component analysis (PCA) showed that clinical course was a strong contributor to the difference in DNA methylation profiles between the two groups. Also, unsupervised hierarchical clustering of the top 5,000 most variable CpG sites revealed three methylation profiles corresponding to, (1) the Reintervention group, (2) the Radiologically stable group, and (3) a mix of the two groups. Although the overall average methylation levels in the two groups did not differ (P = 0.16), 142,521 Differentially Methylated Positions (DMPs) were identified. Of these, 650 showed a difference in beta value methylation larger than 0.2. Interestingly, among the genes harboring the most significantly DMPs was GABRA1, and among the genes containing the largest number of DMPs were GATA2, NUP93, and LGALS1. In addition, the three most hypomethylated and the three most hypermethylated DMPs were all significantly (P <0.05) independently associated with tumor progression free survival (Log-rank test, data not shown). Conclusion: In this novel explorative study, we found a methylation pattern associated with postoperative tumor progression requiring reintervention in patients with NFPA. Among the differently methylated genes we identified, some have previously been described to be involved in pituitary adenoma differentiation or cancer development. Our study is a step towards deciphering epigenetic mechanisms involved in tumor progression and finding epigenetic signatures that might be used to predict postoperative tumor progression in NFPAs.
Hypercoagulability in ACTH-Dependent Cushing Syndrome
Authors: Singh Sumal Amit Kartar , Katznelson Laurence
Published: May 2021

Introduction: ACTH-dependent Cushing syndrome (CS) is associated with hypercoagulability; however, the incidence and timing of thrombosis during evaluation and management of CS is unclear.

Objective: To evaluate the incidence and timing of thrombotic events in patients with ACTH-dependent CS following diagnosis and management.

Methods: We performed a retrospective, longitudinal study of patients with ACTH-dependent CS seen at Stanford University Health Care from 1998 to 2020. Thrombotic events — deep vein thrombosis (DVT), pulmonary embolism (PE), cerebral vascular accident (CVA), and myocardial infarction (MI) — were recorded between diagnosis and 12 months following therapeutic intervention.

Results: Of 108 patients with ACTH-dependent CS, 97 (89.8%) were women, and the mean age at diagnosis was 43.0 years (± 15.7 years). Sixty-eight (63%) patients had hypertension, 38 (35.2%) had diabetes mellitus, and 11 (10.2%) were active smokers. Of the 108 subjects, 97 (89.8%) had Cushing Disease (CD) and 11 (10.2%) had ectopic CS. Of the 97 patients with CD, 38 (39.2%) underwent inferior petrosal sinus sampling (IPSS), 59 (60.8%) underwent transsphenoidal surgery (TSS), 19 required repeat TSS (19.6%), and 15 underwent TSS and bilateral adrenalectomy (BAL) (15.4%). Of the 11 patients with ectopic CS, 3 (27.2%) underwent IPSS, 6 (54.5%) underwent BAL, and 1 (9.1%) underwent TSS and BAL.

There were 10 thrombotic events among 7 (7.2%) CD patients, but no thrombotic events among ectopic CS patients. Of the thrombotic events, there were 7 (70%) DVT/PE, 2 (20%) CVA, and 1 (10%) cortical vein thrombosis. Six (60%) occurred within 30 days after TSS (range 3-25 days), 2 (20%) between 31 days and 1 year after TSS (range 59-165 days), 1 (10%) 26 days after IPSS but prior to TSS, and 1 (10%) in a patient who did not undergo IPSS or surgery. No thrombotic events were noted after BAL. Of the 8 postoperative thrombotic events, 5 (62.5%) occurred while patients received supraphysiologic glucocorticoid replacement (defined as >25mg hydrocortisone or equivalent daily) after curative surgery, 1 (12.5%) occurred after a patient was tapered to physiologic glucocorticoid replacement, and 2 (25%) occurred in patients who had persistent disease despite surgery. The degree of hypercortisolism at baseline was not associated with risk of thrombotic events.

Conclusions: In this retrospective study, 6.5% of ACTH-dependent CS patients had a thrombotic event, all in patients with CD. The majority had venous thromboembolism with DVT/PE, and the highest incidence occurred up to 30 days after surgery. The degree of hypercortisolism at baseline did not correlate with subsequent thrombotic events. Therefore, it is important to monitor all patients with ACTH-dependent CS following surgical intervention for venous thromboembolism.
Markers of Aggressiveness in Craniopharyngiomas
Authors: Del Can-Sanchez Diego Jesús , Sanchez Diego Jesus Del Can , Martínez-Ortega Antonio Jesús , Flores-Martínez Alvaro , Venegas-Moreno Eva , Dios-Fuentes María Elena , Madrazo-Atutxa Ainara , Valdepeñas Eugenio Cárdenas-Ruiz , Kaen Ariel Matías , Márquez-Rivas Francisco Javier , Roldán-Lora Anastasia Florinda , Fajardo-Picó Elena , Cano-González David , Soto-Moreno Alfonso
Published: May 2021

Craniopharyngiomas (CP) are rare tumors that may be locally aggressive. The presence of functional estrogen receptors (ER) has been reported in CP and might be related to risk of recurrence. Our aim is to ascertain if the expression estrogen and progesterone receptor (PR) might be associated with to recurrence in CP. Material and Methods: Descriptive retrospective observational study of patients with confirmed histology of CP and tissue sample available admitted to Virgen Del Rocio University Hospital (Seville, Spain) from January 1967 to October 2020 were included. Estrogen and progesterone receptor expression was analyzed by Immunohistochemistry. Ki-67 levels were also analyzed. Two CP groups were considereded according to Ki67 levels: Group A (Ki67<10%) and group B (Ki67>10%). As all variables followed a non-parametric distribution, U Mann Whitney, Chi-Square, and Z-test with Benjamini-Hochberg correction were used when needed. Results: Our study population includes 80 patients (46 male and 34 female), with a median age at diagnosis of 34 years [10-50.00]. Twenty-six patients were under 18 years old (children) with a median age of 7 years [4.5-10.00], and 54 were adults (aged 18 and above) with a median age of 45 years [33-58.50]. Our data shows higher recurrence rates when Ki67 levels staining were higher than 10%: 8/14 (57.2%) in comparison with Ki67<10% (6/14, 42.9%, p=0.018). In children we found 6 samples with Ki67<10% and 6 samples with Ki67 >10%; recurrences were observed in 2/6 (33,3%) in the first group and in 6/6 (100%) in the second, respectively (p= 0,199). In adults, we found 9 and 3 patients for high and low Ki67 levels, respectively. Recurrences were observed in 4/9 (44,4%) in the group A and in 2/3 (66,7%) in the group B, respectively (p= 0,28). There were no differences between age groups. In patients with positive ER, we observed an increased rate of recurrence: 12/23 (52.17%) versus 2/13 (15,38%) in patients with negative ER stain but it was no significant. (p=0,21). No association between PR and recurrence was observed. Conclusions: In our series, patients with CP with high Ki67 levels are more likely to recur. No clear association between ER, PR expression and recurrence was observed. These findings support the use of Ki67 as a marker of recurrence in CP. Sources of Research Support: Spanish Ministry of Health, ISCIII co-funded with Fondos FEDER (PI16/00175) and Novartis Oncology Spain.
The Change of Bone Turnover Markers and Bone Remodelling Balance in Acromegaly
Authors: Chen Huan , Zhang Yan , Liu Mengsi , Zhang Ziwei , Feng Wenhuan , Li Ping , Zhu Dalong
Published: May 2021

Objectives: The aim of this study was to evaluate the change of bone metabolism and bone remodelling balance in active and biochemical controlled acromegalic patients. Methods: We recruited 68 consecutive patients diagnosed as acromegaly in Nanjing Drum Tower Hospital. Of those, 39 patients after treatments with complete follow-up data were included. 30 sex-, age- and BMI-matched healthy persons were also included as normal controls. Serum calcium, phosphorus, parathyroid hormone (PTH), 25-hydroxy-vitamin D, bone turnover markers [osteocalcin, C-terminal telopeptide of type 1 collagen (CTX) and procollagen type 1 N-terminal propeptide (P1NP)], and bone remodelling index were investigated at baseline and after treatment. Univariate and multivariate linear regression analysis was used to determine the independent predictors of bone turnover markers. Results: At baseline, acromegalic patients had higher serum calcium, phosphorus and lower PTH levels than controls (all P < 0.01). Acromegalic patients had higher serum osteocalcin [45.98(31.98, 60.27) vs 15.83(12.74, 19.96) ng/ml, P < 0.01], P1NP [115.50(72.64, 145.90) vs 36.45(30.45, 44.28) ng/ml, P < 0.01] and CTX than controls [1.12(0.79, 1.56) vs 0.37(0.30, 0.50) ng/ml, P < 0.01], and the bone remodelling index of acromegaly patients is significantly increase compared to healthy controls [4.27(1.31, 5.99) vs 0.03(-0.38, 0.63), P < 0.01]. Multivariate linear regression analysis revealed that serum insulin-like growth factor 1 (IGF-1) was an independent predictor for osteocalcin (r = 0.610, P = 0.000), P1NP (r = 0.601, P = 0.000) and CTX (r = 0.596, P = 0.000). After treatment, serum calcium and phosphorus decreased, while PTH increased in acromegalic patients. Serum levels of bone turnover markers osteocalcin, PINP, CTX and bone remodelling index were also significantly decreased after treatment (all P < 0.05). In patients with biochemical control after treatment, bone turnover markers and bone remodelling index almost decreased to normal level (all P > 0.05). Conclusion: Acromegalic patients exhibited PTH-independent calcium-phosphate alterations, enhanced bone turnover state and positive bone remodeling balance. The abnormal bone metabolism is reversible after biochemical control of acromegaly. [Key words]Acromegaly; Bone turnover markers; Bone remodelling index; Growth hormone; Insulin-like growth factor 1
Vitamin D Metabolism Alteration in Acromegaly and Its Impact on Calcium-Phosphorus Metabolism
Authors: Povaliaeva Alexandra , Rozhinskaya Liudmila , Zhukov ScD, Artem , Pigarova Ekaterina A , Dzeranova ScD, Larisa K , Nikankina ScD, Larisa , Malysheva Natalya , Bogdanov Viktor , Ioutsi Vitaliy
Published: May 2021

Objective: to evaluate metabolism of vitamin D and calcium-phosphorus metabolism in patients with an active phase of acromegaly in comparison with healthy individuals. Materials and Methods: The study included 44 patients with an active acromegaly (IGF-1 788 [521; 963] ng/mL), as well as 49 conditionally healthy individuals. There were more men in the Acromegaly group (41% vs. 20%, p <0.05), patients were older (median age 42.7 [35.5; 26.5] vs. 26.3 [25; 33.5] years, p <0.05) and had a higher BMI (28.4 [25.2; 30.2] vs. 22.2 [20.1; 26.1] kg/m2, p <0.05) in a minor way compared with the control group. All participants were tested for vitamin D metabolites (25(OH)D3, 25(OH)D2, 1,25(OH)2D3, 3-epi-25(OH)D3 and 24,25(OH)2D3) by UPLC-MS/MS, free 25(OH)D and vitamin D-binding protein by ELISA, PTH by electrochemiluminescence immunoassay, as well as routine biochemical parameters of blood serum (calcium, phosphorus, creatinine, albumin, magnesium) and urine (calcium and phosphorus-creatinine ratio in spot urine). Results: In the Acromegaly group, we observed significantly higher levels of serum total calcium (2.46 [2.37; 2.56] vs. 2.38 [2.33; 2.45] mmol/L, p <0.05), albumin-corrected calcium (2.33 [2.28; 2.42] vs. 2.26 [2.21; 2.31] mmol/L, p <0.05) and phosphorus (1.39 [1.25; 1.55] vs. 1.15 [1.06; 1.23] mmol/L, p <0.05) as well as lower levels of serum albumin (45 [44; 47] vs. 46 [45; 48] g/L, p <0.05). The rest of the studied biochemical parameters and PTH levels did not differ significantly between the groups. The IGF-1 level in patients with acromegaly positively correlated with the level of total calcium (r = 0.49, p <0.05), albumin-corrected calcium (r = 0.49, p <0.05) and phosphorus (r = 0.55, p <0.05). The Acromegaly group showed lower levels of 25(OH)D3 (14.8 [11.8; 20.5] vs. 20.5 [14.8; 24.6] ng/mL, p <0.05), 3-epi-25(OH)D3 (1.0 [0.7; 1.4] vs. 1.4 [0.9; 1.8] ng/mL, p <0.05), 24,25(OH)2D3 (0.8 [0.4; 1.2] vs. 1.7 [0.9; 2.6] ng/ml, p <0.05) and free 25(OH)D (4.6 [3.7; 5.6] vs. 5.9 [4.0; 7.5] pg/mL, p <0.05), higher levels of 1,25(OH)2D3 (50 [42; 63] vs. 39 [34; 45] pg/mL, p <0.05), a lower 25(OH)D3/1,25(OH)2D3 ratio (289 [226; 443] vs. 517 [340; 641], p <0.05) and a higher 25(OH)D3/24,25(OH)2D3 ratio (19.3 [15.4; 27.7] vs. 11.9 [9.6; 15.2], p <0.05). Conclusion: Our data suggest that high levels of the active vitamin D metabolite (1,25(OH)2D3) resulting from an increase in 1α-hydroxylase activity may contribute to the elevation of calcium and phosphorus serum levels in patients with acromegaly. Our results also indicate a decrease in 24-hydroxylase activity in patients with acromegaly, which may be due to lower levels of 25(OH)D3 in these patients. The results obtained should be evaluated taking into account the observed differences in age, gender and BMI between groups.
Pituitary Somatotroph Adenoma Cell-Derived Exosomes: Characterization of Novel Non-Hormonal Functions
Authors: Zhou Cuiqi , Shen Stephen , Moran Rosemary , Melmed Shlomo
Published: May 2021

Exosomes, small extracellular vesicles carrying lipids, proteins, DNA and RNA, enable intercellular communication. Pituitary-derived exosomes have not been well validated, and as no human pituitary cell lines are available, we characterized exosomes derived from rat somatotroph tumor cells (GH1 and GH3). Rat FR and H9C2 cells were used as non-pituitary controls. Exosomes were isolated from serum-free culture supernatants by combining ultrafiltration and ultracentrifugation to eliminate hormone contamination. Derived exosomes were analyzed by NanoSight to visualize, size, and count particles. Exosomal proteins were extracted and exosome markers including TSG101, ALIX, CD63, HSP70, HSP90 detected by Western Blot. The exosome inhibitor GW4869 (10 µM, 30 h) reduced exosome release (up to 81%), whereas treating cells with hydrocortisone (0.1 µM, 72 h) increased exosome production (up to 42%) in GH1 and GH3 cells. Exosomal shuttle RNA characterized by RNA-Seq showed distinct pituitary vs non-pituitary exosome RNA profiles. Selected miRNAs assessed in exosomes and corresponding cells by qRT-PCR validated exosomal RNA-seq and suggested that miRNA signatures in exosomes and in respective cells of origin were concordant. Next, we explored downstream signaling of GH1-derived exosomes (GH1-exo) in vitro and in vivo and studied biological actions in normal hepatocytes and in malignant cells. As evidenced by mRNA-seq, GH1-exo distinctly altered signaling pathways in rat primary hepatocytes, vs pathways elicited by GH or PRL (0.5 µg/mL, 24 h). GH1-exo, FR-exo or vehicle were intravenously injected to 4-week-old female Wistar rats twice weekly for 4 weeks (5*109 exo/200 g, n=3), and livers dissected for mRNA-seq. Among GH1-exo specifically regulated genes, EIF2AK/ATF4, involved in cAMP responses and amino acid biosynthesis, were attenuated. In hepatocytes, GH1-exo suppressed up to 65% of nascent protein synthesis and reduced forskolin (10 µM)-stimulated cAMP activity by 19%, while GH (0.01-1 µg/mL) did not affect this pathway. Notably, GH1-exo also attenuated malignant cell motility. Both GH1-exo incubation or GH1 cell co-culture (48 h) suppressed migration, invasion and wound healing of HCT116 cancer cells by up to 70%. In contrast, treatment with rGH (0.5 µg/mL) increased HCT116 motility. Intravenous administration of GH1-exo (1010 exo/mouse, twice a week for 5 weeks) decreased metastatic tumor volume by 40% in nude mice harboring splenic HCT116 implants (5*105 cells/mouse, n=10), and especially abrogated hepatic metastases. mRNA-seq of GH1-exo treated HCT116 cells vs controls indicated dysregulated p53 and MAPK pathways, which may partially explain mechanisms underlying motility attenuation. The results elucidate novel biological actions of somatotroph adenoma cell-derived exosomes and suggest exosomes as non-hormonal messengers produced by pituitary tumors.
Dapagliflozin Exerts Adrenal Sympatholysis via G protein-Coupled Receptor Kinase-2 & Tyrosine Hydroxylase Downregulation
Authors: Lymperopoulos Anastasios , Pollard Celina M , Noa Daniela Pi , Ferraino Krysten E , Cora Natalie , Pereyra Janelle M , Ghandour Jennifer , Valiente Rachel , Maning Jennifer , Brill Ava R , Iacobellis Gianluca
Published: May 2021

The sodium-glucose co-transporter (SGLT)-2 inhibitor dapagliflozin was recently reported to reduce renal tyrosine hydroxylase (TH) and norepinephrine levels, lowering blood pressure and preventing endothelial dysfunction, in a murine model of neurogenic hypertension. This suggested that dapagliflozin may combat sympathetic nervous system (SNS) hyperactivity, which is known to accompany and aggravate chronic heart failure (CHF). Adrenal G protein-coupled receptor kinase (GRK)-2 upregulation is a major driver of circulating catecholamine (CA) elevation and SNS hyperactivity, especially in CHF. GRK2 severely dysregulates adrenal sympatho-inhibitory α 2-adrenergic receptors (ARs), leading to unchecked, chronically elevated CA secretion. Therefore, we hypothesized herein that SGLT2 inhibition with dapagliflozin may lower SNS hyperactivity in the adrenal gland by antagonizing GRK2 actions on α 2ARs in adrenal chromaffin cells. We used the rat pheochromocytoma PC12 cell line expressing human α 2AAR, as well as freshly isolated adrenal glands from adult male Sprague-Dawley rats treated with dapagliflozin in vivo. We measured circulating norepinephrine (NE) in vivo via RIA, GRK2 & TH expression levels via real-time PCR and immunoblotting, adrenal α 2AR density (Bmax) via saturation radioligand binding with [methyl-3H]-rauwolscine, and G protein activation via the GTPγS assay. Dapagliflozin treatment for 7 consecutive days (20 mg/kg/d in drinking water) led to a significant reduction in blood circulating NE levels (217+67 pg/ml, n=6), compared to control, vehicle-treated rats (363+77 pg/ml, n=6, p<.05), suggesting reduced SNS activity. This was accompanied by reduced GRK2 and TH mRNA and protein levels in dapagliflozin-treated rat adrenals vs. vehicle-treated animal-derived glands, indicating reduced adrenal CA synthesis and secretion. Finally, adrenal α 2AR density was higher in dapagliflozin- vs. vehicle-treated rats (51.3+7.3 vs. 26.1+8.1 fmol/mg of protein, respectively; n=12 glands from 6 animals per group, p<.05). These results (i.e. GRK2 and TH downregulation) were completely recapitulated in PC12 α 2AAR-expressing cells in culture, treated with 5 μM dapagliflozin (or vehicle) for 24 hours. Importantly, α 2AR-dependent G protein-mediated signaling towards inhibition of CA secretion was markedly enhanced at 24 hrs post-dapagliflozin application in PC12 cells. This was the result of reduced GRK2-dependent receptor desensitization, since dapagliflozin lacked this effect in cells co-transfected with a GRK2-encoding adenovirus to acutely overexpress GRK2. In conclusion, dapagliflozin exerts a sympatholytic action in the adrenal medulla via downregulation of both TH, which reduces CA biosynthesis, and GRK2, which reduces α 2AR desensitization in favor of enhanced inhibition of CA secretion.
FGF-2-Dependent Signaling Activated in Aged Human Skeletal Muscle Promotes Intramuscular Adipogenesis
Authors: Mathes Sebastian , Fahrner Alexandra , Ghoshdastider Umesh , Rüdiger Hannes A , Leunig Michael , Wolfrum Christian , Krützfeldt Jan
Published: May 2021

Background: A unique feature of muscle during aging, obesity and type 2 diabetes is the appearance of adipose tissue between skeletal muscle fibers, the intramuscular adipose tissue (IMAT). IMAT is generally associated with systemic insulin resistance, decreased muscle strength and, in older adults, impaired mobility. A large body of work over the last years have addressed the origin of IMAT. Skeletal muscle contains preadipocyte progenitors termed fibro/adipogenic progenitors (FAPs) that normally do not form adipocytes, but proliferate during muscle injury to support the commitment of myogenic progenitor cells during muscle repair. However, under the conditions outlined above, FAPs differentiate to adipocytes and give rise to IMAT. The molecular cues that trigger this pathogenesis are unknown and strategies that can modulate the fate of FAPs and their propensity to differentiate to adipocytes in disease states are urgently needed. Methods: We have used molecular cloning and genome editing techniques together with a variety of biochemical, molecular, genomic, transcriptomic and proteomic analyses in pre-adipocyte and muscle cell lines, primary cells isolated from human and mouse skeletal muscle, human skeletal muscle biopsies, and animal models. Results: Using multiple screening assays upstream and downstream of a master regulator of myogenesis, microRNA (miR)-29a, we located the secreted protein and adipogenic inhibitor SPARC to an FGF-2 signaling pathway that is conserved between skeletal muscle cells from mice and humans and that is activated in skeletal muscle of aged mice and humans. We identified FGF-2 as an upstream regulator that increases miR-29a promoter activity and gene expression via MEK1/2/MAPK signaling and consequently decreases the conserved miR-29a target SPARC. FGF-2 induces the miR-29a/SPARC axis through transcriptional activation of FRA-1, which binds and activates an evolutionary conserved AP-1 site element proximal in the miR-29a promoter. CRISPR/Cas9-mediated genomic deletions of miR-29a recognition elements located in the Sparc 3′UTR of muscle cells and AAV-mediated overexpression of FGF-2 or SPARC in mouse skeletal muscle revealed that this axis regulates differentiation of FAPs in vitro and IMAT formation in vivo. Skeletal muscle from human donors aged > 75 years versus < 55 years showed activation of FGF-2-dependent signaling and increased IMAT. Conclusion: FGF-2 not only stimulates muscle growth, but also promotes intramuscular adipogenesis via targeting the fate of FAPs. Thus, our data highlights a disparate role of FGF-2 in adult skeletal muscle and reveals a novel pathway, which has the potential to provide innovative diagnostic and therapeutic approaches to combat fat accumulation in aged human skeletal muscle.
Human GPR17 Nonsynonymous Variants Identified in Individuals with Metabolic Diseases Have Distinct Functional Signaling Profiles
Authors: Conley Jason M , Ren Hongxia
Published: May 2021

GPR17 is a G protein-coupled receptor (GPCR) implicated in the regulation of glucose metabolism and energy homeostasis. Our genetic knockout studies in rodents suggest that GPR17 is a potential therapeutic target for the treatment of metabolic diseases. However, the contributions of GPR17 to human metabolism and metabolic deficits are not well understood. Here, we analyzed the human GPR17 coding sequences of individuals from control and metabolic disease cohorts that were comprised of patients with clinical phenotypes including severe insulin resistance, hypercholesterolemia, and obesity. Across cohorts, 18 nonsynonymous GPR17 variants were identified, including eight variants that were exclusive to the disease cohort. We characterized the protein expression levels, cellular localization, and downstream functional signaling profiles of nine human GPR17 variants (F43L, V96M, V103M, D105N, A131T, G136S, R248Q, R301H, and G354V). We found that the protein expression levels and subcellular localization for each of the nine GPR17 variants were similar to that of the wild type GPR17. As the endogenous GPR17 ligand is still elusive, we used a synthetic GPR17 agonist to quantitatively measure the functional signaling profiles of GPR17 variants. We found some of the variants had distinctly altered signaling profiles. GPR17-G136S lost agonist-mediated cAMP, Ca2+, and beta-arrestin signaling. GPR17-V96M retained cAMP inhibition similar to GPR17-WT but had impaired Ca2+ and beta-arrestin signaling. GPR17-D105N displayed impaired cAMP and Ca2+ signaling but enhanced agonist-stimulated beta-arrestin recruitment. Also, GPR17-G354V retained cAMP and Ca2+ signaling function but had attenuated beta-arrestin recruitment. The identification and functional profiling of naturally occurring human GPR17 variants from individuals with metabolic diseases revealed receptor variants with distinct signaling profiles, including differential signaling perturbations that resulted in receptor signaling bias. These results are expected to contribute to our understanding of the molecular signaling mechanisms underlying GPR17 in metabolic regulation.
12-year Trends in Lipid Levels in Korean Children and Adolescents: A Cross-sectional Study Based on the Korea National Health and Nutrition Examination Survey
Authors: Song Kyungchul , Soyoung Jeon , Lee Hye Sun , Choi Youngha , Oh Jun Suk , Choi Han Saem , Suh Junghwan , Kwon Ahreum , Kim Ho-Seong , Chae Hyun Wook
Published: May 2021

Background: Dyslipidaemia at a young age is one of the risk factors of adulthood cardiovascular disease. However, investigations on the trends in lipid levels among the youth are scarce. Thus, we aimed to analyse the trends in lipid levels in Korean youth from 2007 through 2018. Methods: This cross-sectional study investigated the lipid profiles of 10,734 youths aged 10-18 years using data from the four phases of the Korea National Health and Nutritional Examination Survey: IV (2007-2009), V (2020-2012), VI (2013-2015), and VII (2016-2018). We assessed the age-, sex-, and body mass index (BMI)-adjusted mean levels of lipids at each survey. Findings: The mean levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and non-high-density lipoprotein cholesterol (non-HDL-C) levels increased from phase IV to VII. Among boys, the prevalence of subjects with acceptable levels of TC, LDL-C, and non-HDL-C decreased, whereas that of subjects with adverse levels increased from phase VI to VII. In BMI- and sex-specific analysis, adverse trends in TC, LDL-C, and non-HDL were more apparent among boys with normal BMI than among those who were overweight and obese. Among girls, the mean TC, LDL-C, and triglycerides levels increased in subjects with normal BMI and overweight. Interpretation: Our results suggest worsening trends in lipid levels and dyslipidaemia among Korean youth. These adverse trends were seen even in youth with normal BMI. These findings indicate the need for more concern about lipid levels in youth with normal BMI as well as those with overweight and obesity.
A Novel Graph Based Semi-Supervised Learning Approach to Identify Pathways Contributing to the Development of Diabetes and Obesity
Authors: Lee Garam , Nam Yonghyan , Shivakumar Manu , Joshi Apoorva , Fu Weixuan , Simmons Rebecca Anne , Wang Paul , Kim Dokyoon , Pinney Sara Elizabeth
Published: May 2021

Background: Gestational diabetes (GDM) has profound effects on the intrauterine metabolic milieu, induces marked abnormalities in fetal glucose and insulin secretion and is linked to obesity and diabetes in the offspring, but the mechanisms remain largely unknown. Epigenetic modifications in stems cells may be one mechanism by which an in utero exposure can lead to the development of diabetes and obesity later in life.

Objective: To identify novel pathways contributing to the development of diabetes and obesity in offspring exposed to GDM in utero by integrating data generated from transcriptome and methylome analysis from second trimester human amniocytes exposed to GDM in utero.

Methods: We analyzed RNAseq and genome wide DNA methylation data (ERRBS) generated from second trimester amniocytes obtained from women with GDM (n=14). Amniocytes have stem cells-like characteristics and are derived from the fetus. Expression data of 22,271 genes were retrieved from RNAseq data. CpGs with significant changes in DNA methylation were mapped into 20,028 genes by collapsing methylation probes into promoter and gene regions. To better understand the associations among diverse gene sets or among gene sets and GDM,we first constructed two weighted co-expression networks from RNAseq and DNA methylation data, respectively. Then, two co-expression networks were integrated using a linear combination. With the integrated co-expression network, graph-based label propagation algorithm was employed to prioritize GDM-associated genes.

Results: From the differential expression analysis between GDM and control, the top 20 query genes, including 11 genes and 9 methylated genes, were selected for label propagation. Finally, the top 100 genes were picked up for the pathway enrichment testusing an over-representation analysis approach. Significantly enriched pathways included: Interferon Signaling, N-glycan Antennae Elongation, Sphingolipid Pathway and Metabolism, Classical Complement Pathway, Complement and Coagulation Cascades, Tryptophan Metabolism, Peroxisomal Protein Import, Unsaturated Fatty Acid Metabolism, Complement Activation, Human Innate Immune Response, Ceramide Metabolism, Fertilization Pathway, Keratan Sulfate Biosynthesis Pathway, Transport to the Golgi and Modification Pathway (FDR q<0.05 for all pathways).

Conclusion: Using a novel bioinformatic approach to synthesize transcriptome and methylome data derived from human amniocytes exposed to GDM in utero, we identified potential pathways involved in programming of diabetes and obesity in offspring including pathways involving the immune response, complex lipid metabolism, the complement pathway, and protein transport and processing. Further investigation of these pathways may yield new mechanisms contributing to diabetes and obesity.