Whole body protein metabolism estimated by (15)n tracer experiments and body composition of mice selected for different growth parameters

Abstract Whole body protein synthesis was investigated in growing male mice which were long-time selected for high carcass protein amount (DU-6P, protein line) or for high body weight (DU-6, growth line) and in the unselected randomly bred control (DU-Ks). Six mice/line were housed singly in metabolic cages for the estimation of N balance, whole body protein synthesis (end-product method, single dose of (15)N-labelled amino-acid mixture), and N distribution in the body. Another six mice/line were used for the determination of the body composition. All mice had free access to a commercial stock diet (crude protein 268 g, gross energy 19 MJ/kg dry matter) and to water. Body weight of both selection lines was about twice that of control mice at the same age. Selection for high body weight resulted in higher body fat content. Scaled to the corresponding body protein pools, the protein synthesis rates of selected mice were significantly higher than in controls, but were not significantly different between both selection lines in contrast to the protein deposition rates. The higher protein accretion in the protein line in comparison to the growth line seems to be due to a combination of a lower protein breakdown and an increased protein synthesis rate.     

The Influence of Feeding Frequency on Nitrogen Turnover and Whole Body Protein Synthesis in Adult Rats (Estimation using [14C]- and [15N]- Labelled Leucine)

Abstract

Male Wistar rats (17 wks. old, body weight ～400 g), fitted with an intra gastric cannula and with a catheter in the vena jugularis were divided into 3 groups and given a marginal ration of the feeding solution Nutrison Standard (1g protein and 350 kJ ME per day). Group 1 had ad lib. access to the drinking bottle, the groups 2 and 3 were pair fed by gastric infusion, splitted up into 2 greater meals for group 2 respectively into 6 smaller meals for group 3. After adaptation all animals get an i.p. injection of doubly labelled tracer solution (200μl) containing 2.5mg L-[¹⁵N]leucine (72 atom% ¹⁵N) combined with either [1-¹⁴C]- or [U-¹⁴C] leucine (37 kBq).

The course of ¹⁴CO₂ expiration was estimated by breath test over 4h in intervals of 15 min and the course of urinary ¹⁵N excretion over 24h in intervals of 45 resp. 90 min. An infusion of saline (0.9% 5ml/h) into the vena jugularis was used to provoke sustained urine production of the animals during the experiment.

From the parameters of the excretion curves of breath ¹⁴CO₂ resp. urine ¹⁵N (cumulative end value) and from the N balance the portions of leucine-C and leucine-N used for protein synthesis, transamination decarboxylation and total oxidation as well as the kinetic parameters for whole body protein metabolism were computed.

The following conclusions were drawn:

6 x feeding regime produces a small but measurable amino acid economy effect in comparison to 2 x feeding regime.

Protein gain for 2 x feeding group was significant smaller than for 6 x feeding group, though protein synthesis rate was higher, but was overcompensated by a greater increase of protein breakdown rate for the 2 x feeding group. Energy storage in form of fat and glycogen built from decarboxylation was unaffected by feeding frequency. The amount of leucine oxidized for heat production was 4% higher for the 6 x feeding group. Transamination rate for leucine was estimated to 8–15%. Absolute values for protein flux, protein synthesis and protein breakdown may be overestimated or underestimated because the metabolism of [¹⁵N] leucine does not exactly agree with that of total N; but the proportions of them and therefore also the conclusions will be true. Better results for absolute values will be obtained using a mixture of ¹⁵N labelled AA, ¹⁵N labelled protein or hydrolysate of ¹⁵N labelled protein (yeast) as the tracer source.     

Influence of Food Restriction and Compensatory Growth on Protein Synthesis and Breakdown in Growing Rats

The experiment was carried out with 24 male albino rats (5 weeks old, live weight 79 ± 5.1 g) divided into 2 groups. Gr. 1 (control group) was fed the commercial breeding food Ssniff with 22.4% crude protein ad lib., Gr. 2 (test group) for the first fortnight a mixture of the breeding food/cellulose 60/40 (restriction period) and subsequently for the second fortnight (compensatory period) also the breeding food ad lib. During both of these periods an 8 day N balance trial and a 4 day ¹⁵N tracer trial were included for estimation of growth rate and protein synthesis rate. Protein synthesis was ascertained by the endproduct method (oral application, single dose, mixture of 12 ¹⁵N-labelled amino acids); protein breakdown was calculated as the difference of protein synthesis rate and growth rate. The estimated rates were converted into fractional ones by referring to the body N content of corresponding animals. From the results of this experiment is concluded: The smaller N balance during moderate nutritional restriction can be attributed to a decrease of protein synthesis. The compensatory growth of the animals during the realimentation period is caused by an increase of protein synthesis and the enhanced protein conversion. Protein degradation is nearly unaffected.     

Metabolic Effects of HAY's Diet

Abstract

The aim of the study was, to evaluate the metabolic effect of HAY's diet on protein turnover, fat oxidation, respiratory quotient, body fat and weight loss. Twelve healthy adults received an individually regular diet and thereafter a corresponding isocaloric and isonitrogenous 10-day HAY-diet. Protein turnover and ¹³C-fat oxidation were investigated after administration of [¹⁵N]glycine and an [U-¹³C]algae lipid mixture. The ¹⁵N and ¹³C enrichment in urine and breath were measured by isotope ratio mass spectrometry. The respiratory quotient was measured by indirect calorimetry. Body fat, total body water and lean body mass were estimated by bio-electric impedance analysis. HAY's diet led to a significantly higher ¹³C-fat oxidation (15.4 vs. 22.0% P < 0.01), corresponding to a lower respiratory quotient (0.88 vs. 0.81; P < 0.01), whereas the protein turnover remained constant in both diets (3.06 vs. 3.05 g/kg/day). HAY's diet did not reduce total body water, lean body mass, body fat and body weight (72.2 vs. 71.4 kg).     

Studies on Body Composition in Irradiated Mice

A total of 40 male and 40 female mice of equal body weight were used in the present study. Whole body gamma (⁶⁰Co) irradiation was carried out at 400, 800 and 1200 rad. Life span and several parameters of body composition were measured. Cumulative mortality precentage was lower in irradiated females in comparison to males. Ionizing radiation induced reduction (P < 0.01) in body weight, total body water with its two compartments (extra-and intracellular), fat-free body with its component (body cell mass and extracellular tissue), fat-free dry solids with its constituent (body protein and body ash), body potassium and body fat. While all of the above mentioned body composition parameters decreased (P ≤ 0.01) following whole body irradiation, the rations between the body water to these parameters increased (P ≤ 0.01). Sex difference was noticed either for the parameters or for the ratios.     

Body Water,Lean Body and Fat Mass of Healthy Children as Measured by Deuterium Oxide Dilution

Abstract

Body composition of 165 healthy children was measured using the well-established method of deuterium oxide (²H₂O) dilution. After distribution of an oral load of 2.0 ml ²H₂O/kg body weight body water was estimated from the ²H₂O concentration in urine. Lean body mass was then calculated from body water using previously published age dependent ratios of the water content of the lean body mass. Fat mass was calculated as the difference of body weight and lean body mass. A good correlation was found between body water and body weight. Linear regression revealed TBW = 0.589 BW + 0.728 (r = 0.99). Body water, lean body mass and fat mass were found to change with age. The fat content of the body increases during the first six months of life. It then decreases until four to five years then rising again until 15 years of age.     

The Measurement of Muscle Protein Synthesis in Broilers with a Flooding Dose Technique: Use of 15N-Labelled Phenylalanine,GC-MS and GC-C-IRMS

Abstract

An experiment was carried out to measure fractional muscle protein synthesis rates (k _s ) in broilers with injection of a flooding dose of phenylalanine (1 ml/100 g body weight of 150 mM phenylalanine; 38 atom percent excess (APE) [¹⁵N]phenylalanine). K _s was calculated from the [¹⁵N] enrichment in phenylalanine of tissue-free and protein-bound phenylalanine using both gas chromatography mass spectrometry (GC-MS) and gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) for measurements after a 10 min isotope incorporation period.

The tertiary-butyldimethylsilyl (t-BDMS) derivatives of phenylalanine were used for gas chromatographic separation in both systems. GC-MS and GC-C-IRMS were calibrated for a range of 7 to 37 [¹⁵N]APE and 0 to 0.62 [¹⁵N]APE, respectively, and for sample sizes of 0.45 to 4.5 nmol phenylalanine and 7 to 40 nmol phenylalanine, respectively. Reproducibility of standards as a measure of precision varied from 0.06 to 0.29 [¹⁵N]APE and from 0.0004 to 0.0018 [¹⁵N]APE in GC-MS and GC-C-IRMS, respectively.

K _s was measured in the m. pectoralis major of broilers fed rye based diets (56%) which were provided either unsupplemented (-) or supplemented (+) with an enzyme preparation containing xylanase. K _s in breast muscles was significantly increased from 21.8%/d to 23.9%/d due to enzyme supplementation.

It can be concluded from the study that the measurement of protein synthesis in broilers with the flooding dose technique can be carried out by using [¹⁵N]phenylalanine, GC-MS and GC-C-IRMS.     

Evaluation of the effect of high pressure carbon dioxide-pasteurized food on animal health

ABSTRACT

High pressure-carbon dioxide processing (HPCDP) is used as a non-thermal technology for food pasteurization to produce newer flavors for bread. However, there is no evidence that this process does not produce undesirable toxic materials. To confirm the safety of high pressure carbon dioxide-processed food, we fed high pressure carbon dioxide-processed bread to mice and evaluated their health by monitoring body weight, stress biomarkers, and gut microbiota. There was no significant difference in the body weight of normal and HPCDP bread-fed mice. Similarly, no significant difference in expression of genes important for maintaining the health of the mice was found. Although the mice’s gut microbiota showed a significant difference between the control and the two bread-fed groups, there was no significant difference between the HPCDP and the normal bread-fed groups. Therefore, we found no conclusive evidence to suggest that the processed bread contained toxic materials in comparison to the unprocessed bread.     

Line frequency and line intensity analyses of water vapour

Line position and line intensity analyses of new microwave and infrared data pertaining to water and involving the ground and (010) vibrational states are carried out using the same theoretical approach as in Coudert, L. H., 1997, J. molec. Spectrosc., 181, 246. The line position analysis involves the high J and K_a lines measured by Polyansky, O. L., Zobov, N. F., Viti, S., Tennyson, J., Bernath, P. F. and Wallace, L., 1997, J. molec. Spectrosc., 186, 422 and the ν₂ band transitions published by Toth, R. A., 1998, J. molec. Spectrosc., 190, 379. In contrast to the previous investigation, the maximum K_a value in the data set is 24 for the ground vibrational state and 21 for the (010) state. The data set considered in the line intensity analysis includes mainly the line intensities reported by Toth and is much more complete than that fitted in the present author's previous investigation as it contains many lines within the ground and (010) vibrational states. The theoretical approach accounts correctly for the observed data for both analyses. However, in the line position analysis, a few transitions had to be excluded from the fitting because they involved high lying perturbed rotational levels belonging to the (010) vibrational state.     

Maternal obesity does not influence human milk protein 15N natural isotope abundance

ABSTRACT

Obesity increases protein metabolism with a potential effect on nitrogen isotope fractionation. The aim of this study was to test the influence of obesity on human milk extracted protein ¹⁵N natural isotope abundance (NIA) at one month post-partum and to compare human milk extracted protein ¹⁵N NIA and bulk infant hair ¹⁵N NIA. This cross-sectional observational study involved 16 obese mothers (body mass index (BMI)?≥?30?kg?m^?2 before pregnancy) matched with 16 normal-weight mothers (18.5?kg?m^?2?≤?BMI?<?25?kg?m^?2) for age and pregnancy characteristics. Human milk extracted protein and bulk infant hair ¹⁵N NIA were determined by isotope ratio monitoring by mass spectrometry interfaced to an elemental analyser (IRM-EA/MS). No significant difference was found in human milk protein ¹⁵N NIA values between obese and normal-weight mothers (8.93?±?0.48?‰ vs. 8.95?±?0.27?‰). However, human milk protein ¹⁵N NIA was significantly lower than bulk infant hair ¹⁵N NIA: 8.94?±?0.38?‰ vs. 9.66?±?0.69?‰, respectively. On the basis of these results, it is concluded that human milk protein ¹⁵N NIA measured at one month post-partum is not influenced by maternal obesity. These findings suggest that ¹⁵N NIA may be exploited to study metabolism without considering maternal obesity as a confounder.     

Zum Stand der Stoffwechselforschung mit 15N- und 13C-markierten Tracersubstanzen in der Kinderheilkunde (Protein Turnover Research Using 15N and 13C Labelled Substrates in Pediatrics)

Abstract

Measurements in protein turnover and in metabolism of amino acids and their degradation products by means of stable isotope labelled substrates have been increasingly applied in clinical research over the last years. In spite of numerous studies dealing with this topic, quite a few important insufficiently clarified methodical aspects remain. This refers, for instance, to the choice of suitable tracer substances, the difficulties in the determination of the excretion plateau and the validation of the oxidation rates as measured with individual-labelled amino acids with regard to the whole body protein synthesis. Such problems may become of decisive importance in special subjects, such as preterm infants and critically-ill patients.

Investigations into these issues conducted by our group have revealed that the protein turnover in the very small preterm infant is by no means as intensive as previously claimed. The utilisation of urea nitrogen for the whole body protein synthesis of the infant may assume substantial proportions under the conditions of marginal protein intake and of catchup-growth. Studies conducted by means of ¹⁵N-labelled bifidobacteria have pointed at the intensive substrate exchange existing between microflora and host.

Pediatric research has to be non-invasive. Consequently, methods based on arterio-venous differences in tracer concentrations and on muscle biopsies do not have very high priority in pediatric research. A search for references published in the last five years has shown, that ¹⁵N-glycine is still the most frequently used tracer substance. There is a tendency towards a further increase of cell culture experiments run with stable isotope labelled amino acids.

Clinical research groups increasingly turn their attention to stable isotopes and mass spectrometry. This impressively demonstrates the continuing importance of tracerkinetic methods in all branches of medicine.     

Metabolic effects of HAY's diet

The aim of the study was, to evaluate the metabolic effect of HAY's diet on protein turnover, fat oxidation, respiratory quotient, body fat and weight loss. Twelve healthy adults received an individually regular diet and thereafter a corresponding isocaloric and isonitrogenous 10-day HAY-diet. Protein turnover and 13C-fat oxidation were investigated after administration of [15N]glycine and an [U-13C]algae lipid mixture. The 15N and 13C enrichment in urine and breath were measured by isotope ratio mass spectrometry. The respiratory quotient was measured by indirect calorimetry. Body fat, total body water and lean body mass were estimated by bio-electric impedance analysis. HAY's diet led to a significantly higher 13C-fat oxidation (15.4 vs. 22.0% P < 0.01), corresponding to a lower respiratory quotient (0.88 vs. 0.81; P < 0.01), whereas the protein turnover remained constant in both diets (3.06 vs. 3.05 g/kg/day). HAY's diet did not reduce total body water, lean body mass, body fat and body weight (72.2 vs. 71.4 kg).     

Influence of the Body Position and Emission Number in the Results of the Maximum Phonation Times of Adults without Vocal Complaints

ObjectiveTo analyze if there is an influence of the body position and emission number in the results of the maximum phonation times (MPT) of adults without vocal complaints.Materials and methodsThis is an observational, analytical and cross-sectional study. Sixty subjects participated: 30 men and 30 women without vocal complaints. Subjects were instructed to stay in orthostatic or sitting position, according to the collection's order that was selected. The first 30 subjects were evaluated in order number 1 (sitting and orthostatic) and the other 30 subjects were evaluated in order number 2 (orthostatic and sitting), with a 5-minute interval between the emissions in the two positions. The MPT of vowels /a/, /i/, /u/, fricatives /s/ and /z/, and numbers were collected. After this, the subjects were instructed to do the same emissions mentioned above, but in another position (orthostatic or sitting). The values of MPT were statistically analyzed with both the Analysis of Variance (ANOVA) paired measures test to three criteria and the Tukey Test (P < 0.05).ResultsThere was no difference between the MPT obtained in both positions. Males had all MPT higher than females. Regarding the emission number, there were differences for both sexes in the MPT of the vowel /a/ and the number counting.ConclusionIn general, it is concluded that there is no influence of body position in the results of voice temporal measures, but the sex and the number of the emissions influence the result of the MPT.     

Whole Body Branched-Chain L-Amino Acid Oxidation in Overnight Fasted Human Subjects

Abstract

Catabolism of the natural branched-chain L-amino acids in overnight fasted healthy volunteers was comparatively studied by performing oral loading tests with 1-¹³C-labelled L-leucine, L-valine, and L-isoleucine (38 μmol x kg (body weight)^?1), respectively. On the basis of the ¹³CO₂ exhalation and the ¹³C-isotope enrichment in the plasma branched-chain compounds, whole body branched-chain L-amino acid oxidation rates were estimated applying a seven compartment model and non-linear regression analysis. Mean computed in vivo oxidation rates were in the order L-leucine ? L-valine > L-isoleucine and amounted to 0.32 ± 0.06, 0.22 ± 0.04, and 0.17 ± 0.05 μmol x (kg body wt.)^?1 x min^?1 (n = 5), respectively. The data are discussed with respect to current estimates of human branched-chain L-amino acid requirements.     

Relationships between Nitrogen Translocation and Accumulation in Growing Plant Parts of Grain Legumes

Abstract

Grain legumes absorbed mineral ¹⁵N at every stage of development and transported it directedly (just as symbiotically fixed ¹⁵N₂) into the plant parts growing at the respective stage. The ¹⁵N accumulation in the grains after long-lasting ¹⁵N supply can be ascribed, for the major part, to a secondary ¹⁵N translocation after a temporary incorporation into older plant parts (leaves, stem). Inhibition experiments with antibiotics revealed no direct relation between the accumulation of amino acid ¹⁵N in growing pods and seeds and the protein synthesis in this target organs. It may include, however, processes of (active ?) uptake and transport with a possible contribution of carrier systems specific for distinct amino acids.     

The Study on the Arc Plasma Temperature Measurement by Optical Emission Spectroscopy with Fiber Optical Transmission

Abstract

A method for transmitting radiation of the arc plasma with multimode fused quartz fiber onto the spectrograph has been studied. The plot of the Boltzmann function in emission spectral analysis is used for measuring temperature of the arc plasma. The measured temperature of the arc plasma is 5946.6K from least square linear regression of ln[λI/(gA)] and E_i for a number of the emission line intensities of the excited copper atom. Its regression coefficient and measured precision are ?0.97% and 1.7%, respectively. The advantages of the method of the diagnostic temperature for the arc plasma are absolute measurements of the temperature, remote sensing, precision and suitable for mal-environment, such as high temperature, toxic, explosion, strong magnetic or/and electrical fields.

In addition, we have discussed the effect of the spectroscopic constants, such as transition probability, A , the statistical weight of the upper level, g , and the energy of the upper level, E_i , of copper lines on calculating temperature with a plot of the Boltzmann function in detail. The results show that the accurate measurement of the temperature for the arc plasma is obtained only when the spectroscopic constants are selected correctly.     

Body Temperature Mapping by Magnetic Resonance Imaging

Abstract

There are numerous clinical situations, such as hyperthermia cancer therapy, under which body temperature has to be accurately evaluated. Magnetic Resonance Imaging presently offers the best set of non-invasive methods for body temperature mapping, including diffusion imaging, relaxation time measurement, magnetization transfer contrast and chemical shift spectroscopic imaging. The basic NMR parameters involved in each are first analyzed. The role of temperature in their physical variation is then presented and finally the different practical MRI methods are briefly discussed. In the present slate of available technology and for basic physical reasons, the T 1 method appears the best compromise at low and medium magnetic field in a limited temperature range. Chemical shift could be theoretically more promising but strictly depends from the homogeneity and stability of the magnet and will be available only at high or very high field.

     

SYNTHESIS AND SPECTROSCOPIC PROPERTIES OF HIGHLY PURE PERYLENE FLUORESCENT DYES

Two new perylene dye, N, N′-bis-(aminododecyl)-3, 4, 9, 10-perylenebis (dicarboximide) (1) and N, N′-bis-(4-chlorophenyl)-3, 4, 9, 10-perylenebis (dicarboximide) have been synthesized. Because the solubility of perylene derivatives is very low, their synthesis and purification are difficult. The imides 1 and 2 are easily prepared with high yield. Dye 2 has a lower density than the density of air. They are highly fluorescent and very photostable dyes. With high fluorescent quantum yield they are suggested as new convenient standards for the fluorescence quantum yield measurements in 500–650 nm region. The report includes the electronic absorption and emission spectra, extinction coefficients and fluorescence quantum yields. Applications of the dyes are discussed.     

On the formation of photoinduced high pressure phases in sulfur below 10 GPa

Abstract

We investigate the phase transformations in sulfur for pressures up to 10 GPa by time resolved Raman spectroscopy. The transition to the photosensitive phase p-S is stimulated by the blue laser line between 3 and 9 GPa. The kinetics of this transition as derived from the time evolution of the intensities of characteristic Raman excitations shows the typical features of an activated first order phase transition. This transformation proceeds via a disordered (amorphous) intermediate state.

Above 9 GPa a further phase change to S, is kinetically characterized and follows similar rules i.e. the integral intensities of selected S, Raman lines exhibit a sigmoidal time dependence. In both high pressure phases a broad Raman excitation between 800 and 1000 cm^?1 is observed.     

稀土元素原子发射光谱及其谱线干扰的高分辨率ICP—AES研究： …

本文是系列工作的第四部分。利用光栅刻线为３６００条／ｍｍ的顺序扫描高分辨率ＩＣＰ－ＡＥＳ光谱仪研究了铕和钆分别作为基体时对其他１４个元素共６６条“首选分析线”的光谱干扰情况,获得了相应的光谱干扰轮廓和其他光谱干扰信息,为选择不同稀土基体中的稀土元素最佳分析线提供了有益的参考依据。