Дисертації з теми "T1AM"

Thyroxine (T4) is the predominant form of thyroid hormone (TH). In target tissues, T4 is enzymatically deiodinated to 3,5,3′-triiodothyronine (T3), a high-affinity ligand for the nuclear TH receptors TRα and Trβ. T3 modulates genes transcription via activation of TRα and TRβ. Non-genomic effects have also been described. In 2004 the research groups of professors Scanlan Grandy and Zucchi discovered an endogenous thyroid hormone derivative called 3-iodothyronamine (T1AM). They proved that at nanomolar concentrations it can activate trace amine associated receptors 1 (TAARs)[1] and it may also interact with other targets, such as plasma membrane transporters, mitochondrial proteins and vesicular biogenic amine transporters [2]–[4]. Endogenous T1AM has been detected in human and rodent blood and tissues samples by liquid chromatography coupled mass spectrometry (LC/MS/MS) [5]. Its endogenous levels are a matter of argument due to the challenges that its accurate quantification poses. For this reason, so far a worldwide adopted extraction method has not been established. Circulating T1AM has so far been considered to be largely bound to apolipoprotein apoB100 [6]. The first T1AM functional effect to be discovered was severe hypothermia [7]. This effect is associated to a decrease in oxygen consumption and a reduction of the respiratory quotient (CO2/O2), which reflects the relationship between glucose and fatty acid oxidation, resulting in a shift from carbohydrate to lipid as energy source [8]. The molecular mechanisms underlying T1AM effects are still unknown, however Mariotti and colleagues [9] analyzed gene expression profiles in adipose tissue and liver of T1AM chronically treated rats and found significant transcriptional effects involving sirtuin genes, which regulate important metabolic pathways. Therefore, the first aim of this work was to compare the effect of T1AM and T3 chronic treatment on mammalian sirtuin expression in hepatoma cells (HepG2) and isolated hepatocytes. Isolated rat hepatocytes were obtained by liver in-situ collagenase perfusion. Sirtuin expression was determined by Western Blot analysis in cells treated for 24 h with 1-20 µM T1AM or T3. In addition, cell viability was evaluated by MTT test upon 24 h treatment with 100 nM to 20 µM T1AM or T3. In HepG2, T1AM significantly reduced SIRT1 and SIRT4 protein expression at 20 µM while T3 strongly decreased the expression of SIRT1 (20 µM) and SIRT2 (any tested concentration). In primary rat hepatocytes T1AM, did not affect protein expression whereas T3 decreased SIRT2 at 10 µM. The extent of MTT-staining was moderately but significantly reduced by T1AM, particularly in HepG2 cells, in which the effect occurred at concentration starting from 100 nM. T3 reduced MTT staining in HepG2 but not in isolated hepatocytes. T1AM and T3 differently affected sirtuin expression in hepatocytes. Since SIRT4 is an important regulator of lipid and glucose metabolism, whereas SIRT1 and SIRT2 have a key role in regulating cell cycle and tumorigenesis, our observations are consistent with the shift from carbohydrates to lipids induced by T1AM and indicate a potential new role of T1AM in modulating tumor proliferation. The second part of this project was aimed at clarifying the issue that, so far, every research group working on this molecule has encountered when trying to accurately quantifying T1AM endogenous levels. These difficulties were usually attributed to problems in extraction or other pre‐analytical steps. Most researchers have developed various workaround for this issue. For example, on cell culture experiments, to avoid the presence of serum proteins in the culturing media, experiments have often be performed with unphysiological protein‐free media. The second goal of this project was therefore to evaluate the effect of serum protein on the recovery of exogenous T1AM. Cell culture media (Krebs buffer, DMEM, FBS, DMEM+FBS, used either in the absence or in the presence of NG108‐15 cells) and other biological matrices (rat brain and liver homogenates, human plasma and blood) were spiked with T1AM and/or deuterated T1AM (d4‐T1AM) and incubated for times ranging from 0 to 240 min. Samples were extracted using a liquid/liquid method and analysed using liquid chromatography coupled to mass spectrometry (LC-MS/MS), to assay T1AM and some of its metabolites. For the first time in the history of this molecule, in FBS‐containing buffers, an exponential decrease in T1AM levels was observed over time. T1AM metabolites were not detected, except for minimum amounts of TA1. Notably, d4‐T1AM decreased over time at a much lower rate, reaching 50‐70% of the baseline at 60 min. These effects were completely abolished by protein denaturation and partly reduced by semicarbazide, however, the process could not be reverted. In the presence of cells, T1AM concentration decreased virtually to 0 within 60 min, but TA1 accumulated in the incubation medium, with quantitative recovery. Spontaneous decrease in T1AM concentration with isotopic difference was confirmed in rat organ homogenates and human whole blood. Conclusions. On the whole, these results suggest binding and sequestration of T1AM by blood and tissue proteins, with significant isotope effects. These issues might account for the technical problems complicating the analytical assays of endogenousT1AM.

Thyroid hormones (TH), namely thyroxine (T4) and 3,5,3’-triiodothyronine (T3), are crucial regulators of multiple growth processes and control systems of energy metabolism. T4 and T3 undergo a complex metabolism in vivo, by several enzymes encompassing deiodinases, amine transferases, amine oxidases, decarboxylases and several classes of conjugating enzymes, particularly sulfotransferases and UDP-glucuronosyltransferases. T4 or T3 metabolites can produce significant functional effects when administered via interaction either with Thyroid Hormone Receptor (TR), or with other receptors. They are considered as chemical messengers further enriching TH signaling, and have become known as “novel thyroid hormones” or “active thyroid hormones metabolites”. These novel hormones include: T2; Thyronamines (TAMs), mostly 3-iodothyronamine (T1AM) and non-iodinated thyronamine (T0AM); thyroacetic acids, mostly 3,5,3’,5’-thyroacetic acid (TA4), 3,5,3’-thyroacetic acid (TA3), and 3-thyroacetic acid (TA1). Recently, it emerged that 3-iodothyronamine (T1AM), a derivative of decarboxylation and deiodination of thyroid hormones, has pro-learning and anti-amnestic effects, modulates pain threshold, sleep pattern and food intake. It also counteracts beta-amyloid toxicity in mice. Glutamatergic neurotransmission, the major excitatory system in the brain, plays a key role in regulating neuroplasticity, learning and memory, and it is often compromised in neurological disorders. T1AM reduced availability might results in some disorders associated with thyroid hormones. T1AM binds to the trace amine-associated receptor 1 (TAAR1) a G-protein coupled receptor with a putative role in neurotransmission. In the present work, firstly we characterized the gene expression profile of two different brain cell lines and then we evaluated the effects of T1AM on the expression of proteins involved in the glutamatergic postsynaptic pathway. A hybrid line of cancer cells of mouse neuroblastoma and rat glioma (NG 108-15) and a human glioblastoma cell line (U-87 MG) were used. We first characterized the in vitro model by analyzing gene expression of several proteins involved in the glutamatergic postsynaptic cascade by real time PCR (RT-PCR), and cellular uptake and metabolism of T1AM by HPLC coupled to mass spectrometry (HPLC MS-MS). The cell lines were then treated with T1AM, ranging from 0.1 to 10 μM, alone or in combination with 10 µM resveratrol (RSV) and/or 10 µM amyloid β peptide (25-35). Cell viability, glucose consumption, protein expression, cAMP production and calcium concentration in cell lysates were assessed. Our results indicated that both cell lines expressed receptors implicated in glutamatergic pathway, namely AMPA, NMDA and EphB2, but only U-87 MG cells expressed TAAR1 and they took up T1AM which was catabolized to TA1 and might be used as biochemical model to study its post synaptic signaling cascade. At micromolar concentration T1AM had a slightly but significant cytotoxic effect, that is completely blunted if incubated with RSV and it was able to induce different post-translational modification in neuronal cell lines. T1AM reduced glucose consumption and decreased intracellular calcium concentration in NG 108-15 cell line, while increased cAMP concentration, albeit at different doses. At pharmacological concentrations, the major effect highlighted in both cell lines was an increase in the phosphorylation of proteins involved in the glutamatergic postsynaptic signaling. In the NG 108-15 cells an increase in phosphorylation of ERK extracellular signal-regulated kinases (ERKs) (pERK/ total ERK) and CaMKII Ca-calmodulin-dependent protein kinase (CaMK) II (pCaMKII/total CaMKII). In U-87 MG cells, T1AM induced the phosphorylation of the transcriptional factor cAMP response element-binding protein (CREB) and increase the expression of cFOS. Expression or post-translational modifications of other proteins were not affected. We then extend investigation on the effects of 3-iodothyroacetic acid TA1, a catabolite of T1AM and of thyroid hormone, on brain cell lines focusing on the glutamatergic postsynaptic pathway that we explored by infusion with T1AM, assuming that TA1 may either strengthen T1AM effects or exert parallel actions, especially in brain tissue. First, we assessed uptake and metabolism of TA1. Cell lines were treated with TA1 for 24h, at concentration ranging from 0.1 to 10 μM. Uptake, cell viability, cAMP production and protein expression were assessed. TA1 was taken up by cells, even though only a slight reduction in medium concentration was recorded upon 24h of incubation. Cell viability was significantly increased by TA1 10 µM in U-87 MG cell line, while NG 108-15 cells were unaffected. Western blot analysis indicated that, upon infusion of pharmacological doses of TA1, neither the expression of Sirtuin 1, (p=NS) nor the post-translational modifications of ERK (pERK/total ERK, p=NS) were affected in U-87 MG. Instead TA1 induced the phosphorylation of the transcriptional factor cAMP response element-binding protein (CREB) (pCREB/total). In NG 108-15 cell line, preliminary analysis on protein expression and post-translational modification after TA1 infusion, indicated that no modifications of ERK (pERK/total ERK) were occurred. In conclusion our results indicated that NG 108-15 and U-87 MG cells express receptors implicated in the glutamatergic system and, at pharmacological concentrations, T1AM can affect glutamatergic signaling. Therefore, our preliminary results suggest that, in our experimental models, TA1 does not seem to mimic T1AM effects.

The term “thyroid hormones”, classically referred to both 3,5,3′-triiodothyronine (T3) and thyroxine (T4), seems nowadays to be simplistic; some T3 and T4 metabolites, particularly 3,5diiodothyronine (3,5-T2) and 3-iodothyronamine (T1AM), are independent chemical messengers, with specific metabolic effects. In this PhD thesis we focused on the effects of these derivates on different tissues. In the first project we evaluated the effects of T1AM on the glutamatergic pathway, the main excitatory system in brain. A cancer hybrid cell line of mouse neuroblastoma and rat glioma (NG108-15) and a human glioblastoma cell line (U-87 MG) were used as in vitro model and treated with different T1AM concentration for 24h, alone or in combination with resveratrol 10 µM and/or amyloid β peptide (25-35) 10 µM. Firstly, we characterized cell lines for the expression of receptors implicated in glutamatergic pathway with real time PCR (qRT-PCR) and Western blot. Both cell lines expressed AMPA, NMDAR1 and EphB2, but only U-87 MG expressed TAAR1, the putative T1AM receptor. Using LC-MS-MS we discovered that both lines were able to take up T1AM and, rapidly catabolized it to TA1. In both cell lines, T1AM showed a slightly but significant cytotoxic action starting from 0.1 μM, that increase is presence of β-amyloid (10 µM), but not resveratrol (10 µM) evaluated by MTT test. We then evaluate glucose consumption using a glucose assay and observed in the NG108-15 a metabolic effect mediated by T1AM (p<0.05). For protein expression and post-translation modifications Western blot was used and an increase in the phosphorylation of Ca-calmodulin-dependent protein kinase (CaMK) II (pCaMKII/total CaKMII, p<0.05) in NG108-15 cell line was observed. In association with resveratrol T1AM could increase the expression of PKC (p<0.001 vs RSV) in the same cell line, a synergic effect showed exclusively in presence of both T1AM and resveratrol at all tested thyronamine concentration. In U-87 MG T1AM induce the phosphorylation of the transcriptional factor cAMP response element-binding protein (CREB) (PCREB/total CREB p<0.05). Our results indicated that these two nervous cell lines express receptors implicated in glutamatergic system and might be used as biochemical model to study its post synaptic signaling cascade. T1AM had a minimal cytotoxic effect and it was able to induce different post-translational modification in neuronal cell lines. T1AM might activate mechanisms of action which included increasing CaMKII phosphorylation and PKC expression in NG108-15 while in U-87 MG induced the activation of the transcriptional factor CREB. We then focused on another endogenous thyroid hormone derivate, the 3,5-diiodo-l-thyronine (3,5-T2) and its effect on heart which have been poorly investigated so far. It’s well understood that 3,5-T2 is able to regulate energy expenditure, resting metabolic rate and oxygen consumption with a mechanism that might involve mitochondria. We decided to evaluate the functional metabolic, and toxic effect of 3,5-T2 using both in vitro and ex vivo models of cardiac preparations. As comparison for our results we also evaluated the response to T3 and T4. As cell culture we selected the H9c2 cells (rat cardiomyoblasts) to determine 3,5-T2, T3, and T4 uptake using LC-MS-MS. We treated cells with 3,5-T2 (0.1 to 10 µM) and evaluated cell viability using MTT test and crystal violet staining. We also investigate a possible 3,5-T2 metabolic effect performing a glucose and hexokinase assay. In the end we measured the cardiac functional effects, perfusing isolated working rat hearts with 3,5-T2, T3, or T4 in Krebs-Ringer buffer and recording hemodynamic variables. H9c2 cells took up 3,5-T2, in cell lysate and the analyte levels increased slowly over time. 3,5-T2 significantly decreased MTT staining at 0.5–10 µM concentration, an effect confirmed by the crystal violet staining only at 10 µM T2, while equimolar T3 and T4 did not share this effect. In cells exposed to 0.1 or 1.0 µM of 3,5-T2 glucose uptake increased by 23% or 30% (p < 0.05). On the opposite side, T3 did not affect glucose consumption which was significantly reduced by 1 and 10 µM T4 (−24 and −41%, respectively, p < 0.01 and p < 0.0001). In the isolated perfused rat heart, 10 µM T2 produced a transient and slight reduction in the cardiac output and aortic flow (p<0.05), while thyroid hormone did not induce any hemodynamic change. Our findings demonstrate that 3,5-T2 was taken up by cardiomyoblasts, and in a concentration range between 0.1 µM and 1.0 µM modulated cardiac energy metabolism increasing glucose accumulation. Furthermore, we observed some evidence of cytotoxicity and a transient impairment of contractile performance only at the highest 3,5-T2 concentration tested (10 µM). These effects seem to be specific for 3,5-T2, since they are not reproduced by thyroid hormone. In the end we develop a novel ad-hoc optimized method to quantify T2 isomers using LC-MSMS in human serum. T2 isomers (3,5-T2 and 3,3’-T2) has been detected in human blood using immunological methods, but until now a reliable assay based on mass spectrometry was not available. We obtained 2 mL of serum samples from 28 healthy subjects. The serum was firstly deproteinized with acetonitrile and then exposed to a solid phase extraction-based procedure. Then samples were furtherly cleaned by hexane washing and subjected to another step of deproteinization with acetonitrile to precipitate residual proteins. Both isomers were then analyzed by high performance liquid chromatography coupled to tandem mass spectrometry. We developed a method with 88–104% accuracy, 95–97% precision, 78% recovery and a matrix effect average of +8%. In the serum sample 3,5-T2 was detected with a concentration averaged (mean ± SEM) 41 ± 5 pg/mL and 133±15 pg/mL for 3,3′-T2. Furthermore, we observed a significant correlation between 3,5-T2 and 3,3′-T2 concentrations (r = 0.540. p < 0.01), while no significant relation was observed with thyroid hormone. In conclusion, this method can quantify both T2 isomers in human serum using a reliable assay based on LC-MS-MS. The concentrations of these isomers lie in the subnanomolar range and show a significant correlation in healthy subject.

Abstract Type 1 diabetes mellitus (T1DM) among children is of a particular importance in Finland, where its incidence is the highest in the world and still increasing. However, the aetiology of T1DM is not fully known. According to current knowledge, both genetic and environmental factors operate together, leading to an attack by the immune system on the insulin-producing beta cells. The purpose of this study was to investigate the geographical variation in the incidence of T1DM among children aged up to 14 years in Finland. Geographical Information Systems (GIS) and Bayesian spatial statistics were applied in a search for unusual spatial patterns and risk factor associations. The incidence of T1DM among children aged up to 14 years showed clear geographical variations in Finland. Living in a rural environment increased the risk for T1DM, and the risk was particularly high among children living in rural heartland areas. There was no association between the variation in T1DM incidence and the zinc and nitrate concentrations of drinking water. A male excess in the incidence of T1DM was seen in the low-incidence areas. The geographical variation in the risk of T1DM was marked only among children aged up to 9 years. Because genetics is a necessary but not a sufficient cause of T1DM, it could be hypothesized that there are some thus far unknown environmental risk factors affecting particularly younger children in Finland. Some of those factors may be related to a rural environment. The geographical variation in the M/F ratio of T1DM was a challenging observation and warrants more analytical study.

The purpose of this mixed-methods descriptive study with parents of children newly diagnosed with Type 1 diabetes was to explore their experiences with peer social support following the Social Support to Empower Parents (STEP) intervention and to examine the usefulness of the Family Management Measure (FaMM) in this population. The specific aims were to describe parents' experiences with the STEP social support intervention, describe parents' day-to-day diabetes management as measured by the FaMM, describe the relationship between parental management scores in the six FaMM dimensions and the social support intervention dose used, and explore FaMM scores in relationship to parent satisfaction with the STEP social support intervention. Identified themes of availability, practical tips, and common ground resonated throughout the interviews with parents and reflected Ireys' emotional, informational, and affirmational social support framework. Regardless of the intervention dose, number of parent mentor contacts, or scores on the FaMM scales, all parents interviewed when questioned, gave a 5/5 for satisfaction with the STEP RCT, qualitatively underscoring the positive effect of the intervention.

Introduction: Patients with laryngeal carcinoma often present early due to the change in their voice. The treatment for T1aN0M0 carcinoma varies throughout the world, but whether radiotherapy (RT) or endolaryngeal laser excision is performed both result in excellent local control of the tumour and five year survival rates. There are advantages and disadvantages of either treatment but there are no appropriately powered randomised controlled trials comparing them. Over recent decades external beam RT has become the more popular choice and this is partly due to a perception of poor voice outcomes from surgical excision. However with the development of technology allowing surgical precision, transoral laser microsurgery (TLM) has resulted in low morbidity and good voice outcomes. Objective: This research has three main objectives: a. To describe acoustic parameters of ‘normal’ voice; b. To compare voice outcomes in patients treated with TLM with those treated with radiotherapy for T1a SCC of the glottis; c. To investigate longitudinal changes in voice quality in patients undergoing TLM for T1a SCC of the glottis. Methods: The research was divided into three main parts. The first part was to analyse the acoustic parameters of ‘normal’ voice. To describe the parameters of ‘normal’ voice, adults with no history of voice disorders who scored zero on the voice questionnaire (Voice Handicap Index - 10) were included. The second part comprised a comparative cohort study of 40 patients with T1aN0M0 laryngeal carcinoma, treated with either TLM (20 patients) or RT (20 patients) to compare voice outcomes at least one year following treatment. The third part involved a prospective cohort study of 30 patients with T1aN0M0 laryngeal carcinomas who were treated with TLM, comparing voice qualities before and after treatment. All patients were recruited from those attending the regional Head and Neck centre in Aintree University Hospital. The same methodology was adopted for voice recordings for all three parts of the study. Participants were asked to read a phonetically balanced passage and produce a prolonged vowel sound. In a sound proof room the voice recording included simultaneous audio and electrolaryngograph readings. The voice recordings were scored according to the GRBAS voice scale by an experienced rater. Acoustic analysis was performed form the electrolaryngograph recording using the SpeechStudioTM software. Several objective acoustic parameters were calculated from both sustained vowels and connected speech. These include: fundamental frequency (Fx), jitter, shimmer, harmonics to noise ratio (HNR) and normalized noise energy (NNE). In the comparative study of TLM versus RT and the prospective TLM study, patients were asked to complete voice-specific and quality of life questionnaires. The voice-specific questionnaires were the Voice Symptom Scale (VoiSS) and the Voice Handicap Index-10 (VHI-10). The quality of life questionnaire adopted was the University of Washington Quality of Life (UWQoL) version 4. Results: In the acoustic analysis of sustained vowels in normal speech, females have a statistically significantly higher Fx than males (adjusted p= < 0.05). There is no other statistically significant difference across the domains for sustained vowels in normal speech. In the analysis of connected speech, Fx is again higher in females (p < 0.001). There is no statistically significant difference in amplitude (Ax) or contact quotient (Qx). In the comparison of voice post TLM and RT, there is no statistical difference in voice-specific questionnaires between the groups. The UW-QoL4 found a statistically significantly higher QoL score in the TLM compared with the RT group for appearance (p=0.003), recreation (p=0.048), chewing (p=0.015) and saliva (p=0.016), however these are not statistically significant when adjusted for age. Overall for QoL, the RT group have a statistically significantly lower median score compared to TLM in physical function (p=0.004) and this remains statistically significant when adjusted for age (p=0.036). There is no statistically significant difference for social function (p=0.441). There is no statistically significant difference in perceptual rating (GRBAS score) between RT and TLM groups (total mean 5.49 vs. 5.12, p=0.254). Most domains as part of the acoustic analysis of sustained vowels show no statistically significant difference between RT and TLM. The mean Fx analysis on connective speech is statistically significantly higher in the TLM group (161.2Hz vs. 131.1Hz, adjusted p=0.001). Coherence of frequency is statistically significantly higher in the TLM group (48.6% vs. 36.0%, adjusted p=0.027) and pitch irregularity is statistically significantly higher in the RT group (26.7% vs. 14.9%, adjusted p=0.013). There is no statistically significant difference in mean amplitude between the two groups. Coherence of amplitude is statistically significantly higher in the TLM group (adjusted p=0.006) and amplitude irregularity is statistically significantly higher in the RT group, (12.4% vs. 6.3%, adjusted p=0.005). There is no statistically significant difference in mean contact quotient (p=0.368), coherence (p=0.236) or irregularity (p=0.125) when comparing TLM and RT. In the comparison of voice pre and post TLM, there is no statistical difference in voice-specific questionnaires between the groups. There is no statistically significant difference in the UW-QOLv4 domain scores or composite scores in patients pre- and post- TLM. There was no statistically significant difference in mean score for ‘G’,’R’,’B’ and ‘S’ indicators as part of perceptual rating between pre and post TLM patients, although asthenia was statistically significantly lower post-TLM (0.97 vs. 0.94, adjusted p=0.015). There is no statistically significant difference in any of the domains in the acoustic analysis of sustained vowels pre and post TLM. In the acoustic analysis of connected speech, the mean DFx is statistically significantly higher in the post TLM group (adjusted p=0.001). There is no statistically significant difference in the coherence of frequency or pitch irregularity when comparing pre and post TLM. There is no statistically significant difference in the mean DAx (p=0.121), coherence (p=0.472) or irregularity of amplitude (p=0.184) when comparing pre and post TLM. There is no statistically significant difference in the mean DQx (adjusted p=0.904), coherence (adjusted p=0.293) or irregularity of the contact quotient (adjusted p=0.400) when comparing pre and post TLM. Conclusion: The treatment of T1a laryngeal carcinoma with either TLM or RT has been shown to have comparably good local control. There are advantages and disadvantages of both treatments, however TLM is often preferred by patient and clinician as it is a day case procedure, can provide histological clearance and leaves the option to use RT in the future. However voice outcomes of the procedures have been debated with various reports in the literature. There are challenges when comparing the two treatment modalities due to a number of tumour, patient and surgical factors. It is not surprising that the voice is affected by whatever treatment is performed to treat the glottic carcinoma. This study shows that voice quality is good, however it is measured, for after both TLM and RT.

The incidence of Type 1 Diabetes Mellitus (T1DM) is markedly growing in the past two decades. For the management of this disease, physical exercise has been recommended as a costeffective treatment throughout the global health system. The Non-Obese Diabetic (NOD) mouse represents a well-established experimental model analogous to human T1DM as it is characterized by a progressive autoimmune destruction of pancreatic β-cells. This thesis explored the uses of a mouse motorized treadmill to study the effects of exercise in NOD mice. Body mass, blood glucose level, immunological soluble factors, muscular performance and islets of Langerhans architecture were monitored during 12-week moderate-intensity endurance training in female NOD mice. After 12 weeks of training, no differences were registered as to diabetes incidence (50 vs 45%) and mean glycemia between sedentary controls and mice on exercise (190±34 vs 163±38 mg/dl, mean and SD). Exercise capacity dimished in the exercisingmice with respect to controls (work, distance, VO2max, p<.05). Preliminary data from a morphometric analysis of pancreata indicated the presence of larger infiltrates along with increased endocrine cellareas in the NOD exercising-mice. A higher infiltrate-to-islet ratio was observed in exercising-micewith respect to the controls. An exercise-induced weight loss was also detected. Among key anti- and pro-inflammatory cytokines: TNF-α, MIP-1β and IL-10 resulted to be lower at end of the training in the exercising animals with respect to pre-training values (1353±2 vs 1355±2.3; 984.6±12 vs 1001±37; 396±8.1 vs 407±27 MFI, respectively,p<.05) whereas IL-2P40 was higher in exercising-mice compared baseline (543±12 vs 539±15 MFI, p<.05). Further studies are needed to clarify the utility of the NOD mouse model to mimic and investigate the exercise effects in T1DM, immunomodulation and inflammation. Specifically, dose-response studies in which exercise will be administered to NOD mice at various levels of intensity will be necessary to determine the optimal regimen of physical exercise having clearcut preventive effects on the development of T1DM.

Bakgrund Varje dag insjuknar cirka två barn i diabetes mellitus typ 1 (T1DM) i Sverige. När ett barn blir sjukt är det föräldrarna till barnet som ansvarar för omvårdnaden vardagen. Omvårdnadsansvaret läggs på föräldrarna och för en del kan detta leda till stress och i värsta fall till utbrändhet. Syfte Syftet var att beskriva hur föräldrar upplever och hanterar att deras barn insjuknar i diabetes mellitus typ 1. Metod En kvalitativ semistrukturerad intervjustudie genomfördes. Sju föräldrar till barn med diabetes mellitus typ 1 från Stockholm och Uppsala intervjuades. Samtlig insamlade data analyserades utifrån en kvalitativ innehållsanalys. Resultat De kategorier som framkom under resultatanalysen var; att hantera sitt barns sjukdom, det stora tunga ansvaret, sjukdomens påverkan i det dagliga livet och behov av stöd. Föräldrar upplevde deras barns insjuknande i T1DM som traumatiskt och något som påverkar hela familjens liv. Samtliga föräldrar som deltog i studien uttryckte ett behov av stöd i olika former. Att känna sig ensamma utan stöd från varken anhöriga eller hälso- och sjukvården är en vardag för dessa föräldrar. Stödet som hälso- och sjukvården erbjöd upplevdes bristfälligt och med flera förbättringsområden. Flertalet föräldrar uppgav att de lider av sömnbrist, vilket försvårar det dagliga livet på många sätt. Slutsats Föräldrar till barn med T1DM lär sig, över tid hantera omvårdnaden som sjukdomen medför, men det finns svårigheter kring hanteringen av ansvaret, sorgen och oron. Föräldrarna upplever brister i det långsiktiga stödet som hälso- och sjukvården erbjuder och önskar mer avlastning för att minska riskerna att drabbas av psykisk och fysisk ohälsa. Vi anser att hälso och sjukvården måste erbjuda ett utökat stöd till dessa föräldrar för att tillgodose deras behov innan de riskerar att drabbas av utbrändhet.

Parents of children with a chronic disease are usually highly involved in their child’s treatment and may be affected by the heavy demands and constant stress. This can increase the risk of developing burnout, which is an individual reaction to long-term stress consisting of symptoms associated with emotional exhaustion, as well as physical and cognitive fatigue. The overall aim was to estimate the prevalence of burnout in parents of children with Type 1 Diabetes Mellitus (T1DM) and inflammatory bowel disease (IBD) (paper I), identify the risk factors associated with parenting a child with T1DM (paper II), explore how mothers suffering from burnout describe their mothering of a child with diabetes, with special focus on their need for control and Performance-based self-esteem (PBSE) (paper IV), and to evaluate the effect of a group intervention aimed at reducing stress-related symptoms (paper III). A total of 251 parents of children with T1DM, 38 parents of children with IBD and 124 parents of healthy children participated in a population-based study (I, II). The validated Shirom-Melamed Burnout Questionnaire (SMBQ) was used to assess burnout. 16 parents (SMBQ ≥3.75) participated in a group intervention and were evaluated for changes in SMBQ and PBSE (III). A total of 21 mothers of children with T1DM who scored for clinical burnout (SMBQ) participated in a qualitative study. Semi-structured interviews were conducted and Inductive content analysis was used (IV). In the study group 36.0% parents of children with a chronic disease scored for clinical burnout (SMBQ ≥3.75) compared to 20.2% of the reference parents (p=0.001) with a preponderance of mothers compared to fathers, 42% vs. 20.5% (p=0.001), respectively (I). Less support from the social network, sleep disturbances and lack of personal leisure time and recovery seem to be important risk factors for clinical burnout in parents of children with T1DM, especially mothers (II). Mothers’ experiences of mothering a child with T1DM were interpreted as one theme; Mission impossible, illustrating the extremely difficult circumstances under which they bring up the child with diabetes to adulthood (IV). Parents’ subjective evaluation of the intervention group was mainly positive and SMBQ (p=0.01) and PBSE scale (p= 0.04) measurements were significantly reduced 6 months after completion of the intervention (III). It is important to pay attention to how parents and especially mothers experience their daily life in order to support those who are at risk of developing burnout.

The endoplasmic reticulum (ER) is the organelle responsible for synthesis and folding of secreted and membranous protein and lipid biosynthesis. It also functions as one of the main cellular calcium stores. Pancreatic beta-cells evolved to produce and secrete insulin upon demand in order to regulate blood glucose homeostasis. In response to increases in serum glucose, insulin synthesis represents nearly 50% of the total protein biosynthesis by beta-cells. This poses an enormous burden on the ER, rendering beta-cells vulnerable to agents that perturb ER function. Alterations of ER homeostasis lead to accumulation of misfolded proteins and activation of an adaptive response named the unfolded protein response (UPR). The UPR is transduced via 3 ER transmembrane proteins, namely PERK, IRE-1 and ATF6. The signaling cascades activated downstream of these proteins: a) induce expression of ER resident chaperones and protein foldases. Increasing the protein folding capacity of the ER; b) attenuate general protein translations which avoids overloading the stressed ER with new proteins; c) upregulate ER-associated degradation (ERAD) genes, which decreases the unfolded protein load of the ER. In severe cases, failure by the UPR to solve the ER stress leads to apoptosis. The mechanisms linking ER stress to apoptosis are still poorly understood, but potential mediators include the transcription factors Chop and ATF3, pro-apoptotic members of the Bcl-2 familly, the caspase 12 and the kinase JNK.

Accumulating evidence suggest that ER stress contributes to beta-cell apoptosis in both type 1 and type 2 diabetes. Type 1 diabetes is characterized by a severe insulin deficiency resulting from chronic and progressive destruction of pancreatic beta-cells by the immune system. During this autoimmune assault, beta-cells are exposed to cytokines secreted by the immune cells infiltrating the pancreatic islets. Our group has previously shown that the pro-inflamatory cytokines interleukin-1beta (IL1-beta and interferon-gamma (IFN-gamma), via nitric oxide (NO) formation, downregulate expression and function of the ER Ca2+ pump SERCA2. This depletes beta-cell ER Ca2+ stores, leading to ER stress and apoptosis. Of note, IL1-beta alone triggers ER stress but does not induce beta-cell death, while IFN-gamma neither causes ER stress nor induces beta-cell death. Together, these cytokines cause beta-cell apoptosis but the mechanisms behind this synergistic effect were unknown.

Type 2 diabetes is characterized by both peripheral resistance to insulin, usually as a result of obesity, and deficient insulin secretion secondary to beta cell failure. Obese patients have high levels of circulating free fatty acids (FFA) and several studies have shown that the FFA palmitate induces ER stress and beta-cell apoptosis.

In the present work we initially established an experimental model to specifically activate the ER stress response in pancreatic beta-cells. For this purpose, insulinoma cells (INS-1E) or primary rat beta-cells were exposed to the reversible chemical SERCA pump blocker cyclopiazonic acid (CPA). Dose-response and time course experiments determined the best conditions to induce a marked ER stress without excessive cell death (<25%).

The first goal of the work was to understand the synergistic effects of IL1-beta and IFN-gamma leading to pancreatic beta-cell apoptosis. Our group previously observed, by microarray analysis of primary beta-cells, that IFN-gamma down-regulates mRNAs encoding for some ER chaperones. Against this background, our hypothesis was that IFN-gamma aggravates beta-cell ER stress by decreasing the ability of these cells to mount an adequate UPR. To test this hypothesis, we investigated whether IFN-gamma pre-treatment augments CPA-induced ER stress and beta cell death. The results obtained indicated that IFN-gamma pre-treatment potentiates CPA-induced apoptosis in INS-1E and primary beta-cells. This effect was specific for IFN-gamma since neither IL1-beta nor a low dose CPA pre-treatment potentiated CPA-induced apoptosis in INS-1E cells. These effects of IFN-gamma were mediated via the down regulation of genes involved in beta cell defense against ER stress, including the ER chaperones BiP, Orp150 and Grp94 as well as Sec61, a component of the ERAD pathway. This had functional consequences as evidenced by a decreased basal and CPA-induced activity of a reporter construct for the unfolded protein response element (UPRE) and augmented expression of the pro-apoptotic transcription factor Chop.

We next investigated the molecular regulation of the Chop gene in INS-1E cells in response to several pro-apoptotic and ER stress inducing agents, namely cytokines (IL1-beta and IFN-gamma), palmitate, or CPA. Detailed mutagenesis studies of the Chop promoter showed differential regulation of Chop transcription by these compounds. While cytokines (via NO production)- and palmitate-induced Chop expression was mediated via a C/EBP-ATF composite and AP-1 binding sites, CPA induction required the C/EBP-ATF site and the ER stress response element (ERSE). Cytokines, palmitate and CPA induced ATF4 protein expression and further binding to the C/EBP-ATF composite site, as shown by Western blot and EMSA experiments. There was also formation of distinct AP-1 dimers and binding to the AP-1 site after exposure to cytokines or palmitate.

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Doctorat en Sciences biomédicales et pharmaceutiques
info:eu-repo/semantics/nonPublished

The purpose of this study was to explore the feasibility of using human patient simulation (HPS) to teach Type 1 diabetes (T1DM) management to grandparents of grandchildren with T1DM. Thirty grandparents (11 male, 19 female) of young grandchildren (aged 12 and under) with T1DM were recruited from an urban medical center. Experimental group (n = 14) grandparents received hands-on visual T1DM management education using an HPS intervention, and control group (n = 16) grandparents received similar education using a non-HPS intervention. Post-intervention, researchers interviewed twelve grandparents (50% HPS, 50% non-HPS) who scored highest and lowest on the Hypoglycemia Fear Survey. Using a mixed-method design, researchers integrated study instrument data and post-intervention interview data to describe grandparent’s experience learning T1DM management. Post-intervention, grandparent scores for knowledge, confidence, and fear showed no significant difference by group assignment, however, all grandparent scores showed improvement from Time 1 to Time 2. Grandparents described how taking part in T1DM education heightened their awareness of T1DM risks. GP T1DM knowledge gains aided GPs to make sense of T1DM risks. Newfound T1DM knowledge enhanced GP T1DM management confidence. Improved T1DM knowledge and confidence helped to defuse T1DM management fear. Although study instruments did not measure significant difference between grandparents who received the HPS intervention and those who did not, the consistency of larger HPS-taught grandparent score improvement is suggestive of a benefit for HPS.

Introduction: Type 1 diabetes mellitus (T1DM) is characterized by an absolute deficiency of the insulin-producing beta cells of the islets of Langerhans in the pancreas. Among mechanisms that lead to pathogenesis of T1DM, innate immunity including key cells monocytes are involved. Based on expression of CD14 and CD16 surface markers, monocytes are classified into three subtypes with different functions. In addition to other markers, monocytes express on their surface prolactin receptor (PRLR) and toll-like receptors (TLR), which induce inflammatory responses, and produce extrapituitary hormone prolactin (PRL) that affects immune response. The aim of thesis was to study an effect of exogenous prolactin on the immune responses of monocytes and to try to detect its possible role in the pathogenesis of T1DM. Material and Methods: In vitro cultivation and stimulation of monocytes derived from 10 patients with T1DM and 10 healthy controls. As stimulating agents were used PRL and/or lipopolysacharide (LPS). For determination of mRNA levels of the studied cytokines (TNF-α, IL-6, FOS, IRF-1), total RNA isolated from monocytes acquired by immunomagnetic separation has been quantified by using Real Time PCR. The expression of surface markers (CD14, CD16, PRLR) was detected by flow cytometry. For detection of...

Diabetes mellitus Typ 1 ist eine chronische Autoimmunkrankheit, bei der die Beta-Zellen der Langerhans-Inseln im Pankreas durch autoreaktive T Lymphozyten zerstört werden und somit die Insulinproduktion zum Erliegen kommt. Die vorliegende prospektive Querschnittsstudie untersucht die die Reaktion und Polarisierbarkeit der peripheren T Lymphozyten, die Zytokinproduktion der PBMCs und die Expression der spezifischen Transkriptionsfaktoren Tbet (Th1), FoxP3 (Th17) und RORc (Treg) nach Stimulation mit diabetesspezifischen Antigenen und Candida albicans bei sieben gesunden Kontrollen, neun erstmanifestierten T1DM Patienten und elf langzeiterkrankten T1DM Patienten. Bei der Untersuchung der spezifischen CD4+ T Zellen zeigte sich, dass EM sowohl im unstimulierten Ansatz, aber auch nach Stimulation mit den Antigenen eine größere Proliferationsaktivität aufwies als HD und LS. Interessanterweise war ein vergleichbarer Unterschied bei den CD8+ T Zellen lediglich nach Stimulation mit GAD65 zu sehen. Bei Betrachtung der CD4+ Subpopulationen erkennt man, dass es in allen Kohorten große interindividuelle Unterschiede gibt und man keine signifikanten Unterschiede zwischen den Kohorten beobachten kann. Trotzdem lässt sich sagen, dass sich die Subpopulationen nach den spezifischen Stimulationen in den drei Kohorten teilweise unterschiedlich verschieben und dies Anzeichen dafür ist, dass die T Zellen der T1DM Patienten auf diese Antigene anders verhalten als HD. Bei den CD8+ TEMRAs gibt es mehrere signifikante Unterschiede und es fällt auf, dass EM deutlich weniger TEMRA aufweisen als die anderen beiden Kohorten. Sowohl bei den CD25+ Tregs als auch bei den CD161+ Th17 Zellen zeigen sich keine relevanten Signifikanzen. Die Chemokinrezeptor tragenden weisen sowohl bei den CD4+-T Zellen als auch bei den CD8+ T Zellen Unterschiede und auch Parallelen zwischen den Kohorten auf. Während sich die CD4+CCR5+ Th1 Zellen bei EM durch die Antigene polarisieren lassen, findet bei HD keine Polarisierung statt. Dafür tragen die HD über allen Ansätzen mehr CD4+ und CD8+CXCR3+ Th1 Zellen als EM. Bei den Chemokinrezeptoren CCR6+ und CD25+CCR5+ zeigen sich keine bemerkenswerten Unterschiede oder Polarisierungen durch die Antigene. Im Einklang mit den Ergebnissen der Phänotypisierung der Th1, Th17 und Treg Zellen stehen die der spezifischen Transkriptionsfaktoren. Auch hier waren keine signifikanten Unterschiede zwischen den drei Kohorten vorhanden. Interessanterweise zeigte sich jedoch, dass die relative Transkription nach Stimulation mit den Antigenen in allen drei Kohorten fast ausnahmslos abnahm. Abschließend ist zu erwähnen, dass die Probandenzahl bei dieser Untersuchung sehr klein war und große interindividuelle Unterschiede vorlagen. Bei Betrachtung des Th1 spezifischen Zytokins IFN y fiel auf, dass HD und LS im unstimulierten Ansatz deutlich mehr produzierte als EM, während die Konzentrationen durch Stimulation mit den diabetesspezifischen Antigenen bei HD und LS stark abfiel und bei EM annähernd gleichgeblieben ist. Auffallend war außerdem, dass die durchschnittliche Produktion des Th17 spezifischen Zytokins IL 17 von EM in vielen Ansätzen deutlich größer war als von HD. Das hauptsächlich von den Treg Zellen produzierte IL-10 war bei den T1DM Patienten deutlich kleiner als bei HD. Ebenso wie IFN-y fiel die Konzentration durch die Stimulationen bei HD jedoch stark ab, während sie bei EM und LS gleichblieb oder anstieg. Insgesamt lässt sich sagen, dass es große interindividuelle Unterschiede innerhalb der Kohorten hinsichtlich der Produktion der Zytokine in den verschiedenen Ansätzen gab. Somit ist es von enormem Interesse, die Zytokinproduktion nach Stimulation mit den diabetesspezifischen Antigenen in den verschiedenen Kohorten an einer größeren Anzahl an Probanden zu untersuchen. Zusammenfassend ergeben sich einzelne Hinweise, dass sich die Reaktion der T Zellen auf diabetesspezifische Antigene bei erstmanifestierten T1DM von HD unterscheiden. Inwieweit einzelne Autoantigen-spezifische Subpopulationen, Transkriptionsfaktoren oder proinflammatorische bzw. antiinflammatorische Zytokine eine Rolle in der Pathogenese des T1DM spielen und diese ein Angriffsziel für Therapeutika sein könnten, gilt es weiterhin in humanen Studien herauszufinden
Diabetes mellitus type 1 is a chronic autoimmune disease in which the beta cells of the islets of Langerhans in the pancreas are destroyed by autoreactive T lymphocytes, thus the insulin production is stopped. The present prospective cross-sectional study investigates the response and polarizability of peripheral T lymphocytes, the cytokine production of PBMCs and the expression of the specific transcription factors Tbet (Th1), FoxP3 (Th17) and RORc (Treg) after stimulation with diabetic specific antigens and Candida albicans in seven healthy controls, nine first manifested T1DM patients and eleven long-term T1DM patients. Examination of the specific CD4+ T cells showed that EM had a higher proliferation activity than HD and LS both in the unstimulated approach and after stimulation with the antigens. Interestingly, a comparable difference in CD8+ T cells was only seen after stimulation with GAD65. When looking at the CD4+ subpopulations, it is evident that there are large interindividual differences in all cohorts and no significant differences between the cohorts can be observed. Nevertheless, it can be said that the subpopulations shift differs after specific stimulation in the three cohorts and this is an indication that the T cells of T1DM patients respond differently to these antigens than HD. In CD8+ TEMRAs there are several significant differences and it is noticeable that EM show significantly less TEMRA than the other two cohorts. Both CD25+ Tregs and CD161+ Th17 cells do not show any relevant significance. The chemokine receptor carrying cells show differences and parallels between the cohorts in both CD4+ T cells and CD8+ T cells. While CD4+CCR5+ Th1 cells are polarized by the antigens in EM, no polarization occurs in HD. However, HD carries more CD4+ and CD8+CXCR3+ Th1 cells than EM. The chemokine receptors CCR6+ and CD25+CCR5+ do not show any remarkable differences or polarization by the antigens. The results of phenotyping of Th1, Th17 and Treg cells are consistent with those of the specific transcription factors. Again, there were no significant differences between the three cohorts. Interestingly, however, it was found that relative transcription decreased almost without exception in all three cohorts after stimulation with the antigens. Finally, it should be mentioned that the number of subjects in this study was very small and that there were large interindividual differences. When looking at the Th1 specific cytokine IFN-y, it was noticeable that HD and LS produced significantly more than EM in the unstimulated approach, whereas the concentrations dropped sharply in HD and LS after stimulation with the diabetic-specific antigens and remained approximately the same in EM. It was also remarkable that the average production of the Th17 specific cytokine IL-17 of EM was significantly higher in many approaches than in HD. The IL-10 produced mainly by Treg cells was significantly smaller in T1DM patients than in HD. Like IFN-y, however, the concentration decreased significantly in HD, while it remained the same or increased in EM and LS. Overall, it can be said that there were large interindividual differences within the cohorts regarding the production of cytokines in the different approaches. Therefore, it is of great interest to study the cytokine production after stimulation with the diabetic specific antigens in the different cohorts in a larger number of volunteers. In summary, there is some evidence that the response of T cells to diabetes-specific antigens differs from HD in first manifested T1DM. The extent to which individual autoantigen-specific subpopulations, transcription factors or proinflammatory or anti-inflammatory cytokines play a role in the pathogenesis of T1DM and could be a target for therapeutic agents remains to be determined in human studies

碩士
中原大學
心理學研究所
97
Background and purpose: Metabolic control of diabetes often deteriorates during puberty in adolescents with T1DM. Diabetes management, parent-adolescents relationship and psychological adaptation will also affect the metabolic control as well as puberty. Some studies had studied the relationship between the parent-adolescents conflict and metabolic control, but the results are different. The present study is to examine the relationships among sub-stages of puberty, parent-adolescent conflict, diabetes management, metabolic control, and depressive symptoms in adolescents with T1DM. Methods: The subjects participated in this study were 208 adolescents with T1DM who aged 9 to 20 yr recruits from the medical center in North Taiwan. The instruments are the basic information questionnaire, the Chinese version of Self-care Inventory, the Parent-adolescent Conflict Scale, the Chinese version of Diabetes Family Conflict Scale and the Adolescent Depression Inventory. The statistical analysis included the correlation, one-way ANOVA and multiple regression. Results: The parent-adolescents conflict (include the general parent-adolescents conflict and diabetes-specific family conflict) is positive correlation with depression in adolescents with T1DM, which means higher degree of parent-adolescent conflict, more severity of depression. The diabetes-specific family conflict in early puberty is higher than late puberty; the metabolic control (HbA1c) in early puberty is better than late puberty. The diabetes management is the mediator between parent-adolescent conflict (include the general conflict and diabetes-specific family conflict factor 1) and metabolic control (HbA1c), which means parent-adolescents conflict would influence the metabolic control through diabetes management. The parent-adolescents conflict (the general parent-adolescents conflict and diabetes-specific family conflict) and diabetes management are correlations significant with metabolic control and depression. However, only diabetes management can predict metabolic control. Both parent-adolescents conflict (include the general parent-adolescents conflict and diabetes-specific family conflict) and diabetes management can predict depression in adolescents with T1DM. Conclusion: For the better metabolic control (HbA1c) and reducing the severity of depression, the public health workers and parents should more care about the diabetes management tasks and parent-adolescents conflict in adolescents with T1DM.

Dissertação de Mestrado em Gestão e Economia da Saúde apresentada à Faculdade de Economia
RESUMOIntrodução: A Diabetes Mellitus tipo 1 (DMT1) é uma doença crónica provocada pela redução da secreção de insulina endógena, resultante da destruição autoimune das células β, dos ilhéus de Langerhans, que a produzem. A DMT1 encontra-se frequentemente associada a outras doenças autoimunes (DAI), entre as quais a Tiroideia Autoimune (DTA).Objetivos: Avaliar a incidência de DTA em 2019 e a sua prevalência entre 2002 e 2018, nos doentes com DMT1 seguidos na Unidade de Endocrinologia Pediátrica de Diabetes e Crescimento, do Centro Hospitalar e Universitário de Coimbra (CHUC).Métodos: Realizou-se um estudo observacional analítico retrospetivo para identificar os doentes portadores de DMT1, na Consulta de Diabetologia Pediátrica do CHUC, com monitorização dos marcadores de DTA, através da informação disponibilizada no processo clínico eletrónico.Obtivemos dados sociodemográficos e de natureza clínica. Destacam-se a data de diagnóstico da DMT1 e da DTA, índice de massa corporal (IMC), glicemia média dos últimos 14 dias, hemoglobina glicada (HbA1c), doseamento das hormonas tiroideias, anticorpos anti-pancreáticos, anti-insulina e anti-tiroideus, entre o período de 2002 e 2019. Obtivemos ainda valores séricos do anticorpo AZnT8, que ainda não consta do protocolo de análises de rotina em Portugal.Identificámos a prevalência de DTA entre o período de 2002 e 2018 e a incidência da mesma em 2019.Resultados: Dos 262 doentes inicialmente previstos, 21 foram excluídos, por dados incompletos, sendo que a nossa amostra foi constituída por 241 crianças e jovens com DMT1. A prevalência de DTA, no período de 2002 a 2018, é 21,12% e a incidência em 2019 é 4,17%.Comparando a DMT1 com e sem DTA, a média da idade atual é significativamente superior no grupo que desenvolveu DTA (p < 0,03), são crianças mais altas (p < 0,001) e com um maior IMC (p < 0,001). Verificaram-se ainda diferenças significativas nos níveis séricos de TSH (p < 0,001), na idade das crianças no momento do diagnóstico da DTA e da DMT1 (p < 0,001), sendo o tempo de duração da DTA menor em relação ao da DMT1 (p < 0,001). Discussão e Conclusão: Neste estudo a prevalência de DTA na DMT1 foi de 21,12%, mais elevada do que na população em geral. A incidência em 2019 foi de 4,17%. A Tiroidite Hashimoto (TH) é a DTA mais frequentemente associada à DMT1, como constatámos, o que justifica a realização de um rastreio anual de anticorpos anti-tiroideus. Serão necessários um maior número de resultados, para se ponderar o interesse clínico da inclusão do anticorpo anti ZnT8 nas análises de rotina.Palavras-chave: Diabetes Mellitus Tipo 1, etiopatogénese, doenças autoimunes, tiroidite autoimune, síndromes poliglandulares endócrinos.
ABSTRACTIntroduction: Type 1 diabetes mellitus (T1DM) is a chronic disease that results from a lack of endogenous insulin secretion from the pancreatic cells. Those β- cells are destroyed by a targeted autoimmune process. Type 1 diabetes mellitus is a heterogeneous disease with multiple different features and phenotypic characteristics. Potential triggers of the autoimmune process include genetic and environmental factors. T1DM is associated with other autoimmune diseases, being the most frequent ones the autoimmune thyroid disease.Aim: Evaluate the incidence in 2019 and the prevalence of thyroid disease in patients with T1DM treated at the ambulatory of endocrinology at the Pediatric University Hospital of Coimbra, from 2002 to 2018, was also evaluated.Methods: A retrospective analytical observational study was conducted in order to identify T1DM patients with annual monitoring ATD markers. The medical records of 241 patients with T1DM were reviewed and the collected data on age, weight, height, gender, diagnostic age, disease’s time, thyroid function, antithyroid antibodies, glycosylated hemoglobin, glycemia media of 14 days, Islet cell (ICA), endogenous insulin (IAA), 65 decarboxylase of glutamic acid (GADA), tyrosine phosphatase like (IA2A) and ZnT8 antibody. ZnT8 antibody was evaluated for the first time in these children. The Statistical Package for the Social Science software (SPSS, v. 24) was used to analyze the data.Results and Discussion: Our sample has 262 children, with 21 excluded for incomplete date, leading to 241 patients evaluated (N = 241).The ATD prevalence is 21,12% during the period of 2002-2018. The incidence in 2019 is 4,17%.Children with T1DM and ATD are older than children with only T1DM p<0,03, consequently they have a significant higher p<0,001, a higher IMC p<0,001 and a higher TSH serum levels p<0,0001. There are also significant differences between the two groups concerning children’s age at diagnoses and time of diagnoses. The children are older at the time of ADT diagnoses p<0,0001.The most frequent form of ATD is Hashimoto’s thyroiditis in 50,9% of the children on our samples.Conclusions: ATD prevalence in T1DM between 2002-2018 was 21,12%, higher than in general population and like the literature. ATD incidence has annual increases in many countries, whereby is important to perform national studies to evaluate Portugal’s ATD incidence and compare it with other countries.ATD is the most common autoimmune disease associated with T1DM, which is why it’s important and recommended to make annual screenings.Keys words: T1DM, Etiopathogenesis review, Syndrome Polyglandular, ATD, autoimmune diseases