Five-year follow-up of a cognitive-behavioural lifestyle multidisciplinary programme for childhood obesity outpatient treatment.

OBJECTIVE: Aim of this study is to examine the 5-year follow-up results of MI PIACE PIACERMI (I like to like and please myself), a cognitive-behavioural programme intended to obtain a weight growth regulation over an extended period. DESIGN: Longitudinal observational clinical study. SETTING: Hospital-based programme, through outpatient activities. SUBJECTS: Thirty-one simple obese children, 13 boys and 18 girls, 6-12 years of age on admission. INTERVENTION: The intervention was carried out by a multidisciplinary team (paediatrician, cognitive-behavioural psychologist, physical therapist). It employed cognitive-behavioural techniques, nutrition education, promotion of physical activity, setting a high value on free play in motion. The programme actively involves parents. The following measurements were taken on admission and at 5-year follow-up: height, weight, waist circumference, body mass index (BMI), BMI standardized (BMI-SDS), and adjusted BMI (a-BMI) as actual BMI/BMI (50th percentile)*100. Dietary habits were investigated by interview and 24-h recall. Parents completed the Family Habit Inventory and the Child Behaviour Checklist (CBCL). Motor skills were assessed by using Frostig's test. RESULTS: The dropout rate was 35.5%. In subjects who completed the 5-year follow-up, the mean and s.d. of BMI-SDS and a-BMI were, respectively, 4.23+/-0.71 and 54.7%+/-9.0 at baseline and 2.74+/-0.85 and 43.2%+/-17.3 at the last visit. Waist circumference decreased. Family habits improved significantly. Total energy intake was significantly reduced. Emotional and social aspects of obesity-related behaviours showed positive changes. Motor skills globally improved. CONCLUSIONS: The study provides further evidence that positive persistent results may be obtained in obese children with treatment programmes combining a lifestyle centred approach, parental involvement, nutrition education and cognitive-behavioural strategies.

Bone quality assessed by phalangeal quantitative ultrasonography in children and adolescents with isolated idiopathic growth hormone deficiency.

OBJECTIVE: Some observations indicate that GH deficiency (GHD) may have little impact on bone mineralization in contrast to its effects on bone growth and maturation. The aim of the present study was to evaluate the effects of isolated GHD and GH-replacement therapy on bone quality assessed by a quantitative ultrasound (QUS) technique at the proximal phalanges of the hand. DESIGN: Growth and QUS data of 68 subjects (50 males and 18 females) aged 5-18 yr with isolated GHD were retrospectively examined. A cross-sectional series of 120 observations was collected and compared with data obtained from a control population (1227 healthy children, 641 males and 586 females, aged 3-16 yr). METHODS: QUS variables amplitude- dependent speed of sound (AD-SoS) and bone transmission time (BTT) were assessed by the sonographic device DBM Sonic BP IGEA. Height and weight measurements were performed according to standard techniques. In patients, skeletal age (SA) was determined by Tanner-Whitehouse method (3rd version). RESULTS: Before treatment height, SA, ADSoS and BTT were reduced in patients. Height SD score (SDS), SA SDS, AD-SoS SDS, and BTT SDS improved during treatment. Significant associations of both AD-SoS and BTT with age, SA, height, and therapy duration were observed. Using multivariate regression models the disease state, SA, and height proved to be significant variables in predicting BTT and AD-SoS. CONCLUSIONS: QUS measurements adjusted for body size and skeletal maturity in GHD patients seem to be only slightly reduced. A body size and skeletal maturity adjustment should be incorporated in studies on bone mass in GHD children and adolescents. A non-invasive technique such as QUS technology opens new perspectives.

The ethanol self-administration context as a reinstatement cue: acute effects of naltrexone.

A major difficulty in treating alcohol addiction is the high rate of relapse even after prolonged abstinence. Relapse can be triggered by several factors, including stress, re-exposure to the drug, conditioned discrete stimuli and exposure to the context in which alcohol consumption occurred. The present study investigated the role played by the environmental context on ethanol relapse using an extinction/reinstatement animal model: rats were trained to self-administer ethanol in a distinctive context, and extinction occurred in a setting that differed by visual, tactile and olfactory properties; reinstatement was tested by placing the animals into the ethanol-associated context in the absence of ethanol. We found that re-exposure to the ethanol-associated context significantly increased responses on the ethanol-paired lever. The increase in responding required the presence of the complete configuration of the multimodal context. The non-selective opioid receptor antagonist naltrexone (0.3 mg/kg) administered 20 min prior to the reinstatement test significantly attenuated context-induced reinstatement of lever press responding, compared with saline-treated subjects. These data indicate that the environmental context associated with ethanol availability influences ethanol-seeking behavior in the rat, and that endogenous opioids are involved in this process. Our findings are in accordance with clinical reports demonstrating naltrexone efficacy in the treatment of alcohol relapse in humans, and indicate that the context-induced reinstatement model described here may be useful to investigate the biological mechanisms underlying alcohol relapse.

Structural synaptic remodeling in the perirhinal cortex of adult and old rats following object-recognition visual training.

The ultrastructural features of layer II synapses in the perirhinal cortex of adult (4- to 6-month-old) and old (25- to 27-month-old) rats exposed to a six-session object recognition visual training were investigated by morphometric methods. The comparative analysis showed a higher synaptic numeric density, a lower synaptic average area, and a lower percentage of megasynapses (S > 0.5 microm2) in old trained rats versus controls, and a higher percentage of small (S < 0.15 microm2) junctions in adult trained rats versus controls. The more marked synaptic remodeling underlying memory consolidation in the perirhinal cortex of old rats might reflect a pre-existing lower dynamic status.

Longitudinal assessment of bone quality by quantitative ultrasonography in children and adolescents.

Among the techniques available to assess bone quality, quantitative ultrasonography of the proximal phalanges of the hand (QUS) has emerged as particularly attractive. In this study, amplitude-dependent speed of sound (AD-SoS) and bone transmission time (BTT) were obtained by the sonographic device DBM Sonic BP IGEA in two sessions at two years' interval, in a school-age population (589 subjects, 290 mol/L and 299F, aged 3 to 16 y) with the aim to determine accuracy of QUS measurements, evaluate QUS variable changes during growth, relate these values with age and growth variables. Mean AD-SoS and BTT at age classes from 5 to 12 y as determined at the first and second measurement sessions were not significantly different. A significant increment (p < 0.0001) between the first and the second measurement was observed for both QUS variables. AD-SoS and BTT showed significantly different variations in the various age groups (ANOVA). Correlations were found of AD-SoS and BTT increments with age, height, weight, pubertal stage and with height growth velocity (p < 0.05). AD-SoS and BTT increment curves presented a very similar trend decreasing from 4 to 7 y of age. Thereafter a plateau was reached up to the age of 10 to 11 y in girls and 11 to 12 y in boys, when an increase was observed corresponding to pubertal growth rate acceleration. In conclusion, the present study would confirm that QUS measurements are accurate. Ad-SoS and BTT increment models are similar to most growth velocity curves and follow a strongly age- and growth-dependent pattern.

Bone quality in perinatally HIV-infected children: role of age, sex, growth, HIV infection, and antiretroviral therapy.

Appropriate supportive care and identification of long-term sequels of therapy are of paramount importance in HIV-infected pediatric patients. As low bone mineral quality (BMQ) in patients can be considered a marker of possible degeneration in osteopenia and osteoporosis in adulthood, we evaluated bone features in a pediatric population. Forty-four patients (23 females, 21 males; aged 3-17 years) were compared with a control population (1227 healthy children: 568 females, 641 males; aged 3-18 years). Seven patients were CDC stage C, 18 B, and 18 A. All patients were vertically infected; four were naive to any antiretroviral treatment, seven were taking two NRTIs, and 32 were on HAART. BMQ was assessed by a quantitative ultrasound (QUS) technique. It measures the amplitude-dependent speed of sound (AD-SoS, m/sec) and the bone transmission time (BTT, microsec). QUS values were significantly lower in cases than in controls, even after adjustment for age and body size (AD-SoS: 1924.7 +/- 64.9 and BTT: 0.97 +/- 0.3 in controls; AD-SoS: 1879.7 +/- 57.2 and BTT: 0.80 +/- 0.32 in cases; p < or = 0.001). The associations of AD-SoS and BTT with gender, type of therapy, and CDC stages were not significant. AD-SoS and BTT were significantly associated with age (r = 0.59, p < 0.0001), skeletal age SDS (r = 0.46, p = 0.002), height (r = 0.66, p < 0.0001), and therapy duration (r = 0.31, p = 0.04). Both AD-SoS and BTT values in patients fell below mean values of controls. Follow-up of bone mineral density is important in patients to prevent long-term problems of skeletal status.

Fetal spine ossification: the gender and individual differences illustrated by ultrasonography.

The spatial and temporal pattern of manifestation of ossification nuclei of the spinal column in fetal life have been well established by histologic and radiologic studies. Sonographic evaluation of the fetal spine depends on visualization of the ossification centers, but the sequence of development of ossification centers in the vertebral column obtained by embryologists and sonographers and radiology are conflicting. We carried out a longitudinal study to establish the ultrasonographic appearance and timing of development of primary ossification centers of the fetal spine in the first and second trimesters of pregnancy. A total of 80 mothers were evaluated during their pregnancy with two echographic controls; in the first trimester, the spine length was measured and, in the second trimester of pregnancy, the timing of ossification of the bodies and neural arches of sacral vertebrae and the difference in appearance between the female and male genders were evaluated. Spinal length measurements obtained in the first trimester and percentage of detection of sacral vertebral structures increased progressively with a regular pattern in relation to gestational age. Spinal length at first ultrasound examination was slightly correlated with time of appearance of sacral bodies and arches. Ossification timing was significantly earlier in females than in males. The study has attempted to improve our understanding of the sonographic detection of the spinal ossification. Data presented give some further information on the stages of appearance of sacral vertebrae body centers during intrauterine development. Differences between genders and interindividual variations in ossification timing were observed at a very early stage of development. This could be of value when fetal growth is evaluated. Moreover, further knowledge of spinal development may be useful for early diagnosis of spinal abnormalities and for fetal biometrics.

Noradrenergic inhibition and voltage-dependent facilitation of omega-conotoxin-sensitive Ca channels in insulin-secreting RINm5F cells.

We found that, besides dihydropyridine-sensitive Ca channels, insulin-secreting RINm5F cells also contain a minority (15-25%) of omega-conotoxin (omega-CgTx)-sensitive channels that show a high-affinity binding to [125I] omega-CgTx (Kd 51 pM). Noradrenaline (NA, 10 microM) slows down Ca-channel activation in these cells and produces a sizeable reduction of Ca currents that is relieved by strong pre-conditioning depolarizations (facilitation). The action of NA is mimicked by intracellular application of GTP-gamma-S and is prevented by pertussis toxin (PTX) or by cell pre-incubation with omega-CgTx. This suggests specific noradrenergic inhibition of omega-CgTx-sensitive Ca channels that is modulated by membrane potentials and PTX-sensitive G-protein activation.

Up-regulation of L- and non-L, non-N-type Ca2+ channels by basal and stimulated protein kinase C activation in insulin-secreting RINm5F cells.

We studied the effect of protein kinase C (PKC) inhibition and activation on voltage-dependent Ca2+ channels in rat insulinoma RINm5F cells. PKC down-regulation by chronic (24 h) treatment with the PKC activator phorbol 12-myristate 13-acetate (PMA) reduced by about 60% the Ba2+ currents through L- and non-L, non-N-type high-voltage-activated Ca2+ channels, indicating that PKC tonically up-regulates the two main Ca2+ channel subtypes of RINm5F cells under basal conditions. Consistently, PKC activation by acute PMA application caused only a modest increase (average 23%) of Ba2+ currents in a minority of cells (24%). L- and non-L, non-N-type channels were differentially up-regulated by either basal or stimulated PKC activation. Acute up-regulation was predominant on L-type channels and caused an I/V shift of the Ba2+ currents in the hyperpolarizing direction. Non-L, non-N-type channels were less affected by acute PMA application, possibly reflecting a more effective tonic PKC up-regulatory action. Unexpectedly, the increase of Ba2+ currents during acute PMA application was followed by a progressive current decrease, which was also observed in isolation in another 24% of the cells and could be ascribed to PKC-induced ATP depletion, rather than to a direct effect of PKC on Ca2+ channels. We also provide evidence that PKC-mediated phosphorylation is not involved in the G-protein-mediated noradrenergic modulation of Ca2+ channels in RINm5F cells.

Projections from the Cerebral Cortex to the Red Nucleus of the Guinea-pig. A Retrograde Tracing Study.

The origin and the topographic distribution of corticorubral (CR) projections in the guinea-pig were studied by using the retrograde axonal transport of a tracer, colloidal gold-labelled, enzymatically inactive horseradish peroxidase conjugated to wheat-germ agglutinin (WGAapoHRP - Au), which was injected in the red nucleus (RN). It was found that the bulk of the CR projections arise from layer V neurons of the agranular frontal cortex in both its medial (Agm) and lateral (Agl) subdivisions; in the Agm labelled neurons are preferentially located in the upper part of layer V, whereas in the Agl they are more concentrated in the central band of the layer. Fewer projections originate from areas of the granular parietal and the agranular cingulate and retrobulbar cortices. CR projections have a bilateral origin, with a large ipsilateral predominance. The pattern of retrograde cortical labelling observed after injection of WGAapoHRP - Au in different portions of the RN indicates that CR projections are distributed throughout the entire rostrocaudal extent of the nucleus, but are slightly more concentrated in the rostral parvocellular area. The morphological arrangement of CR projections in the guinea-pig, as demonstrated in the present study, shows several analogies with other mammals. The functional characteristics of the cortical areas in which CR neurons are located indicate that CR projections may play a significant role in the central organization of movement.

Effects of niridazole and 5-nitroimidazoles on heart mitochondrial respiration.

2.3 micromoles/mg protein of MFNI induced a 60% decrease in the heart mitochondrial ADP-stimulated oxygen uptake using glutamate-malate as substrate. The same amount of niridazole, ipronidazole, DA 3851 and ornidazole led to falls of less than 20% in the oxygen uptake, whilst metronidazole was ineffective. State 3 and state 3 mu (uncoupled) respiration were affected to the same extent. Oxygen-uptake using succinate as substrate was not inhibited indicating that the action was exerted at the NADH oxidation level. The relationship between electroreduction potentials of the test compounds and inhibition of respiration has been studied.

Selective cardiodepressant activity of fluodipine, a fluorenone-1,4-dihydropyridine derivative.

The effect of the dihydropyridine derivative, 1,4-dihydro-2,6-dimethyl-4-(fluorenon-4-yl)pyridine-3,5-dicarboxyl ic acid diallyl ester (fluodipine) was studied in vitro in different rabbit, rat and guinea pig preparations and in vivo in the rabbit in order to characterize its pharmacological profile at cardiac and at vascular sites. Compared to nifedipine, fluodipine showed a similar cardiodepressant activity, and a much lower inhibitory activity on vascular contraction. The highest tissue selectivity was observed in guinea pig preparations: fluodipine was about 2-3 times more effective than nifedipine on chronotropism and inotropism in isolated atria, and about 150 times less effective on aortic strip contraction. Accordingly, fluodipine (i) showed high-affinity binding to guinea pig ventricular L-type cardiac Ca2+ channels (Ki=2.57 nM), (ii) was about 80 times less effective than nifedipine to inhibit Ca2+ influx in vascular smooth muscle cells and (iii) induced a significant reduction of heart rate in the anesthetized rabbit (ID25=8.5 mg kg(-1), i.v.) without affecting the blood pressure up to 20 mg kg(-1), whereas nifedipine showed a significant hypotensive effect at very low doses (ID25=0.18 mg kg(-1), i.v.). The pacemaker current If of rabbit sino-atrial node myocytes was not affected by fluodipine. These findings demonstrate that fluodipine exerts selective cardiodepressant activity, likely due to a higher affinity for cardiac than for vascular Ca2+ channels.

Fetal growth velocity: kinetic, clinical, and biological aspects.

With the aim of determining fetal growth kinetics, prenatal data were analysed which had been longitudinally collected in the framework of a perinatal growth survey. The sample comprised 238 singleton normal pregnancies, selected in Genoa and Turin (between 1987 and 1990), and repeatedly assessed by ultrasound scans (five to nine per pregnancy). Five morphometric traits were considered: BPD (biparietal diameter), OFD (occipitofrontal diameter), HC (head circumference), FDL (femur diaphysis length) and AC (abdomen circumference). Growth rate seemed to increase in the early part of the second trimester, and decrease subsequently: velocity peaks were steeper and earlier for head diameters and circumference (about 18 weeks) than for femur length (20 weeks) and abdomen circumference (22 weeks). Velocity standards were traced using a longitudinal two-stage linear model: this ensures unbiased description of the shape of the growth curve, even when growth kinetics are asynchronous, and efficient estimation of the outer centiles--the most useful for diagnostic purposes.

FELS, Greulich-Pyle, and Tanner-Whitehouse bone age assessments in a group of Italian children and adolescents.

Assessments of skeletal maturity are usually made from handwrist radiographs, using either the method of Greulich-Pyle (GP) or Tanner-Whitehouse (TW). Recently the FELS method has been developed, and it represents a potentially valuable approach to skeletal age assessment. The present study evaluates the accuracy and precision of FELS skeletal age assessments compared with ratings by the GP and TW methods in a group of Italian children and adolescents. The hand-wrist radiographs of subjects (171 males and 156 females 1 to 17 years) referred to the "Istituto di Puericultura e Medicina Neonatale" of the University of Genoa in Italy between 1985 and 1990 were assessed according to each method. Two independent observers rated the radiographs and one observer reassessed them after 6 months or more. GP estimates rather closely match chronological age; TW ratings tend to overestimate chronological age by 7-9 months around puberty, mainly in boys; and the FELS method tends to overestimate chronological age by amounts that increase with chronological age. The degree of precision of skeletal age assessments is within the usually accepted limits. Higher levels of repeatability and reproducibility are apparent for the FELS estimates than for GP and TW assessments. Thus, skeletal maturity is likely to be adequately interpreted by the FELS method as well as by the more commonly used GP and TW systems in Italian children and adolescents. © 1992 Wiley-Liss, Inc.

Chemiluminescence imaging in bioanalysis.

The development, analytical performance and applications of chemiluminescence imaging as a tool for quantitative analyte localization in target biological specimens are described. The detection of acetylcholinesterase activity both in array format and on a target surface are described. A proposed application of the method is a 384 well microtiter format assay for high throughput screening of acetylcholinesterase inhibitors such as tacrine, a drug widely used in the treatment of Alzheimer's disease, and two recently developed analogues. The chemiluminescent system in conjunction with optical microscopy allowed localization of acetylcholinesterase in brain tissue sections. We also describe the chemiluminescent immunohistochemical localization of interleukin 8 in Helicobacter pylori infected gastric mucosa cryosections and an in situ hybridization assay for the detection of herpes simplex virus DNA in single cells.

Long-term follow-up of skeletal dysplasia in thalassaemia major.

We report skeletal changes due to deferoxamine (DF) in 15/29 patients with transfusion-dependent thalassaemia major (TM), followed longitudinally for growth assessment. Clinically the earliest signs were decline in height and/or sitting height growth rate, leg and back pain with restricted movement and limb deformity. Radiologically metaphyseal and spinal changes were seen in 5 subjects and vertebral lesions alone in 10. The metaphyseal changes were mild, moderate or severe and affected all long bones, but were most pronounced at wrists and knees. They progressed from widening of the growth plate and defects of metaphyseal margins to appearance of radiolucent pseudocystic areas and, in severe cases, of cupped, rickets-like metaphyses. The spinal changes proceeded from osseous defects of ventral upper and lower edges of vertebrae and biconvex contours of end-plates to platyspondyly with decreased vertebral body height. After DF dose reduction, metaphyseal changes regressed in 2 patients, while they progressed in 3, requiring corrective surgery for severe valgus knee. Spinal abnormalities either remained unchanged or progressed. Final height was very short in patients with spondylometaphyseal lesions, short and disproportionate in patients with only spinal involvement.

Effect of fast moving stimuli and saccadic eye movements on cell activity in visual areas V1 and V2 of behaving monkeys.

Extracellular recordings were carried out in the visual cortex of behaving monkeys trained on a fixation/detection task, during which a target light was displayed stationary or suddenly moving on a tangent translucent screen. The responses of visual cortical cells to fast moving stimuli during steady fixation and those obtained during rapid eye movements (saccades) which moved their receptive field across a stationary stimulus, were studied. Areas V1 and V2 were explored. When tested with rapidly moving stimuli (500 deg/sec) during steady fixation, neurons in each area behaved in almost the same way. About one fourth of them were activated, the remainder showing either no response (little more than a half of them) or a reduction of the spontaneous firing rate. In both areas, some of the neurons activated during steady fixation did not respond or responded very weakly during eye motion at saccadic velocity (500 +/- 50 deg/sec). Neurons of this type, which we refer to as 'real motion' cells, could somehow contribute to the maintenance of visual stability during the execution of large eye movements.

Suppression of epileptiform burst discharges in CA3 neurons of rat hippocampal slices by the organic calcium channel blocker, verapamil.

We studied the effects of the organic calcium channel blocker, verapamil, on spontaneous and bicuculine-induced epileptiform burst discharges in CA3 pyramidal cells of hippocampal slices. A transient increase of burst discharge rate was observed in most cells within 30 min after the addition of verapamil (100 microM) to the perfusing medium. Prolonged verapamil perfusions gradually reduced the rate and duration of burst discharges, then abolished them in all tested slices (over periods of 50-150 min) without blocking synaptic transmission. Responses to intracellular injections of current pulses were also gradually affected by verapamil: Action potential amplitude was decreased, action potential duration increased, frequency adaptation increased, amplitude of the fast hyperpolarization following a single action potential decreased, and amplitude and duration of the slow afterhyperpolarization markedly reduced. The amplitude of calcium spikes elicited in slices perfused with tetrodotoxin-containing medium was not affected by verapamil, but the mean velocity of depolarization near the peak of the calcium spike was decreased. Membrane resting potential and input resistance were not affected by verapamil. These results confirm that verapamil is able to suppress epileptiform activity, but suggest that this effect is rather non-specific, due to inhibition of both postsynaptic sodium and calcium conductances.