Imaging heterogeneity of peptide delivery and binding in solid tumors using SPECT imaging and MRI.

BACKGROUND: As model system, a solid-tumor patient-derived xenograft (PDX) model characterized by high peptide receptor expression and histological tissue homogeneity was used to study radiopeptide targeting. In this solid-tumor model, high tumor uptake of targeting peptides was expected. However, in vivo SPECT images showed substantial heterogeneous radioactivity accumulation despite homogenous receptor distribution in the tumor xenografts as assessed by in vitro autoradiography. We hypothesized that delivery of peptide to the tumor cells is dictated by adequate local tumor perfusion. To study this relationship, sequential SPECT/CT and MRI were performed to assess the role of vascular functionality in radiopeptide accumulation. METHODS: High-resolution SPECT and dynamic contrast-enhanced (DCE)-MRI were acquired in six mice bearing PC295 PDX tumors expressing the gastrin-releasing peptide (GRP) receptor. Two hours prior to SPECT imaging, animals received 25 MBq (111)In(DOTA-(ßAla)2-JMV594) (25 pmol). Images were acquired using multipinhole SPECT/CT. Directly after SPECT imaging, MR images were acquired on a 7.0-T dedicated animal scanner. DCE-MR images were quantified using semi-quantitative and quantitative models. The DCE-MR and SPECT images were spatially aligned to compute the correlations between radioactivity and DCE-MRI-derived parameters over the tumor. RESULTS: Whereas histology, in vitro autoradiography, and multiple-weighted MRI scans all showed homogenous tissue characteristics, both SPECT and DCE-MRI showed heterogeneous distribution patterns throughout the tumor. The average Spearman's correlation coefficient between SPECT and DCE-MRI ranged from 0.57 to 0.63 for the "exchange-related" DCE-MRI perfusion parameters. CONCLUSIONS: A positive correlation was shown between exchange-related DCE-MRI perfusion parameters and the amount of radioactivity accumulated as measured by SPECT, demonstrating that vascular function was an important aspect of radiopeptide distribution in solid tumors. The combined use of SPECT and MRI added crucial information on the perfusion efficiency versus radiopeptide uptake in solid tumors and showed that functional tumor characteristics varied locally even when the tissue appeared homogenous on current standard assessment techniques.

Pubertal maturation characteristics and the rate of bone mass development longitudinally toward menarche.

To assess risks for osteoporosis and to compare bone mass in different groups of healthy children or children with diseases, it is important to have knowledge of their sexual maturation status during puberty. The aim of our study was to evaluate bone mass formation longitudinally in relation to pubertal maturation characteristics in healthy white girls. We investigated the bone mineral content (BMC) and the bone mineral density (BMD) at different skeletal sites in 151 girls with increasing pubertal stages in relation with their chronological age and with an early or late onset of puberty or menarche and with a slow or fast maturation. Bone mass was measured at the onset of puberty, during puberty, and at menarche. We conclude the following: (1) from midpuberty to menarche, the increase in bone mass formation is highest at all skeletal sites in white girls; (2) early mature girls at the onset of puberty have slightly but definitely lower bone masses at all skeletal sites and at all pubertal stages than late mature girls, whereas the average bone mass formation from the onset of puberty to menarche is similar in both groups; (3) girls with a slow rate of pubertal maturation have lower bone mass values 2 years after the onset of puberty, but at menarche bone mass is similar compared with fast maturers; and (4) it cannot be confirmed that there is an effect of menarcheal age on bone mass values at menarche.

Body fat mass, body fat distribution, and pubertal development: a longitudinal study of physical and hormonal sexual maturation of girls.

The rate at which girls progress through the stages of puberty in relation to body fat mass and body fat distribution and its relation to their hormonal profiles was studied. Sixty-eight schoolgirls participated in a longitudinal study during 3 yr. The girls were divided into subgroups with increasing skinfold thicknesses and waist-hip ratio. They were also grouped depending on Tanner's breast development classification (M2 and M3). The age at M2 was only marginally correlated with the menarcheal age, but the age at M2 and the time interval from that age to menarche was negatively correlated. Age at the onset of puberty was not related to body fat mass or distribution. The rate of pubertal development after pubertal stage M3 was negatively related to the body fat mass. Age at M2 was only correlated with estrone (E1), while the rate of pubertal development was associated with higher FSH, E1, estradiol (E2), the fraction of E2 that was not bound to sex-hormone-binding globulin (non-sex-hormone-binding globulin bound E2) and androstenedione plasma levels at the onset of puberty. Body fat distribution, rather than body fat mass was related to the total and the non-sex-hormone-binding globulin bound plasma levels of E2 and testosterone at the onset of puberty. Changes in body fat distribution in early female puberty were chiefly related to the waist circumferences. We found no evidence that body fat mass or body fat distribution triggers the onset of puberty. Body fat distribution was related to early pubertal endocrine activity. Body fat mass was negatively related to the rate of pubertal development toward menarche, but no clear indications for an endocrine-related process is found. We conclude that onset of puberty and menarche are not parallel pubertal events, and that early pubertal plasma E1, E2 and androstenedione levels are predictors for the rate of pubertal development toward menarche. We propose that the control of the onset of puberty and maturation of the hypothalamic-pituitary gonadal axis, with regard to negative feedback control, are at least partially independent. This induces on the average a "catch up" pubertal maturation in girls with a late onset of puberty.

Body fat mass, body fat distribution, and plasma hormones in early puberty in females.

We examined whether there is a relationship between body fat mass or body fat distribution and hormonal profiles in the plasma of early pubertal girls. Thirty-five apparently healthy caucasian schoolgirls were selected for Tanner's breast development stage M2; they had all been classified as being stage M1 6 months earlier. Body fat mass had no relationship with the total plasma sex steroid concentration or gonadotropins. However, body fat mass was correlated with the fraction of testosterone that was not bound to sex hormone-binding globulin and considered the fraction available for biological activity. Body fat distribution, rather than body fat mass, was different in relation to the total concentrations of estrone, estradiol (E2), and testosterone as well as the percentage of available E2 or testosterone. Girls with fat localized predominantly on the hips had the highest levels of sex steroids and gonadotropins. It seems likely that this type of fat distribution is a result of ovarian activity. Girls with predominantly abdominal fat were also more obese and showed increased plasma levels of total E2 and a lower androgen/estrogen ratio in plasma, possibly due to increased aromatization, especially in abdominal adipose tissue. The findings suggest a reciprocal relationship among body fat distribution, plasma sex hormone levels, and availability of sex steroids in early female puberty.

Dietary habits, sexual maturation, and plasma hormones in pubertal girls: a longitudinal study.

The relationships between dietary components and physical or hormonal sexual maturation in 63 pubertal girls were examined. The effects of vegetable protein and dietary fiber on breast development (B = -2.0, P less than 0.05; B = -2.6, P less than 0.05, respectively) became more pronounced in a multivariate analysis, after elimination of the linear effects of body height and energy intake. From the multivariate analysis with combinations of vegetable protein, polysaccharides, and fiber in the equation, fiber appeared to be the most important factor. The gonadotropin and estradiol plasma concentrations were higher in girls who consumed less grain fiber. We conclude that a diet rich in vegetable products, especially fiber, may affect the rate of physical and hormonal sexual maturation, possibly mediated by the hypothalamus-pituitary-gonad system.

Endometrial cancer in a cohort screened for breast cancer.

In a cohort of 25,000 women aged 40-65 years at intake in a periodic screening programme for breast cancer the occurrence of endometrial cancer was studied during a follow-up period of up to 18 years. The authors examined whether they could confirm the existence of a number of relationships between this cancer and some reproductive and anthropometric factors which had been found in several case-control studies and in a few cohort studies. A comparison was made between 147 cases of endometrial cancer occurring during the period of follow-up and a random sample of 900 women taken from the cohort (334 being premenopausal and 566 postmenopausal on the day of intake). An inverse relationship between number of children and endometrial cancer risk was found in the older group; the excess risk among nulliparous women was stronger in married than in single women. Late age at menopause (after age 52) was associated with increased risk. Use of oestrogenic drugs on day of intake for alleviating perimenopausal complaints (reported in 8% of women) increased risk, especially in the younger group. Body weight was positively associated with risk (with an odds ratio of 4 in those over 80 kg) among postmenopausal women. Because there appeared to be a moderately strong risk associated with tall height, especially among postmenopausal women, Quetelet's index performed less well than body weight itself. Nevertheless, subscapular and triceps skinfold thicknesses confirmed an effect of fatness on risk. These results are discussed with reference to other epidemiological studies, in particular two cohort studies from Norway. The effect of height, convincingly shown by Tretli and Magnus in 1990, is explained in a way which does not assume causal mechanisms operating at a young age: absolute fat mass rather than relative weight is regarded as the main determinant of risk in postmenopausal women.

A comparative study of risk factors for hyperplasia and cancer of the endometrium.

A cohort study has been carried out to investigate risk factors for cancer as well as hyperplasia of the endometrium. Over the 13 years for which we followed 25,000 women aged 40-65 (who took part in a population-based screening programme for breast cancer), 111 cases of endometrial cancer and 109 cases of endometrial hyperplasia were diagnosed. A comparison of the outcome between the two disease entities revealed that large body weight among postmenopausal women and the use of oestrogenic drugs at all ages were risk factors for both cancer and hyperplasia of the endometrium. However, reproductive histories and premenopausal steroid profiles differed. Steroid excretion determinations in urine samples collected years before diagnosis provided further evidence in favour of the hypothesis of unopposed action of oestrogens in the aetiology of endometrial cancer. In women who were to develop endometrial hyperplasia or cancer the obesity-oestrogen relationship was stronger than in those who remained free of endometrial disease during the period of follow-up. The possible significance of differences in aromatase activity among the obese is considered.

The onset of puberty and the availability of sex steroids in female gymnasts.

Plasma levels of gonadotropins, (sex) steroids, SHBG bound fractions of 17ß-estradiol (E2) and testosterone (T), and sex-hormone-binding globulin (SHBG) were compared at the onset of puberty in female gymnasts (n = 7) and two groups of schoolgirls with similar anthropometric characteristics. Ten schoolgirls were matched to the gymnasts on the basis of a sum of skinfolds and the waist/hip ratio, while 12 other girls were selected on the basis of the stature and bicristal breadth of the female gymnasts. All girls were in the second stage of breast development (M2) and were classified as being in the first stage (M1) 6 months earlier. When female gymnasts were compared to nonathletic schoolgirls with similar physical characteristics at the same stage of early puberty, the former had significantly lower levels of plasma luteinizing hormone (LH), total and available E2, and T. In conclusion, female gymnasts have significantly lower LH, E2, and T plasma levels than nonathletic schoolgirls in early puberty. The E2 and T plasma levels in early female gymnasts are not related to the individual physical characteristics, i.e., fat mass, short stature, or small bicristal breadth. © 1993 Wiley-Liss, Inc.

Volumetric measurements of bone mineral density of the lumbar spine: comparison of three geometrical approximations using dual-energy X-ray absorptiometry (DXA)

Measurements of bone mineral density using dual-energy X-ray absorptiometry (DXA) gives area values (g cm-2) rather than true volumetric values (g cm-3). To calculate the vertebral volume using planar postero-anterior and lateral DXA values, several different geometrical approximations were used: cubic, cylindrical with a circular cross-section and cylindrical with an elliptical cross-section. The aim of this study was to compare these geometrical approximations with each other and with a reference standard, defined as the volume found on a computed tomographic (CT) scan. L2 and L3 were evaluated in a phantom study. Volume approximations by the cube or cylinder with circular cross-section geometry showed more than a 50% overestimation (range 54-74%). However, the elliptical cylinder approach showed very good agreement: 2.1% and 1.2% for L2 and L3, respectively, when compared to the CT volumes. In addition, we performed four patient studies with both CT and DXA to evaluate the elliptical cylinder estimate in a clinical setting. For L2 and L3, the mean relative difference was less than 2%. We conclude that the elliptical cylinder approach results in the most accurate bone volume estimates in both the phantom and patients.

Changes in body composition during 12 months after discontinuation of growth hormone therapy in young adults with growth hormone deficiency from childhood.

Little is known about the velocity of changes in body composition after discontinuation of GH therapy at final height. The aim of this study was to describe the changes of fat distribution in male and female GH deficient young adults during the first year after discontinuation of GH therapy. Ten Dutch GH deficient young adults who had reached final height were retested and confirmed to be still GH deficient. These preliminary results demonstrate that the greatest gain in subcutaneous fat, measured by skinfold thickness, was observed in the first 3 months after GH withdrawal. Intra-abdominal fat was measured by CT scan at 0 and 12 months study time. The mean gain in intra-abdominal fat after 12 months was dramatically high (48%). We conclude that the subcutaneous and intra-abdominal fat mass increased dramatically in young GH deficient Dutch adults with GH deficiency, after discontinuation of therapy, especially in the first three months. This indicates that GH therapy should be restarted as soon as possible, after reconfirming the diagnosis of GH deficiency in adults. This will reduce the risk for these patients of diseases associated with overweight, such as cardiovascular diseases.

Validation of transrectal ultrasonographic volumetry for orthotopic prostate tumours in mice.

Orthotopic human prostate tumour models in athymic nude mice are regarded as being most suitable for fundamental and pre-clinical research on prostate cancer. The anatomic localization of the tumour in the pelvis, however, provides little possibility for monitoring tumour growth or regression. To assess time-related changes in orthotopic tumour volume, we applied transrectal ultrasonography (TRUS) to the murine prostate. This technique has the advantages of allowing accurate monitoring of tumours during therapeutic manipulations and a reduction of animal use due to a reduction of sacrificing endpoints. To validate the TRUS method, the mouse prostate reconstitution model, RM-9, and the prostate-specific antigen (PSA) producing human prostate cancer xenograft PC-346 were used. Volumetric calliper measurements were performed with a 30 MHz ultrasound probe designed for intra-arterial use in humans. Tumour weight, determined at various time-points, was found to be closely related to actual tumour weight (R = 0.99) and, in the PC-346 model, to the level of PSA in the plasma. Furthermore, the interobserver variation for TRUS was low for tumours above 50 mg. Thus, TRUS for murine prostate tumours proves to be an accurate, reproducible and sensitive method.

Improving radiopeptide pharmacokinetics by adjusting experimental conditions for bombesin receptor-targeted imaging of prostate cancer.

AIM: Prostate cancer (PC) is a major health problem. The Gastrin-Releasing Peptide Receptor (GRPR) offers a promising target for staging and monitoring of PC since it is overexpressed in PC and not in normal prostatic tissue. To improve receptor-mediated imaging we investigated the impact of various experimental conditions on pharmacokinetics using the Indium-111 labelled bombesin (BN) analogue AMBA. Besides frequently used androgen-resistant PC-3 also the clinically more relevant androgen sensitive VCaP celline was used as human PC xenograft in nude mice. METHODS: Non-purified [111In]AMBA was compared with HPLC-purified [111In]AMBA. Effect of specific activity was studied administering 0.1MBq [111In]AMBA supplemented with different amounts of AMBA (1-3000pmol). GRPR was saturated with Tyr4-BN 1 and 4h prior to injection of [111In]AMBA. RESULTS: GRPR-positive tissue showed a significant 2 to 3-fold increase in absolute uptake after HPLC-purification while keeping a stable tumor-to-pancreas ratio. Lowering specific activity resulted in decline in uptake to 43% in tumor, 49% in kidney and 92% in pancreas between 10 and 3000 pmol. Tumor-to-pancreas ratio improved six-fold from 0.1±0 after 10 pmol up to 0.6±0.2 after 3000 pmol (P<0.01). When saturating GRPR 4h prior to [111In]AMBA injection tumor-to-pancreas ratio improved from 0.10±0.3 to 0.22±0.2 (P<0.01) and tumor-to-kidney ratio increased from 0.92±0.16 to 3.45±0.5 (P<0.01). CONCLUSION: Besides specific peptide characteristics also the experimental conditions, such as HPLC-purification, variations in specific activity and saturation of the GRPR prior to [111In]AMBA administration essentially affect radiopeptide pharmacokinetics. Experimental conditions therefore need to be carefully selected in order to compose ideal standardised protocols for optimal targeting.

A transductionally retargeted adenoviral vector for virotherapy of Her2/neu-expressing prostate cancer.

The efficacy of adenovirus (Ad)-based gene therapy of solid tumors, such as prostate cancer, is limited. One of the many problems is that the virus infects many different cell types in the body, resulting in high toxicity, whereas the target cancer cells are often less prone to wild-type Ad infection. Our aim was to develop genetically de- and retargeted Ad vectors to reduce off-target effects and increase target infection for prostate cancer. We have previously reported an Ad5 vector specific for the cancer-associated receptor Her2/neu, created by inserting Her2/neu-reactive Affibody(®) molecules (ZH) into the HI loop of a coxsackievirus and adenovirus receptor binding-ablated fiber (Ad[ZH/1]). In addition to virus retargeting to Her2/neu, this virus was further modified from wild-type Ad by changing the RGD motif in the penton base to EGD and by substitution of the KKTK motif in the third shaft repeat to RKSK, resulting in the vector Ad[ZH/3]. The ZH-containing vectors could be produced to high titers and were specific for their target, resulting in efficient infection and killing of Her2/neu-positive androgen-dependent PC346C prostate cancer cells in vitro. Here we show that the oncolytic Ad[ZH/3] vector significantly prolonged survival time and reduced serum prostate-specific antigen levels in an orthotopic prostate tumor model in nude mice to the same extent as wild-type Ad5. Our results show that Her2/neu targeting using Ad-based vectors for prostate cancer is feasible and may serve as a basis for the development of gene therapy of human prostate cancer as well as other Her2/neu-expressing cancers.

Disconnecting the yin and yang relation of epidermal growth factor receptor (EGFR)-mediated delivery: a fully synthetic, EGFR-targeted gene transfer system avoiding receptor activation.

The epidermal growth factor receptor (EGFR) is upregulated within a high percentage of solid tumors and hence is an attractive target for tumor-targeted therapies including gene therapy. The natural EGFR ligand epidermal growth factor (EGF) has been used for this purpose, despite the risk of mitogenic effects due to EGFR activation. We have developed a fully synthetic, EGFR-targeted gene delivery system based on PEGylated linear polyethylenimine (LPEI), allowing evaluation of different EGFR-binding peptides in terms of transfection efficiency and EGFR activation. Peptide sequences directly derived from the human EGF molecule enhanced transfection efficiency with concomitant EGFR activation. Only the EGFR-binding peptide GE11, which has been identified by phage display technique, showed specific enhancement of transfection on EGFR-overexpressing tumor cells including glioblastoma and hepatoma, but without EGFR activation. EGFR targeting led to high levels of cell association of fluorescently labeled polyplexes after only 30 min of incubation. EGF pretreatment of cells induced enhanced cellular internalization of all polyplex types tested, pointing at generally enhanced macropinocytosis. EGF polyplexes diminished cell surface expression of EGFR for up to 4 hr, whereas GE11 polyplexes did not. In a clinically relevant orthotopic prostate cancer model, intratumorally injected GE11 polyplexes were superior in inducing transgene expression when compared with untargeted polyplexes.

Clinical utility of markers of bone turnover in children and adolescents.

We review the past years' literature on biochemical markers of bone turnover. A general introduction to markers of bone formation and bone resorption is followed by reference values of bone turnover in neonates, infants, and prepubertal and pubertal children. We describe intervention with calcium and physical activity in healthy children and bone turnover in patients. The predictive value of a single measurement of bone markers in individuals is poor, due to the large biologic intraperson variation for bone markers in general. Serious osteoporosis can be diagnosed by the combined results of measurement of several bone formation and resorption markers.

Immunoadsorption assay for bone alkaline phosphatase compared with wheat germ agglutinin precipitation assay in serum from (pre)pubertal girls.

The performance characteristics of two bone alkaline phosphatase (ALP; EC 3.1.3.1) assays, a wheat germ agglutinin (WGA) precipitation assay and a new immunoadsorption assay (IAA), were compared. The within- and between-run imprecision of the IAA (3.6-4.2% and 3.6-7.7%) was comparable with that of the WGA assay. The mean cross-reactivity with liver ALP appeared to be 4% in the WGA assay and 11% in the IAA. The reference ranges in a group of 155 healthy Caucasian (pre)pubertal schoolgirls were: 149-401 U/L (total ALP, 30 degrees C), 105-349 U/L (bone ALP, 30 degrees C, WGA assay), and 58-205 U/L (bone ALP, 25 degrees C, IAA). Comparison of the WGA assay (x) with the IAA (y) demonstrated a correlation coefficient of 0.95 [Deming regression equation: y = (0.56 +/- 0.01)x + (2.0 +/- 1.5); Sy[symbol: see text]x = 5.3 U/L]. Correlation studies of the WGA assay and the IAA results with total ALP demonstrated r = 0.98 and 0.96, respectively.

Neuroendocrine differentiation in human prostatic tumor models.

Neuroendocrine (NE) cells can be identified in benign and malignant prostatic epithelia. Factors regulating their presence and their functions are poorly understood, mainly due to a lack of suitable experimental models. Fifteen in vitro and in vivo prostatic cancer tumor models, including a number of newly established in vivo models, were studied immunohistochemically for the presence of NE cells under different hormonal conditions. None of the in vitro models (PC-3, DU 145, LNCaP, and TSU) contained NE cells. Five of the seven xenograft models established at this laboratory contained NE cells. In three of these, NE cells were found only in the initial mouse passages. In the other two (PC-295 and PC-310), the NE phenotype was stable. NE features were confirmed by transmission electron microscopy and by Western analysis of chromogranin A expression. Immunohistochemical double-labeling experiments confirmed that NE cells in prostate cancer are post-mitotic (no Ki-67 expression) and do not express the androgen receptor. In the PC-295 and PC-310 models, short-term androgen withdrawal resulted in a rapidly increased number of NE cells. A time course experiment with PC-295-bearing mice strongly suggests that this increase occurred by induction of NE differentiation rather than by rapid proliferation and subsequent differentiation or selective persistence. In conclusion, these models are suitable to resolve fundamental questions with regard to the presence and functions of NE cells in human prostate cancer.

Bone metabolism markers and bone mass in healthy pubertal boys and girls.

OBJECTIVE: During puberty, bone growth and mineralization as well as bone turnover increase dramatically. The relation between height velocity and bone turnover is already known, but there are few studies in which both bone metabolism markers and bone mass throughout puberty have been measured. DESIGN: Semi-longitudinal study. In 155 healthy boys (12.0 +/- 1.5 years; range 8.8-15.7 years) and 151 healthy girls (11.2 +/- 1.6 years; range 8.2-14.0 years) markers of bone formation and bone resorption were measured as well as sex steroids, IGF-1 and IGF-BP3, together with bone mineral content (BMC) and bone mineral density (BMD) of the lumbar spine, femur and total body during puberty. All bone measurements were repeated after 1 year. RESULTS: BMC and BMD increased throughout puberty in both sexes. Bone turnover markers increased significantly until maximum values were reached at stage G4 in boys and stage B3 in girls. Height velocity (HV) had a similar changing pattern. Sex steroids and IGF-1 increased and reached adult values at pubertal stage 4. The correlations between bone metabolism markers and BMC were highly significant in boys, while correlations between bone metabolism markers and the increase in BMC over 1 year were significant in both sexes, as was observed for the correlations with HV. CONCLUSIONS: Our data suggest that bone metabolism markers are good predictors of bone mass in boys and of bone mass increase in both sexes. In early puberty, sex steroids stimulate the pubertal growth spurt in conjunction with GH and IGF-1. The fast increase in height gives rise to an increase in bone turnover and bone mineral apposition. It is known that at the end of puberty high levels of oestradiol inhibit chondrocyte proliferation. This leads to a decline in height velocity and bone turnover. Bone mass still increases under the influence of sex steroids and IGF-1. The data in our study confirm previous reports that markers of bone turnover relate positively to height velocity.

Plasma lipids and apoproteins, body fat distribution and body fatness in early pubertal children.

Besides body fatness, the body fat distribution is associated with coronary risk in adults, but little has been reported on this aspect in children. This study describes body fatness, body fat distribution (waist-to-hip ratio, WHR) and the plasma lipid and apoprotein profile (TC, HDL-C, LDL-C, TG, apo A-I and apo B) in 60 boys (age 10.8 +/- 0.1 year; mean +/- s.e.m.) and 64 girls (age 10.2 +/- 0.1 year), all caucasian. To avoid interference by the large changes in plasma sex hormone levels during puberty, only pre- and early pubertal children (Tanner stages of genital c.q. breast development 1 or 2) participated. Physical and sports activity was scored in hours per week using a questionnaire. The boys were taller than the girls (146.2 +/- 0.7 vs 143.2 +/- 0.9 cm; ANOVA, P less than or equal to 0.05) and their WHR was larger (0.88 +/- 0.01 vs 0.83 +/- 0.01; ANOVA, P less than or equal to 0.05). The boys spent 8.0 +/- 0.4 hours weekly on physical and sports activities, the girls 5.5 +/- 0.3 (ANOVA, P less than or equal to 0.05). The plasma lipid and apoprotein profiles were similar in both groups. Body fatness was significantly associated with the lipid and apoprotein profile, although in different ways in boys and girls. In boys there was a relationship with TG (r = 0.49), with apo B (r = 0.33) and with the apo A-I to apo B ratio (r = -0.24); in girls with TG (r = 0.25), HDL-C (r = -0.39), apo A-I (r = -0.28) and with the HDL-C to TC ratio (r = -0.31); P less than 0.05 for all correlations. A regional component of the subcutaneous fatmass, assessed by the partial correlations of the individual skinfold thicknesses with the plasma lipid and apoprotein profile after controlling for body fatness, was lacking in these early and prepubertal children. The WHR was associated with TC (r = 0.35), LDL-C (r = 0.32), apo B (r = 0.36) and with apo A-I/apo B (r = -0.34) in the girls after controlling for body fatness. Although closer investigation into the validity of the WHR as a measure of fat distribution in children is needed, the tentative conclusion is that in pre- and early pubertal girls the WHR has an impact on the plasma lipid and apoprotein profile similar to that seen in adults. It is suggested that in boys these relationships develop later in puberty.