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Biomedical subjects

M F Fuller

Publications and source records attributed to M F Fuller.

At least 37 records · Page 2Linked to original sources

Validation of the doubly labeled water method in growing pigs.

The CO2 production (rCO2) of eight growing pigs was determined by continuous collection of CO2 over 21 days and simultaneously estimated using the doubly labeled water (DLW) method. The aim was to assess the accuracy of the method before and after correction for known sources of error and to test for any residual discrepancy arising from as yet unidentified sources of error. Mass spectrometer accuracy was verified by analyzing serial dilutions of the dose material in the form of an artificial decay curve; no significant bias was detected. The physiological errors were linearly dependent on weight gain. DLW-derived rCO2 (corrected only for fractionated water loss) underestimated the true value by 0.270 l CO2/g wt gain or -8% in the restricted (group R) and -16% in the ad libitum-fed (group AL) groups. Known sources of error accounted for -0.006 (methane), -0.032 (fecal 2H losses), -0.108 (fat synthesis), and -0.146 (changing pool size) l CO2/g wt gain. After correction for these sources of error the DLW-derived rCO2 differed from the true value by -2 +/- 3% in group R and 0 +/- 3% in group AL. Thus there was no significant bias in the DLW method after correction for known sources of error, even during rapid weight gain or at weight stability with or without correction. The precision estimates include both dose and background errors and uncertainty in the correction factors used. Strategies for optimizing precision are presented.

Animals↗

Effects of naloxone, metaclopramide and domperidone on plasma levels of prolactin and LH following suckling in the female rabbit.

Saline, naloxone, domperidone or metaclopramide was injected into lactating rabbits immediately before suckling. Blood samples were taken prior to injection (0 minutes) and then at 15, 30, 45 and 60 minutes after the start of suckling, after which the samples were assayed for plasma prolactin and LH concentrations. In all the does there was a significant increase in prolactin concentration, which was highest 15 minutes after the start of suckling, and which declined exponentially thereafter to levels significantly higher than before suckling. The increase in prolactin concentration was similar in does given saline and naloxone, but it was significantly enhanced in does given metaclopramide; with domperidone the increase was intermediate and not significantly different from that following treatment with saline. In does given saline, domperidone, and metaclopramide plasma LH concentrations declined slowly during the hour after suckling but the concentration was increased significantly in does given naloxone. The inverse correlations between prolactin and LH were low weak and were not significant.

Journal Article↗

Assessment of fermentation in growing pigs given unmolassed sugar-beet pulp: a stoichiometric approach.

In four experiments growing pigs were given a cereal-based diet alone or supplemented with unmolassed sugar-beet pulp (SBP), used as a model substrate for fermentation. The rates of production of methane and gaseous hydrogen were measured and, together with the molar proportions of volatile fatty acids (VFA) in the digesta, used in stoichiometric calculations of fermentation. The resulting estimates were only one-sixth of the observed extent of digestion of SBP. Bacteriostatic levels of antibiotics reduced fermentation by more than half, as judged from the digestion of non-starch polysaccharides: allowing for the incomplete suppression of fermentation it was estimated that the production of methane and VFA could account completely for the digested SBP. The potential contribution of various routes of hydrogen disposal to the error of the stoichiometric calculations is discussed.

Animals↗

The form of response of body protein accretion to dietary amino acid supply.

The mean response of body protein accretion in growing animals to their amino acid intake is sometimes described by a rectilinear ("broken-line") model and sometimes by a curvilinear model. The response of a population may be curvilinear as a result of averaging individual rectilinear responses or because individual responses are themselves curvilinear. This experiment was undertaken to distinguish these possibilities by examining the responses of individual animals. Eighteen pigs with a mean initial weight of 35 kg and a mean final weight of 73 kg were each fed, in a different sequence, six diets providing a daily nitrogen intake of 0.5, 1.5, 1.83, 2.17, 2.5 or 3.5 g/kg body wt0.75. Each diet was given for 10 d, with complete collections of feces over the last 6 d and of urine over the last 4 d. Rates of nitrogen retention (expressed per kg0.75 per day) were related to rates of nitrogen intake (in the same units) using a rectilinear (broken line), an exponential or a saturation kinetics model. Mean square errors were significantly larger for the rectilinear model than for either of the curvilinear models and were slightly but not significantly less for the saturation kinetics model than for the exponential model.

Amino Acids↗

Valine oxidation: the synthesis and evaluation of L-[3-3H]valine as a tracer in vivo.

The suitability of L-[3-3H]valine for measuring valine oxidation was studied by comparing its oxidation rate with that of L-[1-14C]valine in rats and pigs. L-[3-3H]valine was synthesized by removal of the tritium on carbon-2 of L-[2,3-3H]valine by acetylation. The acetyl group was removed enzymatically using pig renal acylase 1 (EC 3.5.1.14) and the product was purified by ion-exchange and paper chromatography. For the first rat experiment L-[3-3H]valine was synthesized in our laboratory; for the subsequent experiments it was produced by Amersham International plc. In the first experiment in rats the two tracers were given by injection and 14CO2 was collected for 2 h. The oxidation of tritiated valine was significantly higher than that of L-[1-14C]valine. In a second experiment there was no difference. This was probably due to the higher purity of the labelled valine which, for the second experiment, was shown by nuclear magnetic resonance to contain only one tritium atom. In a study with pigs in which the two tracers were given by continuous infusion there was no significant difference between them in flux or oxidation. The results of this experiment were used to evaluate a model to estimate amino acid requirements. With pigs given a methionine-limiting diet a reduction in methionine intake, by reducing protein accretion, increased valine oxidation by the same proportion.

Amino Acids↗

Validation of the in vivo measurement of adipose tissue by magnetic resonance imaging of lean and obese pigs.

In vivo quantification of adipose tissue with magnetic resonance imaging (MRI) was validated with pigs. Thirteen transaxial MRI sections were collected, at intervals proportional to body length, from each pig, which was then killed, frozen, and sliced at the locations of the MRI sections. Adipose-tissue quantities were determined by dissecting each slice, and lipid contents of the dissected slices and of the tissue segments between slices were measured. Compared with dissection, MRI underestimated abdominal percent adipose tissue and overestimated cervical percent adipose tissue by less than 6%. When all 13 sections were used, MRI closely predicted percent lipid and dissected percent adipose tissue with small residual SDs (RSD = 1.9 and 2.1, respectively), which increased only slightly if two sections (4, upper thorax and 8, upper abdomen) were used (RSD = 2.3 and 2.6, respectively). In conclusion MRI accurately quantifies adipose tissue in vivo, matching values produced by dissection and chemical analysis.

Adipose Tissue↗

Body fat in lean and overweight women estimated by six methods.

Body fat content of seven lean women (body mass index (BMI) 20.6 (SD 1.8) kg/m2) and seven overweight women (BMI 31.1 (SD 3.3) kg/m2) was estimated by six different methods: underwater weighing (UWW), body-water dilution (BWD), whole-body counting (40K), skinfold thickness (SFT), bioelectrical impedance (BEI) and magnetic resonance imaging (MRI). Using UWW as the reference method, the differences between percentage fat by each other method and the percentage fat by UWW were calculated for each subject. The mean difference was lowest for SFT and highest for BWD. MRI showed the lowest variability in individual results, and 40K the highest. 40K and BWD methods used in combination gave better agreement with UWW results than either 40K or BWD methods alone. There was a weak negative correlation between the difference from the UWW results and percentage fat in the SFT measurements, but not in the BWD, 40K, BEI or MRI measurements, suggesting that for these methods the assumptions involved produced no greater inaccuracy in the overweight women than in the lean women. In all subjects the BEI offered little improvement over the traditional SFT measurements. The agreement between MRI and UWW estimates in both lean and overweight women suggests that MRI may be a satisfactory substitute for the more established methods of body fat estimation in adult women.

Adipose Tissue↗

Total and subcutaneous adipose tissue in women: the measurement of distribution and accurate prediction of quantity by using magnetic resonance imaging.

Total and subcutaneous adipose tissue in seven lean and seven obese women were quantified using magnetic resonance imaging (MRI). The distributions of adipose tissues along the body were closely correlated: subcutaneous with total, both within and between lean and obese groups. Lean women had proportionally less adipose tissue in the lower thorax and upper abdomen than did obese women. Reducing the number of MRI scans from 17 to 4 did not increase the residual SD of predicted body adipose tissue (2.9 percent) when body density was used as the reference measure. MRI gave an estimate of total-body adipose tissue significantly closer to the value for fat percent produced when the results from five other techniques (skinfold thickness, underwater weighing, 40K whole-body counting, isotopic water dilution, and tetrapolar bioelectrical impedance) were averaged than when any other technique was used alone. MRI-determined percent body adipose tissue in women is close to, and proportional to, estimates derived by underwater weighing.

Abdomen↗

Sarcosine kinetics in pigs by infusion of [1-14C]sarcosine: use for refining estimates of glycine and threonine kinetics.

To investigate in vivo the interconversion between glycine (Gly) and its N-methyl product sarcosine (Sar), [1-13C]Gly and [1-14C]Sar were infused into hourly fed pigs receiving diets with low- and high-threonine levels. An open two-pool model was developed to calculate Sar demethylation (DM) and Gly methylation (GM). During [1-14C]Sar infusion, intracellular Gly specific radioactivities (SA) in the liver and kidney were higher than plasma Gly SA, suggesting that demethylation of Sar occurred in those tissues. DM estimated by using hippuric acid (HA) as the production pool had a mean value of 1.55 mumol.kg-1.h-1, similar to the Sar production rate (mean 1.85 mumol.kg-1.h-1). GM was undetectable (less than 0.5 mumol.kg-1.h-1). These results suggest that, in fed pigs, Sar is produced mainly from choline catabolism and is degraded only to Gly in liver and kidney. On the assumption that Sar degradation gave rise only to Gly, the production rate of Gly (Gly PR) was calculated from [1-13C]Gly and [1-14C]Sar infusions using either the primary pools (plasma Gly and HA, respectively) or the secondary pools (HA and plasma Gly, respectively). The results were explained by a liver-plasma Gly exchange model. The whole body Gly irreversible loss, i.e., direct loss from plasma and liver, was calculated from this model to be 832 +/- 58 mumol.kg-1.h-1, showing that the estimation of Gly PR with [1-13C]Gly infusion and plasma Gly enrichment (599 +/- 56 mumol.kg-1.h-1) was a significant underestimate of the true value.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Water hydrogen incorporation into body fat in pigs: effect on double/triple-labeled water method.

A basic assumption of the doubly labeled water (DLW) and triply labeled water (TLW) methods for measuring water flux (rH2O), CO2 production (rCO2), and fractionated water loss (X) is that the H of body water only leaves the body as water. Any loss of isotopes in other products will introduce an error into these techniques. The body fat represents the largest potential sink for water H. 2H sequestration into the carcass fatty acids was investigated in eight pigs labeled with 2H2O for 21 days. rCO2 was measured simultaneously in respiration chambers to allow an accurate assessment of the effect of 2H sequestration on the estimated rCO2. The fat content of the diet (1.63%), level of intake, and stage of maturity were all designed to give the widest possible range of sequestration effects. Four animals were restricted to their estimated maintenance requirement and four were allowed to feed ad libitum giving a range of weight gain from 100 to 650 g/day. This was reflected in the estimated error on rH2O (+0.42% in the restricted group and +2.52% in the fast-growing animals) and on rCO2 (-1.30 and -7.59%, respectively). The error on the calculation of X using TLW was +0.03 units in the restricted group and +0.20 units in the fast-growing animals. The error of +0.2 on X propagates through to an underestimate of rCO2 of approximately 4%, and since this is additive with the error on DLW the ultimate error on rCO2 using TLW would be approximately -12%.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗

Effects of reducing the remating interval after parturition on the fertility and plasma concentrations of luteinizing hormone, prolactin, oestradiol-17 beta and progesterone in lactating domestic rabbits.

Primiparous crossbred does were remated on Day 1 (n = 15) or 14 (n = 25) post partum and killed on Day 10 post coitum to assess their fertility. Blood samples were taken during the pre- (0-12 h post coitum) and post- (1-10 days post coitum) ovulatory periods and plasma was assayed for luteinizing hormone (LH), prolactin, oestradiol-17 beta and progesterone. Ovulation response was significantly greater (P less than 0.01) and ovulation rate significantly lower (P less than 0.001) in does mated on Day 1 than in those mated on Day 14 post partum. Does failing to ovulate on Day 14 post partum exhibited no preovulatory LH surge and had significantly lower (P less than 0.05) premating concentrations of oestradiol-17 beta and prolactin than those ovulating at this time. No significant differences in hormone concentrations were observed during the preovulatory period between does ovulating on Days 1 and 14 post partum, with the exception of oestradiol-17 beta. Concentrations of this hormone were significantly lower (P less than 0.01) in does mated on Day 1, at 1 h post coitum. We conclude that (i) fertility was affected by the remating interval after parturition, (ii) ovulation failure was associated with an absence of the preovulatory LH surge and a reduction in premating concentrations of oestradiol-17 beta and prolactin and (iii) the lower ovulation rate in early lactation was apparently caused by a reduction in ovarian competence to respond to the gonadotrophic stimulus.

Animals↗

A molecular biological approach to reducing dietary amino acid needs.

Rapid developments in transgenic animal technology make it possible to consider introducing new metabolic capabilities into animals, using genes from other species. Lysine and threonine are both essential amino acids in mammals, and are commonly the first and second limiting amino acids, respectively, for protein accretion in pigs and poultry fed cereal based diets. Here we consider the potential for transgenic animals with microbial biosynthetic pathways for these amino acids.

Amino Acids, Essential↗

Quantitative partition of threonine oxidation in pigs: effect of dietary threonine.

Kinetic aspects of threonine (Thr) metabolism were examined in eight pigs fed hourly with a diet containing either 0.68% (LT group) or 0.81% (HT group) of Thr (wt/wt), corresponding to 10 and 30% Thr excess, respectively, compared with an "ideal" diet. Primary production (PR) and disposal (DR) rates were obtained for Thr, glycine (Gly), and 2-keto-butyrate (KB) after a 12-h continuous infusion of L-[U-14C]-Thr together with [1-13C]Gly and a 6-h continuous infusion of [1-14C]KB. Transfer of Thr into secondary pools was also monitored, and from these the rates of Thr oxidation through the catabolic pathways of L-Thr 3-dehydrogenase (DR(Thr-Gly)) and threonine dehydratase (DR(Thr-KB)) were estimated. For the LT group the results were (mumol.kg-1.h-1) PR(Thr) 314 +/- 3, PR(Gly) 551 +/- 24, PR(KB) 41 +/- 3, DR(Thr-Gly) 22 +/- 2, and DR(Thr-KB) 7 +/- 1. For the HT group they were PR(Thr) 301 +/- 23, PR(Gly) 598 +/- 55, PR(KB) 39 +/- 4, DR(Thr-Gly) 32 +/- 2, and DR(Thr-KB) 8 +/- 1. The increase in Thr intake (14 mumol.kg-1.h-1, P less than 0.01) induced a commensurate increase in the sum of DR(Thr-Gly) and DR(Thr-KB) (14 mumol.kg-1.h-1, P less than 0.001) when liver was used as the precursor pool. This was mainly due to the increased DR(Thr-Gly) (13 mumol.kg-1.h-1, P less than 0.01); the change in DR(Thr-KB) was not statistically significant. By comparison of intracellular-to-plasma ratios of specific activities (or enrichments) for different tissues with each type of infusion, liver was shown to be the major site of production of Gly and KB from Thr. These data suggest that in fed growing pigs a 30% excess of Thr in the diet does not alter the partition of Thr oxidation, since 80% of Thr oxidation occurs through the L-Thr 3-dehydrogenase pathway for both LT and HT groups.

Amino Acids↗

The optimum dietary amino acid pattern for growing pigs. 1. Experiments by amino acid deletion.

A series of four nitrogen-balance experiments was carried out with growing pigs to determine the optimum balance amongst the amino acids in the diet. The reduction in N retention when 20% of a single amino acid was removed from the diet was used to calculate a dietary amino acid pattern in which each amino acid would be equally limiting. A mixture of amino acids simulating the amino acid pattern of casein was used with the same efficiency as casein. From two successive deletion experiments an optimum balance amongst the essential amino acids was derived. Expressed relative to lysine = 100 this had threonine 72, valine 75, methionine + cystine 63, isoleucine 60, leucine 110, phenylalanine + tyrosine 120, tryptophan 18. No estimate was made for histidine. Essential amino acids in this pattern were mixed with non-essential amino acids in ratios of 36:64 up to 57:43. The highest efficiency of N retention was achieved with diets having a ratio of at least 45:55. This included (g/16 g N) lysine 6.5, threonine 4.7, valine 4.9, methionine + cystine 4.1, isoleucine 3.9, leucine 7.2, phenylalanine + tyrosine 7.8, tryptophan 12. The N of diets with this amino acid pattern was utilized significantly better than when the pattern proposed by the Agricultural Research Council (1981) was used. The flow of amino acids past the terminal ileum of pigs given the semi-synthetic diet with this amino acid pattern was no greater than that observed with protein-free diets. The proposed pattern thus describes the intrinsic requirements of the growing pig for absorbed amino acids.

Amino Acids↗

The optimum dietary amino acid pattern for growing pigs. 2. Requirements for maintenance and for tissue protein accretion.

Experiments were made to estimate separately the amino acid requirements of growing pigs for maintenance and for protein accretion. The relationship between nitrogen retention and amino acid intake was estimated for each essential amino acid (except histidine) by giving, at rates of N intake of 0.25 and 2.0 g/kg body-weight (W)0.75 per d, diets in which one amino acid was made specifically deficient. From the regression coefficients it was calculated that, for the accretion of 1 g body protein, the dietary amino acid requirements were (mg) threonine 47, valine 53, methionine + cystine 36, methionine 19, isoleucine 43, leucine 78, phenylalanine + tyrosine 84, phenylalanine 41, lysine 68 and tryptophan 12. The daily amino acid requirements for N equilibrium were also estimated. From the relationship between N retention and amino acid intake the daily amino acid requirements for N equilibrium were estimated to be (mg/kg W0.75 per d) threonine 53, valine 20, methionine + cystine 49, methionine 9, isoleucine 16, leucine 23, phenylalanine + tyrosine 37, phenylalanine 18, lysine 36 and tryptophan 11. It was estimated that both for maintenance and for protein accretion tyrosine could provide close to half the total phenylalanine + tyrosine needs. Cystine could supply close to half the total sulphur amino acid needs for protein accretion but 0.8 of the needs for maintenance.

Amino Acids↗

Fertility of the early postpartum, lactating domestic rabbit.

Sixty-four crossbred primiparous lactating does each suckling six pups were allocated at random into four groups and were mated on either Day 1, 2, 3, or 4 post partum (where Day 0 = the day of parturition). They were subsequently killed on Day 10 post coitum (where Day 0 = the day of mating) to assess fertility. There were no significant differences between treatment groups in their mating response (97% overall), ovulation response (77% overall), implantation response (83% overall), implantation rate (8.7 overall), or preimplantation mortality rate (24% overall). Ovulation rate was significantly increased in does mated on Days 3 and 4 (13.3 and 13.1, respectively), compared with those mated on Day 1 (10.2, P<0.05) and Day 2 (9.6, P<0.01) post partum. From these results we conclude that fertility is high throughout the early postpartum period in the lactating rabbit.

Journal Article↗