(Na + K)-ATPase activity of crude homogenates of rat skeletal muscle as estimated from their K-dependent 3-O-methylfluorescein phosphatase activity

A highly sensitive fluorimetric assay using 3-O-methylfluorescein phosphate as substrate was used in the determination of K⁺-dependent phosphatase activity in preparations of rat skeletal muscle. The gastrocnemius muscle was chosen because of mixed fibre composition. Crude, detergent treated homogenate was used so as to avoid loss of activity during purification. K⁺-dependent phosphatase activities in the range 0.19–0.37 μmol · (g wet weight)⁻¹ · min⁻¹ were obtained, the value decreasing with age and K⁺-deficiency. Complete inhibition of the K⁺-dependent phosphatase was obtained with 10⁻³ M ouabain. Using a KSCN-extracted muscle enzyme the intimate relation between K⁺-dependent phosphatase activity and (Na⁺ + K⁺)-activated ATP hydrolysis could be demonstrated. A molecular activity of 620 min⁻¹ was estimated from simultaneous determination of K⁺-dependent phosphatase activity and [³H]ouabain binding capacity using the partially purified enzyme preparation. The corresponding enzyme concentration in the crude homogenates was calculated and corresponded well with the number of [³H]ouabain binding sites measured in intact muscles or biopsies hereof.     

Muscle electrolyte measurements during and after hypokinesia in determining muscle electrolyte depletion during hypokinesia in the rat

Hypokinesia (diminished movement) induces muscle mineral depletion. However, the mechanism of muscle mineral depletion during hypokinesia (HK) remains unknown. Measuring electrolyte retention and electrolyte values in muscle, plasma, and urine during and after HK, the aim of this study was to discover if HK could depress mineral retention and lead to muscle mineral depletion. Studies were done on 204 13-wk-old male Wistar rats (370–390 g) during 10 d pre-HK period, 98 d HK period, and 15 d post-HK period. Rats were equally divided into two groups: vivarium control rats (VCR) and hypokinetic rats (HKR). All hypokinetic rats were kept for 98 d in small individual cages, which restricted their movements in all directions without hindering food and water intakes. All control rats were housed for 98 d in individual cages under vivarium control conditions. Both groups of rats were pair-fed. During the HK period skeletal muscle sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), and water content and electrolyte retention decreased significantly (p < 0.05), while urinary and plasma electrolyte levels increased significantly (p < 0.05) in HKR compared with their pre-HK values and their respective VCR. During the initial days of the post-HK period, mineral retention increased significantly (p < 0.05), plasma and urinary electrolyte level decreased significantly (p < 0.05), while muscle electrolyte and water content remained significantly (p < 0.05) depressed in HKR compared with VCR. Muscle mineral and water content, electrolyte retention, plasma, and urinary electrolyte values did not change in VCR compared with their pre-HK values. It was concluded that during HK decreased muscle mineral content may suggest muscle mineral depletion, while increased urinary electrolyte loss and muscle mineral depletion may demonstrate reduced mineral retention. Reduced electrolyte excretion and depressed muscle mineral content during post-HK may indicate skeletal muscle mineral depletion during HK. Dissociation between electrolyte retention and muscle mineral depletion may demonstrate the presence of decreased electrolyte retention as the mechanism of muscle electrolyte depletion during prolonged HK.     

Dietary arginine supplementation enhances antioxidative capacity and improves meat quality of finishing pigs

The present study was conducted to test the hypothesis that dietary arginine supplementation may improve meat quality of finishing pigs. Beginning at ~60 kg body weight, pigs were fed a corn- and soybean meal-based diet supplemented with 0, 0.5 or 1% l-arginine until they reached a body weight of ~110 kg. On the last day of the experiment, pigs were food-deprived for 16 h before blood samples were obtained for analysis of amino acids, insulin, and other metabolites. Immediately thereafter, pigs were slaughtered for determination of carcass composition, muscle biochemical parameters, and meat quality. The result showed that arginine did not affect pig growth performance or carcass traits. However, 1% arginine decreased drip loss of pork muscle at 48 h postmortem, while increasing intramuscular fat content (P < 0.05). Supplementing 0.5 or 1% arginine to the diet increased arginine concentration and decreased cortisol level in serum, while enhancing antioxidative capacity and glutathione peroxidase activity in serum (P < 0.05). Additionally, 1% arginine increased antioxidative capacity in skeletal muscle (P < 0.05). Furthermore, 0.5 or 1% arginine decreased the cortisol receptor mRNA level in muscle (P < 0.05). Collectively, these results indicate that supplemental arginine improved meat quality and attenuated oxidative stress of finishing pigs.     

滇中喀斯特41种不同生长型植物叶性状研究

该研究以云南石林喀斯特地区41种主要植物(乔木19种、灌木10种、藤本4种、草本8种)为对象,对其叶面积(AR)、叶鲜重(FW)、叶干重(DW)、叶厚度(TH)、叶绿素含量(Chlc)等进行了测定,得到不同植物比叶面积(SLA)、叶干物质含量(DMC)等指标。结果表明:(1)在物种水平上7项植物叶性状值变化范围较大,不同生长型植物差异显著。(2) 41种植物的SLA与DMC之间呈极显著负相关(P0.01),AR与FW、DW呈极显著正相关(P0.01),TH与Chlc呈极显著正相关(P0.01),其他因子间相关性不显著。(3)该研究区植物叶性状变异范围为15.82%～139.14%,多为高变异系数(84.40%～131.01%),叶性状变异中AR与FW贡献率较高,分别为84.40%、90.28%,不同生长型植物在TH上稳定性较好。(4)石林喀斯特地区植物叶片适应特殊生境时表现出较低的TH、SLA、Chlc和较高的DMC等特征。这说明石林喀斯特地区植物通过形成不同的叶性状特征来适应特殊生境,可为喀斯特地区的植被保护与恢复提供理论指导。     

Effects of wet brewers grains on milk yield,milk composition and blood components of dairy cows in hot weather

Twenty multiparous Friesian cows, 60–120 days postpartum, were allotted to two groups of ten cows each according to calving date, lactation number and daily yield, and assigned randomly to one of two diets in a crossover design experiment. The control diet was 45% maize silage (dry basis) and contained ground maize, soya bean meal and wheat bran in proportions which would ensure that the dietary dry matter contained 16.5% crude protein, 3.0 Mcal metabolizable energy kg⁻¹ DM and 14% crude fibre. The treatment diet contained wet brewers grains substituted for maize silage, soya bean meal and wheat bran to change the ruminally undegradable protein from 35% to 39% of crude protein. Ground maize was included in the same quantity as in the control diet. The diets were offered individually, in tie-stalls, as total mixed rations in two equal amounts for ad libitum intakes. The experimental period lasted from 18 June to 12 August 1994. The cows were allowed exercise in an open lot without shade. Dry matter intake, milk protein content and yield, as well as content of milk lactose and non-fat solids were not significantly affected by the diet. In contrast, wet brewers grains supplementation increased actual milk yield (24.8 v. 21.7 kg day⁻¹; P < 0.05), 4% fat-corrected milk yield (25.1 v. 21.1 kg day⁻¹; P < 0.01), milk fat content (4.08 v. 3.82%; P < 0.05), milk total solids content (12.89 v. 12.44%; P < 0.05) and milk fat yield (1.01 v. 0.83 kg day⁻¹; P < 0.05). Blood plasma concentrations of glucose, total protein, albumin, urea, triglycerides, cholesterol, phospholipids, sodium, potassium, calcium, phosphorus and magnesium were not affected by treatment.     

Enhanced endurance in trained cyclists during moderate intensity exercise following 2 weeks adaptation to a high fat diet

These studies investigated the effects of 2 weeks of either a high-fat (HIGH-FAT: 70% fat, 7% CHO) or a high-carbohydrate (HIGH-CHO: 74% CHO, 12% fat) diet on exercise performance in trained cyclists (n = 5) during consecutive periods of cycle exercise including a Wingate test of muscle power, cycle exercise to exhaustion at 85% of peak power output [90% maximal oxygen uptake ( O_2max), high-intensity exercise (HIE)] and 50% of peak power output [60% O_2max, moderate intensity exercise (MIE)]. Exercise time to exhaustion during HIE was not significantly different between trials: nor were the rates of muscle glycogen utilization during HIE different between trials, although starting muscle glycogen content was lower [68.1 (SEM 3.9) vs 120.6 (SEM 3.8) mmol · kg ^–1 wet mass, P < 0.01] after the HIGH-FAT diet. Despite a lower muscle glycogen content at the onset of MIE [32 (SEM 7) vs 73 (SEM 6) mmol · kg ^–1 wet mass, HIGH-FAT vs HIGH-CHO, P < 0.01], exercise time to exhaustion during subsequent MIE was significantly longer after the HIGH-FAT diet [79.7 (SEM 7.6) vs 42.5 (SEM 6.8) min, HIGH-FAT vs HIGH-CHO, P<0.01]. Enhanced endurance during MIE after the HIGH-FAT diet was associated with a lower respiratory exchange ratio [0.87 (SEM 0.03) vs 0.92 (SEM 0.02), P<0.05], and a decreased rate of carbohydrate oxidation [1.41 (SEM 0.70) vs 2.23 (SEM 0.40) g CHO · min^–1, P<0.05]. These results would suggest that 2 weeks of adaptation to a high-fat diet would result in an enhanced resistance to fatigue and a significant sparing of endogenous carbohydrate during low to moderate intensity exercise in a relatively glycogen-depleted state and unimpaired performance during high intensity exercise.     

The effect of a water gradient on the vesicular-arbuscular mycorrhizal status of Lythrum salicaria L. (purple loosestrife)

 The mycorrhizal status of Lythrum salicaria (Lythraceae) was assessed under growth room and field conditions. Growth room studies indicated that L. salicaria is facultatively mycorrhizal and capable of forming vesicular-arbuscular associations with six Glomus species, but not with Gigaspora margarita. Overall, hyphal and arbuscular colonization levels were significantly higher in the wet treatments than in the dry treatments (P<0.0001). However, taken individually, significant increases in arbuscular colonization (P<0.05) were found only in L. salicaria colonized with Glomus clarum, G. aggregatum, and G. versiforme and exposed to the wet treatments compared with the dry treatments, while significant increases in hyphal colonization were found in L. salicaria colonized with G. clarum, and G. versiforme exposed to the wet treatments. There was no overall effect of water availability on levels of vesicular colonization or differences in vesicular colonization levels within species under dry or wet conditions. In contrast, field studies along an existing water gradient revealed that hyphal and arbuscular colonization levels were significantly higher (P<0.05) in the dry and intermediate regions of the gradient than in the wet regions. Vesicular colonization was not significantly affected by the gradient. Total stem height was significantly affected by water availability, plot location and an interaction of the two (P<0.05), and was generally higher in the intermediate and wet plots. Accepted: 20 September 1995     

Quantification of Rat Cerebral Cortex Na+,K+-ATPase: Effect of Age and Potassium Depletion

Na+,K(+)-ATPase concentration in rat cerebral cortex was studied by vanadate-facilitated [3H]ouabain binding to intact samples and by K(+)-dependent 3-O-methylfluorescein phosphatase activity determinations in crude homogenates. Methodological errors of both methods were evaluated. [3H]Ouabain binding to cerebral cortex obtained from 12-week-old rats measured incubating samples in buffer containing [3H]ouabain, and ouabain at a final concentration of 1 x 10(-6) mol/L gave a value of 11,351 +/- 177 (n = 5) pmol/g wet weight (mean +/- SEM) without any significant variation between the lobes. Evaluation of affinity for ouabain was in agreement with a heterogeneous population of [3H]ouabain binding sites. K(+)-dependent 3-O-methylfluorescein phosphatase activity in crude cerebral homogenates of age-matched rats was 7.24 +/- 0.14 (n = 5) mumol/min/g wet weight, corresponding to a Na+,K(+)-ATPase concentration of 12,209 +/- 236 pmol/g wet weight. It was concluded that the present methods were suitable for quantitative studies of cerebral cortex Na+,K(+)-ATPase. The concentration of rat cerebral cortex Na+,K(+)-ATPase showed approximately 10-fold increase within the first 4 weeks of life to reach a plateau of approximately 11,000-12,000 pmol/g wet weight, indicating a larger synthesis of Na+,K+ pumps than tissue mass in rat cerebral cortex during the first 4 weeks of development. K+ depletion induced by K(+)-deficient fodder for 2 weeks resulted in a slight tendency toward a reduction in K+ content (6%, p > 0.5) and Na+,K(+)-ATPase concentration (3%, p > 0.4) in cerebral cortex, whereas soleus muscle K+ content and Na+,K(+)-ATPase concentration were decreased by 30 (p < 0.02) and 32% (p < 0.001), respectively. Hence, during K+ depletion, cerebral cortex can maintain almost normal K+ homeostasis, whereas K+ as well as Na+,K+ pumps are lost from skeletal muscles.     

Characterization of fatty acid interaction with ouabain and vanadate binding to (Na+ + K+)-activated ATPase

The candidateship of unsaturated fatty acids as endogenous ouabain-like factors was studied. Binding of the artificial ligand vanadate at the intracellular phosphorylation epitope of membrane-bound Na+/K+-ATPase was unaffected by linoleic and arachidonic acid. In the (Mg2+ + Pi)-facilitated system for ouabain binding they were characterized as noncompetitive inhibitors of cardiac glycoside binding, however. The ouabain binding capacity as well as the affinity decreased and the ouabain dissociation rate was accelerated by fatty acids. In the presence of vanadate for facilitation of ouabain binding an increase in ouabain affinity was seen. It is concluded that elementary criteria for the characterization of unsaturated fatty acids as ouabain-like factors are not fulfilled. The ratio between E2-subconformations of Na+/K+-ATPase with different ouabain affinities may be changed by incorporation of fatty acids in the lipid membrane.     

Prolonged treatment with the anabolic–androgenic steroid stanozolol increases antioxidant defences in rat skeletal muscle

Testosterone and its synthetic derivatives anabolic–androgenic steroids have been shown to increase skeletal muscle work capacity and fatigue resistance, but the molecular basis for these effects remains uncertain. Since muscle performance has been related to redox status of exercising muscles, this investigation was aimed at testing whether a treatment with suprapharmacological doses of the anabolic–androgenic steroid stanozolol, (2 mg/kg body weight, 5 days/week, for 8 weeks), either alone or in conjunction with treadmill training (12 weeks), enhanced antioxidant defences in rat muscles. Stanozolol treatment did not modify thiobarbituric acid reactive substances and glutathione content in soleus and extensor digitorum longus (EDL) homogenates. In soleus from sedentary rats, superoxide dismutase and glutathione reductase activities were increased by 25% (P < 0.05) and by 40% (P < 0.01) after stanozolol administration, whereas catalase and glutathione peroxidase activities were not modified. This response was similar to that induced by training alone. In EDL from sedentary rats, stanozolol increased only superoxide dismutase activity (20%, P < 0.05). In no case, the effects of steroid administration and training were additive. HSP72 levels were up-regulated in soleus (1.5-fold, P < 0.01) and EDL (threefold, P < 0.001) following training but remained unchanged after stanozolol treatment. Endurance capacity, assessed in a treadmill endurance test, was similar for treated and control rats. We conclude that stanozolol treatment increases antioxidant capacity in selected skeletal muscles from sedentary rats. However, the steroid was not effective in improving endurance capacity or enhancing the training effects on muscle antioxidant defence systems.     

Inhibition of insulin receptors by vanadate and ouabain

Insulin binding studies were performed, using cells from 5 non-obese, non-diabetic subjects, on four separate days: 2 were paired control studies to demonstrate precision, and 2 other sets were binding studies in which one incubation solution was a control and the other contained either vanadate, (10(-4) M) or ouabain (10(-4) M). For both substances tracer binding of 125I insulin was reduced significantly, 27% by vanadate and 30% by ouabain. Furthermore, at all points on the binding curve these substances inhibited binding by 18-98%, in a pattern consistent with reduced receptor number. The concentrations of vanadate or ouabain which we used did not change cell volume or inhibit trypan blue dye exclusion, as an index of cell viability. Because vanadate and ouabain inhibit Na+K+ATPase and have largely dissimilar effects on a variety of cell systems, our observations may reflect specific involvement of Na+K+ATPase in binding or closely related processes.     

Amelioration of altered antioxidant status and membrane linked functions by vanadium andTrigonella in alloxan diabetic rat brains

Trigonella foenum graecum seed powder (TSP) and sodium orthovanadate (SOV) have been reported to have antidiabetic effects. However, SOV exerts hypoglycemic effects at relatively high doses with several toxic effects. We used low doses of vanadate in combination with TSP and evaluated their antidiabetic effects on antioxidant enzymes and membrane-linked functions in diabetic rat brains. In rats, diabetes was induced by alloxan monohydrate (15 mg/100 g body wt.) and they were treated with 2 IU insulin, 0.6 mg/ml SOV, 5% TSP and a combination of 0.2 mg/ml SOV with 5% TSP for 21 days. Blood glucose levels, activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), Na⁺/K⁺ ATPase, membrane lipid peroxidation and fluidity were determined in different fractions of whole brain after 21 days of treatment. Diabetic rats showed high blood glucose (P < 0.001), decreased activities of SOD, catalase and Na⁺/K⁺ ATPase (P < 0.01,P < 0.001 andP < 0.01), increased levels of GPx and MDA (P < 0.01 andP < 0.001) and decreased membrane fluidity (P < 0.01). Treatment with different antidiabetic compounds restored the above-altered parameters. Combined dose ofTrigonella and vanadate was found to be the most effective treatment in normalizing these alterations. Lower doses of vanadate could be used in combination with TSP to effectively counter diabetic alterations without any toxic effects.     

Fish assemblage stability in an intermittently flowing stream from the Brazilian semiarid region

Flooding and drought are important hydrological disturbances occurring in intermittent streams located in semiarid regions. The present study aims to analyse the effects of flood and drought on the diversity and stability of an intermittent stream fish community from the Brazilian semiarid region over the 1996 hydrological cycle. Fourteen fish collections were conducted during the rewetting, wet and drying phases, and 808 individuals were collected. Diversity was measured by using Simpson’s Index (S) and community stability was analysed by using the variation in the abundance, then tested by applying Kendall’s (W) test of concordance over the relative abundance rankings. This test supplies information about the stability of communities by means of the concordance of rankings. A community that is stable or near equilibrium has more constant rankings of its constituent species compared with a fluctuating one. The diversity increased progressively during the rewetting phase (S = 0.480–0.693; coefficient of variation = 0.131), reached its highest values during the wet phase (S = 0.745–0.861; coefficient of variation = 0.072), and remained high during the drying phase, with a lower oscillation than in the other phases (S = 0.699–0.801; coefficient of variation = 0.050). The abundance per collection varied significantly (Kruskal– Wallis 6.94, P = 0.031) throughout the hydrological phases. The stability of the community was higher during the rewetting (W = 0.58, P < 0.001) and drying (W = 0.86, P < 0.001) phases than during the wet phase (W = 0.45, P = 0.1818). In the Brazilian semiarid region, flooding disrupted the stability of the fish community studied and increased the diversity, whereas the absence of floods (expressed by the drying phase) caused a higher stability of the fish community and led to less variation in diversity.     

Quality differences between NILs of wheat variety Long 97-586 possessing HMW-GS 7+8 and 7

The high molecular weight glutenin subunits (HMW-GS) 7+8 were introduced into the Long 97–586 (1,7,2+12) wheat variety (Triticum aestivum) by 5 consecutive backcrosses with biochemical marker–assisted selection.Nearly isogenic lines (NILs) of HMW-GS 7 and 7+8 were obtained,and the NILs were planted in the experimental field at the Crop Breeding Institute of Heilongjiang Academy of Agricultural Science in 2004–2006.The field experiments were designed using the two-column contrast arrangement method with six replicates in 2004–2005 and four replicates in 2006.The result of three years experiments showed that the differences between NILs of Long 97–586 with subunit 7 and those with subunits 7+8 in the quality parameters of flour protein content and dry gluten content were negligible (P0.1).However,the differences in some of the quality parameters were remarkably significant (P0.01),including wet gluten content,ratio of wet gluten/dry gluten,gluten index,Zeleny sedimentation,ratio of sedimentation/dry gluten,and the farinogram parameters of water absorption,development time,stability,breakdown time and degree of softening.The difference between NILs with subunits 7+8 and subunit 7 was significant (P0.05) on the alveogram W value and had a critical value (P=0.05) on the alveogram P value in 2006.The results show that HMW-GS 7+8 is far superior to HMW-GS 7 in terms of baking quality.The possibilities of using subunits 7+8 and subunit 7 in breeding strong and weak gluten wheat varieties are discussed in this paper.     

Microphotometric determination of glycogen in single fibres of human quadriceps muscle

Synopsis A technique for the quantitation of glycogen in single fibres of human skeletal muscle is described. By using microphotometry the loss of glycogen from cryostat sections during a PAS-staining procedure was shown to be negligible. Further, it was found that nearly all the PAS-positive material (98.5%) inside a muscle fibre is glycogen. A significantly higher mean glycogen concentration (P<0.001) was found in type II fibres than in type I fibres in the resting quadriceps muscle of sedentary young males. The coefficient of variation for the glycogen concentration within each fibre type was found to be 17% and 15% for type I and type II respectively. The specificity of the PAS-staining technique for glycogen was confirmed by a statistically significant correlation (r=0.78,P<0.001) between the glycogen concentration measured biochemically and that calculated from microphotometry and area and thickness measurements. With the technique described, it seems possible to measure the glycogen concentration of single muscle fibres in serial sections and to calculate this in standard biochemical terms.     

Determination of intracellular glutathione in human skeletal muscle by reversed-phase high-performance liquid chromatography

A chromatographic method for the specific determination of cellular low molecular mass thiols has been applied to human muscle tissue. The method is based on the derivatisation of thiols using monobromobimane, which is a specific reagent for the sulphydryl group. The glutathione and cysteine bimane adducts were separated by reversed-phase HPLC, whilst quantitation of the cysteine and glutathione adducts was achieved by fluorescence spectroscopy. The method was found to yield a quantitative recovery of glutathione (ca. 96%), to be sensitive (down to 20 pmol glutathione/per injection) and reveal a low intra-individual coefficient of variation (C.V. < 5%) of the glutathione concentrations in human skeletal muscle. The concentrations of reduced and total glutathione were 1320 ± 37 μmol/kg wet weight (mean ± S.E.M.) and 1525 ± 66 μmol/kg wet weight, respectively. The method was also applied to tissues from nine healthy volunteers to determine if fluctuations in glutathione level occurred over a 24-h period. No diurnal variation of glutathione level in human skeletal muscle was observed.     

Resting eggs of conchostracans and their adaptive possibilities

The dimensional specifications and morphology of the resting eggs of ten conchostracan species (Cyzicus tetracerus, Eocyzicus orientalis, Caenestheria davidi, C. perrieri, Leptestheria dahalacensis, Eoleptestheria ticinensis, Limnadia lenticularis, Imnadia yeyetta, Lynceus brachyurus, and L. dauricus) have been studied. The data are presented on the number of eggs produced by females. Unlike other crustaceans, no correlation between the eggs size and their quantity was found. Although the number of eggs can vary one to two times, their size is relatively stable (variation coefficient <5%). The wet (1.5–5.4 μg) and dry (0.41–0.93 μg) weights of eggs and the share of clutch (11.4–21.6%) in the female’s wet weight were determined for eight species. The polyfunctionality of the external envelope of the egg was considered under anhydrobiosis conditions.     

Folic acid rivals methylenetetrahydrofolate reductase (MTHFR) gene-silencing effect on MEPM cell proliferation and apoptosis

Rates of fatty acid oxidation increase rapidly in both rat heart and skeletal muscle in the early postnatal period. Therefore, we examined in heart and soleus muscle, (a) whether there were rapid changes in fatty acid transporter (FAT/CD36, FABPpm) mRNA and protein expression early in life (days 10 –36) and thereafter (days 84, 160, 365), and (b) whether the rates of fatty acid transport and the plasmalemmal content of FAT/CD36 and FABPpm were altered. Protein expression was altered rapidly from day 10–36 in both heart (FAT/CD36 only, +21%, P < 0.05)) and soleus muscle (FAT/CD36 + 100%, P < 0.05; FABPpm −20%, P < 0.05), with no further changes thereafter (P < 0.05). Rates of fatty acid transport (day 10 vs day 160) were increased in heart (+33%, P < 0.05) and muscle (+85%, P < 0.05), and were associated with concomitant increases in plasmalemmal FABPpm (+44%, P < 0.05) and FAT/CD36 (+16%, P < 0.05) in the heart, and only plasmalemmal FAT/CD36 in muscle (+90%, P < 0.05). Therefore, known changes in the rates of fatty acid oxidation in heart and muscle early in life appear to be accompanied by a concurrent upregulation in the rates of fatty acid transport and the expression of FAT/CD36 in heart and muscle, as well as an increase in plasmalemmal FAT/CD36 and FABPpm in the heart, and only plasmalemmal FAT/CD36 in soleus muscle. We speculate that the rapid upregulation of fatty acid transport rates in heart and muscle are needed to support the increased rates of fatty oxidation that have been previously observed in these tissues.     

Vanadate binding to the (Na + K)-ATPase

A particulate (Na + K)-ATPase preparation from dog kidney bound [⁴⁸V]-ortho-vanadate rapidly at 37°C through a divalent cation-dependent process. In the presence of 3 mM MgCl₂ theK _d was 96 nM; substituting MnCl₂ decreased theK _d to 12 nM but the maximal binding remained the same, 2.8 nmol per mg protein, consistent with 1 mol vanadate per functional enzyme complex. Adding KCl in the presence of MgCl₂ increased binding, with aK _0.5 for KCl near 0.5 mM; the increased binding was associated with a drop inK _d for vanadate to 11 nM but with no change in maximal binding. Adding NaCl in the presence of MgCl₂ decreased binding markedly, with anI ₅₀ for NaCl of 7 mM. However, in the presence of MnCl₂ neither KCl nor NaCl affected vanadate binding appreciably. Both the nonhydrolyzable, ,-imido analog of ATP and nitrophenyl phosphate, a substrate for the K-phosphatase reaction that this enzyme also catalyzes, decreased vanadate binding at concentrations consistent with their acting at the low-affinity substrate site of the enzyme; the presence of KCl increased the concentration of each required to decrease vanadate binding. Oligomycin decreased vanadate binding in the presence of MgCl₂, whereas dimethyl sulfoxide and ouabain increased it. With inside-out membrane vesicles from red blood cells vanadate inhibited both the K-phosphatase and (Na + K)-ATPase reactions; however, with the K-phosphatase reaction extravesicular K⁺ (corresponding to intracellular K⁺) both stimulated catalysis and augmented vanadate inhibition, whereas with the (Na + K)-ATPase reaction intravesicular K⁺ (corresponding to extracellular K⁺) both stimulated catalysis and augmented vanadate binding.