Massimo Pellegrini

Gpc3 expression correlates with the phenotype of the Simpson‐Golabi‐Behmel syndrome

Interest in glypican‐3 (GPC3), a member of the glypican‐related integral membrane heparan sulfate proteoglycans (GRIPS) family, has increased with the finding that it is mutated in the Simpson‐Golabi‐Behmel overgrowth syndrome (Pilia et al. [1996] Nat. Genet. 12:241–247). The working model suggested that the membrane‐bound protein acts locally to limit tissue and organ growth and that it may function by interacting with insulin‐like growth factor 2 (IGF2) to limit its local effective level. Here we have tested two predictions of the model. In situ hybridization with the mouse gene cDNA was used to study the expression pattern during embryonic and fetal development. In agreement with predictions, the gene is expressed in precisely the organs that overgrow in its absence; and the patterns of expression of Gpc3 and those reported for Igf2 are strictly correlated. Dev. Dyn. 1998; 213:431–439.

Emx2 developmental expression in the primordia of the reproductive and excretory systems.

Abstract The development of the urogenital system has always attracted many investigators owing to the peculiar aspects of the embryology of the reproductive and excretory organs and to the high number of congenital anomalies related to these structures. It is remarkable because of the common origin of the kidneys, gonads, and genital tracts from the intermediate mesoderm and because differentiation of these organs involves extensive mesenchyme to epithelium transition. Our knowledge about the molecular mechanisms controlling the differentiation of these diverse structures from the same precursor has taken advantage of gene expression data and gene-targeting experiments using genes with a specific expression pattern in the urogenital system. A more detailed function in kidney development has been postulated for transcription factors such as WT-1, Pax-2 or other molecules such as glial cell line-derived neurotrophic factor (GDNF), Wnt-4, c-ret. In the present work we have described the expression pattern of the homeobox-containing gene Emx2 during the development of the urogenital system in mouse embryos. We have found that Emx2 is expressed in the early primordia of the organs that will form the excretory and reproductive systems. In particular we have found that Emx2 is expressed in the epithelial components of pronephros and mesonephros, in Wolffian and Müllerian ducts, in the ureteric buds with their branches and in the early epithelial structures derived from metanephrogenic mesenchyme. Emx2 is also intensely expressed in the ”bipotential” or ”indifferent” gonads and ovaries. These data and the recent finding that Emx2 homozygous mutant mice die soon after birth because of the absence of kidneys indicate an essential role of Emx2 in the morphogenesis of the urogenital system.

Genotype/phenotype correlations of males affected by Simpson-Golabi-Behmel syndrome with GPC3 gene mutations: patient report and review of the literature.

Simpson-Golabi-Behmel syndrome (SGBS) is an X-linked overgrowth syndrome with associated visceral and skeletal anomalies. Deletions or point mutations involving the glypican-3 (GPC3) gene at Xq26 are associated with a relatively milder form of this disorder (SGBS1). GPC3 encodes a putative extracellular proteoglycan, glypican-3, that is inferred to play an important role in growth control in embryonic mesodermal tissues in which it is selectively expressed. It appears to form a complex with insulin-like growth factor-II (IGF-II), and might thereby modulate IGF-II action. We reviewed the clinical findings of all published patients with SGBS1 with GPC3 mutations to confirm the clinical specificity for the SGBS1 phenotype. Moreover, we report on a new patient with a GPC3 deletion and IGF-II evaluation.

Scapula development is governed by genetic interactions of Pbx1 with its family members and with Emx2 via their cooperative control of Alx1

The genetic pathways underlying shoulder blade development are largely unknown, as gene networks controlling limb morphogenesis have limited influence on scapula formation. Analysis of mouse mutants for Pbx and Emx2 genes has suggested their potential roles in girdle development. In this study, by generating compound mutant mice, we examined the genetic control of scapula development by Pbx genes and their functional relationship with Emx2. Analyses of Pbx and Pbx1;Emx2 compound mutants revealed that Pbx genes share overlapping functions in shoulder development and that Pbx1 genetically interacts with Emx2 in this process. Here, we provide a biochemical basis for Pbx1;Emx2 genetic interaction by showing that Pbx1 and Emx2 can bind specific DNA sequences as heterodimers. Moreover, the expression of genes crucial for scapula development is altered in these mutants, indicating that Pbx genes act upstream of essential pathways for scapula formation. In particular, expression of Alx1, an effector of scapula blade patterning, is absent in all compound mutants. We demonstrate that Pbx1 and Emx2 bind in vivo to a conserved sequence upstream of Alx1 and cooperatively activate its transcription via this potential regulatory element. Our results establish an essential role for Pbx1 in genetic interactions with its family members and with Emx2 and delineate novel regulatory networks in shoulder girdle development.

Measured physical activity in anorexia nervosa: features and treatment outcome.

OBJECTIVE To assess the role of measured physical activity (PA) in anorexia nervosa treatment outcome, and to compare the PA of patients with anorexia nervosa with age-matched controls. METHOD PA was assessed by means of Sense Wear Armband before and after a cognitive-behavioral inpatient treatment in 53 consecutive females with anorexia nervosa, and in 53 healthy age-matched controls. RESULTS At baseline, patients with anorexia nervosa exhibited a higher duration of moderate-vigorous PA (MVPA≥3 Metabolic Equivalent Tasks (METs)) than controls (t = 2.91; p = .004). Dropouts had higher duration (sec) and expenditure (kcal·day(-1)) of MVPA than completers. At the end of treatment, completers had a higher number of daily steps, MVPA duration, and expenditure than controls. However, PA was not correlated to eating disorder psychopathology either before or after treatment. DISCUSSION PA is higher in patients with anorexia nervosa than age-matched controls both before and after treatment, and is associated with treatment dropout.

Short-term multidisciplinary non-pharmacological intervention is effective in reducing liver fat content assessed non-invasively in patients with nonalcoholic fatty liver disease (NAFLD)

BACKGROUND Non-alcoholic fatty liver disease (NAFLD) ranges from simple steatosis to steatohepatitis, and cirrhosis in patients with alcohol intake less than 20 g/day, and is usually associated with insulin resistance (IR). AIM Given that no drugs are specifically approved for NAFLD, we tested the efficacy of a non-pharmacological multidisciplinary intervention based on a personalized diet, physical activity and behavior therapy. METHODS In this open non-randomized study, personalized diet, physical exercise and behaviour therapy for 3 months were prescribed in 12 consecutive patients with NAFLD. Lifestyle, including total caloric intake, physical activity and resting energy expenditure was monitored by a SenseWear Armband. Insulin Resistance (IR) was measured by HOMA and oral glucose insulin sensitivity tests (OGIS); fat liver content was estimated by two different semi-quantitative scores and by the Doppler Power Index (DPI). RESULTS Data show that the multidisciplinary intervention produced a significant reduction of total caloric intake, a 8% reduction in body weight, a modest increase in daily physical activity, a significant (P<0.001) reduction of aminotransferases and a decrease of total hepatic fat content. CONCLUSIONS A 3-month multidisciplinary intervention inducing at least 8% of weight loss, improves liver tests and decreases liver fat content.

Body composition, eating disorder psychopathology, and psychological distress in anorexia nervosa: a longitudinal study

BACKGROUND Although the effect of immediate weight restoration on body composition and body fat distribution has previously been studied in anorexia nervosa (AN), its influence in women with AN on eating disorder psychopathology and psychological distress has not previously been investigated to our knowledge. OBJECTIVES We assessed body composition and fat mass distribution before and after body weight restoration and investigated any relation between changes in body fat patterns of patients with AN treated in a specialist inpatient unit and their eating disorder and psychological distress features. DESIGN Body composition was measured by using dual-energy X-ray absorptiometry in 50 female, adult patients with AN before and after complete weight restoration [body mass index (BMI; in kg/m²) ≥18.5] and 100 healthy control subjects matched by age and posttreatment BMI of study group participants. Eating disorder psychopathology and psychological distress were assessed in the AN group before and after weight restoration by using the Eating Disorder Examination interview and the Global Severity Index of the Brief Symptom Inventory (BSI-GSI), respectively. RESULTS After the achievement of complete weight restoration, patients with AN had higher trunk (P < 0.001), android (P < 0.001), and gynoid (P < 0.001) fat masses and lower arm (P < 0.001) and leg (P = 0.001) fat masses with respect to control subjects. No relation was shown between body-composition variables and eating disorder psychopathology in the AN group, and the only significant predictor of change in BSI-GSI was the baseline BSI-GSI score. CONCLUSION The normalization of body weight in patients with AN is associated with a preferential distribution of body fat in central regions, which does not, however, seem to influence either eating disorder psychopathology or psychological distress scores.

Comparison between dual-energy X-ray absorptiometry and skinfolds thickness in assessing body fat in anorexia nervosa before and after weight restoration

BACKGROUND & AIMS The aim of the study was to evaluate the correspondence between body fat mass composition (percentage) measured with dual-energy X-ray absorptiometry (DXA) and estimated by means of skinfold thicknesses (ST) measurement in patients with anorexia nervosa (AN), before and after weight gain. METHODS Percentage body fat (%BF) was measured with DXA and estimated by ST measurements using Siri, Brozek, and Heyward equations in 27 adult patients with AN before and after weight gain (pre- and post-treatment) achieved with inpatient treatment and in 42 healthy age-matched controls. RESULTS Due to Lohman criteria and Bland Altman plot there is no correspondence between the %BF measured with DXA and the %BF estimated by predictive equations based on ST measurements in patients with AN before and after weight gain, with the exception of Brozek equation which showed a mild agreement in pre-treatment AN. However, a correspondence was observed between the two procedures in healthy controls. CONCLUSIONS Our data supporting the use of ST measurements do not appear to be an alternative to DXA in estimating body fat percentage, before and after weight gain in patients with AN.

Developmental overfeeding alters hypothalamic neuropeptide mRNA levels and response to a high-fat diet in adult mice

It has been suggested that nutritional manipulations during the first weeks of life can alter the development of the hypothalamic circuits involved in energy homeostasis. We studied the expression of a large number of the hypothalamic neuropeptide mRNAs that control body weight in mice that were overfed during breastfeeding (mice grown in a small litter, SL) and/or during adolescence (adolescent mice fed a high-fat diet, AHF). We also investigated possible alterations in mRNA levels after 50 days of a high-fat diet (high-fat challenge, CHF) at 19 weeks of age. Both SL and AHF conditions caused overweight during the period of developmental overfeeding. During adulthood, all of the mouse groups fed a CHF significantly gained weight in comparison with mice fed a low-fat diet, but the mice that had undergone both breast and adolescent overfeeding (SL-AHF-CHF mice) gained significantly more weight than the control CHF mice. Of the ten neuropeptide mRNAs studied, only neuropeptide Y (NPY) expression was decreased in all of the groups of developmentally overfed adult mice, but CHF during adulthood by itself induced a decrease in NPY, agouti-related protein (AgRP) and orexin (Orx) mRNA levels. Moreover, in the developmentally overfed CHF mice NPY, AgRP, galanin (GAL) and galanin-like peptide (GalP) mRNA levels significantly decreased in comparison with the control CHF mice. These results show that, during adulthood, hypothalamic neuropeptide systems are altered (NPY) and/or abnormally respond to a high-fat diet (NPY, AgRP, GAL and GalP) in mice overfed during critical developmental periods.

Autophagy activation in COL6 myopathic patients by a low-protein-diet pilot trial.

ABSTRACT A pilot clinical trial based on nutritional modulation was designed to assess the efficacy of a one-year low-protein diet in activating autophagy in skeletal muscle of patients affected by COL6/collagen VI-related myopathies. Ullrich congenital muscular dystrophy and Bethlem myopathy are rare inherited muscle disorders caused by mutations of COL6 genes and for which no cure is yet available. Studies in col6 null mice revealed that myofiber degeneration involves autophagy defects and that forced activation of autophagy results in the amelioration of muscle pathology. Seven adult patients affected by COL6 myopathies underwent a controlled low-protein diet for 12 mo and we evaluated the presence of autophagosomes and the mRNA and protein levels for BECN1/Beclin 1 and MAP1LC3B/LC3B in muscle biopsies and blood leukocytes. Safety measures were assessed, including muscle strength, motor and respiratory function, and metabolic parameters. After one y of low-protein diet, autophagic markers were increased in skeletal muscle and blood leukocytes of patients. The treatment was safe as shown by preservation of lean:fat percentage of body composition, muscle strength and function. Moreover, the decreased incidence of myofiber apoptosis indicated benefits in muscle homeostasis, and the metabolic changes pointed at improved mitochondrial function. These data provide evidence that a low-protein diet is able to activate autophagy and is safe and tolerable in patients with COL6 myopathies, pointing at autophagy activation as a potential target for therapeutic applications. In addition, our findings indicate that blood leukocytes are a promising noninvasive tool for monitoring autophagy activation in patients.