In vivo and in vitro properties of STX2484: a novel non-steroidal anti-cancer compound active in taxane-resistant breast cancer.

BACKGROUND: STX2484 is a novel non-steroidal compound with potent anti-proliferative activity. These studies aimed to identify STX2484's mechanism of action, in vivo efficacy and activity in taxane-resistant breast cancer models. METHODS: Effects of STX2484 and paclitaxel on proliferation, cell cycle and apoptosis were assessed in vitro in drug-resistant (MCF-7(DOX)) and non-resistant cells (MCF-7(WT)). STX2484 efficacy in ßIII tubulin overexpression in MCF-7 cells was also determined. Anti-angiogenic activity was quantified in vitro by a co-culture model and in vivo using a Matrigel plug assay. An MDA-MB-231 xenograft model was used to determine STX2484 efficacy in vivo. RESULTS: STX2484 is a tubulin disruptor, which induces p53 expression, Bcl2 phosphorylation, caspase-3 cleavage, cell cycle arrest and apoptosis. In addition, STX2484 is a potent anti-angiogenic agent in vitro and in vivo. In breast cancer xenografts, STX2484 (20 mg kg(-1) p.o.) suppressed tumour growth by 84% after 35 days of daily dosing, with limited toxicity. In contrast to paclitaxel, STX2484 efficacy was unchanged in two clinically relevant drug-resistant models. CONCLUSIONS: STX2484 is an orally bioavailable microtubule-disrupting agent with in vivo anti-angiogenic activity and excellent in vivo efficacy with no apparent toxicity. Crucially, STX2484 has superior efficacy to paclitaxel in models of clinical drug resistance.

Molecular characterization of avian astroviruses.

Astroviruses are frequently associated with enteric diseases in poultry, being isolated from cases of runting-stunting syndrome (RSS) of broiler chickens, poult enteritis complex (PEC), and poult enteritis mortality syndrome (PEMS) of turkeys. Currently, five types of avian astrovirus have been identified: turkey astroviruses 1 and 2 (TAstV-1, TAstV-2), avian nephritis virus (ANV), chicken astrovirus (CAstV) and duck astrovirus (DAstV). The objective of this study was to molecularly characterize the different types of avian astroviruses circulating in commercial poultry. Sequence analysis of a region of ORF2, which encodes the capsid precursor protein associated with serotype and viral pathogenesis, revealed extensive variation in amino acid sequence within each subtype: TAstV-2 (81.5%-100%), ANV (69.9%-100%), and CAstV (85.3%-97.9%). However, this region was more conserved in TAstV-1's (96.2%-100%). Furthermore, a novel astrovirus was detected in chicken samples and found to be <64% similar to ANV and <30.6% similar to CAstV. The results of this study underline the great genetic variability of avian astroviruses and indicate that there are most likely multiple serotypes of each avian astrovirus circulating in commercial poultry.

17ß-hydroxysteroid dehydrogenase inhibitors.

17ß-hydroxysteroid dehydrogenases (17ß-HSDs) are enzymes which require NAD(P)(H) for activity and are responsible for reduction or oxidation of hormones, fatty acids and bile acids in vivo, regulating the amount of the active form which is available to bind to its receptor. Fifteen 17b-HSDs have been identified to date, and with one exception, 17ß-HSD Type 5 (17ß-HSD5), an aldo-keto reductase, they are all short chain dehydrogenases/reductases. Although named as 17ß-HSDs, reflecting the major redox activity at the 17ß-position of the steroid, overall homology between the enzymes is low and the activities of these fifteen enzymes vary, with several of the 17ß-HSDs able to reduce and / or oxidise multiple substrates at various positions. These activities are involved in the progression of a number of diseases, including those related to steroid metabolism. Many groups are now working on inhibitors specific for several of these enzymes for the treatment of steroid-dependent diseases, including breast and prostate cancer, and endometriosis, with demonstrable efficacy in in vivo disease models, although none have yet reached clinical trials. In this review the recent advances in the development of specific inhibitors of the 17ß-HSD1, 3 and 5 enzymes as targets for the treatment of these diseases and the models used for their evaluation will be discussed.

Parvovirus-associated cerebellar hypoplasia and hydrocephalus in day old broiler chickens.

Cerebellar hypoplasia and hydrocephalus were identified in day old broiler chickens showing nervous signs, impaired mobility, and diarrhea. At postmortem examination, brains of chickens were misshapen and cerebellums were smaller than normal. Microscopically, cerebellar folia were reduced in size and irregularly shaped, and the ventricles were widely distended. Affected cerebellums had focal areas along the base of folia where the internal granular cell layer had been lost, and Purkinje cells were disorganized and located within the molecular layer. Parvovirus DNA was detected by polymerase chain reaction in three of nine brains with oligonucleotide primers designed for amplification of chicken and turkey parvoviruses. On the basis of phylogenetic analyses, the detected virus was most closely related to chicken parvoviruses. These findings suggest that a chicken parvovirus might cause a neurologic disease of young chickens characterized by cerebellar hypoplasia and hydrocephalus; however, its role as the cause of the disease remains to be confirmed.

BCRP expression does not result in resistance to STX140 in vivo, despite the increased expression of BCRP in A2780 cells in vitro after long-term STX140 exposure.

The anti-proliferative and anti-angiogenic properties of the endogenous oestrogen metabolite, 2-methoxyoestradiol (2-MeOE2), are enhanced in a series of sulphamoylated derivatives of 2-MeOE2. To investigate possible mechanisms of resistance to these compounds, a cell line, A2780.140, eightfold less sensitive to the 3,17-O,O-bis-sulphamoylated derivative, STX140, was derived from the A2780 ovarian cancer cell line by dose escalation. Other cell lines tested did not develop STX140 resistance. RT-PCR and immunoblot analysis demonstrated that breast cancer resistance protein (BCRP) expression is dramatically increased in A2780.140 cells. The cells are cross-resistant to the most structurally similar bis-sulphamates, and to BCRP substrates, mitoxantrone and doxorubicin; but they remain sensitive to taxol, an MDR1 substrate, and to all other sulphamates tested. Sensitivity can be restored using a BCRP inhibitor, and this pattern of resistance is also seen in a BCRP-expressing MCF-7-derived cell line, MCF-7.MR. In mice bearing wild-type (wt) and BCRP-expressing tumours on either flank, both STX140 and mitoxantrone inhibited the growth of the MCF-7wt xenografts, but only STX140 inhibited growth of the MCF-7.MR tumours. In conclusion, STX140, a promising orally bioavailable anti-cancer agent in pre-clinical development, is highly efficacious in BCRP-expressing xenografts. This is despite an increase in BCRP expression in A2780 cells in vitro after chronic dosing with STX140.

Myocarditis associated with reovirus in turkey poults.

Myocarditis associated with reovirus was diagnosed in 17-day-old, male turkey poults, based on virus isolation, reverse transcriptase-polymerase chain reaction (RT-PCR), demonstration of reovirus antigen in the cytoplasm of mononuclear inflammatory cells and myocytes in the heart by immunohistochemistry (IHC), and reovirus particles in the endoplasmic reticulum of myocytes by transmission electron microscopy (TEM). Clinical signs in the poults included anorexia, growth depression, and increased mortality. Gross lesions in the six poults examined were increased pericardial fluid, mild-to-moderate dilation of right ventricles, pale-yellow myocardium, and ascites. Other lesions in a few birds included mild pulmonary edema, congestion, and pale serosa of the small intestine that had watery contents in their lumens. Microscopically, in the heart, there was mild-to-severe necrosis of myocytes and infiltration of primarily lymphocytes mixed with a few heterophils, macrophages, and occasionally, plasma cells and multinucleated giant cells. There was mild-to-moderate lymphoid depletion in the bursa of Fabricius. Reovirus was isolated from the heart of the turkey poults in chicken-embryo liver cells and was confirmed by RT-PCR, IHC, and TEM. A retrospective search of the laboratory database for cases of myocarditis associated with reovirus in turkeys revealed that this condition has occurred sporadically in California turkey flocks since 1991. This is the first documentation of myocarditis in turkey poults associated with reovirus.

The therapeutic potential of a series of orally bioavailable anti-angiogenic microtubule disruptors as therapy for hormone-independent prostate and breast cancers.

Therapies for hormone-independent prostate and breast cancer are limited, with the effectiveness of the taxanes compromised by toxicity, lack of oral bioavailability and drug resistance. This study aims to identify and characterise new microtubule disruptors, which may have improved efficacy relative to the taxanes in hormone-independent cancer. 2-Methoxy-3-O-sulphamoyl-17beta-cyanomethyl-oestra-1,3,5(10)-triene (STX641), 2-methoxy-3-hydroxy-17beta-cyanomethyl-oestra-1,3,5(10)-triene (STX640) and 2-methoxyoestradiol-3,17-O,O-bis-sulphamate (STX140) were all potent inhibitors of cell proliferation in a panel of prostate and breast cancer cell lines. STX641 and STX640 significantly inhibited tumour growth in the MDA-MB-231 xenograft model. STX641 inhibited both in vitro and in vivo angiogenesis. Despite good in vivo activity, STX641 was not as potent in vivo as STX140. Therefore, STX140 was evaluated in the prostate hormone-independent PC-3 xenograft model. STX140 had superior efficacy to docetaxel, 2-MeOE2 and bevacizumab. In contrast to vinorelbine, no significant toxicity was observed. Furthermore, STX140 could be dosed daily over a 60-day period leading to tumour regression and complete responses, which were maintained after the cessation of dosing. This study demonstrates that STX641 and STX140 have considerable potential for the treatment of hormone-independent breast and prostate cancer. In contrast to the taxanes, STX140 can be dosed orally, with no toxicity being observed even after prolonged daily dosing.

Pathology and virus tissue distribution of Turkey origin reoviruses in experimentally infected Turkey poults.

The pathogenesis of 4 isolates of turkey-origin reovirus (NC/SEP-R44/03, NC/98, TX/98, and NC/85) and 1 chicken-origin reovirus (1733) was examined by infecting specific pathogen free (SPF) poults. These turkey-origin reovirus (TRV) isolates were collected from turkey flocks experiencing poult enteritis and are genetically distinct from previously reported avian reoviruses. Microscopic examination of the tissues collected from the TRV-infected poults revealed different degrees of bursal atrophy characterized by lymphoid depletion and increased fibroplasia between the bursal follicles. To understand the relationship between virus spread and replication, and the induction of lesions, immunohistochemical staining (IHC) for viral antigen, in situ hybridization (ISH) for the detection of viral RNA, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of apoptosis in affected tissues was performed. Both IHC and ISH revealed viral antigen and RNA in the surface epithelial cells of the bursa, in macrophages in the interstitium of the bursa and, to lesser degree, in splenic red pulp macrophages and intestinal epithelial cells. Increased apoptosis of bursal lymphocytes and macrophages was observed at 2 and 5 days postinoculation. No lesions were found in tissues from poults inoculated with the virulent chicken-origin strain, however viral antigen was detected in the bursa and the intestine. Although all TRVs studied displayed similar tissue tropism, there were substantial differences in the severity of the lesions produced. Poults inoculated with NC/SEP-R44/03 or NC/98 had moderate to severe bursal atrophy, whereas poults inoculated with TX/98 or NC/85 presented a mild to moderate bursal lymphoid depletion. The lymphoid depletion observed in the bursa appears to be the effect of an indirectly induced apoptosis and would most likely result in immune dysfunction in poults infected with TRV.

The regulation and inhibition of 17beta-hydroxysteroid dehydrogenase in breast cancer.

17Beta-hydroxysteroid dehydrogenase Type 1 (17beta-HSD1) has a pivotal role in regulating the synthesis of oestradiol (E2) within breast tumours. In whole body studies in postmenopausal women with breast cancer the conversion of oestrone (E1) to E2 (4.4+/-1.1%) was much lower than the inactivation of E2 to E1 (17.3+/-5.0%). In contrast, an examination of in vivo oestrogen metabolism within breast tumours revealed that whereas little metabolism of E2 occurred, E1 was converted to E2 to a much greater extent in malignant (48+/-14%) than in normal (19+/-6%) breast tissue. Findings from these studies originally suggested that oestrogen metabolism within breast tumours may differ from the mainly oxidative direction found in most other body tissues and that the activity of 17beta-HSD1 might be regulated by tumour-derived factors. Several growth factors (e.g. IGF-I, IGF-II) and cytokines (e.g. IL-6, TNFalpha) have now been identified which can markedly stimulate the activity of 17beta-HSD1 and such a mechanism may account for the high concentrations of E2 found in most breast tumours. Cells of the immune system, which can infiltrate breast tumours, are thought to be a major source of the growth factors and cytokines which can modulate 17beta-HSD1 activity. Given the central role that 17beta-HSD1 has in regulating breast tumour E2 concentrations the development of potent inhibitors of this enzyme has recently attracted considerable attention. Our initial studies in this area explored the use of derivatives of E1 as inhibitors, with 2-ethyl- and 2-methoxy E1 being found to inhibit 17beta-HSD1 activity in T-47D breast cancer cells by 96+/-2 and 91+/-1% respectively at 10 microM, but with a lack of specificity. Using the E1 scaffold a number of potent, selective 17beta-HSD1 inhibitors have now been identified including E1- and 2-ethyl-E1 containing a side chain with a m-pyridylmethylamidomethyl functionality extending from the 16beta position of the steroid nucleus. At 10 microM these compounds both inhibited 17beta-HSD1 activity by >90%, however some inhibition of 17beta-HSD2 activity was exhibited by the E1 derivative (25%) but not the 2-ethyl analogue. It is now apparent that 17beta-HSD1 activity contributes to the high E2 concentrations found in most breast tumours. The identification of potent, selective novel 17beta-HSD1 inhibitors will allow their efficacy to be tested in in vitro and in vivo studies.

The nature of inhibition of steroid sulphatase activity by tibolone and its metabolites.

Tibolone is used for hormone replacement therapy and acts in a tissue-specific manner being oestrogenic on CNS and bone but not on breast tissues or endometrium. The ability of tibolone and its metabolites to inhibit steroid sulphatase (STS) activity has a crucial role in regulating its tissue-specific effects. In this study, we have examined the ability of tibolone and its non-sulphated and sulphated metabolites to inhibit STS activity in different enzyme preparations and in intact cells. For this, we have used an 'extracellular' method, which measures the amount of product released into culture medium, and an 'intracellular' method, which assesses the extent of product formation within cells. In addition, the nature by which tibolone and some of its metabolites inhibit STS activity was investigated using intact cells and an enzyme kinetic method. In MCF-7 and T47D breast cancer cells and JEG-3 choriocarcinoma cells, which have high STS activity, tibolone and its metabolites were relatively potent inhibitors of STS activity (33-57% inhibition at 10 microM) using the extracellular assay method. In HOS-TE-85 osteoblast-like cells, tibolone and its Delta-4 metabolite were relatively inactive whereas the 3alpha/3beta-hydroxy metabolites and their sulphated conjugates inhibited activity by 39-55%. When STS activity was assessed in HOS-TE-85 cells using an 'intracellular' method tibolone and its 3beta-hydroxy metabolite were inactive. Pre-treatment of breast cancer cells and JEG-3 cells, and removal of drugs prior to assaying for STS activity, revealed that in these cells tibolone and its metabolites were acting mainly as reversible inhibitors. This finding was confirmed in an enzyme kinetic study to measure concentration-dependent STS inhibition. In HOS-TE-85 cells, pre-treatment of cells and removal of compounds before assaying for remaining STS activity indicated that some tibolone metabolites appeared to stimulate STS activity. Possible mechanisms by which this might occur are discussed but, if confirmed, this could contribute to the positive oestrogenic effects that tibolone has on bone.

The folded protein modules of the C-terminal G3 domain of aggrecan can each facilitate the translocation and secretion of the extended chondroitin sulfate attachment sequence.

Aggrecan is a multidomain proteoglycan containing both extended and folded protein modules. The C-terminal G3 domain contains a lectin-like, complement regulatory protein-like, and two alternatively spliced epidermal growth factor-like modules. It has been proposed that the lectin module alone has a necessary role in the intracellular translocation and secretion of proteins expressed containing G3. Constructs containing human aggrecan G3 together with 1155 bases of the adjacent chondroitin sulfate attachment region (CS-2) were prepared with different combinations and deletions of the protein modules and transfected into mammalian cells of monkey or hamster origin. The results showed that the products containing only the unfolded protein sequences (CS-2 with or without the C-terminal tail sequence) were translated and accumulated intracellularly but were not secreted. In contrast the constructs containing any of the folded protein modules and the extended CS-2 region were translated and secreted from the cells. The results show that the lectin module was not unique in facilitating the intracellular translocation and secretion of the G3 domain. The conservation of G3-like domains within the aggrecan family of proteoglycans may therefore result from their participation in other extracellular functions.

Effect of topical butyrate on rectal epithelial kinetics and mucosal enzyme activities.

1. This study aimed to determine the effect of luminal butyrate on proliferative kinetics, a differentiation marker (alkaline phosphatase), and a molecule that controls cell-substratum adhesion (urokinase) in histologically normal human rectal mucosa. 2. Ten subjects with a colonoscopically normal colon (seven had previous adenomas) were given either butyrate or saline enemas for 4 days in a double-blind cross-over manner. Rectal biopsies were taken before and after each course of enemas. Epithelial proliferative kinetics were measured immunohistochemically using antibodies to proliferating cell nuclear antigen. Urokinase and alkaline phosphatase activities were measured spectrophotometrically in biopsy homogenates. 3. Both saline and butyrate enemas were well tolerated and induced no histological change except for a significant increase in crypt length (P < 0.05). The number of proliferating cells per crypt also increased significantly after butyrate (P = 0.018). 4. Compared with saline enemas, butyrate did not affect kinetic indices nor alkaline phosphatase activities. However, mucosal urokinase activities were significantly lower in butyrate-treated patients (9.5 +/- 2.0 i.u./g) than in saline-treated patients (12.8 +/- 2.0 i.u./g; P = 0.045). 5. Delivering of extra butyrate to the distal colon in healthy subjects may stabilize cell-substratum adhesion in surface epithelium and therefore offer a potential mechanism by which elevating distal colonic luminal butyrate concentrations might be beneficial in patients with colitis or hyperproliferative large bowel epithelium.

Effect of growth hormone on height, weight, and body composition in Prader-Willi syndrome.

AIMS: To evaluate the effect of the administration of growth hormone on stature, body weight, and body composition in children aged between 4 and 10 years with Prader-Willi syndrome. METHODS: Height, weight, and skinfold thickness were recorded in 25 children using standard anthropometric techniques at recruitment, and six months later, shortly before the start of daily subcutaneous injections of growth hormone. Body composition was assessed via a measurement of total body water using stable isotopes. Measurements were repeated at the end of the six months of growth hormone administration. Measurements of height, weight, and skinfold thickness were expressed as standard deviation scores (SDSs). RESULTS: There was a significant reduction in the percentage of body fat after growth hormone treatment; height velocity doubled during treatment; body weight did not change significantly when expressed as an SDS. Skinfold thickness at both the triceps and subscapular site decreased in absolute terms and when expressed as an SDS. CONCLUSIONS: These results indicate sufficient potential benefit to justify a more prolonged trial of growth hormone treatment and an exploration of different dosage regimens in children with Prader-Willi syndrome.

Energy intake in early infancy and childhood fatness.

OBJECTIVE: To investigate whether aspects of infant energy intake are related to fatness in early childhood. DESIGN: Longitudinal investigation of infants studied at 12 weeks and 2-3.5 y. SUBJECTS: 20 healthy infants, breast-fed or formula-fed, from the general population. MEASUREMENTS: Milk volume intake (MVI) by deuterium turnover, estimated energy intake, weaning status and body composition in infancy, body composition in childhood. RESULTS: MVI was not related to infant skinfolds or percentage fat. Weaning was inversely related to MVI (P < 0.04) at 12 weeks, and inversely related to skinfolds (P = 0.055) and fat mass (P = 0.020) in childhood. MVI and total energy intake were not related to childhood fatness. CONCLUSIONS: Early weaning was associated with a moderate reduction in childhood fatness. Two possible mechanisms are discussed. However, early infant energy intake was not an important determinant of later fatness in this population.

Investigation of the relationship between infant temperament and later body composition.

OBJECTIVE: To investigate the ability of maternally-rated infant temperament to predict fatness and activity patterns in early childhood. DESIGN: Longitudinal investigation of infants studied at 12 weeks and followed up at 2-3.5 y of age. SUBJECTS: Thirty healthy full-term infants from the general population. MEASUREMENTS: Body composition, behavioural activity and temperament at 12 weeks; anthropometry, body composition, diet and behavioural activity at follow-up. RESULTS: Infant temperament predicted later behaviour and fatness. Easily soothable infants had leaner childhood skinfold thicknesses (P < 0.02) and were more active in childhood (P < 0.025). Infant distress was also related to childhood diet composition. CONCLUSIONS: Infant temperament can predict later body composition and behaviour. Both energy intake and energy expenditure may be mechanisms by which the relationship develops.

Total energy expenditure in 9 month and 12 month infants.

OBJECTIVES: To measure the total energy expenditure of 9 and 12 month old infants and compare with current recommendations for energy intake. DESIGN: Cross-sectional study. Total energy expenditure assessed using the doubly labelled water technique over a 10 d period. Classical anthropometric measurements taken. SETTING: Community based study in and around Cambridge, UK. SUBJECTS: Twenty infants aged 9 months of age and 20 infants aged 12 months of age recruited via local hospital birth records. MAIN OUTCOME MEASURES: Total energy expenditure calculated via the doubly labelled water technique. RESULTS: Measurements of total energy expenditure was successful in 34 cases. Mean total energy expenditure was 73.5 kcal/kg, 73.2 kcal/kg, 77.1 kcal/kg and 77.6 kcal/kg in the nine month old boys, nine month old girls, twelve month old boys and twelve month old girls respectively. These measurements are approximately 17% below current recommendations (FAO/WHO/UNU, 1985) at nine months of age and 22% below at one year of age. CONCLUSION: The data are consistent with findings in younger infants and older children in that the measurements of total energy expenditure are about 20-25% below current recommendations. It is unlikely that contemporary infants are being underfed and thus more likely that changes in feeding practices and modification of infant formula composition has led to the reduction in energy intake and energy expenditure in such infants.

Measurement of total body water and urea kinetic modelling in peritoneal dialysis.

Studies of the effect of Kt/V (urea) on prediction of outcome in patients on peritoneal dialysis have shown conflicting results. We performed this study to examine the effects of the measurement of V by varying techniques on the calculation of Kt/V, using body water estimated by deuterium oxide dilution (D2O dilution) as the criterion method for estimation of V. Studies were performed in 20 peritoneal dialysis patients. Kt was calculated from 24-hour dialysate and urine collections and V estimated by D2O dilution, Watson formulae, 58% of body weight, bioelectrical impedance (BIA) and 73% of fat-free mass estimated by DEXA. V was also measured in 35 healthy controls. Hydration, expressed as body water by D2O dilution as a percentage of fat-free mass estimated by DEXA did not differ between peritoneal dialysis patients 71.0 (4.9)% and a healthy control group 71.1 (5.0)%. Mean Kt/V using D2O dilution was 2.14 (0.36). The other techniques resulted in a significantly lower Kt/V; Watson equations 2.01 (0.35), p < 0.005, BIA 1.93 (0.31), p < 0.0001, DEXA 2.06 (0.28), p < 0.05, 58% body weight 1.83 (0.38), p < 0.0001. Limits of agreement of Kt/V by the simpler techniques compared with D2O dilution [mean difference of (other techniques -D2O dilution) as % of mean values +/- 95% limits of agreement] were Watson equation -5.9 +/- 15.3%, BIA -10.1 +/- 15.5%, DEXA -3.4 +/- 13.5% and 58% body weight -9.9 +/- 23.5%. Differences in Kt/V from estimates using D2O dilution were significantly negatively correlated with body fat for 58% body weight (r = -0.80, p < 0.0001) and the Watson formulae (r = -0.49, p < 0.05) but not for BIA or DEXA. We conclude that clinically significant variation in Kt/V may occur due to the estimation of V and may account for the uncertainty of the value of Kt/V as a predictor of outcome in peritoneal dialysis patients. Estimating V by BIA and DEXA did not have any benefit over the Watson formulae in terms of agreement with D2O dilution, though did avoid systematic errors related to body fat. Estimation of V as a fixed proportion of body weight is clearly inferior to the other techniques.

Four-component model of body composition in chronic renal failure comprising dual-energy X-ray absorptiometry and measurement of total body water by deuterium oxide dilution.

1. Assessment of nutrition in patients with chronic renal failure by body composition measurement techniques may be affected by variable hydration. 2. This study aimed to derive a four-component model of body composition (consisting of fat, protein, total body water and body mineral) from a combination of dual-energy X-ray absorptiometry and total body water measured by deuterium oxide dilution, allowing assessment of body protein stores without the effect of variation in hydration. Patients with chronic renal failure on haemodialysis, peritoneal dialysis and conservative treatment and a control group were studied. Patients with chronic renal failure were at an 'ideal' state of hydration on clinical assessment. 3. Hydration was defined by total body water as a percentage of fat-free mass measured by dual-energy X-ray absorptiometry, and no differences were found between chronic renal failure subgroups and control subjects (except in the female undialysed chronic renal failure subgroup). Hydration was significantly correlated with percentage total body fat in the control groups but not in patients with chronic renal failure. 4. Lean tissue measured by dual-energy X-ray absorptiometry was significantly reduced in three of the six chronic renal failure groups compared with control subjects (male and female patients on haemodialysis and female patients on peritoneal dialysis). Protein estimated from the four-component model failed to detect these abnormalities. 5. Lean tissue measured by dual-energy X-ray absorptiometry in normal subjects strongly correlated with fat-free mass measured by total body potassium in normal subjects (male r = 0.91; female r = 0.89, both P < 0.0001). The correlation of protein estimated from the four-component model with fat-free mass measured by total body potassium was far weaker in male control subjects (r = 0.51, P < 0.05) and not significant in female control subjects (r = 0.38, P not significant). In the normal subjects protein estimated from the four-component model showed a much greater variation from protein estimated by total body potassium than did protein estimated simply as 27% of dual-energy X-ray absorptiometry fat-free mass minus total body mineral. 6. Hydration in patients with chronic renal failure in whom fluid balance is believed to be normal on clinical criteria does not differ from that in normal subjects. The combined model of dual-energy X-ray absorptiometry and total body water is not a useful method for the measurement of body protein.

Measurement of total body water by bioelectrical impedance in chronic renal failure.

OBJECTIVE: Abnormalities of body hydration are common in patients with advanced chronic renal failure (CRF) and may be associated with important adverse clinical effects, even in the absence of clinical features of fluid retention or depletion. Bioelectrical impedance analysis (BIA) is a simple, non-invasive method of measuring body water content and thus could be of use in the management and study of patients with CRF. This study was performed to assess the ability of BIA to measure total body water (TBW) in patients with CRF. DESIGN: TBW was measured by two different impedance systems, with comparison of the results with TBW determined by deuterium oxide dilution (D2O). SETTING: Renal Unit, Leeds General Infirmary and Centre for Bone and Body Composition Research, University of Leeds, UK. RESULTS: The range of the 95% limits of method agreement between BIA and D2O expressed as a percentage of the mean for the group was +/- 13.4% for the RJL 101A system and +/- 15.6% for the Holtain system in controls. For the whole CRF group the limits were moderately greater than controls at +/- 17.3% (RJL) and +/- 21.9% (Holtain). Analysis of subgroups of the CRF patients showed the smallest limits in those receiving peritoneal dialysis (+/- 15.5% RJL and 18.2% Holtain). Limits were greater for patients on haemodialysis (+/- 16.0% RJL and +/- 23.8% Holtain) and undialysed patients (+/- 20.1% and +/- 23.0%). CONCLUSIONS: BIA is less accurate for the measurement of TBW in patients with CRF than in healthy subjects, though in some groups of patients the effect of CRF on the validity of this technique may be only mild. Abnormalities of reactance suggest that abnormal variability in the distribution of fluid between intra- and extracellular compartments is the major cause of reduced accuracy of TBW calculated by BIA in CRF patients.

The relationship between components of infant energy expenditure and childhood body fatness.

OBJECTIVE: To investigate whether any component of infant energy expenditure is related to fatness in early childhood, and whether infant fatness is related to childhood variables. DESIGN: Longitudinal investigation of infants studied at 12 weeks and followed up at 2.5 to 3.5 years of age. SUBJECTS: 30 healthy full-term infants selected from the general population. MEASUREMENTS: Sleeping metabolic rate, total energy expenditure, anthropometry and behaviour at 12 weeks; anthropometry, body composition and behaviour in follow-up. RESULTS: Energy expenditure at 12 weeks (minimal metabolism, total energy expenditure, energy expended on physical activity, behaviour) showed no relationship with later fatness. Infant fatness (skinfold thicknesses and percentage fat) showed in contrast a strong relationship with childhood fatness. Infant fatness also predicted childhood behaviour. CONCLUSIONS: These data do not support the theory that reduced energy expenditure in early infancy is related to later fatness. However, infant fatness influences both later fatness and activity patterns.