Effects of a 12-month moderate weight loss intervention on insulin sensitivity and inflammation status in nondiabetic overweight and obese subjects.

Weight loss intervention is the principal non-pharmacological method for prevention and treatment of type 2 diabetes. However, little is known whether it influences insulin sensitivity directly or via its anti-inflammatory effect. The aim of this study was to assess the independent role of changes in inflammation status and weight loss on insulin sensitivity in this population.Overweight and obese nondiabetic participants without co-morbidities underwent a one-year weight loss intervention focused on caloric restriction and behavioral support. Markers of inflammation, body composition, anthropometric para-meters, and insulin sensitivity were recorded at baseline, 6, and 12 months. Insulin sensitivity was assessed with frequently sampled intravenous glucose tolerance test and Minimal Model. Twenty-eight participants (F: 15, M: 13, age 39±5 years, BMI 33.2±4.6 kg/m(2)) completed the study, achieving 9.4±6.9% weight loss, which was predominantly fat mass (7.7±5.6 kg, p<0.0001). Dietary intervention resulted in significant decrease in leptin, leptin-to-adiponectin ratio, hs-CRP, and IL-6 (all p<0.02), and improvement in HOMA-IR and Insulin Sensitivity Index (SI) (both p<0.001). In response to weight loss IL-1ß, IL-2, leptin, and resistin were significantly associated with insulin, sensitivity, whereas sICAM-1 had only marginal additive effect. Moderate weight loss in otherwise healthy overweight and obese individuals resulted in an improvement in insulin sensitivity and in the overall inflammation state; the latter played only a minimal independent role in modulating insulin sensitivity.

Functional thermogenic beige adipogenesis is inducible in human neck fat.

BACKGROUND: Recent studies suggest human neck brown adipose tissue (BAT) to consist of 'brown adipocyte (BA)-like' or beige adipocytes. However, little is known about their thermogenic function. Within the beige adipocyte transcriptome, fibroblast growth factor-21 (FGF21) is a gene whose protein product acts as an adipokine, regulating cold-induced thermogenesis in animals. Here, we explored (i) the adipogenic potential, thermogenic function and FGF21 secretory capacity of beige adipocytes derived from human neck fat and (ii) the role of FGF21 in modulating adipose bioenergetics. METHODS: Progenitors isolated from human cervical fat were differentiated into adipocytes with either a BA-like or white adipocyte (WA) phenotype. FGF21 secretion was measured by enzyme-linked immuosorbent assay. Real-time PCR/western blotting was used to determine cellular mRNA/protein levels. Extracellular flux bioanalyzer was used to quantify adipocyte oxygen consumption and fatty acid oxidation. Adipocyte heat production was measured by infrared thermography. RESULTS: Under hormonal manipulation, primary human neck pre-adipocytes differentiated into adipocytes with either BA-like or WA phenotypes, on gene/protein and functional levels. BA-like cells expressed beige but not classic BA markers. During BA differentiation, FGF21 gene expression and secretion were increased, and were augmented following norepinephrine exposure (a cold mimic in vitro). Differentiated WA expressed ß-klotho, a critical co-factor mediating FGF21 action. Treatment of WA with FGF21-induced UCP1 expression and increased oxygen consumption, respiratory uncoupling, norepinephrine-mediated thermogenesis, fatty acid oxidation and heat production, thus recapitulating the association between cold-induced FGF21 secretion and cold-induced thermogenesis in vivo. CONCLUSION: Beige adipocytes are thermogenic in humans. FGF21 is a beige adipokine capable of promoting a brown fat-like thermogenic program in WAs. SIGNIFICANCE: This study provides first evidence of inducible functional thermogenic beige adipogenesis in human neck fat. FGF21 holds promise as a cold-induced beige adipokine with metabolic benefits of therapeutic relevance through browning of white adipose tissue.

Fibroblast growth factor 21 (FGF21) and bone: is there a relationship in humans?

UNLABELLED: In animals, high fibroblast growth factor 21 (FGF21) states improve insulin resistance but induce bone loss. Whether FGF21 relates to bone mineral density (BMD) is unknown in humans. Contrary to prediction from animal findings, we found higher FGF21 levels associating with greater BMD in women, independent of age and body composition. INTRODUCTION: Recent laboratory studies suggest that FGF21 is involved in reciprocal regulation of bone and energy homeostasis. Systemic administration of FGF21 protects animals from obesity and diabetes but causes severe bone loss, smothering the enthusiasm over FGF21 as a potential antiobesity therapeutic. To date, there is no information on whether FGF21 relates to BMD in humans. We thus studied the relationship between plasma FGF21 levels and BMD in healthy adults. METHODS: Fasting plasma FGF21 levels were measured by enzyme-linked immunosorbent assay and body composition by dual-energy X-ray absorptiometry. RESULTS: Among 40 healthy volunteers (age 32 ± 10 year, 16 women), men had significantly higher lean body mass (p < 0.01) and total BMD (p < 0.05), and lower percent body fat than women (p < 0.01). Median plasma FGF21 levels were not different between the sexes. While there was no association between FGF21 concentrations and body composition in men, FGF21 levels correlated positively with fat mass (p < 0.01) in women. In men, no significant correlation between FGF21 with BMD was observed. However, in women, FGF21 correlated positively with total BMD (R (2) = 0.69, p = 0.003) and spine BMD (R (2) = 0.76, p = 0.001); the correlation remained significant after adjusting for age, ethnicity, and body composition. CONCLUSIONS: This study reveals for the first time a strong positive association between plasma FGF21 levels and BMD in healthy women, suggesting the association between bone loss and high FGF21 states in animals may not be directly translated to humans in physiologic states. We hypothesize that FGF21 may increase bone mass particularly in women through paracrine mechanisms in the bone-adipose interface.

Cold-activated brown adipose tissue is an independent predictor of higher bone mineral density in women.

UNLABELLED: In animals, defective brown adipogenesis leads to bone loss. Whether brown adipose tissue (BAT) mass relates to bone mineral density (BMD) in humans is unclear. We determined the relationship between BAT mass and BMD by cold-stimulated positron-emission tomography (PET) and dual-energy X-ray absorptiometry (DXA) in healthy volunteers. Higher BAT mass was associated with higher BMD in healthy women, but not in men, independent of age and body composition. INTRODUCTION: Contrary to the traditional belief that BAT is present only in infants, recent studies revealed significant depots of BAT present in adult humans. In animals, defective brown adipogenesis leads to bone loss. While white adipose tissue mass is a known determinant of BMD in humans, the relationship between BAT and BMD in humans is unclear. We thus examined the relationship between BAT and BMD in healthy adults. METHODS: BAT volume (ml) and activity (standard uptake value) were determined by 18F-fluorodeoxyglucose PET after overnight mild cold exposure at 19 °C, and BMD was determined by DXA. RESULTS: Among 24 healthy adults (age 28±1 years, F=10), BAT volumes were 82.4±99.5 ml in women and 49.7±54.5 ml in men. Women manifested significantly higher BAT activity, by 9.4±8.1% (p=0.03), than men. BAT volume correlated positively with total and spine BMD (r2=0.40 and 0.49, respectively, p<0.02) in women and remained a significant predictor after adjustment for age, fat, and lean body mass (p<0.05). Total and spine BMD were higher in women who harbored visually detectable BAT on PET images than those without by 11±2% (p=0.02) and 22±2% (p<0.01), respectively. No associations were observed between BAT parameters and BMD in men. CONCLUSIONS: This study demonstrated higher BMD among healthy women with more abundant BAT, independent of age and other body compositional parameters. This was not observed in men. The data suggest that brown adipogenesis may be physiologically related to modulation of bone density.

The IGF axis in prostate cancer.

Prostate cancer, the most frequent non-cutaneous malignancy in men from industrialized countries, is a growing medical problem, representing the second leading cause of male cancer deaths. In the last decade, converging evidence from epidemiological and biological studies suggests that the Insulin-like Growth Factor (IGF) axis is involved in the tumorigenesis and neoplastic growth of prostate cancer. Epidemiological observations indicated that circulating IGF-I levels are positively associated with the increased risk of prostate cancer. The activation of type I IGF receptor (IGF-IR) by IGF-I and/or IGF-II, has mitogenic and antiapoptotic effects on normal and malignant prostate cells. Altered expression of IGF axis components has also been reported in vitro and in animal models of prostate cancer, as well as in human prostate cancer tissue samples. In this review we address and analyze epidemiological studies, in vitro and in vivo cancer models, and human ex vivo prostate cancer researches performed to date supporting the role of IGF axis in prostate cancer.

Circulating adipocytokines in non-diabetic and Type 1 diabetic children: relationship to insulin therapy, glycaemic control and pubertal development.

AIM: To determine the influence of Type 1 diabetes mellitus on circulating adipocytokines in children. METHODS: The circulating concentrations of leptin, adiponectin, resistin and tumour necrosis factor (TNF)-alpha were measured in 91 children, aged 11.1 +/- 2.7 years, with Type 1 diabetes mellitus (T1DM). Ninety-one healthy children were selected as control subjects. RESULTS: Body mass index-adjusted leptin concentrations were higher in the pubertal diabetic children compared with the control children. There was a significant positive correlation between leptin and daily insulin dose in the diabetic group. Circulating adiponectin concentrations were higher in the prepubertal diabetic children and were positively associated with HbA(1c). Resistin concentrations were lower in the prepubertal non-diabetic subjects compared with the pubertal non-diabetic children, whose values were higher than those of the diabetic children. TNF-alpha concentrations were similar in non-diabetic and diabetic children. CONCLUSIONS: Circulating concentrations of adipocytokines are abnormal in Type 1 diabetic children, although the direction of change differs by cytokine. Pubertal development, in addition to insulin treatment and glycaemic control, also influences the concentrations.

Peripheral metabolism of thyroid hormone and glucose homeostasis.

Thyroid hormone action has long been recognized as an important determinant of glucose homeostasis. Recent advances in the knowledge of the physiology of the deiodinases indicate that through tissue-specific regulation of thyroid hormone metabolism, leading to local specificity of thyroid hormone action and target gene transcription patterns, they may have an important function in the modulation of carbohydrate metabolism. This review briefly addresses the role of thyroid hormone action on glucose homeostasis with a specific focus on the significance of the peripheral metabolism of thyroid hormone in the regulation of glucose homeostasis and insulin sensitivity.

Circulating acylated and total ghrelin and galanin in children with insulin-treated type 1 diabetes: relationship to insulin therapy, metabolic control and pubertal development.

OBJECTIVE: To study the circulating levels of two gut-derived peptides in children with type 1 (insulin-dependent) diabetes mellitus (IDDM). RESEARCH DESIGN AND METHODS: Plasma levels of ghrelin, both total ghrelin (TG) and the acylated form (AG), and galanin and their relationships with insulin dosage, metabolic control, IGFBP-1, body mass and pubertal development were evaluated in 91 children, aged 11.1 +/- 2.7 years, affected by IDDM and treated with insulin. Ninety-one healthy children were selected as controls. RESULTS: Body mass index (BMI)-adjusted levels of both forms of ghrelin were reduced in IDDM compared with healthy subjects, with greater values in prepubertal than pubertal IDDM subjects. A negative association was found between AG and fasting insulin serum levels and insulin resistance [measured by using the homeostasis model assessment of insulin resistance (HOMA IR)] among the healthy children. IDDM children showed a negative association of their plasma ghrelin (both acylated and total) with daily insulin dosage, and the three adiposity indices (BMI, skinfold thickness and percentage fat mass). IGFBP-1 levels were higher among the IDDM children without any association with ghrelin serum values. BMI-adjusted plasma levels of galanin were higher among IDDM compared to healthy subjects, irrespective of sex or pubertal development. Greater values for galanin were found among pubertal than prepubertal subjects in both groups without any significant differences between the genders. A positive association was found between galanin and BMI in both groups and between galanin and haemoglobin A1c (HbA1c) among the IDDM children. No relationship existed between either galanin and fasting serum insulin among the healthy subjects or galanin and both insulin dosage or duration of treatment among the IDDM subjects. CONCLUSIONS: The associations found between both ghrelin and galanin with adiposity indices could be considered as an indirect signal of involvement of the two peptides in the development of the nutritional status of the IDDM adolescents. The reduction in both forms of ghrelin could be involved in the development of the body mass increase of IDDM subjects with opposite effects, either influencing insulin sensitivity or exerting a compensatory restraint of feeding.

BMI-conditional standard of leptin serum values in children: proposal of a statistical procedure for the analysis of influencing variables.

In order to define serum leptin values in children, its concentration was assayed in 3,453 children, 5-14 years old, and body mass index (BMI) and pubertal development were recorded. Insulin, testosterone (in males) and 17beta-estradiol (in females), the sum of four skinfold thicknesses (SST), waist-to-hip ratio, and body fat mass were also determined in 1,601 children. Analysis of multicollinearity effects on estimated models demonstrated a quasi-linear correlation between SST and BMI, the former being prevalent. Although other variables were strongly correlated with leptin, assuming only SST as predictor, R2 yielded a value of 0.711 in males and 0.607 in females. When the other variables were added, R2 increased by about 0.03 in both sexes. BMI and SST were the most important of all the predictors and each can act as a sort of proxy for the others. When the z-scores of BMI of all 3,453 children were subdivided into deciles, any correlation with leptin was no more significant inside each BMI z-score range. This study demonstrates that subcutaneous fat mass may be considered the prevalent determinant factor. The adopted statistical procedure furnished results useful for reference values based on BMI z-score as a simple and appropriate evaluation for serum leptin concentrations in children.

Epidemiology of overweight and obesity among school children and adolescents in three provinces of central Italy, 1993-2001: study of potential influencing variables.

OBJECTIVE: The prevalence of overweight and obesity was estimated among the school children and adolescents of three provinces of central Italy, and the role of several possible influencing factors was analysed. DESIGN, SUBJECTS AND MEASUREMENTS: Body mass index (BMI) was measured in 44 231 subjects, age 3-17.5 y, and a household questionnaire was filled out by the parents of 12 143 subjects to collect the following data: subjects, only child or firstborn status, prematurity, birth weight, type of feeding until the fifth month, menarche status in girls; parents, age at the time of the subject's birth; BMI (mean of the two parents) at the time the subject was measured, mother's age of menarche, socioeconomic status. BMI was measured in a subgroup of 10 795 subjects 1 y later to study the yearly sex- and age-related variations from the categories of normal weight to overweight or obesity and vice versa. All females aged 11-14 y were asked if they had their menarche. RESULTS: Striking differences in the proportions of overweight and obesity resulted from the use of two different criteria for defining cutoff points. The overall prevalence of overweight was 13.2 and 20.7% in males, and 13.7 and 18.6% in females, and the overall prevalence of obesity varied between 24.2 and 6.3% in males, and between 22.9 and 6.1% in females, respectively. Parents' BMI, birth weight, firstborn status and post-menarche status in girls showed a significant association with overweight and/or obesity in logistic regression models. CONCLUSIONS: A large prevalence of overweight and obesity was observed in school subjects from three provinces of central Italy. From the comparisons of the prevalence rate, the new internationally agreed criteria seem more appropriate for epidemiological studies in this population. SPONSOR: University of Perugia, Region of Umbria, Commune of Perugia.

Leptin serum levels are involved in the relapse after weight excess reduction in obese children and adolescents.

BACKGROUND: Leptin concentrations as a predictor of weight excess (WE) variations in obese children continue to be controversial. AIM AND DESIGN: To evaluate the relationship between fasting leptin serum concentrations and the ability to maintain loss of WE during obesity treatment, 172 (82 males and 90 females) overweight children and adolescents (OW), 6-16 yr old, were recruited. The subjects were retrospectively selected among those who had demonstrated a reduction of their WE during an initial phase of 12 months of a WE reduction programme (WERP). Fasting serum levels of leptin were assayed, together with insulin, triacylglycerol and cholesterol, before (time 0) and at the end of the first phase of WERP (time 1), and BMI (Z-score) was determined at time 0, time 1 and at the end (time 2) of a subsequent second phase of 12 months. OW were subdivided according to wether their Z-BMI showed a persistent reduction also during the second phase (maintaining WE reduction subjects or MS) or showed a subsequent increase after the reduction observed during the first phase (relapsing WE subjects or RS). RESULTS: A significant reduction in serum levels of leptin, insulin and lipids, paralleling Z-BMI reduction, was observed at the end of the first phase of WERP, during which we found a correlation between the decrease in serum leptin concentrations and the decrease in Z-BMI. The decrease in RS during the first phase ((deltalgL(0-1) was significantly greater when compared to that observed in MS (p < 0.05). In two different multiple logistic regression analyses, where RS = 0 and MS = 1, serum leptin at time 1 [odds ratio (OR) = 1.08; 95% confidence interval (CI) = 1.04-1.13] and deltalgL(0-1) (OR = 0.48; 95% CI = 0.25-0.92), together with final pubertal stage (OR = 0.78; 95% CI = 0.63-0.96), were significantly associated with final subject status. CONCLUSIONS: Leptin serum levels after a previous WE reduction and its parallel decline are related to subsequent adiposity outcome. The lower the leptin serum concentration after previous WE reduction and/or the greater its decrease, the greater was the probability of WE relapse.

Increased expression of thyroid hormone receptor isoforms in end-stage human congestive heart failure.

Thyroid hormone plays an important role on myocardial development and function. The local effects of thyroid hormone are mediated by the receptor isoforms ultimately driving the expression of cardiac-specific genes. Although overt and subclinical thyroid dysfunction causes well-known changes in the cardiovascular system, little is known about local thyroid hormone action in normal and failing human myocardium. With a newly developed multiplex competitive RT-PCR method, we evaluated the expression of thyroid hormone receptor (TR) isoforms alpha-1, alpha-2, and beta-1 in normal human hearts and in end-stage congestive heart failure. A statistically significant difference in the expression of all three TR isoforms was observed among samples from normal subjects, ischemic heart disease (IHD), and dilated cardiomyopathy (DCM). In DCM, compared with normal, the studied TR isoforms were significantly increased. In IHD, the increased expression was found significant only for alpha-1 and alpha-2 isoforms. No differences were observed between the pathologic groups. In conclusion, a coordinated increment in the expression of the TR isoforms was observed in both DCM and IHD by multiplex competitive RT-PCR. The observed changes could represent a compensatory mechanism to myocardial failure or to locally altered thyroid hormone action.

Age-related prevalence of platelet-associated immunoglobulins G in nonthrombocytopenic patients with autoimmune thyroid disease and autoimmune polyglandular syndrome.

OBJECTIVE: The prevalence of platelet-associated IgG (paIgG) in nonthrombocytopenic patients with autoimmune thyroid disease (AITD) alone or associated with autoimmune polyglandular syndrome (APS) has been studied. SUBJECTS: A total of 164 individuals were enrolled in this study: 81 patients with AITD alone, 33 patients with APS, and 50 healthy controls. RESULTS: The presence of paIgG was recorded in 41 of 81 patients with AITD (51%) as compared with 2 of 50 control subjects (4%, p < 0.0001). The prevalence of paIgG in patients with APS was higher even when compared with patients with AITD alone (25/33, 76%; p = 0.02). The presence of paIgG was not related to the functional thyroid parameters. The prevalence of paIgG was higher in the older than in the younger patients (75 vs. 47%, p = 0.0037). CONCLUSIONS: The results indicate that the prevalence of paIgG in patients with AITD is higher than previously thought, namely in elderly patients and in patients with APS, and not related to the thyroid function.

Leptin serum concentrations predict the responsiveness of obese children and adolescents to weight excess reduction program.

BACKGROUND: A role for leptin to predict weight gain is still controversial. OBJECTIVE AND DESIGN: To determine the relationship between baseline serum leptin values and responsiveness to an educational-based weight excess reduction program (WERP), 418 (241 males and 185 females) obese subjects, aged 9-15 y, were recruited. WERP required 2 y of follow-up. Body mass index (BMI) was evaluated at baseline and at each semester of follow-up. The obese subjects were subdivided into responsives and non-responsives, according to reduction or not of their BMI Z-scores during the WERP. Leptin concentrations were assayed at baseline and were included together with other independent variables in statistical multiple regression analysis. RESULTS: At a preliminary multiple regression analysis, a significant positive correlation between leptin values and BMI Z-score reduction at the second, third and fourth semester of follow-up was registered. To determine the odds ratio of the subjects who were responsive or non-responsive at the various semesters of WERP follow-up, a stepwise logistic regression was used incorporating the same predictors, with the serum leptin values subdivided into quintiles and responsiveness and non-responsiveness as a binary outcome variable. The model offered a satisfying goodness of fit as shown by the sensitivity and specificity. The odds ratio of being responsive were significantly increased by greater quintiles of leptin serum concentrations. Furthermore, such odds ratios were much higher in pubertal than in prepubertal subjects. CONCLUSIONS: These findings support a significant role for serum leptin concentration in predicting BMI changes as a response to an educational excess weight reduction program.

Molecular scanning for mutations in the insulin receptor substrate-1 (IRS-1) gene in Mexican Americans with Type 2 diabetes mellitus.

BACKGROUND: Insulin receptor substrate-1 (IRS-1) is an endogenous substrate for the insulin receptor tyrosine kinase, which plays an important role in insulin signaling. Mutations in the IRS-1 gene are associated in some populations with obesity and Type 2 diabetes. METHODS: To determine whether variation in the IRS-1 gene contributes to genetic susceptibility to insulin resistance and Type 2 diabetes in Mexican Americans, the entire coding region of the IRS-1 gene was screened for variation in 31 unrelated subjects with Type 2 diabetes using single-stranded conformational polymorphism analysis (SSCP) and dideoxy sequence analysis. Variants encoding amino acid substitutions were genotyped in 27 unrelated nondiabetic Mexican Americans and in all family members of subjects containing these variants, and association analyses were performed. To trace the ancestral origins of the variants, Iberian Caucasians and Pima Indians were also genotyped. RESULTS: Eight single base changes were found: four silent polymorphisms and four missense mutations (Ala94Thr, Ala512Pro, Ser892Gly and Gly971Arg). Allele frequencies were 0.009, 0.017, 0.017 and 0.043, respectively. There were no significant associations of any of these variants with diabetes, glucose or insulin levels during an oral glucose tolerance test, or with body mass index (BMI) in Mexican American families except for a modest association between the Ala94Thr variant and decreased BMI (30.4 kg/m(2) vs 24.0 kg/m(2); p=0.035). None of these four missense mutations were detected in Pima Indians. In Iberian Caucasians, neither Ala94Thr nor Ser892Gly were detected, and Ala512Pro was detected in only 0/60 diabetic patients and 1/60 nondiabetic controls. Gly971Arg was relatively more common in Iberian Caucasians with 12/58 diabetic patients and 7/60 nondiabetic controls being heterozygous for this variant (p=0.21 for comparison between diabetic and nondiabetic subjects). CONCLUSIONS: Ala94Thr, Ala512Pro and Ser892Gly mutation are rare in the populations studied. Gly971Arg, is more common in Mexican Americans and Caucasians, but is not a major contributor to genetic susceptibility to Type 2 diabetes.

Structural organization and chromosomal localization of the human type II deiodinase gene.

OBJECTIVE: The selenoenzyme type 2 iodothyronine 5' deiodinase (DII) catalyzes the conversion of thyroxine into its active form tri-iodothyronine (T3), modulating thyroid hormone homeostasis in a local, tissue-specific manner. The amphibian, rodent and human cDNAs encoding this enzyme have been recently cloned and expressed. At present, little information regarding the genomic structure of mammalian DII is available. DESIGN AND METHODS: The complete structure, including intron-exon junctions, of the human DII (hDII) gene was obtained by long PCR and rapid amplification of cDNA ends (RACE). Chromosomal assignment of the hDII gene was performed by fluorescence in situ hybridization using a highly specific probe. RESULTS AND CONCLUSIONS: Our data demonstrated that hDII is a single copy gene located on chromosome 14, position 14q24.3. The gene spans over 15 kb, and the 7 kb transcript is encoded by three exons of 149 bp, 273 bp and 6.6 kb separated respectively by two 274 bp and 7.4 kb introns. A restriction map of the hDII gene is also reported. These data will help in further studies of the role of DII in the maintenance of peripheral thyroid hormone homeostasis.

Isolation of human type 2 deiodinase gene promoter and characterization of a functional cyclic adenosine monophosphate response element.

We analyzed the structure and function of the 5' flanking region of the human type 2 deiodinase (hD2) gene. Two major transcription start sites were identified at -470/-474 from the ATG. The 5' flanking region of hD2 gene efficiently directed transcription in transient transfection studies, using luciferase as reporter gene, in HEK 293 cells. Basal transcriptional activity was significantly reduced by deleting the region containing a canonical cAMP-responsive element (CRE) located -766/-759 from ATG. Forskolin treatment significantly increased luciferase activity in cells transfected with CRE-containing constructs. This effect was abolished in constructs that did not contain CRE or contained the mutagenized CRE. Northern blot analysis in JEG-3 cells revealed that the hD2 messenger RNA was markedly increased after stimulation with cAMP agonist. The electrophoretic mobility shift assay with hD2-CRE probe and HEK 293 nuclear extract showed the occurrence of a DNA-protein complex, which was competed by specific unlabeled oligonucleotides and supershifted by the anti-CREB and anti-CRE modulator-1 antibodies. A-CREB, a dominant negative inhibitor of CREB, completely inhibited forskolin induction of the hD2 promoter. CREB protein, once cotransfected with hD2 promoter construct and pKA in F9 teratocarcinoma cells, which are unresponsive to cAMP, was able to stimulate the hD2 gene transcription. These results indicate the existence of a functional promoter within the 5' flanking region of hD2 gene which is characterized by the presence of a CRE. The specific involvement of CREB in the cAMP-mediated hD2 gene promoter induction also has been demonstrated.

Body mass index in children and adolescents according to age and pubertal stage.

OBJECTIVE: To evaluate the dependence of body mass index (BMI) values on pubertal stage in subjects similar in age. DESIGN, SUBJECTS AND MEASUREMENTS: Height and weight were recorded cross-sectionally in school subjects from three provinces in central Italy. The subjects were subdivided into three groups: (1) 4271 school subjects (2125 males and 2146 females; 8.5-15.5 y old), in whom the pubertal development was also recorded, were selected to subdivide BMI values according to pubertal stage and age; (2) 6345 females (10.5-14.5 y old), who were asked whether or not they had had their first menstrual period, were selected to subdivide BMI values according to age in pre-menarche and post-menarche girls, separately; and (3) 1919 females (10.5-14.5 y old), who had presented their menarche within the previous 6 months, were selected to subdivide short-term post-menarche BMI values according to age. RESULTS: The medians and interquartile ranges of BMI varied according to age and pubertal stage. Kruskall-Wallis test performed in subjects similar in age demonstrated that significant differences existed among the medians of BMI values of subjects at different pubertal stages in 12-14-y-old males (P<0.05), and in 11-14-y- old females (P<0.001). The difference also proved to be significant between stage I and stage II (P<0.05) in 10-y-old females, but not in 10-11-y-old males. The Kruskal-Wallis test performed in subjects similar in pubertal stage demonstrated that significant differences among the medians of BMI at different ages existed only in females at stages II and III. A significant positive trend was observed in both genders according to pubertal stage for BMI values of subjects similar in age (z-test for trend, P<0.01). On the contrary, a negative age trend proved to be significant in females at stages I (P<0.01), II (P<0.01) and III (P<0.001), but not in males when the subdivision of BMI was made according to age in subjects similar in pubertal stage. BMI values were significantly higher in post-menarche girls as compared to pre-menarche girls similar in age (P<0.001). However, at partial regression analysis BMI values were influenced by pubertal stage and, to a lesser extent, by age, but not by menarcheal status. An inverse association between short-term post-menarche BMI and age was observed, with the highest values in girls presenting menarche at 11 y of age (P<0.05). The negative trend was demonstrated at the z-test for trend (P<0.001). CONCLUSIONS: BMI values depend on pubertal degree of maturation, especially in girls. This influence should be taken into account when BMI is evaluated in adolescents. SPONSOR: University of Perugia, Region of Umbria.

Using obese-specific charts of height and height velocity for assessment of growth in obese children and adolescents during weight excess reduction.

OBJECTIVE: The influence of weight excess reduction on height and height velocity of obese subjects should be evaluated on the basis of appropriate standards, since the pattern of growth of obese subjects is different from that of normal weight subjects. DESIGN, SUBJECTS AND MEASUREMENTS: Height, weight and triceps skinfold thickness were recorded from 17987 school subjects (9256 males and 8731 females), 3-18 y of age, from three provinces of central Italy, and a growth reference curve of height was constructed. Using BMI (as computed using the tables of Rolland-Cachera et al) and triceps skinfold thickness, normal-weight subjects (NWS) and obese subjects (OS) were identified and specific reference curves (mean+/-s.d. every sixth month of age) were developed for both groups. Centiles of height were also calculated for OS. Various (2-4) measurements of height in school subjects were performed and a graph of height velocity (HV) was constructed in NWS and in OS using the JPPS method. The yearly mean +/-s.d. of HV was also calculated, based on square root transformed data (in order to realise a Gaussian distribution), deriving from successive measurements in total subjects, in NWS and in OS. The z-scores of height and of the square root of HV were calculated in 217 obese subjects (125 males and 92 females) before and during a weight excess reduction programme (WERP). Obese subjects in WERP who showed a reduction of z-score of BMI were considered as 'responsive'; those who either maintained or showed an increase of z-score of BMI were considered as 'non-responsive'. Obese subjects in WERP were followed for 1-4 y, giving the following results: 0-1 y, 142 responsives and 75 non-responsives; 0-2 y, 76 responsives and 33 non-responsives; 0-3 y, 35 responsives and 30 non-responsives; 0-4 y, 24 responsives and 18 non-responsives. RESULTS: Compared to NWS, OS showed a significantly greater HV in 4-9y males and in 4-8y females, but in older children the pubertal spurt was reduced and more precocious. As a result, the height of OS, which was greater in 3-13 year-old males and in 3-11.5 year-old females, subsequently showed a reduction, as compared to that of NWS, in 16-18 year-old males and in 13-18 year-old females. In both responsive and non-responsive groups of obese subjects in WERP, the z-scores of height showed a reduction during WERP when evaluated using the reference curve of the total school population. In contrast, when their growth was evaluated according to the obese-specific reference curve, no significant variation was observed comparing both z-scores before and during the WERP. CONCLUSIONS: More appropriate information on the growth of obese subjects may be obtained when evaluating the height and HV according to obese-specific reference standards from the same population of origin. Adopting this modality, no significant variation of height resulted during WERP in obese children.