Vallo Tillmann

The Dynamics of the Human Infant Gut Microbiome in Development and in Progression toward Type 1 Diabetes

Colonization of the fetal and infant gut microbiome results in dynamic changes in diversity, which can impact disease susceptibility. To examine the relationship between human gut microbiome dynamics throughout infancy and type 1 diabetes (T1D), we examined a cohort of 33 infants genetically predisposed to T1D. Modeling trajectories of microbial abundances through infancy revealed a subset of microbial relationships shared across most subjects. Although strain composition of a given species was highly variable between individuals, it was stable within individuals throughout infancy. Metabolic composition and metabolic pathway abundance remained constant across time. A marked drop in alpha-diversity was observed in T1D progressors in the time window between seroconversion and T1D diagnosis, accompanied by spikes in inflammation-favoring organisms, gene functions, and serum and stool metabolites. This work identifies trends in the development of the human infant gut microbiome along with specific alterations that precede T1D onset and distinguish T1D progressors from nonprogressors.

Serum leptin through childhood and adolescence

Leptin is the protein product of the recently cloned ob gene, that has been implicated in the control of body weight and thermogenesis, but also independently stimulates the reproductive axis. As major changes in body composition and gonadal function occur during human adolescence, we have assessed serum leptin concentration through childhood.

Biochemical tests in the diagnosis of childhood growth hormone deficiency

GH stimulation tests are widely used in the diagnosis of GH deficiency (GHD), although they are associated with a high false positive rate. We have examined, therefore, the performance of other tests of the GH axis [urinary GH excretion, serum insulin-like growth factor I(IGF-I), and IGF-binding protein-3 (IGFBP-3) levels] compared with GH stimulation tests in identifying children defined clinically as GH deficient. Group I comprised 60 children (mean age, 10.3 +/- 4.8 yr) whose diagnosis of GHD was based on a medical history indicative of pituitary dysfunction (n = 43) or on the typical phenotypic features and appropriate auxological characteristics of isolated GHD (n = 17). Group II comprised 110 short children (mean age, 9.8 +/- 4 yr) in whom GHD was not suspected, but needed exclusion. The best sensitivity for a single GH test was 85% at a peak GH cut-off level of 10 ng/mL, whereas the best specificity was 92% at 5 ng/mL. The sensitivities of IGF-I, IGFBP-3, and urinary GH, using a cut-off of -2 SD score were poor at 34%, 22%, and 25%, respectively, with specificities of 72%, 92%, and 76% respectively. Only 2 of 21 pubertal children in group I and none of the 27 subjects with radiation-induced GHD had an IGFBP-3 SD score less than -1.5. We devised a scoring system based on the positive predictive value of each test, incorporating data from the GH test and the IGF-I and IGFBP-3 levels. A specificity of 94% could be achieved with a score of 10 or more (maximum 17) (sensitivity 34%). The latter could not be improved above 81% with a score of 5 points or more (specificity, 69%). A high score was, therefore, highly indicative of GHD, but was achieved by few patients. A normal IGFBP-3 level, however, did not exclude GHD, particularly in patients with radiation-induced GHD and those in puberty. A GH test with a peak level more than 10 ng/mL was the most useful single investigation to exclude a diagnosis of GHD.

Green areas around homes reduce atopic sensitization in children

Western lifestyle is associated with high prevalence of allergy, asthma and other chronic inflammatory disorders. To explain this association, we tested the ‘biodiversity hypothesis’, which posits that reduced contact of children with environmental biodiversity, including environmental microbiota in natural habitats, has adverse consequences on the assembly of human commensal microbiota and its contribution to immune tolerance.

Arterial stiffness, carotid artery intima-media thickness and plasma myeloperoxidase level in children with type 1 diabetes

OBJECTIVE The present study investigated the functional-structural changes of the arteries along with a new biochemical marker of atherosclerosis, plasma myeloperoxidase level, in children with type 1 diabetes (T1DM). METHODS We studied 30 children with T1DM, aged 4.7-18.6 years (mean T1DM duration 5.4+/-3.4 years, mean HbA(1c) 9.8%) and 30 healthy subjects, matched by sex, age and body mass index. Fasting blood samples were obtained for myeloperoxidase (MPO). Ultrasonography and pulse wave analysis were used to measure carotid intima-media thickness (IMT), augmentation index corrected to a heart rate of 75 (AIx@75) and pulse wave velocity (PWV). RESULTS Children with diabetes had significantly higher plasma MPO levels (p=0.006), increased AIx@75 (p=0.02), IMT (p=0.005) and IMT standard deviation scores (IMT SDS) (p=0.02), compared to the control group. IMT SDS correlated positively with HbA(1c) (r=0.39, p=0.05). PWV, adjusted for age and mean arterial blood pressure, correlated with diabetes duration (r=0.49, p=0.02). CONCLUSIONS Children with T1DM have substantially elevated plasma levels of myeloperoxidase as well as atherosclerosis-related structural and functional changes of the arterial wall.

Constitutional delay in growth and puberty (CDGP) is associated with hypoleptinaemia

Serum leptin concentrations are higher in early adolescence compared with childhood and may play a facilitatory role in pubertal development. Constitutional delay in growth and puberty (CDGP) is a disorder of the tempo of physical maturation and may be associated with relative hypoleptinaemia. We have therefore compared serum leptin concentrations in normal boys with those in boys exhibiting constitutional delay of growth, controlling for pubertal status, age and body mass index (BMI).

Increased FOXP3 expression in small-bowel mucosa of children with coeliac disease and type I diabetes mellitus

Objective. To determine whether the expression of FOXP3 is changed in small-bowel mucosa in coeliac disease (CD). Material and methods. The study comprised 52 patients (mean age 8.01±6.14 years) who had undergone small-bowel biopsies. CD only was diagnosed in 16 patients, and CD with type I diabetes mellitus (T1D) in 7. These 23 patients and 4 others without CD had partial or subtotal villous atrophy (PVA, SVA). Twenty-five persons without CD had normal mucosa. The transcription level of the FOXP3 gene (Hs00203958_m1) was evaluated in biopsy samples (small bowel) using TaqMan gene expression assays. FOXP3 protein in mucosal cells was evaluated with mouse anti-human FOXP3 antibodies and CD25+, and CD4+ T cells were evaluated by mouse monoclonal antibodies. Results. Expression of FOXP3 mRNA was higher in both PVA and SVA compared to normal mucosa (p=0.007). Patients with CD and T1D had higher expression of FOXP3 mRNA than patients with CD alone (p=0.02). The number of FOXP3+ cells in intestinal mucosa was higher in patients with CD, especially those with coexisting T1D, than in those with normal mucosa (p=0.01). The results of double staining showed that, among all positive cells, FOXP3 expression alone was revealed in 25.6% of the cells, CD25 positivity in 18% and both markers simultaneously were found in 56.5% of lymphocytes (p=0.03). Double staining for CD4 and FOXP3 showed that 87.5% of cells were CD4+, 2.8% were FOXP3+ and 9.7% of cells simultaneously expressed the CD4 and FOXP3 markers. Conclusions. A more pronounced expression of FOXP3 mRNA and also the number of FOXP3+ cells (with simultaneous expression of CD25 and CD4 markers) were found in the small-bowel biopsy specimens obtained from children with CD, particularly those with coexisting T1D, compared with the FOXP3 expression in normal mucosa.

Elevated plasma adiponectin and decreased plasma homocysteine and asymmetric dimethylarginine in children with type 1 diabetes

Objective. Type 1 diabetes has a bad prognosis concerning the pathogenesis of cardiovascular diseases (CVD). The purpose of this study was to evaluate different possible new risk indices for CVD in children with type 1 diabetes. Material and methods. The present study included 30 children with diabetes (mean HbA1C 9.8%), aged between 4.7 and 18.6 years and with no clinical evidence of vascular complications, and 30 healthy subjects matched by sex, age and body mass index. Blood pressure was measured and blood samples were obtained for lipid profile, creatinine, glucose, high sensitive C‐reactive protein (hsCRP), intercellular adhesion molecule‐1 (ICAM‐1), asymmetric dimethylarginine (ADMA), adiponectin and homocysteine. Results. Children with diabetes had significantly higher blood pressure, plasma hsCRP, ICAM‐1, adiponectin levels and lower homocysteine, ADMA concentrations than their control subjects. In multivariate regression analysis, the best predictors for systolic blood pressure were diabetes group, plasma homocysteine concentration and BMI (Adj R2 = 0.38, p<0.0001), and for diastolic blood pressure diabetes group and triglycerides level (Adj R2 = 0.27, p<0.0001). Conclusions. Children with diabetes, in view of their higher future risk of CVD, are characterized by a higher concentration of protective adiponectin and paradoxically lower blood concentrations of some other possible risk markers of atherosclerosis, i.e. ADMA and homocysteine compared to healthy children.

The relationship between stature, growth, and short-term changes in height and weight in normal prepubertal children

Human growth is a nonlinear process with marked variation in growth rate during the short-term. It is not known how long-term height gain or stature is influenced by short-term changes in height and weight. This study has addressed these issues by using thrice weekly height and weight measurements during 1 year in 43 normal prepubertal children (aged 5.7-7.7 y) to construct individual height and weight velocity curves by regression analysis. The former were comprised of 3 to 6 growth spurts separated by stasis, whereas the latter were characterized by 2 to 5 periods of weight gain separated by periods of weight loss. Stepwise regression analysis to determine characteristics of these curves that influence stature and growth showed that height SD score was correlated to the mean absolute weight velocity amplitude (+), the mean lenght of height velocity peaks (-), and the number of periods of weight gain (-) (r2 = 38%). In contrast, change in height SD score (Δheight SD score) was correlated to the number (+) and mean amplitude (+) of the periods of weight gain and the mean height velocity peak amplitude (+) (r2 = 44%). Examination of changes in height relative to weight during the year in the whole group revealed that height increased relative to weight in autumn and spring, whereas the reverse occurred during the winter months. We conclude that 1) both height and weight velocities during 1 year show a biphasic pattern, 2) there is seasonal variation in the short-term change in height relative to weight, and 3) prepubertal stature and the amount grown through the year are related to short-term changes in height and weight. Our data indicate that large but infrequent changes in weight with growth spurts of short duration are found in tall children. Good growth during the year was related to large but frequent gains in weight and large individual spurts in height.

Elevated Serum IL-6, IL-8, MCP-1, CRP, and IFN-γ Levels in 10- to 11-Year-Old Boys with Increased BMI

Background/Aims: Many inflammation parameters are associated with obesity, but few comparable data are found in youth. This study aims to characterize the differences in serum levels of 13 biochemical inflammatory markers between boys with increased BMI and boys with normal BMI, and examine the relationships between inflammation markers, skinfold thicknesses, and body composition. Participants/Methods: The participants were 38 boys (BMI above 85th percentile) and 38 boys (normal BMI) at the age of 10–11 years. Measurements included BMI, 9 skinfold thicknesses, waist and hip circumferences, and total body and trunk fat mass and percentage as indices of obesity, fasting insulin, glucose, and serum concentrations of IL-2, IL-4, IL-6, IL-8, IL-10, VEGF, IFN-γ, TNF-α, IL-1α, IL-1β, monocyte chemoattractant protein-1 (MCP-1), epidermal growth factor, and CRP. Results: Overweight boys (OWB) were taller and more frequently in puberty than normal-weight boys (NWB). Skinfold thicknesses and body composition parameters were higher in OWB. They had significantly higher serum IL-6, IL-8, IFN-γ, MCP-1, and CRP values compared to NWB. Conclusions: Six of 13 measured biochemical markers were significantly increased in OWB, indicating that many low-grade inflammatory processes are already involved in the development of obesity in childhood.