Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Background and Aims: To describe and compare characteristics and metabolic control of a population with dyslipidemia prior to and during the COVID19 pandemic. Methods: We included all the patients with dyslipidemia followed at an Endocrinology Department. We analysed age, gender, number of appointments, lipid profile (triglycerides (TG), total cholesterol (TC), LDL cholesterol, HDL cholesterol and non-HDL cholesterol), fasting blood glucose, glycated hemoglobin (HbA1c) and uric acid. We compared the results between the years 2019 and 2020 in the period of March to December. Data were obtained from clinical records. Descriptive statistics were applied and the results are presented by the mean and standard deviation. Statistical significance was accepted for p <0.05. Results: We included 755 patients, 446 (59,1%) were male, age at diagnosis was 63.1±13.1 years and follow up time was 12.3±7.8 years. Five-hundred thirty two patients (70.5%) had type 2 Diabetes Mellitus. Coronary artery disease was found in 11.1% of the patients, peripheral arterial disease in 8.9% and cerebrovascular disease in 7.9%. In 2020 patients had less appointments (2.1 vs 3.4; p<0.001). The mean TC was lower in 2020 (175.3±42.4 vs 179.8±44.9mg/dL; p<0.01) as well as LDL cholesterol (93.8±35.9 vs 98.3±30.2mg/dL; p<0.01), non-HDL cholesterol (122.2±41.5 vs 128.7±43.7mg/dL; p<0.01), HbA1c (7.4±1.2 vs 7.6±1.3%; p<0.01) and fasting blood glucose (140.46±49.2 vs 145.3±57.7mg/dL; p=0.026). The mean HDL cholesterol, TG and acid uric were similar between both years. Conclusions: In our sample, we found that despite a lower number of consultations during the pandemic we obtained lower values of TC, LDL cholesterol, non–HDL cholesterol, HbA1c and fasting blood glucose.

Background Type 1 Diabetes Mellitus (T1DM) is the most common metabolic disease in children. Growth parameters are important indicators of child´s health. Objective To evaluate final height of patients with T1DM correlating the metabolic control and disease duration with growth and puberty. Subjects and methods Retrospective analysis of a cohort of adolescents, aged between 15 and 18 years, with T1DM, followed up to final height at a tertiary Hospital clinic. The variables collected were: age, sex, height at diagnosis, final height, parents’ height, pubertal height gain, metabolic control during puberty (mean A1cHB). Statistical analysis was performed using SPSS®v20; results are presented as mean ± SD. Results Forty six adolescents were included [59% male (M), 41% female (F)]. Mean age at diagnosis was 9.3 ± 3.5 years. Mean A1cHB was 8.15 ± 1.4. In 26 patients, T1DM was diagnosed before puberty; in these, the age at the onset of puberty was 10.8 ± 1.5 (M) and 9.2 ± 0.6 SD years (F). Height SDS at diagnosis was 0.5 ± 1.5 (M) and 0.35 ± 1.2 (F). Final height was -0.2 ± 1 (M) 0.08 ± 0.9 (F). Target height was -0.29 ± 1.1 (M) -0.02 ± 1 (F). Patients were significantly taller than their parents at diagnosis (p = 0,03), and lost height during follow up to final height (p = 0,004) yet final height was within target height (p = 0,3). There was no correlation between final height and metabolic control (p = 0,9) or duration of diabetes (p = 0,4). Conclusion In spite of a taller stature at diagnosis and variable metabolic control, final height was not compromised, arguing against growth compromise being a major hallmark of deficient metabolic control.

Tese de doutoramento, Medicina (Fisiopatologia), Universidade de Lisboa, Faculdade de Medicina, 2011

Pancreatic islet transplantation has demonstrated that long-term insulin independence may be achieved in patients suffering from diabetes mellitus type 1. However, because of limited availability of islet tissue, new sources of insulin producing cells that are responsive to glucose are required. Development of pancreatic beta-cell lines from rodent or human origin has progressed slowly in recent years. Current experiments for ex vivo expansion of beta cells and in vitro differentiation of embryonic and adult stem cells into insulin producing beta-cell phenotypes led to promising results. Nevertheless, the cells generated to date lack important characteristics of mature beta cells and generally display reduced insulin secretion and loss of proliferative capacity. Therefore, much better understanding of the mechanisms that regulate expansion and differentiation of stem/progenitor cells is necessary. Here, we review recent advances in the identification of potential cellular sources, and the development of strategies to regenerate or fabricate insulin producing and glucose sensing cells that might enable future cell-based therapies of diabetes mellitus type 1.

Background: Krueppel-like transcription factor 11 (KLF11, TIEG2) is a member of the Sp1/KLF transcription factor family which plays an important role in mammalian gene regulation via binding to GC-rich promoter elements comprising CGCCC (GC) or CACCC box core sequences. Previously we demonstrated that human KLF11 (hKLF11) inhibits human proinsulin promoter activity in rodent beta-cell lines (INS-1, beta-TC3). At insulin gene upstream sequences hKLF11 binds to a GC box but not to a proximal CACCC box. Surprisingly, deletion and mutation of the GC box did not alter hKLF11-mediated inhibition of proinsulin promoter activity. Here we present results from ongoing experiments which were designed to clarify the underlying molecular mechanisms of hKLF11 proinsulin promoter regulation and the functional role of the CACCC box.

The Krüppel-like factor 11 (KLF11; TIEG2), a pancreas-enriched Sp1-like transcription factor, is a known negative regulator of pancreatic exocrine cell growth. A recent study indicated KLF11-induced activation of the human proinsulin promoter (hInsP).

p8 protein expression is known to be upregulated in the exocrine pancreas during acute pancreatitis. Own previous work revealed glucose-dependent p8 expression also in endocrine pancreatic beta-cells. Here we demonstrate that glucose-induced INS-1 beta-cell expansion is preceded by p8 protein expression. Moreover, isopropylthiogalactoside (IPTG)-induced p8 overexpression in INS-1 beta-cells (p8-INS-1) enhances cell proliferation and expansion in the presence of glucose only. Although beta-cell-related gene expression (PDX-1, proinsulin I, GLUT2, glucokinase, amylin) and function (insulin content and secretion) are slightly reduced during p8 overexpression, removal of IPTG reverses beta-cell function within 24 h to normal levels. In addition, insulin secretion of p8-INS-1 beta-cells in response to 0-25 mM glucose is not altered by preceding p8-induced beta-cell expansion. Adenovirally transduced p8 overexpression in primary human pancreatic islets increases proliferation, expansion, and cumulative insulin secretion in vitro. Transplantation of mock-transduced control islets under the kidney capsule of immunosuppressed streptozotocin-diabetic mice reduces blood glucose and increases human C-peptide serum concentrations to stable levels after 3 days. In contrast, transplantation of equal numbers of p8-transduced islets results in a continuous decrease of blood glucose and increase of human C-peptide beyond 3 days, indicating p8-induced expansion of transplanted human beta-cells in vivo. This is underlined by a doubling of insulin content in kidneys containing p8-transduced islet grafts explanted on day 9. These results establish p8 as a novel molecular mediator of glucose-induced pancreatic beta-cell expansion in vitro and in vivo and support the notion of existing beta-cell replication in the adult organism.

Objectives: Strategies for cell based-therapy of type 1 diabetes mellitus are based on pancreatic islet replacement and islet regeneration. Human islet-derived precursor cells (hIPC) expressing nestin and c-met have been investigated as an additional source of beta-cells. These cells have been demonstrated to differentiate in vitro into insulin producing cells and are assumed to be of endodermal origin. In continuation of previous work we provide further evidence that hIPCs share a common phenotype with human bone marrow derived mesenchymal stem cells (hMSC) and, in addition, bear the potential to differentiate along mesenchymal maturation pathways.

KLF11 (TIEG2) is a member of the Sp1/KLF transcription factor family which plays an important role in mammalian gene regulation. KLF11 is a pancreas-enriched transcription factor that has elicited significant attention because of its role as negative regulator of exocrine cell growth in vitro and in vivo. However, its functional role in the endocrine pancreas remains to be established. Here we investigated endogenous expression of KLF11 in pancreatic beta-cells and analyzed the effects of KLF11 overexpression on human proinsulin promoter activity. RT-PCR revealed endogenous KLF11 mRNA expression in whole rat pancreas, human pancreatic islets and rat INS-1 beta-cells. Glucose stimulation does not seem to modulate KLF11 expression in INS-1 beta-cells. Transient co-transfection of human proinsulin promoter driven secreted alkaline phosphatase (SEAP) reporter gene vectors with KLF11 expression plasmids in INS-1 and betaTC3 beta-cells demonstrates dose-dependent, but glucose-independent KLF11-mediated repression of insulin promoter activity. 5((prime))-deletion analysis of the human proinsulin promoter sequence and gel-shift assays (EMSA) of protein/DNA binding indicate specific GC and CACCC boxes within the human proinsulin promoter as potential KLF11 binding sites. Further, KLF11 may interfere with SP1 binding sites at these response elements. In summary, we have identified Krueppel-like transcription factor 11 (KLF11) as a novel negative transcriptional regulator of human proinsulin gene expression in pancreatic beta-cells. Further we have characterized specific KLF11 DNA-response elements within the human proinsulin promoter. Impairment of KLF11-dependent transcriptional regulation of proinsulin expression may contribute to beta-cell dysfunction seen in the pathogenesis of diabetes mellitus.

The proliferation associated protein p8 is highly expressed within pancreatic acinar cells during experimentally induced pancreatitis and improves pancreatic resistance by enhanced expression of the anti-inflammatory protein PAP I. In previous studies we demonstrated glucose-dependent expression of p8 also in pancreatic beta-cells, and enhanced expansion and growth of beta-cells by p8-overexpression. Bone marrow derived adult mesenchymal stem cells (BMSC) are investigated as an alternative source for beta-cell replacement therapy, but their proliferation potential in vitro is limited. Here we investigated the human BMSC line hMSC-TERT to clarify the question whether p8-induced tissue regeneration is mediated by enhanced proliferation and/or reduced apoptosis. p8 expression was analyzed by RT-PCR, Western blotting and immunocytochemistry in hMSC-TERT cells. Further, cells were stably transfected with either a CMV promoter driven IRES-plasmid which expresses both p8 and GFP (p8-hMSC-TERT) or a control vector expressing only GFP (mock-hMSC-TERT). Numbers of cells positive for BrdU (proliferation), GFP (expansion) and annexin V (early apoptosis) were measured by FACS analysis. Annexin V data were corrected for necrosis by paralleled propidium iodide staining. p8 is highly expressed in proliferating hMSC-TERT cells. hMSC-TERT subclones overexpressing human p8 demonstrate durable significant 3 fold augmentation of cell proliferation and 1.75 fold expansion of cell numbers as compared to mock controls. In parallel, p8 overexpression remarkably reduced basal apoptosis rate in p8-hMSC-TERT versus mock control cells. These results characterize p8 as a potent molecular mediator of BMSC growth and expansion acting through both induction of cell proliferation and inhibition of apoptosis. This underlines the presumed regenerative properties of p8. The proliferation-inducing and apoptosis-inhibiting properties of p8 may be useful in the development of BMSC or beta-cell based tissue regeneration strategies.

Aim: To synthesise published evidence regarding the effectiveness of training and procedural interventions aimed at improving the identification and management of child abuse and neglect by health professionals.Methods: Systematic review for the period 1994 to 2005 of studies that evaluated child protection training and procedural interventions. Main outcome measures were learning achievement, attitudinal change, and clinical behaviour.Results: Seven papers that examined the effectiveness of procedural interventions and 15 papers that evaluated training programmes met the inclusion criteria. Critical appraisal showed that evaluation of interventions was on the whole poor. It was found that certain procedural interventions ( such as the use of checklists and structured forms) can result in improved recording of important clinical information and may also alert clinical staff to the possibility of abuse. While a variety of innovative training programmes were identified, there was an absence of rigorous evaluation of their impact. However a small number of one-group pre- and post-studies suggest improvements in a range of attitudes necessary for successful engagement in the child protection process.Conclusion: Current evidence supports the use of procedural changes that improve the documentation of suspected child maltreatment and that enhance professional awareness. The lack of an evidence based approach to the implementation of child protection training may restrict the ability of all health professionals to fulfil their role in the child protection process. Formal evaluation of a variety of models for the delivery of this training is urgently needed with subsequent dissemination of results that highlight those found to be most effective.