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Impact of SiO2 NP-allergen interaction on dendritic cell function altering the immune response, eventually resulting in a harmless, beneficial outcome in terms of allergic reactivity.

The utility of decision-making tools for the risk governance of nanotechnology is at the core of this paper. Those working in nanotechnology risk management have been prolific in creating such tools, many derived from European FP7 and H2020-funded projects. What is less clear is how such tools might assist the overarching ambition of creating a fair system of risk governance. In this paper, we reflect upon the role that tools might and should play in any system of risk governance. With many tools designed for the risk governance of this emerging technology falling into disuse, this paper provides an overview of extant tools and addresses their potential shortcomings. We also posit the need for a data readiness tool. With the EUs NMP13 family of research consortia about to report to the Commission on ways forward in terms of risk governance of this domain, this is a timely intervention on an important element of any risk governance system.

Nanosafety assessment has experienced an intense era of research during the past decades driven by a vivid interest of regulators, industry, and society. Toxicological assays based on in vitro cellular models have undergone an evolution from experimentation using nanoparticulate systems on singular epithelial cell models to employing advanced complex models more realistically mimicking the respective body barriers for analyzing their capacity to alter the immune state of exposed individuals. During this phase, a number of lessons were learned. We have thus arrived at a state where the next chapters have to be opened, pursuing the following objectives: (1) to elucidate underlying mechanisms, (2) to address effects on vulnerable groups, (3) to test material mixtures, and (4) to use realistic doses on (5) sophisticated models. Moreover, data reproducibility has become a significant demand. In this context, we studied the emerging concept of adverse outcome pathways (AOPs) from the perspective of immune activation and modulation resulting in pro-inflammatory versus tolerogenic responses. When considering the interaction of nanomaterials with biological systems, protein corona formation represents the relevant molecular initiating event (e.g., by potential alterations of nanomaterial-adsorbed proteins). Using this as an example, we illustrate how integrated experimental-computational workflows combining in vitro assays with in silico models aid in data enrichment and upon comprehensive ontology-annotated (meta)data upload to online repositories assure FAIRness (Findability, Accessibility, Interoperability, Reusability). Such digital twinning may, in future, assist in early-stage decision-making during therapeutic development, and hence, promote safe-by-design innovation in nanomedicine. Moreover, it may, in combination with in silico-based exposure-relevant dose-finding, serve for risk monitoring in particularly loaded areas, for example, workplaces, taking into account pre-existing health conditions. This article is categorized under: Toxicology and Regulatory Issues in NanomedicineToxicology of Nanomaterials.

Two training programs were developed and training materials were assembled for teaching early career researchers on deeper understanding of nanorisk governance. The here provided teaching materials are part of the deliverables of NANORIGO project, grant agreement no. 814530, call H2020-NMBP-TO-IND-2018. The training programs and the related materials are described in the document 0_D5.2 Report CS1. 0_D5.2 Report CS1: This document reports about the activities and results of the NANORIGO Case Study 1 CS1 (task T5.1) “Case study on training”, which aims (i) at testing whether the NANORIGO nanorisk governance framework (NRGF) can improve training towards safe working practices and particularly in nanorisk governance, and (ii) at demonstrating the usefulness of the NRGF for preparing the training by trainers. In this context the document comprises two training programs for early career researchers on deeper understanding and practical application of the NRGF, including working safely with nanomaterials (NM) with respect to hazard and exposure as well as to accountability and responsibility. 1_The TiO2 case - a perspective on the history and how it developed: Introduction into the TiO2 case about classification and the EFSA conclusion that E 171 can no longer be considered as safe when used as a food additive, its history, public involvement and how MNBP-13 projects see their contribution to the topic. 2_The TiO2 Case - background and instructions for group work: Introduction in the TiO2 case and group work instructions for two groups: Group 1: ”Carcinogenic Classification/Labeling Request, 2016, Inhalation Route, French ANSES”; Group2: “TiO2‘s (Food additive E171) Genotoxicity cannot be ruled out, EFSA” 3_The TiO2 case - background and instructions for role play: Introduction in the TiO2 case, summarized results of the most relevant different studies concerning TiO2 as food additive and instructions for the role play 4_TiO2 as food additive - documents in a nutshell: The most relevant statements and abstract from EFSA report - Safety assessment of titanium dioxide (E171) as a food additive 5_EFSA report - Safety assessment of titanium dioxide (E171) as a food additive: Safety assessment of titanium dioxide (E171) as a food additive from 25 March 2021, doi: 10.2903/j.efsa.2021.6585 6_ANSES CLH report Proposal for Harmonised Classification and Labelling: CLH report; Proposal for Harmonised Classification and Labelling; Based on Regulation (EC) No 1272/2008 (CLP Regulation), Annex VI, Part 2; Substance Name: Titanium dioxide; EC Number: 236-675-5; CAS Number: 13463-67-7; May 2016

ZusammenfassungDieses Positionspapier gibt eine umfassende Orientierung im Umgang und in der Betreuung von Kindern und Jugendlichen mit Diabetes in Schulen, um bestehende Informationsdefizite zu kompensieren und das Problembewusstsein für die Thematik zu erhöhen. Die steigende Zahl an Schulkindern mit Diabetes sowie die längere Aufenthaltsdauer in Schulen und schulnahen Einrichtungen machen eine offizielle Stellungnahme der Österreichischen Diabetesgesellschaft und der Österreichischen Gesellschaft für Kinder- und Jugendheilkunde notwendig, um eine standardisierte Betreuung von Kindern und Jugendlichen mit Diabetes bundesweit an allen Schulen in Österreich zu gewährleisten.

Background Pharmaceutical intervention in the CNS is hampered by the shielding function of the blood–brain barrier (BBB). To induce clinical anesthesia, general anesthetics such as isoflurane readily penetrate the BBB. Here, we investigated whether isoflurane can be utilized for therapeutic drug delivery. Methods Barrier function in primary endothelial cells was evaluated by transepithelial/transendothelial electrical resistance, and nanoscale STED and SRRF microscopy. In mice, BBB permeability was quantified by extravasation of several fluorescent tracers. Mouse models including the GL261 glioma model were evaluated by MRI, immunohistochemistry, electron microscopy, western blot, and expression analysis. Results Isoflurane enhances BBB permeability in a time- and concentration-dependent manner. We demonstrate that, mechanistically, isoflurane disturbs the organization of membrane lipid nanodomains and triggers caveolar transport in brain endothelial cells. BBB tightness re-establishes directly after termination of anesthesia, providing a defined window for drug delivery. In a therapeutic glioblastoma trial in mice, simultaneous exposure to isoflurane and cytotoxic agent improves efficacy of chemotherapy. Conclusions Combination therapy, involving isoflurane-mediated BBB permeation with drug administration has far-reaching therapeutic implications for CNS malignancies.

COVID-19, predominantly a mild disease, is associated with more severe clinical manifestation upon pulmonary involvement. Virion-laden aerosols and droplets target different anatomical sites for deposition. Compared to droplets, aerosols more readily advance into the peripheral lung. We performed in silico modeling to confirm the secondary pulmonary lobules as the primary site of disease initiation. By taking different anatomical aerosol origins into consideration and reflecting aerosols from exhalation maneuvers breathing and vocalization, the physicochemical properties of generated respiratory aerosol particles were defined upon conversion to droplet nuclei by evaporation at ambient air. To provide detailed, spatially-resolved information on particle deposition in the thoracic region of the lung, a top-down refinement approach was employed. Our study presents evidence for hot spots of aerosol deposition in lung generations beyond the terminal bronchiole, with a maximum in the secondary pulmonary lobules and a high preference to the lower lobes of both lungs. In vivo, initial chest CT anomalies, the ground glass opacities, resulting from partial alveolar filling and interstitial thickening in the secondary pulmonary lobules, are likewise localized in these lung generations, with the highest frequency in both lower lobes and in the early stage of disease. Hence, our results suggest a disease initiation right there upon inhalation of virion-laden respiratory aerosols, linking the aerosol transmission route to pathogenesis associated with higher disease burden and identifying aerosol transmission as a new independent risk factor for developing a pulmonary phase with a severe outcome.

The emergence of nanoinformatics as a key component of nanotechnology and nanosafety assessment for the prediction of engineered nanomaterials (NMs) properties, interactions, and hazards, and for grouping and read-across to reduce reliance on animal testing, has put the spotlight firmly on the need for access to high-quality, curated datasets. To date, the focus has been around what constitutes data quality and completeness, on the development of minimum reporting standards, and on the FAIR (findable, accessible, interoperable, and reusable) data principles. However, moving from the theoretical realm to practical implementation requires human intervention, which will be facilitated by the definition of clear roles and responsibilities across the complete data lifecycle and a deeper appreciation of what metadata is, and how to capture and index it. Here, we demonstrate, using specific worked case studies, how to organise the nano-community efforts to define metadata schemas, by organising the data management cycle as a joint effort of all players (data creators, analysts, curators, managers, and customers) supervised by the newly defined role of data shepherd. We propose that once researchers understand their tasks and responsibilities, they will naturally apply the available tools. Two case studies are presented (modelling of particle agglomeration for dose metrics, and consensus for NM dissolution), along with a survey of the currently implemented metadata schema in existing nanosafety databases. We conclude by offering recommendations on the steps forward and the needed workflows for metadata capture to ensure FAIR nanosafety data.

Type I hypersensitivity reactions against environmental entities, i.e. allergens, are an ever growing problem, with increasing numbers of affected individuals in Western countries. In parallel, nanotechnology has been growing during the past decades, entering more and more areas of application in everyday life. Exposure to allergens may occur in a combined way with environmental particulate matter or (nano-)particles potentially abundant at the workplace. Such combinations may thus affect workers and consumers alike. The present study conceived a methodological and mechanistic approach based on recombinant allergen–SiO2 nanoparticle conjugates and the novel hAELVi cell line as two well-defined model systems to investigate the effects of co-exposure towards allergens and nanoparticles. We applied pristine nanoparticles, allergen–nanoparticle conjugates and unbound allergen under submerged and air–liquid interface conditions. The uptake kinetics and mechanisms, cellular localization and resulting immune responses were studied in hAELVi cells, a cell line derived from human alveolar epithelium showing the characteristics of type I epithelial cells. Modification in the uptake kinetics as well as an increase in pro-inflammatory cyto-/chemokine expression was observed under submerged conditions, but much less under air–liquid interface conditions. While for mechanistic investigations the submerged culture system still proved suitable, more robust and cost-effective, these culture conditions showed considerable deviations in cellular susceptibility observed under more realistic conditions.

The immune system is professional in recognizing and responding to non-self, including nanomaterials. Immune responses by professional and nonprofessional immune cells are thus nearly inevitable upon exposure of cells and organisms to such materials. The state of research into taking the immune system into account in nanosafety studies is reviewed and three aspects in which further improvements are desirable are identified: 1) Due to technical limitations, more stringent testing for endotoxin contamination should be made. 2) Since under overdose conditions immunity shows unphysiological responses, all doses used should be justified by being equivalent to tissue-delivered doses. 3) When markers of acute inflammation or cell stress are observed, functional assays are necessary to distinguish between homeostatic fluctuation and genuine defensive or tolerogenic responses. Since immune activation can also indicate that the immune system considers a stimulus to be harmless and induces tolerance, activation markers by themselves do not necessarily imply a danger to the body. Guidelines such as these are necessary to approach the point where specific nanomaterials are classified as safe based on reliable testing strategies.

Nanotechnology is a fast growing field and already a multi-billion dollar market with numerous products available for consumers. TiO2 and SiO2 are by mass the most produced nanomaterials and, thus, of particular interest regarding their biological effects upon inhalation – especially in combination with other inhalable biomolecules such as allergens. We investigated the protein-binding capacity of these two nanomaterials and present detailed uptake profiles of protein-conjugated TiO2 or SiO2 nanoparticles (NPs) in A549 lung epithelial and THP-1 macrophage-like cells. TiO2 and SiO2 NPs (both with a hydrodynamic diameter of about 150 nm) are able to bind bovine serum albumin (BSA), green fluorescent protein (GFP) and the major birch pollen allergen Bet v 1 in substantial amounts that suggest protein monolayers around the particles. GFP-conjugated TiO2 and SiO2 NPs were taken up in A549 and THP-1 cells in a time-dependent manner as demonstrated by confocal laser scanning microscopy, flow cytometry and life cell imaging. By the aid of fluorescent proteins (GFP or fluorescently labeled Bet v 1) we found that both, the NPs and the conjugated proteins entered the cells. Using A549/THP-1 co-cultures, we showed that the majority of SiO2 NPs is taken up by THP-1 cells; however, also A549 cells take up significant amounts of particles when co-cultured with THP-1 macrophage-like cells. The here discussed data provide valuable insights into protein (allergen) delivery by NPs at the lung epithelial barrier.

The aim of this study was to develop and evaluate a clinically feasible approach to diffusion-weighted (DW) MRI of the prostate without susceptibility-induced artifacts. The proposed method relies on an undersampled multi-shot DW turbo-STEAM sequence with rotated radial trajectories and a multi-step inverse reconstruction with denoised multi-shot phase maps. The total acquisition time was below 6 min for a resolution of 1.4 × 1.4 × 3.5 mm3 and six directions at b = 600 s mm-2 . Studies of eight healthy subjects and two patients with prostate cancer were performed at 3 T employing an 18-channel body-array coil and elements of the spine coil. The method was compared with conventional DW echo-planar imaging (EPI) of the prostate. The results confirm that DW STEAM MRI avoids geometric distortions and false image intensities, which were present for both single-shot EPI (ssEPI) and readout-segmented EPI, particularly near the intestinal wall of the prostate. Quantitative accuracy of the apparent diffusion coefficient (ADC) was validated with use of a numerical phantom providing ground truth. ADC values in the central prostate gland of healthy subjects were consistent with those measured using ssEPI and with literature data. Preliminary results for patients with prostate cancer revealed a correct anatomical localization of lesions with respect to T2 -weighted MRI in both mean DW STEAM images and ADC maps. In summary, DW STEAM MRI of the prostate offers clinically relevant advantages for the diagnosis of prostate cancer compared with state-of-the-art EPI-based approaches. The method warrants extended clinical trials.

Introduction: Posttransplantation diabetes mellitus (PTDM) increases the risk of cardiovascular disease, graft loss, and decreased survival. Follow-up treatment after solid organ transplantation (SOT) needs to focus on, inter alia, maintaining balanced glucose metabolism. This study aimed to ascertain the prevalence of PTDM and describe patient characteristics in the large DPV (Diabetes Patienten Verlaufsdokumentation) pediatric diabetes database. Methods: DPV data of 71902 patients from the January 01, 1995 to January 04, 2015 period were analyzed for patients with and without cystic fibrosis (CF) after SOT (kidney, liver, heart, and lung). Multivariable analysis served to assess differences between SOT patient groups at risk for developing diabetes. Results: Out of 109 SOT patients, 51 had CF;72.5% received steroids and 62% were additionally given tacrolimus. PTDM developed in 45% of CF patients and 12% of non-CF patients. SOT patients were older at diabetes onset (mean age, 12.50 +/- 3.98 years), shorter (height z-score, -1.67 +/- 1.25), and lighter (weight z-score, -1.59 +/- 1.57) than non-SOT diabetes patients (P < 0.01). With transplantation, glycated hemoglobin (HbA1c) was significantly lower and treatment for hypertension and dyslipidemia was increased. Among SOT patients, weight and body mass index (BMI) z-scores were significantly lower in patients with CF-related diabetes (P < 0.05). Conclusions: SOT was present in 6.6% of children with diabetes, and this might aggravate the risk of cardiovascular disease in populations with already increased rates of hypertension and dyslipidemia. Dystrophy and short stature were also present, particularly in transplant recipients with CF and diabetes. Comorbidities and long-term consequences call for multidisciplinary collaboration.

Background: The assessment of carotid artery flow by neurovascular ultrasound (nvUS) may be complemented by real-time phase-contrast (RT-PC) flow MRI which apart from quantitative flow parameters offers velocity distributions across the entire vessel lumen. Methods: The feasibility and diagnostic potential of RT-PC flow MRI was evaluated in 20 healthy volunteers in comparison to conventional nvUS. RT-PC flow MRI at 40 ms temporal resolution and 0.8 mm in-plane resolution resulted in velocity maps with low phase noise and high spatiotemporal accuracy by exploiting respective advances of a recent nonlinear inverse model-based reconstruction. Peak-systolic velocities (PSV), end-diastolic velocities (EDV), flow volumes and comprehensive velocity profiles were determined in the common, internal and external carotid artery on both sides. Results: Characteristics in pulsatility and laminar flow, including individual flow abnormalities shown on nvUS, could be reproduced and visualized in detail by RT-PC flow MRI. Fig. 1 shows a representative color-coded duplex sonography with doppler spectrum (A), flow volume measurement by RT-PC flow MRI (B) and a three-dimensional representation of through-plane velocities at peak systole (C) and end diastole (D) in the internal carotid artery. PSV to EDV differences revealed good agreement between both techniques, mean PSV and EDV were significantly lower and flow volumes were higher for MRI. Conclusions: Our findings suggest that RT-PC flow MRI may add to clinical diagnostics, e.g. by alterations of dynamic velocity distributions in patients with carotid stenosis. Lower PSV and EDV values than for nvUS mainly reflect the longer MRI acquisition time which attenuates short peak velocities, while higher flow volumes benefit from a proper assessment of the true vessel lumen.

Background. In T1DM, delayed pubertal development and reduced final height are associated with inadequate metabolic control. Objective. To assess whether T1DM affects pubertal growth spurt and whether metabolic control during puberty is gender-related. Methods. Using a large multicentre database, longitudinal data from 1294 patients were analysed. Inclusion criteria: complete records of height and HbA1c from the age of seven to 16 years. Exclusion criteria: other significant chronic diseases and medications, T1DM duration less than three months, and initial BMI 97th percentile. Results. Growth velocity (GV) was impaired with a significant reduction of peak GV by 1.2 cm in boys. HbA1c increase during male puberty was lower except for a period of 1.5 years. The highest HbA1c increase in boys coincided with maximum growth spurt. In girls, the highest HbA1c increase was observed during late puberty. Even though there is impaired GV, both sexes reach a height at 16 years of age which corresponds to the background population height. Conclusion. Worsening of metabolic control is sex-discordant and associated with gender-specific alterations of GV. However, the vast majority of boys and girls with T1DM seems to reach normal height at the age of 16 years.

Lipoprotein-associated phospholipase A2 (Lp-PLA2) was identified as a strong predictor for cardiovascular events. Furthermore, it is highly associated with obesity. The role of Lp-PLA2 in diabetes mellitus is controversial and analyses, especially in adolescents with type 2 diabetes (T2D), are missing. Therefore, we compared Lp-PLA2 activity between two obese age-, sex-, and BMI-matched cohorts of adolescents with and without T2D. Relationships between Lp-PLA2 activity and age, BMI, hemoglobin A1c, lipids, and adipokines were evaluated. Lp-PLA2 activity was analyzed in serum of 72 obese adolescents without T2D (mean age 15.2 ± 1.6 years) and in 65 obese adolescents with T2D (mean age 15.5 ± 1.8 years). Clinical data were obtained from the Diabetes-Patienten-Verlaufsdokumentation (DPV) registry. Surprisingly, obese adolescents with T2D had lower levels of Lp-PLA2 activity than obese children without T2D (160.2 ± 45.0 versus 180.9 ± 35.6 nmol/min/ml, p = 0.003), but this decrease could only be detected in male (158.8 ± 45.3 versus 190.8 ± 31.3 nmol/min/ml, p

OBJECTIVE: The aim of this study was to develop a rapid diffusion-weighted (DW) magnetic resonance imaging (MRI) technique for whole-brain studies without susceptibility artifacts and measuring times below 3 minutes. MATERIALS AND METHODS: The proposed method combines a DW spin-echo module with a single-shot stimulated echo acquisition mode MRI sequence. Previous deficiencies in image quality due to limited signal-to-noise ratio are compensated for (1) by radial undersampling to enhance the flip angle and thus the signal strength of stimulated echoes; (2) by defining the image reconstruction as a nonlinear inverse problem, which is solved by the iteratively regularized Gauss-Newton method; and (3) by denoising with use of a modified nonlocal means filter. The method was implemented on a 3 T MRI system (64-channel head coil, 80 mT · m gradients) and evaluated for 10 healthy subjects and 2 patients with an ischemic lesion and epidermoid cyst, respectively. RESULTS: High-quality mean DW images of the entire brain were obtained by acquiring 1 non-DW image and 6 DW images with different diffusion directions at b = 1000 s · mm. The achievable resolution for a total measuring time of 84 seconds was 1.5 mm in plane with a section thickness of 4 mm (55 sections). A measuring time of 168 seconds allowed for an in-plane resolution of 1.25 mm and a section thickness of 3 mm (54 sections). Apparent diffusion coefficient values were in agreement with literature data. CONCLUSIONS: The proposed method for DW MRI offers immunity against susceptibility problems, high spatial resolution, adequate signal-to-noise ratio and clinically feasible scan times of less than 3 minutes for whole-brain studies. More extended clinical trials require accelerated computation and online reconstruction.

Background/Aims: The ATP12A gene codes for a non-gastric H+/K+ ATPase, which is expressed in a wide variety of tissues. The aim of this study was to test for the molecular and functional expression of the non-gastric H+/K+ ATPase ATP12A/ATP1AL1 in unstimulated and butyrate-stimulated (1 and 10 mM) human myelomonocytic HL-60 cells, to unravel its potential role as putative apoptosis-counteracting ion transporter as well as to test for the effect of the H+/K+ ATPase inhibitor SCH28080 in apoptosis. Methods: Real-time reverse-transcription PCR (qRT-PCR) was used for amplification and cloning of ATP12A transcripts and to assess transcriptional regulation. BCECF microfluorimetry was used to assess changes of intracellular pH (pHi) after acute intracellular acid load (NH4Cl prepulsing). Mean cell volumes (MCV) and MCV-recovery after osmotic cell shrinkage (Regulatory Volume Increase, RVI) were assessed by Coulter counting. Flow-cytometry was used to measure MCV (Coulter principle), to assess apoptosis (phosphatidylserine exposure to the outer leaflet of the cell membrane, caspase activity, 7AAD staining) and differentiation (CD86 expression). Results: We found by RT-PCR, intracellular pH measurements, MCV measurements and flow cytometry that ATP12A is expressed in human myelomonocytic HL-60 cells. Treatment of HL-60 cells with 1 mM butyrate leads to monocyte-directed differentiation whereas higher concentrations (10 mM) induce apoptosis as assessed by flow-cytometric determination of CD86 expression, caspase activity, phosphatidylserine exposure on the outer leaflet of the cell membrane and MCV measurements. Transcriptional up-regulation of ATP12A and CD86 is evident in 1 mM butyrate-treated HL-60 cells. The H+/K+ ATPase inhibitor SCH28080 (100 µM) diminishes K+-dependent pHi recovery after intracellular acid load and blocks RVI after osmotic cell shrinkage. After seeding, HL-60 cells increase their MCV within the first 24 h in culture, and subsequently decrease it over the course of the next 48 h. This effect can be observed in the overall- and non-apoptotic fraction of both untreated and 1 mM butyrate-treated HL-60 cells, but not in 1 mM butyrate-stimulated phosphatidylserine-positive cells. These cells do not shrink from 24 h to 72 h and have finally a higher MCV than untreated cells unless they are exposed to SCH28080. 10 mM butyrate induces apoptosis within 24 h. Conclusion: In summary we show that in HL-60 cells ATP12A is a functionally active H+/K+ ATPase that may counteract events during early apoptosis like intracellular acidosis, loss of intracellular K+ ions and apoptotic volume decrease. Its expression and/or susceptibility to the H+/K+ ATPase inhibitor SCH28080 becomes most evident in cells exposing phosphatidylserine on the outer leaflet of the cell membrane and therefore during early apoptosis.

This position statement is based on the current evidence available on the safety and benefits of continuous subcutaneous insulin pump therapy (CSII) in diabetes with an emphasis on the effects of CSII on glycemic control, hypoglycaemia rates, occurrence of ketoacidosis, quality of life and the use of insulin pump therapy in pregnancy. The current article represents the recommendations of the Austrian Diabetes Association for the clinical praxis of insulin pump treatment in children, adolescents and adults.

This position statement represents the recommendations of the Austrian Diabetes Association regarding the clinical diagnostic and therapeutic application, safety and benefits of continuous subcutaneous glucose monitoring systems in patients with diabetes mellitus, based on current evidence.

ObjectiveTo identify risk factors for the development and progression of untreated persistent microalbuminuria in children and adolescents with type 1 diabetes.Design and methodsA total number of 683 children and adolescents with type 1 diabetes recruited from the prospective nationwide German and Austrian diabetes survey (DPV) were included in the analysis. Inclusion criteria were onset of type 1 diabetes under the age of 11 years, diabetes duration of more than 1 year and continuous follow-up over 5 years with at least two documented urine analyses per year. Subjects treated with angiotensin-converting enzyme inhibitors were excluded. Risk factors such as sex, body mass index SDS, diabetes duration, HbA1c, total cholesterol, HDL-cholesterol, LDL-cholesterol, systolic and diastolic blood pressure, and immigrant status were analysed by logistic regression.ResultsAt baseline (age 10.5±0.1 years, diabetes duration 4.6±2.4 years and HbA1c 7.4±1.1%), 75.6% of children had normoalbuminuria, 15.7% had intermittent microalbuminuria, 8.6% had persistent microalbuminuria and 0.1% had macroalbuminuria. After a follow-up of 5 years, 59.4% of adolescents continued to have normoalbuminuria, 18.4% had progression, 15.2% had regression of microalbuminuria, and in 6.9% of the subjects, microalbuminuria remained unchanged. We found significant associations between persistent microalbuminuria at baseline and during each year of follow-up (PP=0.009 andP=0.009).ConclusionsThe survey in a real-world setting shows that diabetes duration and immigrant status are risk factors for the development and progression of untreated microalbuminuria in children and adolescents with type 1 diabetes.

OBJECTIVE To investigate the temporal trend of metabolic control and potential predictors in German and Austrian children and adolescents with type 1 diabetes. RESEARCH DESIGN AND METHODS This study is based on a large, multicenter database for prospective longitudinal documentation of diabetes care in Germany and Austria. Data from 30,708 patients documented in 305 diabetes centers between 1995 and 2009 were analyzed. Generalized linear mixed regression models were used to adjust trend analysis for relevant confounders. RESULTS Unadjusted mean HbA1c decreased from 8.7 ± 1.8% in 1995 to 8.1 ± 1.5% in 2009. In multiple regression analysis, treatment year, age, sex, diabetes duration, migration background, BMI-SDS, and daily insulin dose were significant predictors of metabolic control (P < 0.001). After multiple adjustment, mean HbA1c decreased significantly by 0.038% per year (95% CI 0.032–0.043%), average odds ratio (OR) per year for HbA1c >7.5% (>9.0%) was 0.969 (95% CI 0.961–0.977) (0.948, 95% CI 0.941–0.956). Intensified insulin regimen was associated with lower frequency of poor metabolic control (HbA1c >9%; P = 0.005) but not with average HbA1c (P = 0.797). Rate of severe hypoglycemia and hypoglycemic coma decreased significantly (relative risk [RR] per year 0.948, 95% CI 0.918–0.979; RR 0.917, 95% CI 0.885–0.950) over the study period. Diabetic ketoacidosis rate showed no significant variation over time. CONCLUSIONS This study showed a significant improvement in metabolic control in children and adolescents with type 1 diabetes during the past decade and a simultaneous decrease in hypoglycemic events. The improvement was not completely explained by changes in the mode of insulin treatment. Other factors such as improved patient education may have accounted for the observed trend.

Objective: We investigated influences of a 12-h fast, age, gender, body mass index (BMI), hemoglobin A1c (HbA1c) on total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) to provide reference percentiles for TC, LDL-C, and HDL-C of patients with good diabetes control (HbA1c

The fibers of the pyramidal tract (PT) that connect the precentral and postcentral gyrus with the spinal cord are organized in a topological manner. However, their exact arrangement and orientation in the internal capsule is still a mat- ter of debate. Here, we applied magnetic resonance diffusion tensor imaging and tract tracing techniques to determine and compare the pyramidal fibers from the primary motor and somatosensory system in humans and rhesus monkeys in vivo. The results demonstrate that the pyramidal systems of the human and monkey brain differ in their orientation at the level of the internal capsule, whereas track orientations in the gyrus and brainstem appear similar. In the monkey internal cap- sule the somatotopic arrangement of PT fibers from the upper and lower extremities form an angle with the left-right axis of 146° ± 23° (n=4). Thus, fibers from lateral areas of the gyrus are located more anterior-medial to fibers from medial cortical areas. In contrast, in humans the angle is only 50° ± 16° (n=9) yielding a more anterior-lateral orientation of PT fibers along the short axis of the internal capsule. These species dependencies may possibly be due to structural con- straints of the smaller and differently shaped monkey brain.

Introduction Proton magnetic resonance spectroscopy (MRS) and diffusion tensor imaging (DTI) yield different parameters for characterizing the evolution of a demyelinating white matter disease. The purpose was to elucidate biochemical and microstructural changes in Balo’s concentric sclerosis lesions and to correlate the findings with the clinical course. Methods Localized short-echo time MRS and DTI were performed over 6 years in a left occipital lesion of a female patient (age at onset 13.8 years) with Balo’s concentric sclerosis. A right homonym hemianopsia persisted. Results Metabolite patterns were in line with initial active demyelination followed by gliosis and partial recovery of neuroaxonal metabolites. Fractional anisotropy and mean diffusivity of tissue water remained severely altered. Fiber tracking confirmed a disruption in the geniculo-calcarine tract as well as involvement of the corpus callosum. Conclusion MRS and DTI depict complementary parameters, but DTI seems to correlate better with clinical symptoms.

The study of complex fiber systems in relation to the cognitive abilities of humans is a long-standing challenge for neuroscientists. With the development of diffusion tensor imaging (DTI) it is now possible to visualize large fiber bundles non-invasively. The existing knowledge of the white matter architecture largely stems from either lesion studies of human patients or, in more detail, tracer injection studies of non-human primates. Hence, it seems mandatory to com- pare DTI results with histochemical findings obtained for the same species. Using a geometrically undistorted DTI tech- nique and fiber tractography, we examined the fiber anatomy of the macaque brain in vivo and related the results to fiber pathways previously identified in monkeys with conventional tract tracing. The approach identified multiple fiber tracts including the main association and projection pathways as well as fibers of the limbic system, commissural system, optic system, and cerebellar system. In conclusion, in vivo fiber tractography based on current-state DTI allows for a compre- hensive analysis of major fiber pathways in the intact macaque brain.

DesignThe purpose of this study was to generate insulin dose (ID) percentiles for children and adolescents with type 1 diabetes mellitus (DM1) having the opportunity to assess this important parameter in relation to age and sex.MethodsDaily IDs per weight (ID/kg) were recorded in 22 177 patients with DM1 (3–25 years of age, DM1 duration of more than 2 years, 48% female) and ID percentiles (ID-Perc) were created statistically. The ID-Perc were compared between male and female, and between multiple insulin injection therapy (MIT) and continuous s.c. insulin infusion (CSII). A multivariate regression analysis was performed for ID in the third year of DM1 with ID/kg, body weight, age, gender, and insulin delivery regimen as variables.ResultsThe 50th ID-Perc (P50) varied among 0.67 IU/kg (age 3 years), 0.93 IU/kg (13 years), and 0.70 IU/kg (23 years) increasing from early childhood to adolescence and decreasing toward adulthood. Highest P50 ID was found at 12 years in females (0.94 IU/kg) and at 14 years in males (0.92 IU/kg). Using ICT, the ID was significantly higher compared with CSII (P50: 0.94 IU/kg versus 0.79 IU/kg at 13 years). In multivariate regression analysis, ID was significantly (P>0.001) associated with age, gender, and insulin delivery regime.ConclusionThe ID-Perc were significantly different during various periods of childhood and were influenced by gender, body weight, and insulin injection regimes. Therefore, the presented data 1) provide evidence to interpret individual ID in children and adolescents with DM1 and 2) more specifically identify children with unusually high (insulin resistance and non-compliance) or low (MODY and persistent remission) insulin requirement.

OBJECTIVE— In type 1 diabetes, plantar fascia, a collagen-rich tissue, is susceptible to glycation and oxidation. Paraoxonase-1 (PON1) is an HDL-bound antioxidant enzyme. PON1 polymorphisms have been associated with susceptibility to macro- and microvascular complications. We investigated the relationship between plantar fascia thickness (PFT) and PON1 gene variants, p.Leu54Met, p.Gln192Arg, and c.-107C>T, in type 1 diabetes. RESEARCH DESIGN AND METHODS—This was a cross-sectional study of 331 adolescents with type 1 diabetes (162 male and 169 female). PFT was assessed by ultrasound, PON1 was assessed by genotyping with PCR and restriction fragment–length polymorphism, and serum PON1 activity was assessed by rates of hydrolysis of paraoxon and phenylacetate. RESULTS—Median (interquartile range) age was 15.4 (13.5–17.3) years, and diabetes duration was 7.6 (4.9–10.6) years. The distribution of p.Leu54Met genotypes was LL 135 (40.8%), ML 149 (45%), and MM 47 (14.2%). PFT was abnormal (>1.7 mm) in 159 adolescents (48%). In multivariate analysis, predictors of abnormal PFT were ML/LL versus MM p.Leu54Met polymorphism (odds ratio 3.84 [95% CI 1.49–9.82], P = 0.005); BMI (percentile) (1.02 [1.01–1.03], P = 0.007); systolic blood pressure (percentile) (1.01 [1.00–1.02], P = 0.03); and male sex (3.29 [1.98–5.46], P < 0.001). CONCLUSIONS—Thickening of the plantar aponeurosis occurs predominantly in overweight and male adolescents with type 1 diabetes. The MM genotype at PON1 p.Leu54Met is associated with a reduced risk of abnormal PFT.