Search

Growth failure is a well recognised consequence of severe congenital heart disease. Total daily energy expenditure (TDEE) was investigated in eight infants with severe congenital heart disease to determine whether an increase in this parameter is an important factor in their failure to thrive, and to estimate the energy intake that would be required to allow normal growth. The infants were studied over a seven day period before surgery using the doubly labelled water method. Growth failure was evident; their mean age standardised body mass index was 80% of the expected value. Mean TDEE was 425 kJ/kg, significantly greater than in healthy infants (mean TDEE/kg SD score = +1.4; 95% confidence interval +0.27 to +2.57). In contrast, their energy intake was only 82% of the estimated average requirements. It was estimated that in early infancy a gross energy intake of 600 kJ/kg/day is required for normal growth in patients with congenital heart disease. This is unlikely to be achieved by energy supplements alone and early recourse to nasogastric feeding should be considered. [ABSTRACT FROM AUTHOR]

Few studies report the individual effect of 9c,11t- and 10t,12c-CLA on human energy metabolism. We compared the postprandial oxidative metabolism of 9c,11t- and 10t,12c-CLA and oleic acid (9c-18:1) in 22 healthy moderately overweight volunteers. After 24 weeks supplementation with 9c,11t-, 10t,12c-CLA or 9c-18:1 (3 g/day), subjects consumed a single oral bolus of the appropriate [1-(13)C]-labeled fatty acid. 8 h post-dose, cumulative oxidation was similar for 9c-18:1 and 10t,12c (P = 0.66), but significantly higher for 9c,11t (P < 0.01).

Stable isotope ratio analysis of light elements (including C, N, and S) is a powerful tool for inferring the production and geographic origins of animals. The objectives of this research were to quantify experimentally the isotopic turnover of C, N, and S in bovine skeletal muscle (LM and psoas major) and to assess the implications of the turnover for meat authentication. The diets of groups (n = 10 each) of beef cattle were switched from a control diet containing barley and unlabelled urea to an experimental diet containing maize, (15)N-labeled urea, and seaweed for periods of up to 168 d preslaughter. The feeding of the experimental diet was clearly reflected by the delta(13)C, delta(15)N, and delta(34)S values of the LM and psoas major muscles, but isotopic equilibrium was not reached in either muscle for C, N, or S after 168 d of feeding the experimental diet. The slow turnover in skeletal muscle was reflected by the C and N half-lives of 151 and 157 d for LM and 134 and 145 d for psoas major, respectively, and by an S half-life of 219 d in LM. It is concluded that the turnover of light elements (C, N, and S) in bovine skeletal muscles is a slow process; therefore, skeletal muscles contain isotopic information on dietary inputs integrated over a long period of time (months to years).

We present the results of a series of controlled feeding experiments with sheep, designed to investigate the effects of age and level of food intake on the kinetics of incorporation of the dietary carbon signal into wool. Four different groups of three sheep each, ranging in age from 6 to 78 months, were fed a C(3) diet and switched to a C(4) diet for up to 250 days. Different quantities of the same C(4) diet were provided to each group, in order to achieve different growth rates (high, low, and no growth). Wool was repeatedly shorn from each animal and processed for delta(13)C analyses. Results show that newly grown wool does not start recording the isotope composition of the new diet immediately after the diet-switch. The time-lag varies according to the age of the animal, from 6 +/- 1 days in lambs to up to 15 +/- 4 days in the older ewes. Wool from fast-growing lambs approached equilibrium faster than that from slow-growing lambs and young ewes, with old ewes being the slowest. However, 3 weeks after the diet-switch, the differences in wool delta(13)C values between the four different groups of animals were relatively small and represented less than 15% of the isotopic difference between the two diets. These results suggest that a single equation can be used to reconstruct previous diets for animals of different age, provided that the diet is similar and all individuals are in positive protein balance., (Copyright (c) 2008 John Wiley & Sons, Ltd.)

Trace elements often accumulate in keratin-rich tissues. Hair, nails, and horns grow steadily but once formed are metabolically inactive and provide an archive of trace element exposure when analyzed in segments. Here we demonstrate the use of laser ablation ICP-MS for the high-resolution monitoring of trace elements in the horns of seaweed-eating sheep from North Ronaldsay, which live on grass only during lambing time. Due to this peculiar husbandry/dietary pattern and the fact that seaweed is rich in arsenic and iodine, we hoped to use iodine and arsenic as markers for seaweed ingestion. Cross sections and scans along the growing axis (representing the first 8-10 months of the sheep's life) revealed that these elements were not homogeneously distributed in the horn, with arsenic representing the amount of seaweed intake. The scans show the periods in which the lambs were fed on milk and grass and the change to seaweed ingestion with the successive replacement of milk with seaweed; this was supported by the carbon and nitrogen isotope signatures (delta13C and delta15N) of the horn and the arsenic speciation in the horn. The period of low arsenic accumulation in the horn had terrestrial isotope signatures and accumulated arsenic of mainly inorganic origin. The period of high arsenic accumulation was characterized by isotope signatures of marine origin, and the majority of accumulated arsenic in the horn was the main arsenosugar metabolite dimethylarsinic acid. Although we have investigated only four different horns of individual sheep, this study shows that arsenic is not significantly transported with milk. However, the high concentration of arsenic in the oldest part of the horn, which was formed in utero, points to a relatively high placental transport of arsenic while the ewe was eating seaweed. In contrast to arsenic, iodine is transported not only through milk ingestion but also through the placenta in large quantities.

The objective of this study was to ascertain whether sequential sampling and isotopic analysis of bovine hooves could be used to reconstruct the dietary history of cattle. A controlled, on-farm experiment was conducted in which cattle were switched from a barley-based diet to an isotopically different diet incorporating maize, urea and seaweed (the isotopic spacing between diets was 13.6 per thousand for delta(13)C and 8.0 per thousand for delta(15)N) and maintained on that diet for 168 days. Postmortem sampling of the cleaned anterior wall of the lateral, left front claw was carried out on five individuals using a micro-drilling technique. From the first 60 mm of each claw, up to 41 samples with a spacing between them of less than 1 mm were collected. Bands were less than 1 mm deep and had a mean width of 1.2 mm. The hoof keratin showed a rapid increase followed by a slower increase in its delta(13)C and delta(15)N values following the diet switch, suggesting that C and N in hoof keratin originate from more than one pool. However, the response of the N isotope composition of the hoof was somewhat delayed compared with that of C. Estimated mean hoof growth rates for these cattle were 10.5 +/- 2.3 mm per month and 6.7 +/- 1.0 mm per month (+/-SD, n = 5) when receiving the barley-based transition diet and the maize-based experimental diet, respectively. These values are considerably higher than previous estimates obtained by visual methods and they suggest that diet may have a greater influence on hoof growth rates than seasonality. These results demonstrate that hooves are a suitable incremental tissue for high-resolution isotopic reconstruction of the dietary history of bovine animals., (Copyright (c) 2007 John Wiley & Sons, Ltd.)

To investigate the usefulness of bovine hooves as incremental tissues, the objective of this research was to gain a better understanding of hoof growth in three dimensions. In a controlled experiment, cattle were switched from a barley-based diet to a maize-based diet (C isotopic spacing between diets was 13.6 per thousand) and maintained on this experimental diet for 168 days. To compare growth rates along the hoof wall, three slices were sampled post-mortem from one bovine claw. In addition, one claw from each of three different animals was sampled at different depths from the surface to determine any possible time lag ('offset') in the laying down of keratin tissue layers. From each hoof as many as 41 superficial samples were taken over the first 60 mm, starting at the periople, and up to 12 samples were taken sequentially at increasing depths to a depth of 6 mm at five particular points on the surface. The growth rate of the abaxial wall of the bovine claw increased from the anterior to the posterior region of the bovine hoof. Analysis of the deep samples revealed that the deeper layers were younger than the surface layers. This offset was inversely related to the hoof growth rate, i.e. hooves with a high hoof growth rate showed a smaller offset. Observed offsets ranged between 9.2 +/- 1.8 days per mm in depth for a high and 14.0 +/- 2.8 days per mm in depth for a low hoof growth rate and were significantly different (t > or = 3.92, p < 0.0005, n = 19 or 27). The results of this study demonstrate that when sampling hooves or hoof fragments for applications such as diet reconstruction, careful consideration needs to be given to sample location., (Copyright (c) 2007 John Wiley & Sons, Ltd.)

The impact of ancient fertilization practices on the biogeochemistry of arable soils on the remote Scottish island of Hirta, St Kilda was investigated. The island was relatively unusual in that the inhabitants exploited seabird colonies for food, enabling high population densities to be sustained on a limited, and naturally poor, soil resource. A few other Scottish islands, the Faeroes and some Icelandic Islands, had similar cultural dependence on seabirds. Fertilization with human and animal waste streams (mainly peat ash and bird carcases) on Hirta over millennia has led to over-deepened, nutrient-rich soils (plaggen). This project set out to examine if this high rate of fertilization had adversely impacted the soil, and if so, to determine which waste streams were responsible. Arable soils were considerably elevated in Pb and Zn compared to non-arable soils. Using Pb isotope signatures and analysis of the waste streams, it was determined that this pollution came from peat and turf ash (Pb and Zn) and from bird carcases (Zn). This was also confirmed by (13)C and (15)N analysis of the profiles which showed that soil organic matter was highly enriched in marine-derived C and N compared to non-arable soils. The pollution of such a remote island may be typical of other 'bird culture' islands, and peat ash contamination of marginal arable soils at high latitudes may be widespread in terms of geographical area, but less intense at specific locations due to lower population densities than on Hirta.

Nitrogen translocation was measured in Cladonia portentosa during 2 yr growth in Scottish heathland. Translocation was predicted to occur if N is resorbed from senescent basal tissue and recycled within the thallus. (15)N was introduced into either the lower (TU thalli) or upper (TD thalli) 25 mm of 50-mm-long thalli as (15)N-NH(4) (+), (15)N-NO(3) (-) or (15)N-glycine. Labelled thalli were placed within intact lichen cushions, either upright (TU) or inverted (TD). Vertical distribution of label was quantified immediately following labelling and after 1 and 2 yr. Independently of the form of introduced label, (15)N migrated upwards in TU thalli, with new growth being a strong sink. Sink regions for (15)N during year 1 (including new growth) became sources of (15)N translocated to new growth in year 2. Upward migration into inverted bases was minimal in TD thalli, but was again marked in new growth that developed from inverted apices. Relocation of N to regions of growth could facilitate internal N recycling, a process postulated to explain the ecological success of mat-forming lichens.

Rhizosphere microorganisms play an important role in soil carbon flow, through turnover of root exudates, but there is little information on which organisms are actively involved or on the influence of environmental conditions on active communities. In this study, a 13CO2 pulse labelling field experiment was performed in an upland grassland soil, followed by RNA-stable isotope probing (SIP) analysis, to determine the effect of liming on the structure of the rhizosphere microbial community metabolizing root exudates. The lower limit of detection for SIP was determined in soil samples inoculated with a range of concentrations of 13C-labelled Pseudomonas fluorescens and was found to lie between 10(5) and 10(6) cells per gram of soil. The technique was capable of detecting microbial communities actively assimilating root exudates derived from recent photo-assimilate in the field. Denaturing gradient gel electrophoresis (DGGE) profiles of bacteria, archaea and fungi derived from fractions obtained from caesium trifluoroacetate (CsTFA) density gradient ultracentrifugation indicated that active communities in limed soils were more complex than those in unlimed soils and were more active in utilization of recently exuded 13C compounds. In limed soils, the majority of the community detected by standard RNA-DGGE analysis appeared to be utilizing root exudates. In unlimed soils, DGGE profiles from 12C and 13C RNA fractions differed, suggesting that a proportion of the active community was utilizing other sources of organic carbon. These differences may reflect differences in the amount of root exudation under the different conditions.

The influence of liming on rhizosphere microbial biomass C and incorporation of root exudates was studied in the field by in situ pulse labelling of temperate grassland vegetation with (13)CO(2) for a 3-day period. In plots that had been limed (CaCO(3) amended) annually for 3 years, incorporation into shoots and roots was, respectively, greater and lower than in unlimed plots. Analysis of chloroform-labile C demonstrated lower levels of (13)C incorporation into microbial biomass in limed soils compared to unlimed soils. The turnover of the recently assimilated (13)C compounds was faster in microbial biomass from limed than that from unlimed soils, suggesting that liming increases incorporation by microbial communities of root exudates. An exponential decay model of (13)C in total microbial biomass in limed soils indicated that the half-life of the tracer within this carbon pool was 4.7 days. Results are presented and discussed in relation to the absolute values of (13)C fixed and allocated within the plant-soil system.

This study investigated the effect of substituting grass silage (C3 photosynthetic plant product) with maize silage (C4 photosynthetic plant product) on the natural abundance carbon (delta13C) and nitrogen (delta15N) stable isotope composition of bovine muscle tissue. Forty-five continental crossbred heifers were assigned to one of three diets consisting of 3 kg of a barley-based concentrate plus grass silage, maize silage or an equal mixture (dry matter basis) of grass silage and maize silage, fed ad libitum, for 167 days. Substitution resulted in less negative delta13C values (P<0.001) in lipid-free muscle and in lipid, and also a lower delta15N (P<0.001) in lipid-free muscle. Feeding of maize silage was clearly reflected in the delta13C of muscle, with each 10% difference in the dietary C4 carbon intake resulting in a 0.9 to 1.0 per thousand shift of delta13C in lipid-free muscle and a 1.0 to 1.2 per thousand in lipid. Minimum detectable mean differences (95% confidence, power 0.80, n=15) in this experiment were about 0.5 per thousand and 1.0 per thousand for delta13C of lipid-free muscle and lipid, respectively, and about 0.5 per thousand for delta15N of lipid-free muscle. The power analysis presented here is useful for estimating minimum isotopic differences that can be detected between any two groups of beef samples with a given number of replicates. It is concluded that carbon stable isotope ratio analysis of meat can be used to quantify C3/C4 dietary constituents in beef production., (Copyright (c) 2005 John Wiley & Sons, Ltd.)

Responses of Rhizoctonia solani to spatial heterogeneity in sources of carbon, and associated translocation of carbon (C), were studied in a simple microcosm system comprising two discrete domains of agar gels separated on a glass slide and overlain with a porous membrane. Two arrangements of the gel pairs were used, one containing two equally large resources (representing 'homogeneous' conditions) and one containing a large and a negligible resource (representing 'heterogeneous' conditions). The nutrient sources were a standard mineral salt medium with or without glucose as sole C source. The fungus was inoculated onto one domain and growth responses determined by direct measurement of biomass. Translocation of C was quantified by use of 13C-enriched glucose. This substrate was either added to the agar at the outset, when studying newly developing colonies, or as a pulse into already established colonies. When growing in heterogeneous conditions, the fungus actively translocated C from a glucose-containing domain to sustain growth in the adjacent region lacking such a resource. In homogeneous conditions there was evidence of passive translocation (diffusion), but the fungus preferentially used local resource to maintain growth. Active translocation was only observed in newly growing colonies, whereas passive translocation occurred in both growing and established colonies. When the fungus was pulsed with a 13C-enriched glucose solution after 10 d growth, 2.5 times more 13C was taken up by the fungus grown in heterogeneous than homogeneous conditions, suggesting uptake exceeded local demands. In heterogeneous conditions, the total amount of 13C enriched glucose taken up by the fungus was independent of the location of the enriched glucose in the underlying medium. When the nylon membrane was replaced by Cellophane (an additional C source), degradation of the membrane and an increase in biomass occurred only in the heterogeneous system. The possible implications for these results in soil systems are discussed.

Natural abundance of (15)N and [N] was studied in thalli of mat-forming lichens collected from tundra and heathland sites in the northern and southern hemispheres. The study includes samples of British Cladonia portentosa from sites in regions of high and low N-loading and in heathland growing both directly on peat and independently of the soil substratum, in a canopy of prostrate gorse ( Ulex minor). In the mat-forming lichens examined, a non-random pattern in [N] and delta(15)N was characterised by a minimum in delta(15)N, which occurred most frequently at 20-40 mm below the thallus apex. Nitrogen concentration increased above this point, towards the apex, though remained invariably low towards the thallus base. We discuss the significance of the pattern in [N] and delta(15)N for current theories describing the uptake and recycling of nitrogen by mat-forming lichens in oligotrophic habitats. Our data are incompatible with the suggested uptake of soil organic-N depleted in (15)N, though are consistent with possible internal recycling and the development of a structural necromass. The study emphasises the internal fractionation of nitrogen isotopes and provides a caveat against the assumption that values of delta(15)N provide an unequivocal indicator of source-sink relationships in nitrogen cycling.

• Plant root mucilages contain powerful surfactants that will alter the interaction of soil solids with water and ions, and the rates of microbial processes. • The lipid composition of maize, lupin and wheat root mucilages was analysed by thin layer chromatography and gas chromatography-mass spectrometry. A commercially available phosphatidylcholine (lecithin), chemically similar to the phospholipid surfactants identified in the mucilages, was then used to evaluate its effects on selected soil properties. • The lipids found in the mucilages were principally phosphatidylcholines, composed mainly of saturated fatty acids, in contrast to the lipids extracted from root tissues. In soil at low tension, lecithin reduced the water content at any particular tension by as much as 10 and 50% in soil and acid-washed sand, respectively. Lecithin decreased the amount of phosphate adsorption in soil and increased the phosphate concentration in solution by 10%. The surfactant also reduced net rates of ammonium consumption and nitrate production in soil. • These experiments provide the first evidence we are aware of that plant-released surfactants will significantly modify the biophysical environment of the rhizosphere.

The conversion of ammonium (NH(4) (+)) to 1-sulfonato-iso-indole has been examined as a method for natural abundance measurement of delta(15)N of NH(4) (+). The reaction is complete within 2 h and is based on the derivatisation of NH(4) (+) by o-phthaldialdehyde and sodium sulfite at a high pH, 11.2. The product is readily concentrated from dilute solutions by reverse-phase solid-phase extraction (SPE). The method is compound-specific despite partial derivatisation of potentially interfering amino acids, as their derivatives are not extracted by SPE. delta(15)N values of NH(4) (+) in KCL soil extracts can be measured within 48 h by automated continuous-flow IRMS with a precision of 0.23 per thousand (1 SD). Parallel measurements of NH(4) (+) standards of known delta(15)N are made to allow correction for the isotopic dilution by non-sample NH(4) (+). The practicality of this method is demonstrated by measuring the changes in NH(4) (+) concentration and delta(15)N following the addition of urea as a nitrogen source to inorganic N-depleted soil., (Copyright 2003 John Wiley & Sons, Ltd.)

Much evidence suggests that life originated in hydrothermal habitats, and for much of the time since the origin of cyanobacteria (at least 2.5 Ga ago) and of eukaryotic algae (at least 2.1 Ga ago) the average sea surface and land surface temperatures were higher than they are today. However, there have been at least four significant glacial episodes prior to the Pleistocene glaciations. Two of these (approx. 2.1 and 0.7 Ga ago) may have involved a 'Snowball Earth' with a very great impact on the algae (sensu lato) of the time (cyanobacteria, Chlorophyta and Rhodophyta) and especially those that were adapted to warm habitats. By contrast, it is possible that heterokont, dinophyte and haptophyte phototrophs only evolved after the Carboniferous-Permian ice age (approx. 250 Ma ago) and so did not encounter low (

Barley traits related to salt tolerance are mapped in a population segregating for a dwarfing gene associated with salt tolerance. Twelve quantitative trait loci (QTLs) were detected for seven seedling traits in doubled haploids from the spring barley cross Derkado x B83-12/21/5 when given saline treatment in hydroponics. The location of QTLs for seedling growth stage (leaf appearance rate), stem weight prior to elongation, and tiller number are reported for the first time. In addition, four QTLs were found for the mature plant traits grain nitrogen and plot yield. In total, seven QTLs are co-located with the dwarfing genes sdw1, on chromosome 3H, and ari-e.GP, on chromosome 5H, including seedling leaf response (SGa) to gibberellic acid (GA(3)). QTLs controlling the growth of leaves (GS2) on chromosomes 2H and 3H and emergence of tillers (TN2) and grain yield were independent of the dwarfing genes. Field trials were grown in eastern Scotland and England to estimate yield and grain composition. A genetic map was used to compare the positions of QTLs for seedling traits with the location of QTLs for the mature plant traits. The results are discussed in relation to the study of barley physiology and the location of genes for dwarf habit and responses to GA.

The literature, and previously unpublished data from the authors' laboratories, shows that the δ 13 C of organic matter in marine macroalgae and seagrasses collected from the natural environment ranges from -3 to -35‰. While some marine macroalgae have δ 13 C values ranging over more than 10‰ within the thallus of an individual (some brown macroalgae), in other cases the range within a species collected over a very wide geographical range is only 5‰ (e.g. the red alga Plocamium cartilagineum which has values between -30 and -35‰). The organisms with very negative δ 13 C (lower than -30‰) are mainly subtidal red algae, with some intertidal red algae and a few green algae; those with very positive δ 13 C values (higher than -10‰) are mainly green macroalgae and seagrasses, with some red and brown macroalgae. The δ 13 C value correlates primarily with taxonomy and secondarily with ecology. None of the organisms with δ 13 C values lower than -30‰ have pyrenoids. Previous work showed a good correlation between δ 13 C values lower than -30‰ and the lack of CO2 concentrating mechanisms for several species of marine red algae. The extent to which the low δ 13 C values are confined to organisms with diffusive CO2 entry is discussed. Diffusive CO2 entry could also occur in organisms with higher δ 13 C values if diffusive conductance was relatively low. The photosynthesis of organisms with δ 13 C values more positive than -10‰ (i.e. more positive than the δ 13 C of CO2 in seawater) must involve HCO3 - use.

Background: Epoxy fats in the diet may adversely affect human health. There are no data on the absorption of these fats in humans., Methods: Triglycerides were synthesized containing two U-13C-labelled monoepoxy or diepoxy stearic acid molecules. Apparently healthy women consumed a standardized fatty meal (30 g fat) containing either 20 mg monoepoxy or 25 mg diepoxy fat (n = 6 and n = 7, respectively). Plasma lipid [U-13C]monoepoxy and diepoxy stearate concentrations were determined (0-24 h) by gas chromatography-mass spectrometry., Results: Plasma triglycerides increased from 1.05 +/- 0.12 to 1.83 +/- 0.13 mmol L-1 (n = 6) and from 1.10 +/- 0.19 to 1.41 +/- 0.27 mmol L-1 (n = 7) (both P < 0.001). Plasma [U-13C]monoepoxy and diepoxy stearate levels increased to 0.18 +/- 0.07 micromol L-1 (n = 6) and to 0.08 +/- 0.03 micromol L-1 (n = 7), respectively. Monoepoxy triglyceride was better absorbed than diepoxy triglyceride: 17 +/- 4 vs. 8 +/- 1% of dose (determined from area under curve (plasma 13C) normalized to that of absorbed triglycerides (plasma 12C); P < 0.02 after log transformation). The absorption of monoepoxy- and diepoxy-labelled triglycerides was related to that of normal triglycerides (r = 0.80, P < 0.05 and r = 0.91, P < 0.001, respectively)., Conclusions: Monoepoxy fats are better absorbed than diepoxy fats in women (17 +/- 4 vs. 8 +/- 1% of dose, P = 0.02). This difference in absorption is important when considering the relative toxicity of epoxidized material in the food chain.

The linear hexitol altritol has only been identified in six genera, all of them in the order Fucales of the brown algae. Five of these genera are closely related according to molecular phylogenetic and other data, while the sixth (Notheia) is an obligate epiphyte on two other altritol-containing genera with which it is symphanic. The possibility that Notheia obtains altritol from the algae on which it is epiphytic rather than by synthesizing altritol independently was investigated by supplying 13C-inorganic carbon in the light followed by mass spectrometric and nuclear magnetic resonance analysis. Notheia separated from the phorophyte Hormosira during exposure to 13C showed 13C enrichment in both altritol and mannitol, while the Hormosira only showed significant labelling of mannitol. These data show that altritol can be synthesized by Notheia, with implications for the number of gains and losses of the capacity to synthesize altritol in the evolution of the Fucales.

Human lipid intake contains various amounts of trans fatty acids. Refined vegetable and frying oils, rich in linoleic acid and/or alpha-linolenic acid, are the main dietary sources of trans-18:2 and trans-18:3 fatty acids. The aim of the present study was to compare the oxidation of linoleic acid, alpha-linolenic acid, and their major trans isomers in human volunteers. For that purpose, TG, each containing two molecules of [1-(13)C]linoleic acid, alpha-[1-(13)C]linolenic acid, [1-(13)C]-9cis,12trans-18:2, or [1-(13)C]-9cis,12cis,15trans-18:3, were synthesized. Eight healthy young men ingested labeled TG mixed with 30 g of olive oil. Total CO(2) production and (13)CO(2) excretion were determined over 48 h. The pattern of oxidation was similar for the four fatty acids, with a peak at 8 h and a return to baseline at 24 h. Cumulative oxidation over 8 h of linoleic acid, 9cis,12trans-18:2, alpha-linolenic acid, and 9cis,12cis,15trans-18:3 were, respectively, 14.0 +/- 4.1%, 24.7 +/- 6.7%, 23.6 +/- 3.3%, and 23.4 +/- 3.7% of the oral load, showing that isomerization increases the postprandial oxidation of linoleic acid but not alpha-linolenic acid in men.

New evidence, obtained using a robust method for measuring the delta(15)N of NO(3)(-)-N in soil, is consistent with denitrification being the major determinant in the vertical distribution of NO(3)(-)-delta(15)N in soil profiles. These data also suggest that varying moisture regimes result in different effects of soil NO(3)(-)-N leaching on residual whole soil delta(15)N., (Copyright 2001 John Wiley & Sons, Ltd.)

The effect of interactions between Casuarina species, Frankia strains and AMF on nitrogen isotope fractionation within the plant were determined under conditions where changes in source nitrogen were minimized by growing plants in mineral nitrogen-deficient conditions and without added organic N. Casuarina cunninghamiana, C. equisetifolia, C. glauca, and C. junghuniana were inoculated singly with three Frankia strains or were dual inoculated with Frankia and Glomus fasciculatum. The %N and delta 15N of separated parts of plants inoculated with the three Frankia strains or with Frankia + Glomus were not significantly different within Casuarina species. However, the slow-growing C. junghuniana differed in several variables from the other three species. There was a highly significant, linear relationship between the natural logarithms of cladode N content and delta 15N of plants of the four Casuarina species when inoculated with Frankia or with Frankia + Glomus, showing that nitrogen supply and the correlated variable, plant growth rate, were major determinants of delta 15N. Provision of small quantities of (NH4)2SO4 or KNO3 increased several-fold the growth of three of the Casuarina species when inoculated with Frankia alone or with Frankia + Glomus. Within species, mycorrhizal and non-mycorrhizal plants receiving supplementary soluble phosphate were of similar dry weights at harvest. delta 15N values for cladodes of C. cunninghamiana, C. equisetifolia and C. glauca were similar, but values for the poor growing C. junghuniana were more variable and, with the exception of plants receiving KNO3, were lower than those of the other three species. Reduced growth due to suboptimal availability of N or P had a major influence on delta 15N and, in these conditions where plants could not access significant amounts of organic N, outweighed any effects on cladode delta 15N of colonization by Glomus. delta 15N values of nodules were higher than other parts of Frankia or Frankia + Glomus inoculated Casuarinas, conceivably due to retention in nodules of fixed N, with delta 15N close to zero.

To integrate the complex physiological responses of plants to stress, natural abundances (delta) of the stable isotope pairs 15N/14N and 13C/12C were measured in 30 genotypes of wild barley (Hordeum spontaneum C. Koch.). These accessions, originating from ecologically diverse sites, were grown in a controlled environment and subjected to mild, short-term drought or N-starvation. Increases in total dry weight were paralleled by less negative delta 13C in shoots and, in unstressed and droughted plants, by less negative whole-plant delta 13C. Root delta 15N was correlated negatively with total dry weight, whereas shoot and whole-plant delta 15N were not correlated with dry weight. The difference in delta 15N between shoot and root varied with stress in all genotypes. Shoot-root delta 15N may be a more sensitive indicator of stress response than shoot, root or whole-plant delta 15N alone. Among the potentially most productive genotypes, the most stress-tolerant had the most negative whole-plant delta 15N, whether the stress was drought or N-starvation. In common, controlled experiments, genotypic differences in whole-plant delta 15N may reflect the extent to which N can be retained within plants when stressed.

It has long been evident that plant (15)N chiefly reflects the processes which fractionate (15)N/(14)N rather than the (15)N of plant N source(s). It has emerged recently that one of the most important fractionating processes contributing to the whole plant (15)N is the presence/absence, type or species of mycorrhiza, especially when interacting with nutrient deficiency. Ecto- and ericoid mycorrhizas are frequently associated with (15)N-depleted foliar (15)N, commonly as low as -12 per thousand. As shown by the present study, plants having no mycorrhiza, or those infected with various species of arbuscular mycorrhiza (AM)-forming fungi, interact with varying concentrations of soil nitrogen [N] and moisture to enrich plant (15)N by as much as 3.5 per thousand. Hence the lack of a mycorrhiza, or variation in the species of AM-forming fungal associations, can account for about 25% of the usually reported variations of foliar (15)N found in field situations and do so by (15)N enrichment rather than depletion. Copyright 1999 John Wiley & Sons, Ltd.

The conversion of nitrate (NO(3)(-)) to 1-phenylazo-2-naphthol (Sudan-1) has been examined as a method for natural abundance measurement of delta(15)N of NO(3)(-). The reaction results in dilution of NO(3)(-)-N with only one reagent-derived N and the product is readily concentrated from dilute samples by reverse phase chromatography. There is systematic isotopic fractionation during the reaction, but this can be allowed for by analysing known NO(3)(-) standards along with each sample set. Sudan-1 prepared from surface water samples containing approximately 50 &mgr;g NO(3)(-)-N can be analysed by automated continuous flow isotope ratio mass spectrometry with a precision of 0.2 per thousand (one standard deviation) and the accuracy is not affected by interference from other nitrogenous species in the sample or reagents. Copyright 1999 John Wiley & Sons, Ltd.

We used natural abundance stable isotope techniques to estimate carbon and nitrogen turnover rates in body tissue and mucus of earthworms. Isotope ratios of carbon (delta(13)C) and nitrogen (delta(15)N) were monitored simultaneously in body tissue and mucus for up to 101 days in feeding or fasting Lumbricus festivus kept in an artificial substrate. When the diet of the earthworms was switched from clover (C(3) plant, legume) to maize (C(4), non-legume), the new dietary delta(13)C signature manifested itself much more rapidly in the mucus than in the body tissue of the animals, causing a delta(13)C shift of about 4 per thousand in mucus and 1 per thousand in tissue after 13.5 days. Turnover of earthworm body tissue carbon, unlike that of mucus carbon, was described adequately by an exponential, single-pool model. Nitrogen turnover could not be assessed because the delta(15)N difference between sources was too small. Fasting for 56 days did not result in the expected whole-body (15)N or (13)C enrichment, but it caused a significant decrease in mucus and tissue C:N ratios and in the ratio (mucus C:N ratio):(tissue C:N ratio). We conclude that the separate analysis of body tissue and mucus has great potential for studying the ecophysiology, feeding ecology and role in elemental cycling of earthworms and other invertebrates.

We report the first isotopic study of an animal host-parasite system. Parasitic, intestinal nematodes, Graphidium strigosum and Passalurus ambiguus, were 15N-enriched relative to their host, the European rabbit Oryctolagus cuniculus, while parasitic cestodes, Cittataenia denticulata and Mosgovoyia pectinata, were 15N-depleted, suggesting different trophic relationships. Host embryos were more similar in their delta 13C and delta 15N values to maternal muscle than were any of the parasites. Coprophagy, the direct recycling of food by the rabbit eating its own faeces, did not lead to isotopic differences between stomach contents and faeces, suggesting that the major point for isotopic discrimination in lagomorph nitrogen metabolism is in the animal rather than in the gut. We conclude that bulk delta 13C and delta 15N can reveal valuable new information about host-parasite relationships, and these could be explored further at the biochemical level using compound-specific isotopic analyses.

This work explored whether the natural abundances of carbon and nitrogen isotopes could be used to describe the movement of C and X within wheat plants; we also considered whether isotopic analyses of aphids or their honeydew would substitute for direct analysis of phloem exudate. The δ 13 C of ears and roots (sinks) most closely matched those of the sugars + organic acids fraction (sources) in both growth stages; phloem δ 13 C matched that of leaf blade sugars. Xylem exudate δ 13 C matched no other putative (and measured) source in the ear-forming stage and matched that of whole roots and ears in the grain-filling stage. The δ 15 N of grain and roots (sinks) resembled that of leaf amino acids (sources) in the ear-forming stage. In the gram-filling stage, ear δ 15 N continued to resemble that of leaf amino acids, and δ 15 N of roots most closely resembled that of whole leaves. In the grain-filling stage, phloem δ 15 N fell between that of leaf blade amino acids and that of whole leaves and was 15 X-depleted relative to internal and external NO, -N. In both growth stages, xylem exudate δ 15 N was less than that of soil NO 3 - -N and more than that of residual soil N after mineral N extraction. The isotopic values are generally in agreement with data from other approaches, such as isotope labelling; they show NO 3 - -N reduction in both shoots and roots of wheat and significant N recycling (root-shoot-phloem-root) and C movement. Aphids might serve as a substitute for isotopic analysis of phloem δ 15 N. having the same value as their food source. Their excreta was 15 N-enriched relative to phloem.

Abstract We report the first isotopic study of an animal host-parasite system. Parasitic, intestinal nematodes, Graphidium strigosum and Passalurus ambiguus, were (15)N-enriched relative to their host, the European rabbit Oryctolagus cuniculus, while parasitic cestodes, Cittataenia denticulata and Mosgovoyia pectinata, were (15)N-depleted, suggesting different trophic relationships. Host embryos were more similar in their δ(13)C and δ(15)N values to maternal muscle than were any of the parasites. Coprophagy, the direct recycling of food by the rabbit eating its own faeces, did not lead to isotopic differences between stomach contents and faeces, suggesting that the major point for isotopic discrimination in lagomorph nitrogen metabolism is in the animal rather than in the gut. We conclude that bulk δ(13)C and δ(15)N can reveal valuable new information about host-parasite relationships, and these could be explored further at the biochemical level using compound-specific isotopic analyses.

Thirty-nine genotypes of Hordeum spontaneum were selected from three geographically separated areas (southwestern, northern, and southeastern) of the Fertile Crescent. The lines were subject to AFLP (amplified fragment length polymorphism) analysis from which a similarity matrix was produced. A dendrogram of the data showed, with two exceptions, that the genotypes grouped together according to area of origin. This was confirmed by principal coordinate analysis in which the first principal coordinate separated the genotypes of the southwestern area from the other two areas, which in turn could be separated by the second principal coordinate. While genotypes from the same site of origin can exhibit very similar AFLP profiles, sharp genetic differences were detected between genotypes separated by relatively short distances. Thirty of the 39 genotypes were subjected to hydroculture salt tolerance tests. These were analysed for shoot Na+ content and carbon isotope composition (&delta;13C) after 4 weeks of treatment (100 mol&#x2219;m&minus;3 NaCl). Shoot Na+ content and &delta;13C were highly correlated. Twelve AFLP markers were found to be associated with both shoot Na+ content and shoot &delta;13C and were also associated with site of origin ecogeographic data, particularly longitude. The most salt tolerant genotype came from Ilam in the southeastern area and the most salt sensitive genotype originated in the southwestern area. The 12 markers were partitioned into groups that showed significant associations within groups but no significant association between groups. In a multiple regression analysis, three AFLP markers, from separate groups, accounted for more than 60% of the variation for shoot Na+ content and &delta;13C. The results demonstrate the effectiveness of AFLP fingerprinting in genetic studies of complex traits at the wild species and (or) population level.

The energy requirements of people doing physical work in hot climates are not clearly understood. In particular, we know little about the combined effects of heat stress and muscular work on energy requirements. During military exercises in the African bush, soldiers are supplied with standard rations, the adequacy of which is unknown. We have now assessed the adequacy of these food and water rations in 12 male Zimbabwean soldiers during 12 days of strenuous, heat-stress exercise in the field. We used two methods to measure energy expenditure: the double-labelled water method (DLW) and the energy balance method (i.e. comparing dietary energy with changes, if any, in body energy stores). Two groups were studied: one group (eight subjects) carried out field exercises; the control group consisted of four soldiers doing normal work. Mean daily energy expenditure as assessed by the DLW method was [mean (SE)] 23 (1.5) MJ x day(-1) for the field group and 14 (0.5) MJ x day(-1) for the control group (P < 0.001). By the energy balance method, daily energy expenditure was calculated to be 26 (0.7) MJ x day(-1) and 15.5 (0.4) MJ x day(-1) for the field group and control group, respectively. Body mass loss was 3 (0.1) kg [4.6 (0.3)% of body mass] for the field group, but the control group gained 1.1 (0.1) kg. Mean daily fluid intake was 11 (0.5) 1 x day(-1), suggesting that the standard ration supplied was inadequate. Body mass loss was caused by both the energy deficit and total body water loss. These results suggest strenuous work in hot, dry field conditions imposes extra energy requirements.

In order to determine which of maize syrup solids, glucose and sucrose were more readily oxidised during exercise and least readily oxidised afterwards, the rates of oxidation of three almost identical isoenergetic solutions of carbohydrates (330 ml of 18.5% w/v solutions of glucose, maize syrup solids and sucrose, 989-1050 kJ total energy) naturally enriched with 13C were examined at rest and during and after 1 h uphill walking at 75% maximum oxygen uptake (VO2max) in nine subjects [mean (SEM) VO2max, 45.4 (0.9) ml.kg-1.min-1]. Rates of production of expired 13CO2 were used to estimate rates of oxidation of each exogenous substrate. Energy expenditure and the contributions from total carbohydrate and fat oxidation were calculated from whole-body gas exchange. At rest, maize syrup solids were oxidised less than sucrose during the 1st h [glucose 2.7 (0.2) g.h-1, maize syrup solids 1.9 (0.3) g.h-1, sucrose 3.7 (0.2) g.h-1; maize syrup solids vs sucrose P < 0.01], but this difference disappeared after a further 3 h at rest [glucose 8.3 (0.5) g.h-1, maize syrup solids 7.7 (0.5) g.h-1, sucrose 8.1 (0.4) g.h-1]. During exercise, all the carbohydrates were oxidised to the same extent [glucose 23.0 (2.8) g.h-1, maize syrup solids 23.9 (3.4) g.h-1, sucrose 27.5 (2.6) g.h-1) but during 4 h of recovery after exercise, maize syrup solids were oxidised least [glucose 4.6 (0.1) g.h-1, maize syrup solids 3.7 (0.1) g.h-1, sucrose 6.4 (0.1) g.h-1; P < 0.05] suggesting that it may be stored to a greater extent. The results suggest that 18.5% glucose, maize syrup solids and sucrose solutions were equally well oxidised during exercise. During recovery from exercise maize syrup solids were oxidised less than glucose, which in turn was oxidised less than sucrose.

Insulin and sulphonylurea therapies have both been reported to cause weight gain in Type 2 diabetic patients whereas metformin does not have this adverse effect. The mechanism for this difference is unclear. We have investigated in a cross-over study the effect of sulphonylurea and metformin therapy on energy expenditure and body composition in 10 Type 2 diabetic patients (7 females, 3 males) of various weights (mean body mass index 33.4 (SD 7.6 kg m-2)). Free living total energy expenditure was measured over 14 days by the doubly labelled water method adjusted for urinary glucose energy losses and resting energy expenditure by ventilated hood indirect calorimetry. Overall, total energy expenditure (12.88 +/- 4.17 vs 13.1 +/- 3.69 MJ 24 h-1) and resting metabolic rate (7.30 +/- 1.75 vs 7.23 +/- 1.74 MJ 24 h-1) were similar on metformin and sulphonylurea therapy, respectively. When adjusted for differences in fat free mass, resting metabolic rate on sulphonylurea therapy was slightly but significantly lower (mean difference -5.5 kJ 24 h-1 kg-1, 95% CI -1.2, -9.9 kJ 24 h-1 kg-1, p < 0.05). Fat free mass also increased significantly by 1.3 kg (95% CI 0.4, 2.4 kg, p < 0.05) when on sulphonylurea therapy, thus compensating for the lower resting metabolic rate per kg fat free mass to leave overall resting metabolic rate unchanged compared to metformin therapy. We also investigated the effect of adding metformin to six Type 2 diabetic patients already on insulin. This did not lead to any measurable changes in any of the components of energy expenditure.(ABSTRACT TRUNCATED AT 250 WORDS)

To test the hypothesis that acidosis contributes to the insulin resistance of chronic renal failure (CRF) and impairs the action of insulin to decrease protein degradation, eight CRF patients were studied using the combined L-[1-13C]leucine-euglycemic clamp technique before (acid) and after (NaHCO3) 4 wk treatment with NaHCO3 (pH: acid 7.29 +/- 0.01 vs. NaHCO3 7.36 +/- 0.01, P < 0.001). Protein degradation (PD) was estimated sequentially from the kinetics of a primed continuous infusion of L-[1-13C]leucine in the basal state and during a hyperinsulinemic euglycemic clamp. Insulin sensitivity was measured during the clamp. The correction of acidosis significantly increased the glucose infusion rate necessary to maintain euglycemia (acid 6.44 +/- 0.89 vs. bicarbonate 7.38 +/- 0.90 mg.kg-1.min-1, P < 0.01) and significantly decreased PD in the basal state (acid 126.4 +/- 8.1 vs. bicarbonate 100.1 +/- 6.9 mumol.kg-1.h-1, P < 0.001). Hyperinsulinemia decreased PD in both studies (acid basal 126.4 +/- 8.1 vs. clamp 96.5 +/- 7.7, P < 0.001; bicarbonate basal 100.1 +/- 6.9 vs. clamp 88.2 +/- 5.5 mumol.kg-1.h-1, P = 0.06), its effect being unaltered by acidosis, with a reduction of 24% before and 12% after the correction of acidosis. In conclusion, acidosis contributes to the insulin resistance of CRF but does not affect the action of insulin on PD.

In order to investigate the effects of muscular work and preceding exercise on the retention of exogenous labelled bicarbonate, we studied the effects of oral administration of [13C]bicarbonate (0.1 mg kg-1) in five subjects at rest before exercise and during and after 1 h of treadmill walking at 73% VO2max on three separate occasions. Elimination of CO2 from labelled bicarbonate was 62.6 +/- 8.1% at rest, 103.6 +/- 11.3% during exercise (P < 0.01) and 43.0 +/- 4.7% during recovery from exercise (P = 0.01). During exercise mean residence time (MRT) was shorter than at rest (35 +/- 7 min vs. 54 +/- 9 min, P < 0.02) and CO2 pool size was larger (998 +/- 160 ml CO2 kg-1, vs. 194 +/- 28 ml CO2 kg-1, P < 0.001). Compared to values obtained at rest, during recovery from exercise, MRT and CO2 pool size were reduced (34 +/- 5 min, P < 0.05; 116 +/- 19 ml CO2 kg-1, P < 0.02, respectively). In an additional five subjects acidosis and alkalosis were induced prior to administration of oral [13C]bicarbonate at rest. Elimination of bicarbonate was lower during acidosis (46.1 +/- 5.6%, P < 0.01) but was unaltered (50.9 +/- 5.6%, NS) during alkalosis, compared to the values obtained at resting pH. During acidosis MRT and CO2 pool size decreased (37 +/- 3 min, P < 0.01 and 123 +/- 10 ml CO2 kg-1, P < 0.01, respectively) whereas in alkalosis MRT was unchanged (65 +/- 8 min NS) but CO2 pool size was increased (230 +/- 23 ml CO2 kg-1, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

1. The effect of euglycaemic hyperinsulinaemia on the recovery of 13C in expired CO2 has been assessed in six normal subjects. Each was studied on three occasions: once with a 6 h primed constant infusion of NaH13CO3 combined with a euglycaemic hyperinsulinaemic clamp for the last 3 h (study 1), once with a 6 h primed constant infusion of NaH13CO3 alone (study 2) and once with a 6 h infusion of normal saline combined with a hyperinsulinaemic clamp for the last 3 h (study 3). Measurements of 13C enrichment of expired CO2 were made in the third and sixth hour of each infusion. 2. There was no significant increase in enrichment during study 3 (3 h 0.00047 +/- 0.00016 versus 6 h 0.00069 +/- 0.00028 atom per cent excess) with potato-starch-derived D-glucose used to maintain euglycaemia. 13C recovery increased in the sixth hour of both study 1 and 2 (study 1: 3 h 74.4 +/- 2.0 versus 6 h 85.5 +/- 2.6%, P < 0.01; study 2: 3 h 72.1 +/- 2.4 versus 6 h 81.7 +/- 1.4%, P < 0.01). There was no significant difference in recovery between studies 1 and 2. 3. These results suggest that increased recovery during a sequential euglycaemic clamp is predominantly time-dependent. Studies which use this technique to examine the effect of insulin on substrate oxidation should take this into account.

The 13 C/ 12 C ratio (expressed as δ 13 C) of benrhic photolithotrophs. in the Dighn Water (= Burn) were measured fur comparison with that of the potential inorganic carhun sources. CO 2 and HCO 3 - , in the Burn. The Burn water contains an average of 65.7 mmol m -3 CO 2 with δ 13 C of -14.7% and 1600 mmol m -3 HCO 3 - with δ 13 C of -4.%. δ 13 C values of riparian vegetation were also measured as contributors, after respiration in the soil or the Burn, to the δ 13 C of inorganic carbon in the Burn. The potential range of differences in 13 C/1 2C between dissolved CO 2 and plant organic C is set by the intrinsic 13 c/ 12 C discrimination (α value) in CO 2 fixation by Rubisco. Main results and conclusions are. as follows, (i) A literature survey suggests that there is no convincing evidence that the α, Values (rate constant for 12 CO 2 fixation relative to that for 13 CO 2 fixation by Rubisco in the absence of CO 2 transport limitation) for the'lower plants'in the Burn (diatoms, green and red algae, mosses) are significantly different from the well-established α p values for the flowering plum enzyme. (ii) In confirmation of earlier work, the semi-erect 'streamer'gametophytes of the red alga Lemanea mamillosa and the moss Fontinalis antipyetica have δ 13 C values which can only be interpreted in terms of diffusive CO 2 entry with minimal limitation of photosynthesis by CO- diffusion, (iii) The serui-erect grren alga Cladophora glomerata and the flowering plant Ranunculus penicillatus ssp. pseudofluitons (formerly var. calcareus) are- both able to use HCO 3 - . Their δ 13 C values indicate that, if the HCO 3 - -use system does not (as is likely) discriminate significantly between 13 C and 12 C, then a substantial fraction of the inorganic C made available to Rubisco must return to the medium, carrying 13 C-inorganic C not fixed by Rubisco. (iv) Two sets of δ 13 C data from different hydrodynamic regimes distance from leading edge of a flat stone; different size of thalli) show that the attainable differences in situ in thickness of the diffusion boundary layer do not alter the fractional limitation of photosynthesis of Cladophora by external diffusion of inorganic C, considered with HCO 3 use. (vi) The entrusting red alga Hildenbrandia rivularis has a δ 13 C value suggestive of CO 2 as the inorganic C source, but not entirely ruling nut HCO 3 - . Marine species of both Hildenbrundia and Cladophora have δ 13 C values which, even when corrected for source inorganic C δ 13 C values, are 10%, more positive than the freshwater species. (vii) Mats of pennate diatoms were shown by pH-drift to by able to use HCO 3 - ; the relatively high (i.e. not very negative) δ 12 C value of these mats could relate to a relatively'non-leaky'HCO 3 - aequisition mechanism and/or to limitation by external diffusion (e.g. through the mat).

Objectives: Weight gain had previously been thought to be due to increased calorie intake alone though no information on its effect on total energy expenditure is available in humans. We therefore assessed whether weight gain associated with glucocorticoids is due to a reduction in energy expenditure., Design: We performed an open study with 1 mg of betamethasone given orally twice a day for 21 days., Subjects: Seven healthy female volunteers, age range 26-55 years, body mass index 19 to 40, mean 27 kg/m2., Measurements: Total free living energy expenditure was measured by the doubly labelled water method (D2 18O), resting metabolic rate by ventilated hood indirect calorimetry and fat free mass from the dilution volume of oxygen-18 labelled water. Body composition and components of energy expenditure were assessed before and during the final 14 days of betamethasone administration., Results: Weight increased by a mean of 1.2 kg (P < 0.05) because of a significant rise in fat mass (1.5 kg) with no change in fat free mass. Resting metabolic rate remained unaltered on betamethasone but total energy expenditure increased in all subjects with a significant mean rise of 26% from 11.7 to 14.7 MJ/24 h (P < 0.05). The energy component of physical activity with thermogenesis increased on average 52% (from 5.8 to 8.9 MJ/24 h; P < 0.05). The rise in energy expenditure was still apparent after correction for the increase in body weight. Fasting respiratory quotient (RQ) increased from 0.81 to 0.86 with no change in fasting blood glucose. Betamethasone did not result in an energy sparing effect on the two components of energy expenditure studied., Conclusions: Body weight increased on betamethasone entirely due to an increase in fat mass. This occurred despite a rise in total energy expenditure which involved specifically that component accounted for by physical activity plus thermogenesis. The most likely explanation is that betamethasone increased dietary energy intake significantly in excess of expenditure. We estimate that an average extra energy intake of 2.8 MJ/day would have had to be consumed for this rise in fat mass to occur even before taking into account the energy intake cost of the rise in expenditure.

Recently, we demonstrated increased incorporation of [13C]valine tracer into muscle protein after administration of a flooding dose of L-leucine. We have now investigated the possibility of a similar effect on albumin synthesis in the same group of volunteers. We gave L-[1-13C]leucine (20 atom%, 0.05 g/kg) during the final 90 min of a 7.5-h primed constant infusion of L-[1-13C]valine (99 atom%, 1.5 mg/kg prime constant infusion of 1.5 mg.kg-1.h-1) in healthy male volunteers in the postabsorptive state. Blood samples, taken at 0.5- to 1-h intervals during the constant infusion and at 5- to 30-min intervals during the application of the flooding dose, were analyzed for the concentration and 13C enrichment of leucine, valine, and their ketoacids. Albumin was isolated and hydrolyzed, and the enrichments of incorporated valine and leucine were compared with the mean enrichment of various possible precursor pools to calculate the apparent rate of albumin protein synthesis according to the standard procedures. During constant infusion of [13C]valine tracer the rate of albumin synthesis (measured using alpha-ketoisovalerate labeling as a surrogate for the true precursor) was 0.250 +/- 0.041%h (SD), a value identical to that routinely obtained using constant leucine tracer infusion and alpha-ketoisocaproate labeling. During the application of the flooding dose of leucine, the rate of incorporation of tracer [13C]valine into albumin increased by 73% to 0.433 +/- 0.129%/h (P < 0.05); the apparent protein synthetic rate calculated from the incorporation of leucine applied during the flood was 0.402 +/- 0.057 (P < 0.001). These results raise further doubts about the validity of the flooding dose method for the measurement of rates of human protein synthesis.

Objectives: We assessed whether the obesity observed in growth hormone deficient adults is maintained by a reduction in energy expenditure. We studied the effects of exogenous growth hormone on energy expenditure and body composition., Design: We performed an open study with growth hormone administered at 0.5 units per kilogram ideal body weight per week for 3 months., Patients: Seven growth hormone deficient adults were studied. Thirty-eight healthy volunteers had their resting metabolic rate measured, with seven of them proceeding to have their total energy expenditure assessed., Measurements: Total energy expenditure was measured by the doubly labelled water method (D2O18), resting metabolic rate by ventilated hood indirect calorimetry, and fat free mass from the dilution volume of oxygen-18. Body composition and components of energy expenditure were assessed before, at 2 weeks and at the end of the 3-month treatment period on exogenous growth hormone., Results: Growth hormone deficient adults did not have a low total energy expenditure compared to healthy controls (13.12 vs 12.75 MJ/24 h) with only one patient expending less than 10 MJ/24 h. None had a resting metabolic rate lower than the 95% confidence limits of normality. The amount of energy expended on physical activity and thermogenesis was significant (6.54 MJ/24 h) and was similar to healthy controls (6.47 MJ/24 h). Resting metabolic rate increased by 15.9% after 14 days on exogenous growth hormone and was elevated 12.1% after 3 months treatment but the ratio to fat-free mass remained unaltered. Total energy expenditure increased by 13.4% after 14 days therapy. Fat-free mass increased significantly after 3 months treatment by (mean) 4.5 kg with no change in fat mass and no loss in body weight., Conclusions: Obesity maintenance in growth hormone deficient adults is not a consequence of reduced total energy expenditure or a reduced exercise energy output. There was also no evidence for an energy sparing mechanism. Energy expenditure was increased by exogenous growth hormone but was not associated with a loss in fat mass or body weight suggesting the need for dietetic advice for those already obese at the outset of therapy.

Obesity is a common problem among Type 2 diabetic patients. To investigate the role of energy expenditure in the maintenance of obesity in diabetic subjects, total energy output was measured during weight stability in 23 diabetic patients: 8 lean, 5 overweight, and 10 obese. Free living total energy expenditure was measured over 14 days using doubly labelled water method, resting metabolic rate by indirect calorimetry, and urinary energy losses were assessed. Total energy output was higher in the obese (13.66 +/- SD 3.18 MJ 24 h-1) than normal weight patients (10.84 +/- 2.02 MJ 24 h-1; p < 0.05); 11.96 +/- 2.51 MJ 24 h-1 in the overweight. None of the lean but four of the obese had total energy output > 16 MJ 24 h-1. Urinary energy losses accounted for only 0.6% of total energy output in lean, 2.8% in overweight, and 3.1% in obese. Resting metabolic rate was significantly higher in obese (7.47 +/- 1.69 MJ 24 h-1) compared to lean (5.87 +/- 1.07; p < 0.05) and resting metabolic rate correlated with lean body mass (r = 0.8, p < 0.001). Thermogenesis plus physical activity was substantial and not lower in the obese (5.77 versus lean 4.97 MJ 24 h-1). The mean ratio of total energy expenditure to resting metabolic rate was in the moderate exercise category and similar in lean (1.87) and obese (1.80). Resting metabolic rate, total energy expenditure, and thermogenesis and physical activity were similar in all three groups when corrected for differences in lean body mass.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of correction of acidosis in chronic renal failure (CRF) was determined from the kinetics of infused L-[1-13C]leucine. Nine CRF patients were studied before (acid) and after two 4-wk treatment periods of sodium bicarbonate (NaHCO3) and sodium chloride (NaCl) (pH: acid 7.31 +/- 0.01, NaHCO3 7.38 +/- 0.01, NaCl 7.30 +/- 0.01). Leucine appearance from body protein (PD), leucine disappearance into body protein (PS) and leucine oxidation (O) decreased significantly with correction of acidosis (PD: acid 122.4 +/- 6.1, NaHCO3 88.3 +/- 6.9, NaCl 116.2 +/- 9.1 mumol.kg-1.h-1, acid vs. NaHCO3 P < 0.01, NaHCO3 vs. NaCl P < 0.01, acid vs. NaCl NS; PS: acid 109.4 +/- 5.6, NaHCO3 79.0 +/- 6.3, NaCl 101.3 +/- 7.7 mumol.kg-1.h-1, acid vs. NaHCO3 P < 0.01, NaHCO3 vs. NaCl P < 0.01, acid vs. NaCl NS; O: acid 13.0 +/- 1.2, NaHCO3 9.2 +/- 0.9, NaCl 15.0 +/- 1.9 mumol.kg-1.h-1, acid vs. NaHCO3 P < 0.05, NaHCO3 vs. NaCl P < 0.01, acid vs. NaCl NS). There were no significant changes in plasma amino acid concentrations. These results confirm that correction of acidosis in chronic renal failure removes a potential catabolic factor.

Before elective cesarean delivery (4 h), we infused L-[1-13C]leucine and L-[15N]phenylalanine into the maternal circulation and measured enrichment and concentration of amino acids and carbon dioxide in cord blood of six normal human fetuses at delivery. There were net fetal uptakes of leucine (2.22 +/- 0.29 mumol.kg-1.min-1) and phenylalanine (0.80 +/- 0.11 mumol.kg-1.min-1) with net outputs of CO2 (6.11 +/- 1.12 ml.kg-1.min-1) and the transamination product of leucine, alpha-ketoisocaproate (1.04 +/- 0.32 mumol.kg-1.min-1). Fetal amino acid oxidation accounted for a substantial proportion of the flux from the mother (leucine, 0.36 +/- 0.09 mumol.kg-1.min-1 and phenylalanine, 0.18 +/- 0.04 mumol.kg-1.min-1). Fetal whole body accretion of leucine carbon (0.82 +/- 0.21 mumol.kg-1.min-1) was 69% of the umbilical uptake, and that of phenylalanine (0.62 +/- 0.08 mumol.kg-1.min-1) was 78%. Fetal whole body protein synthesis was approximately 13 g.kg-1.day-1, i.e., much faster than in adults but similar to that in the newborn. Net protein accretion was 2-4 g.kg-1.day-1. The placental supply of leucine and phenylalanine exceeds the fetal demand for protein synthesis by only a small amount, suggesting that the safety margin of placental transfer may be small for these amino acids. The results suggest that the method could be applied safely to studies of fetal growth retardation.

Equilibration of hydrogen gas with the water in a variety of biological sample materials was carried out in Vacutainers using platinum-on-alumina catalyst physically isolated from the liquid water. The equilibration takes 3 days at room temperature--much slower than with catalysts which float on the water surface--but this reduces the short-term temperature sensitivity of the procedure, and the inexpensive materials used allow convenient disposal of biologically contaminated samples after analysis. The slow equilibration also allows time for complete exchange with water contained in a complex sample matrix such as plant stems or soil without the need for prior isolation of the water sample. The method has particular application in studies of human total body water where repeated studies can be carried out over a short time period without the need to wait for the previous dose to wash out. Water turnover rates from total diurnal energy expenditure studies measured by this method were not significantly different from those obtained by the zinc reduction method.

1. Whole body protein turnover was measured using a primed-constant infusion of L-[1-13C]leucine with measurement of breath 13CO2 production and plasma 13C alpha-ketoisocaproate enrichment. Ten fasting patients, requiring mechanical ventilation and suffering from multiple organ failure, and six healthy control subjects were studied. 2. Protein breakdown and leucine removal from the plasma for protein synthesis were significantly higher in the patients than in the control subjects (P < 0.01). In addition, leucine oxidation was almost 75% higher in the patients than in the healthy control subjects (P < 0.05). 3. Plasma concentrations of glucose, insulin and growth hormone were not different between the two groups, but those of glucagon (not significant), noradrenaline (P < 0.05) and cortisol (P < 0.01) were almost two- and three-fold higher in the patients than in the control subjects. 4. Mean energy expenditure, measured by indirect calorimetry, was 30% higher in the patients than in the healthy control subjects (P < 0.01). 5. Combining the data from both groups of subjects and using multiple regression analysis, cortisol was found to be the most significant predictor of (i) protein breakdown (48% of variance explained), (ii) leucine oxidation (69%) and (iii) hourly energy expenditure (54%). 6. The present investigation using [13C]leucine tracer methods demonstrated, in patients with multiple organ failure, that whole body protein breakdown and synthesis increased concomitantly and were twice as high as rates measured in healthy control subjects. Of the hormones measured in the present study, cortisol appears to have the most significant effect on whole body protein turnover.

Fourteen children receiving one year of recombinant human growth hormone (rhGH) treatment underwent measurement of serial changes in body composition (measured by skinfold thickness, bioelectrical impedance, and H2(18)O dilution), resting energy expenditure (REE, estimated by ventilated hood indirect calorimetry), and total free living daily energy expenditure (TEE, measured by the doubly labelled water technique). Mean height velocity increased from 4.9 to 8.6 cm/year after six months of treatment. Fat free mass (FFM) increased more during the first six weeks (24.4 g/day) than from six to 26 weeks of treatment (6.8 g/day); fat mass decreased by 7.2 g/day and 1.1 g/day respectively. The six week increase in REE (kJ/day) was maintained after six months of treatment, though expressed per kilogram FFM (kJ/kgFFM/day), returned to pretreatment values by three months. Height velocity increases at six months correlated with six week changes in fat mass measured by skinfold thickness and REE, though use of this relationship to predict growth response in individuals is limited by the wide 95% prediction intervals. No significant changes in growth, body composition, or energy expenditure were observed between six and 12 months of treatment, in either patients who had initially responded well to treatment or those who were poor initial responders to treatment and who had their dose of rhGH doubled after six months.