Aims: Better understanding of the timeline and risk factors for the appearance of complications in pediatric Type-1-diabetes is key for developing prevention strategies. We studied endothelial markers and their determinants in adolescents with Type-1-diabetes at different time points from diagnosis. Methods:A cross-sectional study of 58 adolescents, mean age 15.0 ± 2.4 years; 20 with recent-onset Type-1-diabetes, 20 with over 7 years of Type-1-diabetes and 18 controls. Clinical and biochemical data were collected. Fingertip arterial reactive hyperemia (EndoPAT) and carotid intima-media-thickness (cIMT) were measured to assess endothelial function and structure.Results: Compared to controls, individuals with prolonged Type-1-diabetes had higher mean cIMT (0.49 ± 0.07 mm vs. 0.43 ± 0.05 mm p = 0.021) and maximal cIMT (0.61 ± 0.08 mm 0.52 ± 0.08 mm, p = 0.025). Endothelin-1 levels were significantly lower in subjects with prolonged Type-1-diabetes (1.2 ± 1.0 pg/ml) compared to controls (3.0 ± 1.7, p = 0.008 pg/ml); they negatively correlated with the mean cIMT (c = À 0.291, p = 0.031) and mean 6 months hemoglobin A1c (c = À 0.301, p = 0.022) and positively correlated with mean c-peptide levels (c = 0.356, p = 0.006) and the weekly exercise time (c = 0.485, p < 0.001). Endothelin-1 levels did not correlate with EndoPAT results.Conclusions: Our results suggest that the early years after the diagnosis of Type-1-diabetes are an important window for prevention of arterial damage in the pediatric population. The trajectories of relationships of Endothelin-1 with metabolic and vascular measures were opposite from the anticipated, yet consistent. Endothelin-1 related indirectly to adverse measures and directly to favorable measures. Decreased Endothelin-1 levels might reflect early stages in endothelial impairment in Type-1-diabetes, yet its' exact role in the development of complications is yet to be unraveled.

Visible light irradiation is an emerging area in regenerative medicine research. We hypothesized that low-intensity-pulsed LED light irradiance may exert photobiomodulatory effects on cultured osteoblast-like cells. To test this hypothesis, we investigated cell proliferation and markers of cell maturation and metabolic activity following pulsed LED irradiance. Monolayer explant cultures of human osteoblast-like cells were exposed four times in 24-h intervals to 2 min of pulsed white LED irradiance of 2.4-2.5 mWÁcm À2 and its different spectra of 0.2-0.5 mWÁcm À2 (frequency range of 10-40 Hz). Cell proliferation was estimated from microscopic cell counting and cell death by lactate dehydrogenase activity in culture media (measured by a colorimetric method). The early markers of osteoblast maturation and metabolic activity, that is, cellular alkaline phosphatase activity and osteocalcin content, were measured using a colorimetric method and ELISA, respectively. Irradiance of 40 Hz caused the highest increase in cell number (P < 0.01). Osteocalcin content in cells decreased following 40 Hz and 10 Hz irradiance (P < 0.05). The 40 Hz blue range irradiance (diffuse transmittance 420-580 nm, maximal cell irradiance 0.5 mWÁcm À2) caused a decrease in alkaline phosphatase cellular activity (P < 0.001) and an increase in media osteocalcin content (P < 0.05). The 40 Hz green range (diffuse transmittance 560-650 nm, maximal cell irradiance 0.4 mWÁcm À2) irradiance caused an increase in the number of cells and in cell death. In summary, pulsed (40 Hz) white light irradiance has photomodulatory effects, with its green range spectrum affecting cell proliferation and cell death, and its blue range spectrum affecting cellular maturation and metabolism. The results indicate a low-intensity threshold of photobiomodulation of osteoblast-like cells in vitro.

Background & Objective Patients with type-1-diabetes (T1D) are at risk of long-term micro and macrovascular complications causing significant morbidity and mortality. Overt complications are not common in childhood; however, subclinical impairments in endothelial function, may be found. Better understanding of the timeline for the appearance of diabetic complications and identifying individuals at increased risk is key for developing prevention strategies. We aimed to study endothelial function and it’s determinants in adolescents with T1D at different time points from diagnosis. Methods Forty adolescents 11-20 years of age with T1D followed at our pediatric diabetes clinic and 18 healthy control subjects were included. Two groups of patients were recruited based on time from T1D diagnosis; 20 individuals were diagnosed 2-4 months prior to the study visit and 20 at least 7 years prior to the visit. Investigations included: i) medical and demographic data ii) anthropometrics iii) fasting blood samples iv) EndoPAT testing of endothelial function and heart rate variability (Itamar Medical Ltd., Israel) v) Carotid intima media thickness (CIMT). Results Mean age differed slightly between groups being 14.1±2.0years in individuals with recent-onset T1D, 16.2±2.5 in those with prolonged T1D, and 14.8±2.3 in the control group (p=0.02). There were no significant differences in pubertal stage or in BMI z-score between groups. Thirty-three (57%) females participated. No patient suffered from diabetic complications. Mean CIMT was significantly higher in individuals with prolonged T1D (0.49±0.07mm) compared to control subjects (0.43±0.05mm; p=0.013) and did not differ significantly between patients with recent-onset T1D (0.45±0.07mm) and controls. This difference remained significant when age and sex were included in the model. EndoPAT measures of endothelial function and heart rate variability did not differ significantly between groups. Mean HbA1c at the time of the visit differed between groups (6.7%±0.7, 9.6%±1.8, 5.4%±0.3, p&lt;0.001). However, the average of HbA1c reflecting the 6-7 months prior to the visit did not differ significantly between subjects with recent onset T1D (9.8%±1.3) and those with prolonged T1D (9.5%±1.7). LDL was higher in subjects with prolonged T1D (114±28mg/dl) compared to either controls (93±26mg/dl) or recent onset T1D (88±19mg/dl),p=0.002. Diastolic blood pressure was higher in subjects with prolonged T1D (70±6mmHg) than in controls (61±6, p=0.007). Conclusions Our results demonstrate disease duration to be an important factor in the development of subclinical arterial damage in the pediatric age group. Early in the course of T1D, CIMT results were similar in patients and control subjects, suggesting an important window for prevention. Larger studies could shed light on the precise timeline of endothelial impairment.

ContextThe 250µg-cosyntropin stimulation test (CST) is used to diagnose non-classic congenital adrenal hyperplasia (NCCAH). The current recommendation is to perform CST when follicular 17-hydroxyprogesterone (17OHP) is 6-30 nmol/L, a cutoff derived from radioimmunoassay (RIA). Recently, enzyme-linked immunosorbent assay (ELISA) has replaced RIA.ObjectivesWe aimed to (1) determine the RIA and ELISA-based 17OHP cutoffs at which CST should be performed, (2) identify predictors of NCCAH.MethodsA retrospective study at an Israeli Health Maintenance Organization. Data were retrieved from women with suspected NCCAH, referred for CST during 2001–2020. NCCAH was defined as a stimulated 17OHP &gt;30 nmol/L. Serum 17OHP levels were assayed by RIA from 1/2000-3/2015, and by ELISA from 4/2015-12/2020. ROC curves were generated and optimal 17OHP thresholds were determined. Multivariate analysis was performed.ResultsCST was performed in 2409 women (1564 in RIA, 845 in ELISA). NCCAH was diagnosed in 4.7% of the RIA group and 7.5% of the ELISA group. The optimal basal 17OHP cutoff values predicting NCCAH were 6.1 nmol/L in RIA (sensitivity=93.2%, specificity=91.7%) and 8.2 nmol/L in ELISA (sensitivity=93.7%, specificity=92.3%). In multivariate analysis, higher basal 17OHP, lower LH: FSH ratio, and oligomenorrhea were predictors of NCCAH in RIA. Higher basal 17OHP, androstenedione, and total testosterone were predictors of NCCAH in ELISA. A lower LH: FSH ratio showed similar trend in ELISA.ConclusionsOptimal RIA-based basal 17OHP cutoff was comparable with that recommended in guidelines. The results suggest adopting a higher 17OHP cutoff when using ELISA. LH : FSH ratio improves the negative predictive value of basal 17OHP.