Biomedical subjects

A J Hulbert

Publications and source records attributed to A J Hulbert.

At least 55 records · Page 3Linked to original sources

PubMed1998

Endurance in high-fat-fed rats: effects of carbohydrate content and fatty acid profile.

The purpose of this experiment was to study endurance performance and substrate storage and utilization in fat- or carbohydrate-fed rats. Ninety-nine rats were randomly divided into three groups and over 4 wk were fed either a carbohydrate-rich [CHO; 10% total energy content in the diet (E%) fat, 20 E% protein, 70 E% carbohydrate] diet or one of two fat-rich diets (65 E% fat, 20 E% protein, 15 E% carbohydrate) containing either saturated (Sat) or monounsaturated fatty acids (Mono). Each dietary group was randomly assigned to a trained (6 days/wk, progressive to 60 min, 28 m/min at a 10% incline) or a sedentary group. Rats were killed either before or after a treadmill endurance run to exhaustion. Training increased endurance (206%), but diet composition did not affect endurance in either trained or sedentary rats. beta-Hydroxyacyl-CoA dehydrogenase activity was increased in fat-fed but not carbohydrate-fed rats (P < 0.05). Respiratory exchange ratio during the initial phase of exercise was lower after the Mono compared with the Sat diet (P < 0. 05) and higher after the CHO than the Sat diet (P < 0.05). Thus adaptation to a high-fat diet containing a moderate amount of carbohydrates did not induce enhanced endurance in either trained or untrained rats; however, substrate utilization was modulated by both amount and type of dietary fat during the initial stage of exercise in trained and sedentary rats.

3-Hydroxyacyl CoA Dehydrogenases↗

PubMed1997

The proton permeability of liposomes made from mitochondrial inner membrane phospholipids: no effect of fatty acid composition.

The proton permeability of the mitochondrial inner membrane has been shown to correlate with the fatty acid composition of its phospholipids. In this paper, we test the hypothesis that the proton permeability of the phospholipid bilayer portion of the membrane depends on phospholipid fatty acid composition. We measured the proton permeability of liposomes made from the mitochondrial inner membrane phospholipids of eight vertebrates, representing a ten-fold range of mitochondrial proton leak and a three fold range of unsaturation index. At a membrane potential (delta psi) of 160 mV at 37 degrees C, the liposomes all had the same proton leak rate, about 30 nmol protons min-1 mg-1 phospholipid. There was no correlation between liposome proton permeability and phospholipid fatty acid composition.

Animals↗

PubMed1997

Dietary fatty acid profile affects endurance in rats.

Typically athletes are advised to increase their consumption of carbohydrates for energy and, along with the general population, to reduce consumption of saturated fats. It is now recognized that fats are not identical in their influence on metabolism, and we argue that the composition of the polyunsaturated fat component should not be ignored. The aim of this study was to manipulate the dietary fatty acid profile in a high-carbohydrate diet in order to investigate the effect of dietary polyunsaturates on submaximal endurance performance in rats. Rats were fed one of three isoenergetic diets containing 22 energy percentage (E%) fat for 9 wk. The diets comprised an essential fatty acid-deficient diet (containing mainly saturated fatty acids); a diet high in n-6 fatty acids, High n-6; and a diet enriched with n-3 fatty acids, High n-3. Submaximal endurance in rats fed the High n-3 diet was 44% less than in rats fed the High n-6 diet (P < 0.02). All rats were then fed a standard commercial laboratory diet for a 6-wk recovery period, and their performances were reevaluated. Although endurance in all groups was lower then at 9 wk, it was again significantly 50% lower in the High n-3 group than the High n-6 group (P < 0.005). Although n-3 fats are considered beneficial for cardiovascular health, they appear to reduce endurance times, and their side effects need to be further investigated.

Animals↗

PubMed1996

Dietary fatty acid profile influences the composition of skeletal muscle phospholipids in rats.

Changes in dietary fatty acid composition alter phospholipid fatty acid composition in a variety of tissues, but little attention has been paid to skeletal muscle. In this study, rats were fed for 9 wk one of three isoenergetic diets: an essential fatty acid-deficient diet, a diet high in (n-6) fatty acids, and a diet enriched with (n-3) fatty acids. Some rats from each group were fed a nonpurified diet for a further 2 or 6 wk. Neither body mass nor food consumption varied among the dietary groups at any stage. Analyses of total phospholipids in soleus (a "slow" twitch muscle) and extensor digitorum longus (a "fast" twitch muscle) revealed that after 9 wk of test diet consumption, muscle phospholipids from rats fed the essential fatty acid-deficient diet were deficient in essential polyunsaturated fatty acids (triene:tetraene ratio >0.5), whereas the polyunsaturated fatty acids in muscle phospholipids from rats fed the high (n-6) fatty acid and high (n-3) fatty acid diets reflected the composition of their respective diets. Nevertheless, phospholipid fatty acid compositions of seemed to be selectively dynamic. After recovery, although the phospholipid fatty acid compositions of all groups were similar, they all contained a much higher proportion of (n-3) fatty acid than provided in the diets. Overall, these results demonstrate that in rats, the fatty acid profile of skeletal muscle phospholipids is strongly influenced by dietary changes, with most effects being reversible after short periods of adequate dietary intake.

Analysis of Variance↗

PubMed1996

Allometry of mitochondrial proton leak: influence of membrane surface area and fatty acid composition.

We investigated why liver mitochondria from small mammals are leakier to protons than those from larger mammals. Sixty-nine percent (+/-23%) of the proton leak differences appeared to relate to membrane area (less inner membrane surface area in larger animals); any residual differences must reflect differences in membrane properties. There were differences in phospholipid fatty acid composition; unsaturation index, monounsaturates, palmitate (16:0), stearate (18:0), docosahexaenoate [22:6(n-3)], and the 22:6(n-3)/22:5(n-3) ratio all correlated with body mass. Proton flux per square centimeter did not correlate significantly with body mass or, in general, with phospholipid fatty acid composition, suggesting little role for fatty acid composition in determining proton leak in mammals of different body mass. However, unsaturation index and n-3 polyunsaturated fatty acid content correlated significantly with proton leak per milligram phospholipid when literature data from reptiles and rats in different thyroid states were included, giving some support to suggestions of a general role for phospholipid fatty acid composition in determining mitochondrial proton leak.

Animals↗

PubMed1996

Effects of changes in dietary fatty acids on isolated skeletal muscle functions in rats.

The effects of manipulating dietary levels of essential polyunsaturated fatty acids on the function of isolated skeletal muscles in male Wistar rats were examined. Three isoenergetic diets were used: an essential fatty acid-deficient diet (EFAD), a diet high in essential (n-6) fatty acids [High (n-6)], and a diet enriched with essential (n-3) fatty acids [High (n-3)]. After 9 wk, groups of rats on each test diet were fed a stock diet of laboratory chow for a further 6 wk. Muscle function was examined by using a battery of five tests for soleus (slow twitch) and extensor digitorum longus (EDL; fast twitch). Tests included single muscle twitches, sustained tetanic contractions, posttetanic potentiation, sustained high-frequency stimulation, and intermittent low-frequency stimulation. Results for muscles from the High (n-6) and High (n-3) groups were very similar. However, the EFAD diet resulted in significantly lower muscular tensions and reduced response times compared with the High (n-6) and High (n-3) diets. Peak twitch tension in soleus muscles was 16-21% less in the EFAD group than in the High (n-6) and High (n-3) groups, respectively [analysis of variance (ANOVA), P < 0.01). During high-frequency stimulation, EDL muscles from the EFAD rats fatigued 32% more quickly (ANOVA, P < 0.01)]. Also, twitch contraction and half-relaxation times were significantly 5-7% reduced in the EFAD group (ANOVA, P < 0.01). During intermittent low-frequency stimulation, soleus muscles from the EFAD group generated 25-28% less tension than did the other groups (ANOVA, P < 0.01), but in EDL muscles from the EFAD group, endurance was 20% greater than in the High (n-6) group (ANOVA, P < 0.05). After 6 wk on the stock diet, there were no longer any differences between the dietary groups. Manipulation of dietary fatty acids results in significant, but reversible, effects in muscles of rats fed an EFAD diet.

Animals↗

PubMed1995

Membrane fatty acid composition of tissues is related to body mass of mammals.

Phospholipids were extracted from tissues (heart, skeletal muscle, kidney cortex, liver and brain) of mammals representing a 9,000-fold range in body mass (mouse, rat, rabbit, sheep and cattle) and their fatty acid composition was determined. In heart, skeletal muscle and kidney cortex, there were significant allometric decreases in the Unsaturation Index (UI; average number of double bonds per 100 fatty acid molecules) with increasing body mass. There were significant inverse allometric relationships between body mass and the proportion of docosahexaenoic acid (22:6 omega 3) in heart and skeletal muscle. In heart, skeletal muscle and kidney cortex, larger mammals also had shorter fatty acid chains in their phospholipids and a higher proportion of monounsaturates. In liver, smaller mammals had a higher UI than larger mammals (except the rabbit, which had the lowest UI and very low proportions of omega 3 fatty acids). The brain of all mammals maintained a high UI with similar levels of polyunsaturated fatty acids, especially 22:6 omega 3. Our results suggest that in heart, skeletal muscle and kidney cortex the activity of the elongases and desaturases are reduced in large mammals compared to small mammals. The allometric trends in membrane composition may be involved in modifying membrane permeability. It is proposed that the elevated degree of polyunsaturation in the membranes of several tissues from small mammals is related to their higher metabolic activity.

Animals↗

PubMed1995

Wallaby transthyretin.

A cDNA library was constructed from liver RNA of the Australian diprotodont marsupial Macropus eugenii, the Tammar wallaby. A cloned full-length transthyretin cDNA was sequenced. The derived amino-acid sequence showed 68% overall similarity to that of human transthyretin, with 86% similarity in the thyroxine binding site. Comparisons of nucleotide and amino acid sequences from several vertebrate species indicated that the greatest differences were in the region corresponding to the disordered N-terminus of mature human transthyretin. The evolutionary trees deduced from parsimony analyses of amino acid and nucleotide sequences of transthyretins, are consistent with that derived from fossil records.

Amino Acid Sequence↗

PubMed1995

Relationship between body mass, tissue metabolic rate, and sodium pump activity in mammalian liver and kidney.

The allometric relationship between body mass and tissue metabolism was examined in liver and kidney cortex slices from mouse, rat, rabbit, sheep, and cattle, representing an approximately 12,000-fold difference in body mass and an 11-fold difference in mass-specific basal metabolic rate. Larger mammals have lower tissue metabolic rates (TMR; mumol O2.g wet wt-1.min-1) at 37 degrees C, yielding the equations TMR = 3.6 M-0.21 for liver slices and TMR = 3.2 M-0.11 for kidney cortex slices, where M is body mass in grams. This appears to be an intrinsic property of the tissue and is not due to differences in extracellular space or tissue protein content, because these are relatively constant in all mammalian species examined. The allometric relationships remain when tissue metabolism is expressed relative to "active cell mass" in tissue slices. Potassium uptake rate (KUR; mumol K+.g wet wt-1.min-1) was also measured (as 86Rb+ uptake) and was also lower in larger mammals, yielding the equations KUR = 1.2 M-0.14 in liver slices and KUR = 3.4 M-0.13 for kidney cortex slices. The energetic costs of sodium pump activity were estimated to be < 10% of TMR for liver and kidney cortex from all five mammalian species.

Animals↗

PubMed1995

Dietary omega 6 fatty acids and the effects of hyperthyroidism in mice.

The influence of the type of dietary fat on the effects of thyroid hormones was investigated in mice. Hyperthyroidism was achieved by providing thyroid hormones (T3 and T4) in the drinking water. Both hyperthyroid and euthyroid mice (Mus musculus) were fed isoenergetic diets containing 18% (w/w) total lipid but differing in fatty acid composition. Diets were either low in the polyunsaturated linoleic acid (18:2, omega 6) and high in saturated fatty acids (SFAs) or low in saturated fats and high in the polyunsaturated fatty acid (PUFA), linoleic acid. Treatments were maintained for 21-22 days. Plasma thyroid hormone levels, standard metabolic rate (SMR), changes in body mass, specific activities of malic enzyme (ME), Na-K-ATPase and glycerolphosphate dehydrogenase (GPDH) of the liver were measured. Fatty acid composition of the liver phospholipids was also determined. Levels of T3 (15-17 nM) and T4 (250-255 nM) were significantly higher in the respective hyperthyroid groups. There was no significant influence of the diet on hormone levels. Hyperthyroidism increased the SMR 37-44% above the euthyroid levels. A significant body weight loss of 14-18% was observed in hyperthyroid mice on the PUFA diet but not in those on the SFA diet. PUFA diet significantly reduced the activity of ME but had no effect on Na-K-ATPase or GPDH activity. Activities of Na-K-ATPase and GPDH were significantly elevated in all hyperthyroid groups. Mice on T4 and PUFA diet showed a highly significant 399% increase in GPDH activity above the euthyroid level.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Characteristics of mitochondrial proton leak and control of oxidative phosphorylation in the major oxygen-consuming tissues of the rat.

Maintenance of an electrochemical proton gradient across the mitochondrial inner membrane against the significant proton permeability of the membrane accounts for 25-30% of resting oxygen consumption in hepatocytes. It has been proposed that proton leak could be a significant contributor to resting metabolic rate in mammals if it were present in other tissues. Mitochondria were isolated from the major oxygen-consuming tissues (liver, kidney, brain and skeletal muscle) of the rat. In each tissue, the mitochondria showed significant proton leak with the same characteristic non-linear dependence on membrane potential. Liver and kidney mitochondria showed similar membrane proton permeability per mg of mitochondrial protein; brain and muscle permeabilities were greater when expressed in this way. Differences in the kinetic response of the substrate oxidation and phosphorylating systems to membrane potential were observed. The substrate oxidation system was more active in kidney, brain and skeletal muscle mitochondria than in liver mitochondria per mg of mitochondrial protein. Liver and kidney phosphorylating systems were less active than brain and skeletal muscle per mg of mitochondrial protein. The control of oxidative phosphorylation was also assessed. The distribution of control in mitochondria isolated from the four tissue types was found to be similar.

Animals↗

PubMed1994

Liposomes from mammalian liver mitochondria are more polyunsaturated and leakier to protons than those from reptiles.

Liposomes were prepared from phospholipids extracted from liver mitochondria of the rat (Rattus norvegicus) and an agamid lizard, the bearded dragon (Amphibolurus vitticeps) and liposome proton conductance was measured at an imposed membrane potential of 160 mV as well as the fatty acid composition of the liposomes. Despite presumed changes in fatty acid composition during liposome preparation, the mammalian liposomes had a significantly lower content of the monounsaturated oleic acid and a significantly greater content of the omega-3 polyunsaturated docosahexaenoic acid. There were significant direct correlations between the liposome arachidonic and docosahexanoic acid content and bilayer proton flux and a significant inverse correlation between liposome oleic acid content and bilayer proton flux. "Apparent valinomycin-catalysed proton flux" was significantly directly correlated with liposome docosahexaenoic acid content and inversely correlated with oleic acid content. It is suggested that the high content of long-chain polyunsaturates in the mammalian mitochondrial membrane is responsible for an increased proton leak across the mitochondrial inner membrane and thus partly responsible for the high metabolic rate in endothermic mammals compared to their ectothermic reptilian predecessors.

Animals↗

PubMed1994

Dietary fats, membrane phospholipids and obesity.

Modifications in dietary fat profile have been shown to affect body weight gain and adiposity. This may occur through changes in the partitioning between oxidation and storage and/or alterations in membrane structure, which may in turn influence metabolic rate. All the dietary fat classes are substrates for the biosynthetic elongase and desaturase enzymes. Obesity is associated with increased delta 9 desaturase activity, reduced delta 5 desaturase activity and perhaps reduced delta 6 desaturase activity. Dietary lipid profile can affect the activity of each of these enzymes. A number of possible mechanisms linking dietary fat subtypes with development of obesity are discussed, including modification of sodium potassium pump activity and alterations in mitochondrial proton leakage.

Animals↗

PubMed1991

Evolution of energy metabolism. Proton permeability of the inner membrane of liver mitochondria is greater in a mammal than in a reptile.

Standard metabolic rate is 7-fold greater in the rat (a typical mammal) than in the bearded dragon, Amphibolurus vitticeps (a reptile with the same body mass and temperature). Rat hepatocytes respire 4-fold faster than do hepatocytes from the lizard. The inner membrane of isolated rat liver mitochondrial has a proton permeability that is 4-5-fold greater than the proton permeability of the lizard liver mitochondrial membrane per mg of mitochondrial protein. The greater permeability of rat mitochondria is not caused by differences in the surface area of the mitochondrial inner membrane, but differences in the fatty acid composition of the mitochondrial phospholipids may be involved in the permeability differences. Greater proton permeability of the mitochondrial inner membrane may contribute to the greater standard metabolic rate of mammals.

Adenosine Triphosphate↗

PubMed1991

Development of mammalian endothermic metabolism: quantitative changes in tissue mitochondria.

The development of energy metabolism of mammalian tissues was assessed in the tammar wallaby Macropus eugenii by the measurement of mitochondrial parameters in the liver, heart, kidney, and brain. Tissues taken from wallabies (n = 27) ranging from 10-day-old pouch young (weighing approximately 4 g) to adults (averaging 6.2 kg) were weighed and fixed, and mitochondrial volume and mitochondrial membrane surface area (MMSA) were determined by quantitative electron microscopy techniques. Developmental changes in these parameters were analyzed chronologically and allometrically. Relative growth rates of all four tissues decreased during development. Liver and heart showed constant allometric growth throughout development, whereas kidney and brain showed biphasic allometric growth. Tissue metabolic intensity assessed by MMSA (m2/cm3 tissue) was constant in liver, showed a threefold increase in brain during pouch life, showed a fourfold increase in the heart between 100 and 200 days of age, and showed a twofold increase in the kidney at the end of pouch life. In all tissues, adult levels of tissue metabolic capacity were present at pouch exit. In all four tissues, total MMSAs were at "reptilian" levels at birth and gradually increased to "mammalian" levels. Each tissue exhibited a different developmental timetable. When the total MMSAs for all four tissues were summed there was a similar pattern of allometric development between summed MMSA and whole animal metabolic rate.

Aging↗

PubMed1990

Membrane fatty acid changes during the cell cycle of CV-1 cells.

Monolayers of CV-1 cells were synchronized at the G1/S boundary of the cell cycle by a 24-h 2 mM thymidine blockade. Uptake of tritiated thymidine indicated that the peak DNA synthesis occurred 6-8 h after release from the block and that cell cycle time was 18-20 h. The fatty acid composition of phospholipids extracted from cells at 0, 7, and 18 h postblockade was measured by gas chromatography. The results indicate cyclic changes in membrane fatty acids with a significant increase in long-chain polyunsaturated fatty acids during the DNA synthesis phase (S phase) of the cell cycle.

Animals↗

PubMed1990

Thyroid hormones during development of a marsupial, the tammar wallaby, Macropus eugenii.

The levels of thyroid hormones in the plasma and the activities of 5'-deiodinase activity in liver and kidney were determined in the tammar wallaby, Macropus eugenii, from early pouch life to adulthood. The total concentration of plasma thyroxine (T4) was below 15 nmol/l before day 75 of pouch life, rose to about 75 nmol/l at day 160, and then decreased to about 12 nmol/l in the adult. The total concentration of plasma tri-iodothyronine (T3) was below 0.4 nmol/l before day 120, increased to 3 nmol/l by about day 220 and then decreased to 1.0 nmol/l in adults. Concentrations of free T4 and free T3 followed a similar pattern but peaked at 45 and 160 pmol/l respectively. Concentrations of reverse T3 (rT3) were extremely variable, ranging from 0 to 1 nmol/l at day 100, and from 0 to greater than 2 nmol/l at day 180. After about day 230, rT3 levels fell rapidly and were below 0.3 nmol/l in adults. Liver and kidney 5'-deiodinase activities, which were undetectable before day 80, reached adult levels by day 220. Half-maximal activity of both these enzymes occurred at about day 205, mid-way between the peaks of T4 and T3. These findings suggest that the systems supporting synthesis and release of hormones from the thyroid gland are probably mature by about day 160 of pouch life in the tammar, while peripheral deiodinase activity, which is a major factor in the production of T3 in the plasma, matures by about day 220. These events thus precede the development of physiological independence of the young tammar from its mother.

Animals↗

PubMed1989

Evolution of mammalian endothermic metabolism: mitochondrial activity and cell composition.

Body composition was measured and compared in Amphibolurus vitticeps and Rattus norvegicus (a reptile and a mammal with the same weight and body temperature). Homogenates were prepared from liver, kidney, brain, heart, lung, and skeletal (gastrocnemius) muscle, and mitochondria were isolated. Cytochrome oxidase activities of both tissue homogenates and isolated mitochondria were measured (at 37 degrees C) as was protein content. Phospholipids were extracted from liver and kidney, and the fatty acid composition was determined. The brain, liver, kidney, heart, and skeletal muscle were significantly larger in the mammal, whereas the skin, reproductive organs, lung, and digestive tract showed no significant difference in size. All mammalian tissues examined contained approximately 50% more protein and phospholipid than the respective reptilian tissue. Although the mammalian phospholipids contained significantly less total unsaturated fatty acids, these unsaturated fatty acids were significantly more polyunsaturated than in the reptilian tissues. Tissue cytochrome oxidase activity was significantly greater in mammals when expressed on a wet weight basis but not when expressed on a tissue protein basis. Mitochondrial cytochrome oxidase activity (on a protein basis) was the same in both species in liver, kidney, and brain, but in heart, lung, and skeletal muscle mammalian mitochondria were twice as active as reptilian mitochondria. The implications of these differences in tissue composition were discussed relative to the evolution of mammalian endothermy.

Animals↗