Association of circulating miRNAS in patients with Alstrom and Bardet-Biedl syndromes with clinical course parameters.

Background: Patients with the rare syndromic forms of monogenic diabetes: Alström syndrome (ALMS) and Bardet-Biedl syndrome (BBS) have multiple metabolic abnormalities, including early-onset obesity, insulin resistance, lipid disorders and type 2 diabetes mellitus. The aim of this study was to determine if the expression of circulating miRNAs in patients with ALMS and BBS differs from that in healthy and obese individuals and determine if miRNA levels correlate with metabolic tests, BMI-SDS and patient age. Methods: We quantified miRNA expression (Qiagen, Germany) in four groups of patients: with ALMS (n=13), with BBS (n=7), patients with obesity (n=19) and controls (n=23). Clinical parameters including lipids profile, serum creatinine, cystatin C, fasting glucose, insulin and C-peptide levels, HbA1c values and insulin resistance (HOMA-IR) were assessed in patients with ALMS and BBS. Results: We observed multiple up- or downregulated miRNAs in both ALMS and BBS patients compared to obese patients and controls, but only 1 miRNA (miR-301a-3p) differed significantly and in the same direction in ALMS and BBS relative to the other groups. Similarly, 1 miRNA (miR-92b-3p) was dysregulated in the opposite directions in ALMS and BBS patients, but diverged from 2 other groups. We found eight miRNAs (miR-30a-5p, miR-92b-3p, miR-99a-5p, miR-122-5p, miR-192-5p, miR-193a-5p, miR-199a-3p and miR-205-5p) that significantly correlated with at least of the analyzed clinical variables representing an association with the course of the diseases. Conclusions: Our results show for the first time that serum miRNAs can be used as available indicators of disease course in patients with ALMS and BBS syndromes.

Metabolic bone markers can be related to preserved insulin secretion in children with newly diagnosed type 1 diabetes.

INTRODUCTION: Type 1 diabetes (T1D) may be associated with numerous complications including bone metabolism disorders. The aim of the study was to evaluate the bone metabolism markers twice in children with a newly diagnosed T1D and after an average of seven months of its duration in relation to parameters of the clinical course of diabetes. MATERIAL AND METHODS: In 100 T1D patients and 52 control subjects, the following bone turnover markers were evaluated: osteocalcin - OC, osteoprotegerin - OPG, sRANKL, and deoxypyridoline in urine - DPD and DXA examination was also performed. RESULTS: Lower OC concentration at T1D onset in comparison to controls (p < 0.001) and its increase during follow-up (p < 0.001) was ob-served. The OPG concentration was elevated at T1D onset as compared to the control group (p = 0.024) and decreased thereafter (p < 0.001). The s-RANKL level increased during follow-up (p < 0.001) and was lower than in controls (p < 0.001). Urine DPD con-centration also increased during follow-up in the T1D patient group (p < 0.001) and was higher in comparison to the control group (p = 0.021). BMD-TBLH was higher in the control group as compared to patients both at T1D onset (p = 0.025) and in follow-up ob-servation (p = 0.034). Moreover, OPG correlated positively with glycated haemoglobin (HbA1c) (p = 0.004) and negatively with fasting C-peptide level (p = 0.046) and BMI Z-score (p = 0.003), whereas s-RANKL correlated positively with both fasting (p < 0.001) and stimulated C-peptide levels (p < 0.001). CONCLUSIONS: Bone metabolism disorders observed at T1D onset in children and modified after reaching the metabolic control of the disease seem to be most strongly associated with preserved insulin secretion.

Alcohol and cigarette use among adolescents with type 1 diabetes.

In this study, we compare the prevalence of alcohol and cigarette use among Polish adolescents with type 1 diabetes mellitus (T1DM) (n = 209), aged 15-18 years, with that of a large cohort of their healthy peers, using standardized questionnaire used in the European School Survey Project on Alcohol and Drugs (ESPAD). The lifetime, previous year, and past 30-day prevalence of alcohol consumption was high among adolescents with T1DM but lower than in the controls (82.8 vs 92.0%, 71.7 vs 85.6%, and 47.5 vs 69.7%, respectively, p < 10-5). The lifetime and 30-day prevalence of cigarette use was also lower among patients than the controls (54.6 vs 65.5%, p = 0.001 and 27.3 vs 35.9%, p = 0.012, respectively). Patients who admitted smoking exhibited worse metabolic control than non-smokers (p < 0.0001) and had a higher chance of developing diabetic ketoacidosis. The incidence of severe hypoglycemia was higher among those who reported getting drunk in the previous 30 days (p = 0.04) and lifetime smoking (p = 0.01). CONCLUSIONS: Although alcohol and cigarette consumption is lower than in controls, it is common among teenagers with type 1 diabetes, effecting metabolic control and causing the risk of acute diabetes complications. Better prevention strategies should be implemented in this group of patients in their early teen years. What is Known: • Substance use remains a significant cause of morbidity and mortality among teenagers with type 1 diabetes. • Current medical literature contains inconsistent data on the prevalence of alcohol and cigarette use among adolescents with type 1 diabetes, mostly due to methodological problems with conducting such surveys. What is New: • Methodological approach: we used a validated questionnaire from the European School Survey Project on Alcohol and Other Drugs (ESPAD) and compared the results to a large national control group of 12,114 healthy students who took part in ESPAD in 2011.

Factors affecting long-term efficacy of T regulatory cell-based therapy in type 1 diabetes.

BACKGROUND: Recent studies suggest that immunotherapy using T regulatory cells (Tregs) prolongs remission in type 1 diabetes (T1DM). Here, we report factors that possibly affect the efficacy of this treatment. METHODS: The metabolic and immune background of 12 children with recently diagnosed T1DM, as well as that of untreated subjects, during a 2-year follow-up is presented. Patients were treated with up to 30 × 106/kg b.w. of autologous expanded CD3+CD4+CD25highCD127- Tregs. RESULTS: The disease progressed and all patients were insulin-dependent 2 years after inclusion. The ß-cell function measured by c-peptide levels and the use of insulin were the best preserved in patients treated with two doses of Tregs (3/6 in remission), less so after one dose (1/6 in remission) and the worst in untreated controls (no remissions). Increased levels of Tregs could be seen in peripheral blood after their adoptive transfer together with the shift from naïve CD62L+CD45RA+ to memory CD62L+CD45RA- Tregs. Increasing serum levels of proinflammatory cytokines were found: IL6 increased in all subjects, while IL1 and TNFα increased only in untreated group. Therapeutic Tregs were dependent on IL2, and their survival could be improved by other lymphocytes. CONCLUSIONS: The disease progression was associated with changing proportions of naïve and memory Tregs and slowly increasing proinflammatory activity, which was only partially controlled by the administered Tregs. The therapeutic cells were highly dependent on IL2. We conclude that the therapy should be administered at the earliest to protect the highest possible mass of islets and also to utilize the preserved content of Tregs in the earlier phases of T1DM. Trial registration http://www.controlled-trials.com/ISRCTN06128462 ; registered retrospectively.

Differential regulation of serum microRNA expression by HNF1ß and HNF1α transcription factors.

AIMS/HYPOTHESIS: We aimed to identify microRNAs (miRNAs) under transcriptional control of the HNF1ß transcription factor, and investigate whether its effect manifests in serum. METHODS: The Polish cohort (N = 60) consisted of 11 patients with HNF1B-MODY, 17 with HNF1A-MODY, 13 with GCK-MODY, an HbA1c-matched type 1 diabetic group (n = 9) and ten healthy controls. Replication was performed in 61 clinically-matched British patients mirroring the groups in the Polish cohort. The Polish cohort underwent miRNA serum level profiling with quantitative real-time PCR (qPCR) arrays to identify differentially expressed miRNAs. Validation was performed using qPCR. To determine whether serum content reflects alterations at a cellular level, we quantified miRNA levels in a human hepatocyte cell line (HepG2) with small interfering RNA knockdowns of HNF1α or HNF1ß. RESULTS: Significant differences (adjusted p < 0.05) were noted for 11 miRNAs. Five of them differed between HNF1A-MODY and HNF1B-MODY, and, amongst those, four (miR-24, miR-27b, miR-223 and miR-199a) showed HNF1B-MODY-specific expression levels in the replication group. In all four cases the miRNA expression level was lower in HNF1B-MODY than in all other tested groups. Areas under the receiver operating characteristic curves ranged from 0.79 to 0.86, with sensitivity and specificity reaching 91.7% (miR-24) and 82.1% (miR-199a), respectively. The cellular expression pattern of miRNA was consistent with serum levels, as all were significantly higher in HNF1α- than in HNF1ß-deficient HepG2 cells. CONCLUSIONS/INTERPRETATION: We have shown that expression of specific miRNAs depends on HNF1ß function. The impact of HNF1ß deficiency was evidenced at serum level, making HNF1ß-dependent miRNAs potentially applicable in the diagnosis of HNF1B-MODY.

How does autoimmune thyroiditis in children with type 1 diabetes mellitus influence glycemic control, lipid profile and thyroid volume?

AIM: To investigate whether autoimmune thyroiditis (AIT) in children with type 1 diabetes mellitus (DM1) has any influence on glycemic control, lipid profile or thyroid volume. METHODS: A total of 330 patients with DM1 and AIT (DM1+AIT group) were compared with 309 children with DM1 without AIT (control group). Patients were treated in four Polish academic pediatric diabetes centers from 2008 to 2012: Warsaw, Lodz, Katowice and Gdansk. All patients underwent measurements of thyroid-stimulating hormone (TSH), free thyroxine, anti-thyroid peroxidase (anti-TPO) antibody, anti-thyroglobulin (anti-TG) antibody and HbA1c levels, and thyroid ultrasound examination. RESULTS: Among AIT+DM1 patients, 62% (n=205) were female, whereas in the control group 60.8% (n=188) were male (p<0.0001). Children with AIT+DM1 had lower a BMI-SDS (mean difference of -0.5, 95% CI -0.68 to -0.33; p<0.0001), had a higher SDS thyroid volume (0.27, 95% CI 0.03-0.51; p=0.014) and needed less insulin (-0.15, 95% CI -0.20 to -0.11 U/kg body weight per day; p<0.0001) in comparison with the control group. AIT patients had higher HbA1c levels (0.66, 95% CI 0.36%-0.96%, p<0.0001), lower HDL-cholesterol levels (-3.68, 95% CI -1.41 to -5.94 mg/dL, p=0.002) and higher triglyceride levels (7.16, 95% CI 1.22-13.10 mg/dL, p=0.02). Patients with positive anti-TPO and anti-TG antibodies were older (by 1.95 years, 95% CI 0.98-2.92 years, p=0.006) and had longer DM1 duration (by 1.64 years, 95% CI 0.76-2.52 years, p=0.006). Presence of anti-TPO antibodies was associated with higher TSH levels (odds ratio 2.34, 95% CI 1.36-4.04; p=0.007). CONCLUSION: AIT accompanying DM1 is associated with worse glycemic control and lipid profile as well as a lower daily insulin requirement. The female gender is more likely to develop AIT and hypothyroidism.

The prevalence of Wolfram syndrome in a paediatric population with diabetes.

INTRODUCTION: Wolfram syndrome (WFS) is the most frequent syndromic form of monogenic diabetes coexisting with optic atrophy and many other disorders. The aim of this study was to estimate the prevalence of Wolfram syndrome among children with diabetes in Poland. MATERIAL AND METHODS: These calculations were performed among Polish diabetic children, aged 0-18 years, from three administrative regions between January 2005 and December 2011. Epidemiological data was obtained by matching the results from the EURO-WABBPoland Project and the PolPeDiab Registry. RESULTS: Throughout the study period, we confirmed genetic diagnosis of Wolfram syndrome in 13 patients from Poland. Three patients originated from the studied regions with complete epidemiological data on paediatric diabetes. The total number of patients with diagnosed diabetes in the study equalled 2,568 cases. The prevalence of Wolfram syndrome among Polish children with diabetes is 0.12% (95% Confidence Interval 0.04-0.34%). CONCLUSIONS: We estimate that Wolfram syndrome is: 26 to 35 times less frequent than monogenic diabetes (MODY and neonatal diabetes) in the Polish paediatric population.

Management of familial hypercholesterolemia in children and adolescents. Position paper of the Polish Lipid Expert Forum.

Familial hypercholesterolemia (FH) affects on average 1 in 500 individuals in European countries, and it is estimated that FH in Poland may affect more than 80,000 people. However, in Poland, only about 20% of the population is estimated to have been diagnosed with FH, of which only a small number receive adequate treatment. FH results in more rapid development of atherosclerosis and is associated with a high risk of cardiovascular events. Atherosclerosis develops beginning in childhood in patients with FH and reaches advanced stages before clinical manifestations develop. Inadequate diagnostics and treatment of FH in Polish children suggests a need for raising the level of awareness and understanding of the condition in both society and among health professionals. These recommendations present the current epidemiological status, guidelines for diagnosing FH in Polish children and adolescents, and effective treatment options.

L-thyroxine stabilizes autoimmune inflammatory process in euthyroid nongoitrous children with Hashimoto's thyroiditis and type 1 diabetes mellitus.

OBJECTIVE: To investigate if L-thyroxine (T4) treatment may influence the clinical course of autoimmune thyroiditis (AIT) or prevent progression to subclinical or overt hypothyroidism in euthyroid nongoitrous pediatric patients with type 1 diabetes mellitus (T1DM) and AIT. METHODS: The study was performed in four Polish pediatric diabetes centers. Of 330 children with T1DM and AIT followed between 2008 and 2012, 101 received L-T4 and 160 underwent clinical observation for 24 months. Thyroid stimulating hormone (TSH), free T4 (fT4), anti thyroid peroxidase antibody (anti-TPO), anti thyroglobulin antibody (anti-TG), glycosylated hemoglobin (HbA1c) levels, and lipid profile were assessed in all patients. Ultrasonographic evaluation was also performed in all children at each examination. RESULTS: Patients treated with thyroid hormones had higher TSH levels (3.99; interquantile 3.5 to 4.52 vs. 2.09 mIU/L; interquantile 1.55 to 3.06; pp<0.0001). A fall in TSH level (0.87 mIU/L 95% CI 0.43-1.30; pp<0.0001) was documented after the first year of treatment. FT4 level did not differ between the groups at baseline (p=0.7434), but rose in the treatment group and fell in the control group [mean difference 0.78 95% CI-0.22-1.53 pmol/L (p=0.02) after 12 months and 0.98 95% CI 0.04-1.76 (p=0.005) after 24 months]. Higher levels of anti-TPO were initially found in the treated patients (pp<0.0001) and significantly decreased over the 24-month period (pp<0.0001). Children in the treatment group had higher anti-TG levels (pp<0.0001), which showed a borderline decrease (p=0.08) in time. In the control group, anti-TG levels rose marginally (p=0.06) during the study. CONCLUSIONS: The data demonstrate that treatment with L-T4 in euthyroid pediatric patients with T1DM and AIT stabilizes autoimmune inflammation in the thyroid gland and is to be recommended as soon as the diagnosis is established.

Efficacy of metabolic and psychological screening for mood disorders among children with type 1 diabetes.

OBJECTIVE: To compare the diagnostic accuracy and time expenditure of screening models based on glycated hemoglobin (HbA(1c)) level and psychometric measures for mood disorder (MD) among children with type 1 diabetes. RESEARCH DESIGN AND METHODS: With semistructured clinical interviews (Schedule for Affective Disorders and Schizophrenia for Children-Present and Lifetime version, 120 min/patient) as a reference for diagnosing MD, including major depressive disorder (MDD), we tested 163 subjects, aged 8 to 18 years, with type 1 diabetes. We evaluated four screening approaches: 1) Children's Depression Inventory (CDI) at 30 min/patient, 2) HbA(1c) level, 3) HbA(1c) level plus CDI, and 4) HbA(1c) level plus Children's Depression Rating Scale (CDRS) at 40 min/patient. These tests were conducted with all participants, and the total time expenditure for all four approaches was calculated as the total time needed to implement successfully the screening for MD or MDD in the center. RESULTS: HbA(1c) performed on par with individual psychometric tests in diagnosing MD or MDD. The HbA(1c) plus CDRS model was the best screening procedure for both MD and MDD, with diagnostic thresholds for HbA(1c) established at 8.7% and 9.0%, respectively. Cutoff points for CDRS assessed after filtering by HbA(1c) were 26 (MD) and 30 (MDD) points. Center-wide application of this procedure would result in an 83% reduction of the examination time necessary for the psychiatrist for MD screening and a 91% reduction for MDD screening, as compared with standard screening with CDI. CONCLUSIONS: Use of HbA(1c) level followed by CDRS is a time-efficient procedure to screen for MD in children with type 1 diabetes.

Hypoglycemia and glycemic variability among children with acute lymphoblastic leukemia during maintenance therapy.

Symptomatic, chemotherapy-related hypoglycemia is a rare complication associated with the administration of purine analogs. The aim of the study was to evaluate 24 h glucose variability and frequency of hypoglycemia among patients with acute lymphoblastic leukemia (ALL) during maintenance therapy (MT). Eighteen children with ALL underwent continuous glucose monitoring (CGM). The number of episodes of hypoglycemia and glucose variability were analyzed. Serum alanine aminotransferase, asparagine aminotransferase, and γ-glutamyl transferase levels were measured as liver function markers. The mean glucose level in CGM equaled 105 ± 13 mg/dL, with standard deviation (SD) 13.8 ± 6.1 mg/dL, and the mean amplitude of glycemic excursions (MAGE) equaled 44.7 ± 19.9 mg/dL. Eight patients had at least one measurement below 70 mg/dL while four patients had measurements below 50 mg/dL. Children with hypoglycemia in CGM examination had a lower median body mass index standard deviation score (BMI Z-score) (-0.65 [-0.94 to -0.27] vs. -0.14 [-0.29 to 0.35]; p = 0.05) and shorter duration of MT (6.5 [4-15] vs. 22.5 [16.5-28] weeks; p = 0.004). Glucose variability parameters were strongly correlated with BMI Z-score and liver function enzymes. Hypoglycemia, particularly at night-time, may develop as a complication of MT in children with ALL. The risk factors for low glucose level are low BMI Z-score and initiation of MT.

[Optimization of monogenic diabetes screening programme--initial report on recruitment efficacy of the TEAM project]. / Optymalizacja programu poszukiwania cukrzyc monogenowych - wstepne wyniki dzialan rekrutacyjnych projektu TEAM.

INTRODUCTION: Due to the lack of precise diagnostic criteria, current search strategy for monogenic diabetes is predominantly based on atypical clinical course of diabetes and intuition of the attending physician. Yet another issue is the common view that monogenic diabetes is rare. It discourages from performing deepened diagnostics and makes it difficult to gain experience necessary to select appropriate patients for genetic examination. AIM OF THE STUDY: Estimating the true incidence of patients with a high probability of monogenic background of the disease and compare their search strategies based on clinical practice or structured databases. MATERIAL AND METHODS: The authors compared the current strategy of selecting candidates for screening with a directed search strategy based on immunologic (lack of islet autoantibodies), functional (presence or complete lack of c-peptide at onset and follow-up) and familial (dominant pattern of inheritance) criteria. The number of patients selected for the screening was chosen as efficacy measure selected among 1281 diabetic patients diagnosed and treated between 1983-2009. RESULTS: Screening based on clinical assessment yielded 37 patients (2.9%) chosen for genetic screening. Criteria used by the physicians were based on up-to-date guidelines and unusual clinical course. Active search of the database according to predefined criteria resulted in selecting: 121 patients (9.4%) with likely monogenic background of diabetes (71 - lack of autoantibodies, 8 - normal C-peptide, 6 - lack of both c-peptide and autoantibodies, 36 - diabetes in at least one parent). The difference in screening efficacy was statistically significant (p <0.0001). CONCLUSIONS: Periodic reevaluation of patients' data allows a significant increase in the number of candidates subjected to genetic screening and potentially achieving beneficial therapeutic effects by means of pharmacogenetics.

Parametric clearance kidney scintigrams; diagnostic potential in diabetes.

BACKGROUND: The diagnostic usefulness of parametric clearance kidney images was studied in the early diagnosis of diabetic nephropathy, juxtaposed with conventional dynamic urinary investigation (renoscintigraphy) combined with deconvolution procedure of renal and blood time activity curves and determination of plasma clearance of (99m)Tc-ethylenedicysteine ((99m)Tc-EC). MATERIAL AND METHODS: The investigation was performed on a group of 70 individuals (41 males, 29 females) in whom diabetes type 1 was diagnosed (age 10 to 30 y.; mean 19 y.) and on a control group of 35 healthy individuals (15 males, 20 females) in the age-bracket of 18-25 years (mean 19 y.). In all subjects studied, renoscintigraphy was performed after administration of (99m)Tc-EC (activity 40-120 MBq) combined with determination of urinary clearance (ERPF) of the radiopharmaceutical. The renographic curves were evaluated taking into account their shape and individual share of each kidney, and the clearance function was calculated (RClr). From analysis of the time-activity, kidney curves T(max) and T(1/2) were assessed. In addition, the mean (99m)Tc-EC transport time through the complete kidney (MTT) and organ's parenchyma (PTT) were calculated from results of deconvolution of the curve. From the dynamic urinary system study, conventional images of radiopharmaceutical distribution in the kidneys in the secretion phase were obtained. The parametric clearance images were also computed on the basis of relative clearance values in all the pixels of both kidney regions of interest. The disturbances in kidney function were assessed separately by means of conventional scintigram analysis and of corresponding parametric images. A three-stage classification was used in both cases for the evaluation of abnormal findings in the kidneys RESULTS AND CONCLUSIONS: In all studied individuals, the (99m)Tc-EC (ERPF) clearance values were within the normal range. When renographic time activity curves were considered the flattening of the curves (III phase) was more frequent in diabetic individuals than in the controls (39.3% vs. 15.7%; p = 0.001). The shape of the curves in phases I and II were normal in all studied individuals of both groups. There were no differences observed between mean values of T(max), T(1/2) and PTT in diabetics and controls. However, mean MTT values were significantly higher in diabetics than in controls (p = 0.02). In conventional summation images (phase II of the renograms), there were no significant differences in frequency of defects in kidney parenchyma diabetics and controls (4.3% vs. 2.9%). In contrast, analysis of parametric kidney clearance images revealed that parenchyma defects were found with significantly greater frequency in diabetic individuals (35.7%) than in control subjects (8.6%; p < 0.001). Summarizing the findings, it appears that parametric clearance kidney images reveal local deviations of renal uptake and secretory function while conventional indicators of renal function are still in the normal range. This observation points to the fact that clearance parametric images may have potential value in the early diagnosis of diabetic nephropathy, and perhaps in other types of renal damage. Incorporation of parametric images into the dynamic study of the urinary system may be promising when early detection of kidney damage seems vital.

[Post-exercise microalbuminuria in newly diagnosed type 1 diabetic children - preliminary report]. / Mikroalbuminaria powysilkowa u dzieci ze swiezo rozpoznana cukrzyca typu 1 - doniesienie wstepne.

THE AIM: of this study was the evaluation of some factors having an influence on post-exercise albuminuria level in children with newly diagnosed type 1 diabetes as a prognostic factor of developing nephropathy. MATERIAL AND METHODS: 24 newly diagnosed type 1 diabetic children, aged 5.5-16.9 years, mean 10.2+/-3.2, were examined. In order to provoke albuminuria the patients underwent standardized exercise test using treadmill ramp according to the Bruce protocol. Pre- and post-exercise albuminuria and C-peptide levels by radioimmunoassay were evaluated. In urine of patients the albumin / creatinine ratio (ACR) was determined. RESULTS: the tendency towards an increase in ACR ratio in children after exercise (10.5 mg/g (4-27.5) in comparison to the value in pre-exercise urine (7 mg/g (2.5-13), p=0.17) was observed. In 67% of patients (16/24) the ACR ratio was higher in post-exercise urine. In 25% of children (6/24) the ACR ratio was above 30 mg/g which was considered as post-exercise microalbuminuria. Next, parameters of metabolic control of type 1 diabetes were compared between patients with and without post-exercise microalbuminuria. No differences in the serum glucose, HbA1c, BMI, triglycerides, total cholesterol, insulin dose, frequency of ketoacidosis and C-peptide level between these groups of children were noted. CONCLUSIONS: we concluded that the post-exercise microalbuminuria as a marker of diabetic nephropathy risk may be observed in some patients already at the clinical onset of type 1 diabetes but further evaluation is needed to verify this hypothesis.