Nevin Manimala

J Ethnopharmacol. 2022 Jan 14:114991. doi: 10.1016/j.jep.2022.114991. Online ahead of print.

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Platycodon grandiflorus (Jacq.) A.DC. (PG) is a common natural medicine with a history of thousands of years. The processing products were mainly recorded as raw, honey-processed, wine-fried, yellow-fried, and bran-fried PG, which were respectively used for different clinical purposes. Therefore, it is necessary to study the chemical composition and pharmacological activity of PG after processing.

AIM OF THE STUDY: To explore the effects of different processing methods on the composition and biological activity of PG using metabonomics and pharmacologic design.

MATERIALS AND METHODS: UPLC-QTOF-MS combined with multivariate statistical analysis was used to identify different metabolites before and after the processing of PG. Network pharmacology was used to construct the metabolite-target-disease network. CCK-8 assay, flow cytometry, and western blotting were used to detect cell viability, apoptosis, and the expression of related proteins, respectively.

RESULT: A total of 43 differentially expressed metabolites (VIP >10) were detected and identified in the analyzed groups. Based on their chemical nature, these metabolites were divided into five categories, namely, saccharolipids, flavonoid glycosides, alkynes, saponins, and lipids (including fatty acids, phospholipids, fatty aldehydes, and sterols). The content of lipids in the five processed groups (CH, FC, JZ, MZI, and MZG) was found to be higher than that in raw PG. In particular, the processing approaches explored herein increased the contents of many phospholipids, namely, glycerophosphoinositols, phosphatidic acids, and lysophosphatidyle·thanolamines. In total 8 metabolites were found by venn diagram to distinguish different processed products (metabolites 2, 6, 19, 20, 21, 26, 28, and 38). The results of network pharmacology analysis showed that the primary anti-cancer targets of 43 metabolites of PG processing products are PIK3CA, Akt, and STAT3, and based on CCK-8 assay, MZI-processed PG has a significant killing effect on A549 cells, compared to other processing techniques. Moreover, flow cytometry analysis showed that the cells treated with MZI-processed PG exhibit significantly increased cell apoptosis, and that the effect is dose-dependent. Finally, the western blots performed herein demonstrated that the MZI-processed PG effectively inhibits the expression of p-Akt and p-STAT3, which is consistent with the network pharmacology results.

CONCLUSION: Depending on the processing technique, the contents of 43 different metabolites in PG vary significantly. Specifically, the contents of phospholipids and fatty acids increase, whereas the contents of large Mw saponins decrease. Compared to the other investigated processing methods, MZI increases the potential of PG in inducing cell apoptosis and inhibiting cell proliferation by affecting the Akt and STAT3 signaling pathways. The increased levels of 3-O-β-glucopyranosyl polygalacic acid and platycoside F after honey-frying confirm these results.

PMID:35038566 | DOI:10.1016/j.jep.2022.114991

Contraception. 2022 Jan 14:S0010-7824(22)00004-X. doi: 10.1016/j.contraception.2022.01.003. Online ahead of print.

ABSTRACT

OBJECTIVE(S): The Medicaid consent policy has been identified as a major barrier to desired permanent contraception, particularly for low-income communities and communities of color. As each state may modify their state Medicaid sterilization consent form, variation in the form has been reported. This study aims to characterize state-level variation in Medicaid Title XIX consent form interpretation and application.

STUDY DESIGN: We aimed to collect primary data from Medicaid officials in all 50 United States from January to May 2020 via a 25-question electronic survey regarding state-level consent form implementation. Questions targeted consent form details and definitions, insurance and billing, clinician correspondence, and administrative processes. We used Qualtrics XM® to collect survey responses. We performed descriptive statistics on the survey responses. There were no exclusion criteria.

RESULTS: We had 41 responses from 36/50 states (72% participation rate). Heterogeneity existed in the key definitions of “Premature Delivery” and “Emergency Abdominal Surgery.” One in five respondents reported the consent form was only available in English. Variation among Current Procedural Terminology codes covered in each state’s sterilization policy were noted. Nearly a quarter of respondents did not know how Medicaid informed healthcare providers of consent form denials. Most participants (90%) were unaware of differences between state sterilization policies.

CONCLUSION: This study demonstrates variation in terms of consent form definitions, procedures covered, correspondence with clinicians, and administrative review processes among state Medicaid offices regarding the sterilization consent form. Greater transparency is necessary in order to reduce administrative barriers to desired permanent contraception.

PMID:35038447 | DOI:10.1016/j.contraception.2022.01.003

Microvasc Res. 2022 Jan 14:104316. doi: 10.1016/j.mvr.2022.104316. Online ahead of print.

ABSTRACT

INTRODUCTION: Frailty is highly prevalent in heart failure (HF) patients. HF is associated with oxidative stress and chronic inflammation, which impair oxygen use by skeletal muscles. Little is known about the influence of frailty on vascular responsiveness and tissue oxygenation.

OBJECTIVE: Analyze the influence of frailty on vascular responsiveness and muscle oxygenation in elderly individuals with and without HF.

METHODS: Individuals aged ≥60 years, with or without HF, were evaluated for frailty (phenotype). Near-infrared spectroscopy (NIRS) was used to assess muscle oxygenation at rest (oxygen saturation – StO₂ and deoxyhemoglobin) and during handgrip exercise (minimum StO₂ and maximum deoxyhemoglobin), and oxygenation variables.

STATISTICAL ANALYSIS: Results were grouped according to the frailty phenotype: non-frail, pre-frail, and frail. Shapiro-Wilk test was used to assess normality. Data were compared using a two-way analysis of variance (ANOVA). Bonferroni post hoc test was applied to determine the influence of frailty or HF on NIRS variables. SPSS software was used in the analyses; p < 0.05 was considered significant.

RESULTS: 55 elderly participants (61.8% female; 70.4 ± 7.2 years old; 28 HF patients) participated in the study. 32.7% (n = 18) were classified as non-frail, 43.3% (n = 24) as pre-frail, and 23.6% (n = 13) as frail. The analysis of vascular responsiveness (n = 52) identified an influence (p < 0.05) of frailty on the reperfusion rate (slope 2 and ∆StO₂ of nadir-peak) and desaturation during occlusion (area under the curve of StO₂) in HF patients. There was no influence of frailty or HF on muscle oxygenation at rest and during exercise (n = 54; p > 0.05).

CONCLUSION: The coexistence of frailty and HF seems to impair vascular responsiveness, as frail elderly participants with HF presented lower reperfusion rates and higher desaturation levels during the arterial occlusion test. However, the presence of frailty or HF alone had no influence on muscle oxygenation at rest or during exercise.

PMID:35038445 | DOI:10.1016/j.mvr.2022.104316

Lancet Oncol. 2022 Jan 14:S1470-2045(21)00718-X. doi: 10.1016/S1470-2045(21)00718-X. Online ahead of print.

ABSTRACT

BACKGROUND: Trastuzumab is a monoclonal antibody against HER2 (also known as ERBB2). The primary objective of the NRG Oncology/RTOG-1010 trial was to establish whether trastuzumab improves disease-free survival when combined with trimodality treatment (paclitaxel plus carboplatin and radiotherapy, followed by surgery) for patients with untreated HER2-overexpressing oesophageal adenocarcinoma.

METHODS: NRG Oncology/RTOG-1010 was an open label, randomised, phase 3 trial for which patients were accrued from 111 NRG-affiliated institutions in the USA. Eligible patients were adults (aged ≥18 years) with newly diagnosed pathologically confirmed oesophageal adenocarcinoma, American Joint Committee on Cancer 7th edition T1N1-2 or T2-3N0-2 stage disease, and a Zubrod performance status of 0-2. Patients were stratified by adenopathy (no vs yes [coeliac absent] vs yes [coeliac present ≤2 cm]) and randomly assigned (1:1) to receive weekly intravenous paclitaxel (50 mg/m² intravenously over 1 h) and carboplatin (area under the curve 2, intravenously over 30-60 min) for 6 weeks with radiotherapy 50·4 Gy in 28 fractions (chemoradiotherapy) followed by surgery, with or without intravenous trastuzumab (4 mg/kg in week one, 2 mg/kg per week for 5 weeks during chemoradiotherapy, 6 mg/kg once presurgery, and 6 mg/kg every 3 weeks for 13 treatments starting 21-56 days after surgery). The primary endpoint, disease-free survival, was defined as the time from randomisation to death or first of locoregional disease persistence or recurrence, distant metastases, or second primary malignancy. Analyses were done by modified intention to treat. This study is registered with Clinicaltrials.gov, NCT01196390; it is now closed and in follow-up.

FINDINGS: 606 patients were entered for HER2 assessment from Dec 30, 2010 to Nov 10, 2015, and 203 eligible patients who were HER2-positive were enrolled and randomly assigned to chemoradiotherapy plus trastuzumab (n=102) or chemoradiotherapy alone (n=101). Median duration of follow-up was 2·8 years (IQR 1·4-5·7). Median disease-free survival was 19·6 months (95% CI 13·5-26·2) with chemoradiotherapy plus trastuzumab compared with 14·2 months (10·5-23·0) for chemoradiotherapy alone (hazard ratio 0·99 [95% CI 0·71-1·39], log-rank p=0·97). Grade 3 treatment-related adverse events occurred in 41 (43%) of 95 patients in the chemoradiotherapy plus trastuzumab group versus 52 (54%) of 96 in the chemoradiotherapy group and grade 4 events occurred in 20 (21%) versus 21 (22%). The most common grade 3 or worse treatment-related adverse events for both groups were haematological (53 [56%] of 95 patients in the chemoradiotherapy plus trastuzumab group vs 55 [57%] of 96 patients in the chemotherapy group) or gastrointestinal disorders (28 [29%] vs 20 [21 %]). 34 (36%) of 95 patients in the chemoradiotherapy plus trastuzumab group and 27 (28%) of 96 patients in the chemoradiotherapy only group had treatment-related serious adverse events. There were eight treatment-related deaths: five (5%) of 95 patients in the chemoradiotherapy plus trastuzumab group (bronchopleural fistula, oesophageal anastomotic leak, lung infection, sudden death, and death not otherwise specified), and three (3%) of 96 in the chemoradiotherapy group (two multiorgan failure and one sepsis).

INTERPRETATION: The addition of trastuzumab to neoadjuvant chemoradiotherapy for HER2-overexpressing oesophageal cancer was not effective. Trastuzumab did not lead to increased toxicities, suggesting that future studies combining it with or using other agents targeting HER2 in oesophageal cancer are warranted.

FUNDING: National Cancer Institute and Genentech.

PMID:35038433 | DOI:10.1016/S1470-2045(21)00718-X

Lancet Oncol. 2022 Jan 14:S1470-2045(21)00650-1. doi: 10.1016/S1470-2045(21)00650-1. Online ahead of print.

ABSTRACT

BACKGROUND: PD-1 inhibitor plus chemotherapy had been shown to be an effective first-line treatment for patients with metastatic non-small-cell lung cancer (NSCLC). However, there was no robust evidence showing a PD-L1 inhibitor combined with chemotherapy benefited patients with squamous and non-squamous NSCLC. GEMSTONE-302 aimed to evaluate the efficacy and safety of a PD-L1 inhibitor, sugemalimab, plus chemotherapy for patients with metastatic squamous or non-squamous NSCLC.

METHODS: This randomised, double-blind, phase 3 trial was done in 35 hospitals and academic research centres in China. Eligible patients were aged 18-75 years, had histologically or cytologically confirmed stage IV squamous or non-squamous NSCLC without known EGFR sensitising mutations, ALK, ROS1, or RET fusions, no previous systemic treatment for metastatic disease, and an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. Patients were randomly assigned (2:1) to receive sugemalimab (1200 mg, intravenously, every 3 weeks) plus platinum-based chemotherapy (carboplatin [area under the curve (AUC) 5 mg/mL per min, intravenously] and paclitaxel [175 mg/m², intravenously] for squamous NSCLC, or carboplatin [AUC 5 mg/mL per min, intravenously] and pemetrexed [500 mg/m², intravenously] for non-squamous NSCLC; sugemalimab group) or placebo plus the same platinum-based chemotherapy regimens for squamous or non-squamous NSCLC as in the sugemalimab group; placebo group) for up to four cycles, followed by maintenance therapy with sugemalimab or placebo for squamous NSCLC, and intravenous sugemalimab 500 mg/m² or matching placebo plus pemetrexed for non-squamous NSCLC. Randomisation was done by an interactive voice-web-response system via permuted blocks (block size was a mixture of three and six with a random order within each stratum) and stratified by ECOG performance status, PD-L1 expression, and tumour pathology. The investigators, patients, and the sponsor were masked to treatment assignment. The primary endpoint was investigator-assessed progression-free survival in the intention-to-treat population. Safety was analysed in all patients who received at least one treatment dose. Results reported are from a prespecified interim analysis (ie, when the study met the primary endpoint) and an updated analysis (prespecified final analysis for progression-free survival) with a longer follow-up. This study is registered with ClinicalTrials.gov (NCT03789604), is closed to new participants, and follow-up is ongoing.

FINDINGS: Between Dec 13, 2018, and May 15, 2020, 846 patients were assessed for eligibility; 367 were ineligible, and the remaining 479 patients were randomly assigned to the sugemalimab group (n=320) or placebo group (n=159). At the preplanned interim analysis (data cutoff June 8, 2020; median follow-up 8·6 months [IQR 6·1-11·4]), GEMSTONE-302 met its primary endpoint, with significantly longer progression-free survival in the sugemalimab group compared with the placebo group (median 7·8 months [95% CI 6·9-9·0] vs 4·9 months [4·7-5·0]; stratified hazard ratio [HR] 0·50 [95% CI 0·39-0·64], p<0·0001]). At the final analysis (March 15, 2021) with a median follow-up of 17·8 months (IQR 15·1-20·9), the improvement in progression-free survival was maintained (median 9·0 months [95% CI 7·4-10·8] vs 4·9 months [4·8-5·1]; stratified HR 0·48 [95% CI 0·39-0·60], p<0·0001). The most common grade 3 or 4 any treatment-related adverse events were neutrophil count decreased (104 [33%] of 320 with sugemalimab vs 52 [33%] of 159 with placebo), white blood cell count decreased (45 [14%] vs 27 [17%]), anaemia (43 [13%] vs 18 [11%]), platelet count decreased (33 [10%] vs 15 [9%]), and neutropenia (12 [4%] vs seven [4%]). Any treatment-related serious adverse events occurred in 73 (23%) patients in the sugemalimab group and 31 (20%) patients in the placebo group. Any treatment-related deaths were reported in ten (3%) patients in the sugemalimab group (pneumonia with respiratory failure in one patient; myelosuppression with septic shock in one patient; pneumonia in two patients; respiratory failure, abdominal pain, cardiac failure, and immune-mediated pneumonitis in one patient each; the other two deaths had an unspecified cause) and in two (1%) patients in the placebo group (pneumonia and multiple organ dysfunction syndrome).

INTERPRETATION: Sugemalimab plus chemotherapy showed a statistically significant and clinically meaningful progression-free survival improvement compared with placebo plus chemotherapy, in patients with previously untreated squamous and non-squamous metastatic NSCLC, regardless of PD-L1 expression, and could be a newfirst-line treatment option for both squamous and non-squamous metastatic NSCLC.

FUNDING: CStone Pharmaceuticals.

TRANSLATION: For the Chinese translation of the abstract see Supplementary Materials section.

PMID:35038432 | DOI:10.1016/S1470-2045(21)00650-1

J Am Med Dir Assoc. 2022 Jan 14:S1525-8610(21)01104-X. doi: 10.1016/j.jamda.2021.12.030. Online ahead of print.

ABSTRACT

OBJECTIVE: To test the effect of a personalized music intervention on agitated behaviors and medication use among long-stay nursing home residents with dementia.

DESIGN: Pragmatic, cluster-randomized controlled trial of a personalized music intervention. Staff in intervention facilities identified residents’ early music preferences and offered music at early signs of agitation or when disruptive behaviors typically occur. Usual care in control facilities may include ambient or group music.

SETTING AND PARTICIPANTS: The study was conducted between June 2019 and February 2020 at 54 nursing homes (27 intervention and 27 control) in 10 states owned by 4 corporations.

METHODS: Four-month outcomes were measured for each resident. The primary outcome was frequency of agitated behaviors using the Cohen-Mansfield Agitation Inventory. Secondary outcomes included frequency of agitated behaviors reported in the Minimum Data Set and the proportion of residents using antipsychotic, antidepressant, or antianxiety medications.

RESULTS: The study included 976 residents with dementia [483 treatment and 493 control; mean age = 80.3 years (SD 12.3), 69% female, 25% African American]. CMAI scores were not significantly different (treatment: 50.67, SE 1.94; control: 49.34, SE 1.68) [average marginal effect (AME) 1.33, SE 1.38, 95% CI -1.37 to 4.03]. Minimum Data Set-based behavior scores were also not significantly different (treatment: 0.35, SE 0.13; control: 0.46, SE 0.11) (AME -0.11, SE 0.10, 95% CI -0.30 to 0.08). Fewer residents in intervention facilities used antipsychotics in the past week compared with controls (treatment: 26.2, SE 1.4; control: 29.6, SE 1.3) (AME -3.61, SE 1.85, 95% CI -7.22 to 0.00), but neither this nor other measures of psychotropic drug use were statistically significant.

CONCLUSIONS AND IMPLICATIONS: Personalized music was not significantly effective in reducing agitated behaviors or psychotropic drug use among long-stay residents with dementia. Barriers to full implementation included engaging frontline nursing staff and identifying resident’s preferred music.

PMID:35038407 | DOI:10.1016/j.jamda.2021.12.030

Colorectal Dis. 2022 Jan 17. doi: 10.1111/codi.16054. Online ahead of print.

ABSTRACT

AIM: To determine safety of performing an anastomosis after rectal cancer (RC) resection in patients with a previously treated prostate cancer (PC).

METHODS: Patients with a previously treated PC who underwent rectal resection from 2008 to 2018 were retrospectively included. Outcomes were compared between patients who underwent rectal resection with anastomosis (restorative surgery, RS+ group) and those with a definitive stoma (RS- group). In the RS+ group, anastomotic leak (AL) rates were assessed according to the type of reconstruction.

RESULTS: 126 patients underwent rectal surgery for mid-low RC after a previously PC treated by RT and/or radical prostatectomy. Overall, 80 patients (63%) underwent a RS and 46 patients (37%) underwent rectal surgery with a definitive stoma. There was no statistical difference between the two groups in terms of intraoperative data, except for the type of resection with more multivisceral resection in the RS- group (p<0.01). In the RS+ group, a diverting stoma was performed in 74% of cases. No difference between the two groups in terms of overall morbidity was found. In the RS+ group (n=80), 17 patients (21%) experienced AL. Of these none was observed when delayed coloanal anastomosis was performed (p=0.16). Long-term permanent stoma in the RS+ group was 16% (n=13).

CONCLUSION: Restorative surgery after resection for RC in patients with a previous history of RT and/or radical prostatectomy for PC is safe without additional morbidity. In selected patients for restorative surgery, performing delayed coloanal anastomosis may represent a promising option.

PMID:35038368 | DOI:10.1111/codi.16054

J Clin Endocrinol Metab. 2022 Jan 17:dgac021. doi: 10.1210/clinem/dgac021. Online ahead of print.

ABSTRACT

PROPOSE: Obesity-related metabolic risk factors in adolescents with overweight/obesity may be associated with systemic low-grade inflammation, and therefore we investigated whether a 6-month exercise training altered markers of inflammation.

METHODS: Secondary analyses of a randomized controlled exercise-based intervention trial (September 2017 to December 2018). Adolescents aged 11 to 17 years (Tanner stage II to V), 70% girls, with a body mass index (BMI) z-score at or above the 85 th percentile, and/or with excess of adiposity (body fat >30%). The participants were randomly assigned to the following 4 groups for 6 months: (1) standard physical education lessons, as a control (CTRL); (2) high-intensity physical education class (HIPE); (3) low-to-moderate intensity physical education class (LIPE); (4) a combined group (PLUS group). Inflammatory markers and immune molecules including chemokines, cytokines, and growth factors (n=65 biomarkers) were determined by cytokine antibody array.

RESULTS: Of the 120 randomly assigned participants, 95 were included in the analysis. Considering these 22 proteins, the LIPE group shows statistical significance in 9 proteins with logfold-change (logFC) and p<0.05 was found in BLC, Eotaxin, FGF-6, GCP-2, I-309, IGFBP-4, MCP-4, NAP-2, and PARC), followed by the PLUS group in 9 proteins (BLC, EGF, Eotaxin, FGF-6, MCP-4, NAP-2, Osteopontin, PARC, and RANTES), the HIPE group in 7-proteins (FGF-4, FGF-7, GCP-2, IGF-1, IGFBP-1, IGFBP-4, and MIP-1 delta), the CTRL group in 6 proteins (FGF-4, IP-10, Leptin, MCP-1, MIG, and MIP-1 delta). However, sub-analysis performed to detect differentially expressed proteins at baseline and post-intervention, with significance at an adjusted p value ≤ 0.05 and absolute (logFC) ≥ 1.0, shown three down-regulated proteins in the LIPE group (BLC (logFC)= 1.27, Eotaxin (logFC)= 1.18 and MCP-4 (logFC)= 1.14), and four proteins in the HIPE group (BLC (logFC)= 1.45, FGF-6 (logFC)= 1.20, MCP-4 (logFC)= 1.50, and PARC ​(logFC)= 1.33), supporting that the changes that we observed in the exercise groups were not time-related changes but occurred in response to exercise.

CONCLUSIONS: Implementing a 6-month physical exercise program in overweight/obese adolescents, based on LIPE and PLUS groups, significantly change several circulating inflammatory levels. Interventions involving supervised physical exercise may reduce the associated effects of systemic low-grade inflammation, thus preventing the development of obesity-related metabolic diseases in adolescents with overweight/obesity.

PMID:35038337 | DOI:10.1210/clinem/dgac021

J Trop Pediatr. 2022 Jan 7;68(1):fmab112. doi: 10.1093/tropej/fmab112.

ABSTRACT

OBJECTIVE: This study was conducted to determine the relationship between their serum zinc levels and the CD4% in a cohort children living with HIV.

METHODS: One hundred asymptomatic, anti-retroviral Therapy (ART) naïve children living with HIV (participants) aged 5-60 months who were enrolled into the Paediatric HIV clinic of The University of Nigeria Teaching Hospital were recruited in the study over a 10-month period. Blood samples were collected in the morning from non-fasting participants and serum zinc levels were analysed using Atomic Absorption Spectrophotometer. The CD4% was ascertained using the CD4% easy count kit on the Partec® Cyflow Counter machine. Data were analysed using Statistical Package for the Social Sciences version 19.

RESULT: The median (IQR) serum zinc level for the participants was 55.5 µg/dl (49.75) while their median (IQR) CD4% was 27.79% (18.67). Males had a median (IQR) CD4% of 24.29% (19.10) which was significantly lower than those of females [32% (20.59) (p = 0.047)]. No significant relationship was found between CD4% and zinc levels among the subjects (r = -0.061, p = 0.557).

CONCLUSION: Serum zinc levels of asymptomatic ART naïve children living with HIV have no relationship with their CD4%.

PMID:35038323 | DOI:10.1093/tropej/fmab112

Eur J Endocrinol. 2022 Jan 1:EJE-21-1056.R1. doi: 10.1530/EJE-21-1056. Online ahead of print.

ABSTRACT

Background The UICC/AJCC TNM staging system for differentiated thyroid cancer (DTC) involves a single age cutoff as a prognostic criterion. Because a single cutoff is a dichotomization of what might be a sliding scale, using multiple age cutoffs might result into a better stage definition. The aim of our study was to investigate if using a two-step age-based cutoff would improve the TNM staging system regarding disease specific survival (DSS). Methods We retrospectively studied two cohorts of adult DTC patients from The Netherlands and Germany. DSS was analyzed for papillary (PTC) and follicular thyroid cancer (FTC) separately, investigating several two-step age-based cutoffs for those with distant metastases; below lower threshold classified as stage I, between lower and upper threshold as stage II, and above upper threshold as stage IV. Results We included 3074 DTC patients (77% PTC). For PTC, an age cutoff of 45 with 50 years had the best statistical model performance, while this was 25 with 40 years for FTC. However, differences with the optimal single age cutoffs of 50 years for PTC and 40 years for FTC were small. Conclusions The optimal two-step age-based cutoff to predict DSS is 45 with 50 years for PTC, and 25 with 40 years for FTC, rather than 55 years currently used for DTC. Although, these two-step age-based cutoffs were marginally better from a statistical, from a clinical point of view the recently defined optimal single age cutoffs of 50 years for PTC, and 40 years for FTC might be preferable.

PMID:35038308 | DOI:10.1530/EJE-21-1056