Influence of the time of intake of a high fat diet on gluconeogenesis

Male Wistar rats aged 75 and 150 days were given high fat diet (36.5 weight % and 30 weight % fat) over a period of 14 days. The growth (PER, NPR) and utilization (NPU, LPU) parameters of protein biological value and liver phosphoenolpyruvate carboxykinase (PEPCK) activity were determined. In another experiment, the time dependence of liver gluconeogenesis enzyme (PEPCK and fructose-1,6-diphosphatase /FDP-ase/) and transaminase (alanine and aspartate aminotransferase /ALT, AST/) activities during 24 days' administration of the diet were determined. A 14 days' high fat intake had a negative effect on protein utilization in the organism of 75- and 150-day-old animals, which was more pronounced in the younger age group (a bigger drop in net protein utilization /NPU/ and greater stimulation of PEPCK activity). In 150-day-old animals the negative effect of a high fat intake was already manifested on the 6th to 10th day of the diet to the same degree as in the younger animals on the 14th day, as seen from the increase in all the enzyme activities. The paper presents findings on differences in the degree of the negative effect of a high fat intake on protein utilization with reference to age.     

A study of the changes in protein composition of mouse brain during ontogenetic development

Of the total protein in an adult mouse brain, 35 per cent is water-soluble and 65 per cent water-insoluble. Using an extraction scheme of sequential treatment with water, Triton X-100, and sodium lauryl sulphate, it was possible to solubilize at least 90 per cent of this total in distinct groups. Qualitative analysis of the extracts was achieved by electro-phoresis in porosity gradient polyacrylamide gels. In this way, each fraction was resolved into 20-50 protein bands. This sequential extraction-fractionation procedure was employed in a study of the onto-genetic changes in mouse brain proteins. The most pronounced accumulation of protein and alteration of protein composition occurred during the first few weeks after birth. Beyond that period, both quantitative and qualitative changes were much less dramatic, except in the 1 % SLS fraction, which continued to increase in size throughout the mouse's lifespan. Whereas the aqueous-soluble proteins predominated during the very early stages of development, the detergent-soluble constituents accounted subsequently for a steadily increasing proportion of the total protein.     

Age-correlation of protein utilization to saccharide intake

Male Wistar rats aged 30, 90, 150 and 360 days were fed ad libitum on diets with an optimum protein and fat content for their respective ages and an increasing saccharide content. Net protein utilization (NPU) was determined from the body nitrogen and protein intake values and the course of gluconeogenesis in the liver was measured by specific phosphoenolpyruvate carboxykinase (PEPCK) activity. According to the growth curve for the standard diet, animals aged 30 and 90 days have a high growth rate (3.245 g/day), 150-day-old rats grow more slowly (1.856 g/day) and 360-day-old animals put on scarcely any weight at all (277 mg/day). In 30-day-old rats, NPU attains maximum values in the presence of a 36% saccharide content in the diet, in 3- and 5-month-old animals in the presence of 51% saccharides and in one-year-old animals in the presence of 41% saccharides in their food. The course of gluconeogenesis also corresponds to these values. PEPCK activity in the youngest age group is greatest in the presence of 31% saccharides in the food, at 90 days it is stimulated in the presence of 31-46% saccharides, at 150 days the decisive concentration is 41 and 46% and at one year proteins are used for saccharide synthesis in diet with a 31 and 36% saccharide concentration. For optimum saccharide values, PEPCK activity is reduced in every age group; together with the maximum NPU values, this indicates that proteins are used for growth and building of the organism at an early age and for the renewal of tissues and organs and maintenance of the organism in adulthood.     

Protein utilization in correlation to protein intake

In a 14-day experiment, weaned and adult rats were given ad libitum isocaloric diets with a mounting casein content (5, 10, 15, 25 and 40% by weight) and growth parameters of protein biological value, PER and NPR, and the utilization parameters NPU (body protein) and LPU (liver protein) were determined together with phosphoenolpyruvate carboxykinase (gluconeogenetic enzyme) and pyruvate kinase (glycolytic enzyme) activity in the animals' liver. The decrease in all the biological value parameters in weaned rats on 25% and 40% casein diets and in adult rats on 15%, 25% and 40% casein diets shows that these concentrations are too high for the organism. The decrease in PER and diminished weight and body and liver nitrogen increments in both age groups in animals with a low protein intake is evidence that 5% casein is an inadequate concentration. The optimum diet for weaned rats is thus a 15% casein diet and for adult rats a 10% casein diet, as confirmed by the linear correlation between weight increments, body and liver nitrogen and protein intake and also by gluconeogenetic enzyme activity. Under the given experimental conditions the study is a contribution to the determination of optimum physiological doses of proteins.     

Effect of feed intake on heat production and protein and fat deposition in milk-fed veal calves

Energy requirements for veal calves have not been updated recently despite the increased age at slaughter and the predominance of the Prim'Holstein breed in Europe. The objectives of this study were to determine the effects of four feeding levels (FLs) on protein and fat deposition and heat production in milk-fed calves at three stages of fattening and to determine energy requirements of calves. At each stage, 16 Prim'Holstein male calves (mean body weight (BW): 73.4, 151.6 and 237.4 kg) were fed a milk replacer at 79%, 87%, 95% or 103% of a reference FL. Measurements for one stage were conducted over 4 successive weeks in two open-circuit respiration chambers and consisted of a 6-day nitrogen and energy balance followed by a fasting day for estimating fasting heat production (FHP) of the calves. Heat production (HP) measurements were analyzed using a modeling approach to partition it between HP due to physical activity (AHP), feed intake (thermic effect of feeding (TEF)) and FHP. There was no effect of FL and stage on apparent digestibility coefficients, except for a tendency for increased digestibility coefficient of fat as animals got older. The metabolizable energy (ME)/digestible energy (DE) ratio did not depend on FL but decreased (P < 0.01) as animals got older in connection with marked increases in urinary glucose and urea excretion. The AHP and TEF components of HP were not affected by stage or FL and averaged 8.4% and 7.8% of ME intake, respectively. The FHP, expressed per kg BW0.85, increased with increasing FL, suggesting that also ME requirement for maintenance (MEm) may depend on FL. For an average intake of 625 kJ ME/kg BW0.85 per day (95% of the reference FL), FHP was 298 kJ/kg BW0.85 per day. Energy retention as protein and fat increased with increasing FL resulted in higher BW gain. But the rate of increase depended on stage of growth. The slope relating protein deposition to FL was lower in the finishing phase than in the growing phase, while the slope for lipid deposition was greater. Protein and fat contents of BW gain were not affected by FL but increased as animals got older. From these results, the energy requirements of veal calves are proposed according to a new approach, which considers that MEm (expressed per kg BW0.85) depends on ME intake (kJ/kg BW0.85) according to the following relationship: MEm = 197 + 0.25 × ME intake. The corresponding marginal efficiencies of ME utilization for protein and fat deposition are then 82% and 87%, respectively.     

Effect of a protein preload on food intake and satiety feelings in response to duodenal fat perfusions in healthy male subjects

The control of food intake and satiety requires a coordinated interplay. Oral protein and duodenal fat inhibit food intake and induce satiety, but their interactive potential is unclear. Our aim was therefore to investigate the interactions between an oral protein preload and intraduodenal (ID) fat on food intake and satiety feelings. Twenty healthy male volunteers were studied in a randomized, double-blind, four-period crossover design. On each study day, subjects underwent one of the following treatments: 1) water preload plus ID saline perfusion, 2) water preload plus ID fat perfusion, 3) protein preload plus ID saline perfusion, or 4) protein preload plus ID fat perfusion. Subjects were free to eat and drink as much as they wished. An oral protein preload significantly reduced caloric intake (19%, P < 0.01). Simultaneous administration of an oral protein preload and ID fat did not result in a positive synergistic effect with respect to caloric consumption, rejecting the initial hypothesis that the two nutrients exert a positive synergistic effect on food intake. An oral protein preload but not ID fat altered the feelings of hunger and fullness. These data indicate that the satiety effect of an oral protein preload is not amplified by ID fat; indeed, the effect of a protein preload does not seem to be mediated by cholecystokinin, glucagon-like peptide-1, or peptide YY. Much more information is necessary to understand the basic physiological mechanisms that control food intake and satiety.     

Effect of garlic on high fat induced obesity

The present study was performed to examine the effects of garlic on obesity and blood lipid profiles in high-fat induced obesity mice model, and to elucidate the molecular mechanisms responsible for such effect. C57BL/6 mice were fed a standard diet (STD) or high-fat diet (HFD) for 5 weeks to induce obesity. Mice were then randomly divided into four groups with 10 mice per group, and fed experimental diet for 4 weeks; STD group, HFD group, HFD containing 2% or 4% garlic group (HFD + G2 or HFD + G4, respectively). Administration of garlic significantly reduced HFD-induced body weight, epididymal fat accumulation, hyperlipidemia and hypercholesterolemia. Consequently, the atherogenic indexes were reduced by 83% and 91%, respectively, in 2% and 4% garlic supplemented group. Liver steatosis induced by HFD was ameliorated by garlic supplementation. Furthermore, garlic affected the down regulation of expression patterns of epididymal adipose tissue genes such as peroxisome proliferator-activated receptor γ (PPARγ), acetyl CoA carboxylase (ACC), adipose specific fatty acid binding protein (aP2), and glycerol-3-phosphate dehydrogenase (GPDH). These results suggest that garlic may have a potential benefit in preventing obesity.     

Hypophysis: a new scheme of ontogenetic development

It is usually accepted that hypophysis originates from two ecotodermal primordiums that give rise to its two main parts. The transformation of initial primordiums into definitive hypophysis is regarded as a result of mutual inductive influences between hypothalamus and Rathke's pouch. This scheme was remaining unchanged during many decades. Based upon the arguments that Spemann's organizer, prechordal mesoderm (PCM) and hypophysis are consecutive derivatives that replace each other during ontogeny, the author proposes that the two novel stages preceding the former starting point should be added to traditional scheme of pituitary development. Besides, according to new scheme the PCM (successor of Spemann's organizer) plays the main role in hypophysis development by active stimulation of morphogenesis in adjacent tissues. This leads to formation of Rathke's pouch and infundibulum, to their approaching each other, and to reprogrammin of Rathke's pouch ectoderm into hormonal cells of adenohypophysis. According to new hypothesis role of Rathke's pouch is accessorial and consists in provision of cell material.     

Regenerative capability in the hindlimb of Xenopus laevis during ontogenetic development

The regenerative capacity of hindlimb of Xenopus laevis was investigated by amputating the limbs at four levels in various developmental stages including younger postmetamorphosed froglets. Amputations of limbs were performed at the base of limb in stages 50, 51, 52, 53, 54, 55, 58, and 60 (Nieuwkoop and Faber's table), at the middle of limb bud in stages 50, 51, 52 and 54, and at mid-thigh and mid-shank in stages 58 and 60, and the froglets in 2 and 3 cm in snout-vent length. In the present experiments the regenerative capacity of limbs was expressed by the rate of regeneration and morphogenesis. Tadpoles in the stages after 55 failed to regenerate when the limbs were amputated at base level, but individuals in all the other experimental series exhibited regeneration in various rates irrespective of the level of amputation and the stage. The regenerative capacity increased distally along the proximo-distal axis of the limb when amputated at the same stage, while regeneration was better in younger stages than that in older stages when amputations were made at the same levels. The regenerates obtained by amputation of limbs in stages between 50 and 54, were mainly digitated in that they had 5 toes with 3 claws which is the same pattern with the normal limb, 4 toes with 2 claws, 3 toes with 2 claws or one, and 2 toes with one claw etc. Tadpoles at stage 50 could regenerate toes and claws without defect, but in the later the regenerative capacity gradually declined by reducing the number of toes and claws and accompanied by malformation of skeleton as the stage proceeded. The tadpoles in stages after 58, and the froglets of 2 and 3 cm, produced various types of heteromorphic regenerates of shapes such as cone, spike or rod of which the centra were occupied with cartilage rods. However these regenerates showed no morphological differences according to the developmental stages. These heteromorphic regenerates continued their growth even after one year without any sign of development of digitated feet.     

Structure-function relationships in the integument of Salamandra salamandra during ontogenetic development

Morphological, cytological and transport properties of the integument of Salamandra salamandra were investigated during natural ontogenetic development, from birth to adult. Three stages were operationally defined: I, larvae, from birth to metamorphosis; II, metamorphosis (judged externally by the colour change and loss of the gills); and III, post-metamorphosis to adult. Pieces of skin were fixed at various stages for immunocytochemical examinations, and the electrical properties were investigated on parallel pieces. Distinct cellular changes take place in the skin during metamorphosis, and lectin (PNA, WGA and ConA) binding indicates profound changes in glycoprotein composition of cell membranes, following metamorphosis. Band 3 and carbonic anhydrase I (CA I) were confined to mitochondria-rich (MR)-like cells, and were detected only in the larval stage. CA II on the other hand, was detected both in MR-like and in MR cells following metamorphosis. The electrical studies show that the skin becomes more tight (transepithelial resistance increases) upon metamorphosis, followed by manifestation of amiloride-sensitive short-circuit current (I(SC)) indicating that functional Na+ uptake has been acquired. The skin of metamorphosed adults had no finite transepithelial Cl- conductance, and band 3 was not detected in its MR cells. The functional properties of MR-like and MR cells remain to be established.     

Functional impact of high protein intake on healthy elderly people

Decline in muscle mass, protein synthesis, and mitochondrial function occurs with age, and amino acids are reported to enhance both muscle protein synthesis and mitochondrial function. It is unclear whether increasing dietary protein intake corrects postabsorptive muscle changes in aging. We determined whether a 10-day diet of high [HP; 3.0 g protein x kg fat-free mass (FFM)(-1) x day(-1)] vs. usual protein intake (UP; 1.5 g protein x kg FFM(-1) x day(-1)) favorably affects mitochondrial function, protein metabolism, and nitrogen balance or adversely affects insulin sensitivity and glomerular filtration rate (GFR) in 10 healthy younger (24+/-1 yr) and 9 older (70+/-2 yr) participants in a randomized crossover study. Net daily nitrogen balance increased equally in young and older participants, but postabsorptive catabolic state also increased, as indicated by higher whole body protein turnover and leucine oxidation with no change in protein synthesis. Maximal muscle mitochondrial ATP production rate was lower in older people, with no change occurring in diet. GFR was lower in older people, and response to HP was significantly different between the two groups, with a significant increase occurring only in younger people, thus widening the differences in GFR between the young and older participants. In conclusion, a short-term high-protein diet increased net daily nitrogen balance but increased the postabsorptive use of protein as a fuel. HP did not enhance protein synthesis or muscle mitochondrial function in either young or older participants. Additionally, widening differences in GFR between young and older patients is a potential cause of concern in using HP diet in older people.     

Effect of inflammation stimulation on energy and nutrient utilization in piglets selected for low and high residual feed intake

Selection of animals for improved feed efficiency can affect sustainability of animal production because the most efficient animals may face difficulties coping with challenges. The objective of this study was to determine the effects of an inflammatory challenge (using an intravenous injection of complete Freund’s adjuvant – CFA) in piglets from two lines of pigs divergently selected during the fattening period for a low (RFI−) or a high (RFI+) residual feed intake (RFI; difference between actual feed intake and theoretical feed requirements). Nitrogen and energy balances (including heat production – HP – and its components: activity-related HP – AHP, thermic effect of feeding, and resting HP) were measured individually in thirteen 20-kg BW castrated male piglets (six and seven from RFI+ and RFI− line, respectively) fed at the same level (1.72 MJ ME/kg BW^0.60 per day) from 3 days before to 3 days after CFA injection. Dynamics of dietary U-¹³C-glucose oxidation were estimated from measurements of ¹³CO₂ production on the day before and 3 days after the CFA injection. Oxidation of dietary nutrients and lipogenesis were calculated based on HP and O₂ consumption and CO₂ production. The data were analyzed as repeated measurements within piglets in a mixed model. Before CFA injection, RFI− piglets had a lower resting energy expenditure than RFI+ piglets, which tended to increase energy retention because of a higher energy retention as fat. The CFA injection did not affect feed intake from the day following CFA injection onwards but it increased energy retention (P=0.04). Time to recover 50% of ¹³C from dietary glucose as expired ¹³CO₂ was higher in RFI+ piglets before inducing inflammation but decreased after to the level of RFI− piglets (P<0.01). Oxidation of U-¹³C-glucose tended to slightly increased in RFI− piglets and to decreased in RFI+ piglets (P=0.10) because of CFA. Additionally, RFI− piglets had a lower respiratory quotient during the 1st day following the CFA injection whereas RFI+ piglets tended to have a higher respiratory quotient. In conclusion, selection for RFI during the fattening period also affected the energy metabolism of pigs during earlier stages of growth. The effects of CFA injection were moderated in both lines but the most efficient animals (RFI−) exhibited a marked re-orientation of nutrients only during the 1st day after CFA, and seemed to recover thereafter, whereas the less efficient piglets expressed a more prolonged alteration of their metabolism.