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Variation and limitations in fiber enzymatic and size responses in hypertrophied muscle.

The present study was designed to determine whether the degree and kind of adaptation of a muscle fiber to a functional overload (FO) are determined by properties that are intrinsic to that fiber. The study also addresses the question of the capability of fibers to maintain a normal level of coordination of proteins per fiber as fiber volume changes dramatically. The plantaris muscle of six adult female cats was overloaded for 12 wk by bilateral synergist removal. Plantaris muscle fiber mean size doubled after FO, although some very small fibers that stained dark for adenosinetriphosphatase (ATPase) were observed in some of the FO muscles. There appeared to be no change in total succinate dehydrogenase activity per fiber. A reduction in succinate dehydrogenase activity per unit volume was observed in a substantial number of fibers, reflecting a disproportionate increase in fiber volume relative to mitochondrial volume. In contrast, total alpha-glycerophosphate dehydrogenase activity and actomyosin ATPase activity increased as fiber size increased, whereas there was no change in alpha-glycerophosphate dehydrogenase and ATPase activities per unit volume. Control and FO muscle fibers generally expressed either a fast or slow myosin heavy chain type, but in some cases FO muscle fibers expressed both fast and slow myosin heavy chains. The persistence of variability in fiber sizes and enzyme activities in fibers of overloaded muscles suggests a wide range in the adaptive potential of individual fibers to FO. These data indicate that a severalfold increase in cell size may occur without significant qualitative changes in the coordination of protein regulation associated with metabolic pathways and ATP utilization.

Adenosine Triphosphatases↗

Comparative histochemical localization of some hydrolytic enzymes in mammalian epididymides.

The activity of enzymes hydrolyzing sodium-beta-glycerophosphate, adenosine triphosphate, adenosine monophosphate, and glucose-6-phosphate in the epididymides of adult ram, rabbit, rat and hamster has been investigated histochemically. The enzymes hydrolyzing sodium-beta-glycerophosphate, adenosine triphosphate and adenosine monophosphate were rather similar in their distribution in the intertubular connective tissue of the epididymides of the four species studied. However, the distribution and activity of these enzymes in the stereocilia varied. There was a high activity in the stereocilia of the middle segments of ram, rabbit and rat and in the proximal part of the terminal segment of hamster epididymides. Glucose-6-phosphatase was distributed homogeneously in the epithelial cells of the epididymides throughout the duct. The possible functions of these enzymes in the mammalian epididymis are briefly discussed.

Adenosine Monophosphate↗

Characteristics of plasmalemma alkaline phosphatase of rat mesenteric artery.

General characteristics of alkaline phosphatase activity of the plasma membrane-enriched fraction isolated from rat mesenteric arteries were investigated. The vascular smooth muscle plasmalemma alkaline phosphatase is a metalloenzyme which is strongly inhibited by chelating agents and this inhibition can be completely overcome by addition of Mg2+ or Ca2+. Zn2+ only partially reactivates the enzyme in the presence of low concentrations of EDTA. The enzymatic hydrolysis of p-nitrophenyl phosphate, beta-glycerophosphate, alpha-glycerophosphate, or 3'-adenosine monophosphate showed an optimal activity in the alkaline region between pH 9 and 11. The alkaline phosphatase activity is distinctly different from the plasmalemma ATPase and 5'-nucleotidase activities with respect to their pH dependence, influence by added divalent metal ions and stability against heat inactivation. Vanadate ion, being structurally similar to the transition state analog of the phosphoryl group, potently inhibits alkaline phosphatase with an apparent Ki of 1.5 microM. The altered alkaline phosphatase activity of vascular smooth muscle in relation to its possible physiological function and pathophysiological manifestation associated with hypertensive disease are discussed.

Alkaline Phosphatase↗

Correlation between distal nephron enzyme activity, structure and function in rats during lithium and lithium plus neuroleptic treatment.

The histochemical activities of nonspecific acid and alkaline phosphatases, NADH- and NADPH-tetrazolium reductases, alpha-glycerophosphate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase and glucose-6-phosphate dehydrogenase were investigated in kidneys from rats treated with lithium and lithium plus neuroleptics. During the first 8 weeks of lithium treatment the activity of NADH-tetrazolium reductase, succinate dehydrogenase and alpha-glycerophosphate dehydrogenase activity in the collecting ducts increased. The other enzymes did not change. After 8 weeks of treatment no further changes in enzyme activity occurred. Withdrawal of lithium caused normalization of enzyme activity after 8 weeks. A decrease in concentration ability was found in parallel with the increase in enzyme activities (p less than 0.001). The changes in enzyme activity were not significantly correlated to morphological changes in the collecting ducts. Treatment with neuroleptics alone caused no change in enzyme activity. During combined lithium plus neuroleptic treatment the enzyme activities changed in a similar way as during lithium therapy, but the changes were less pronounced. In parallel, a less pronounced decrease in concentration ability was found during this treatment.

Animals↗

Development of enzymes of energy metabolism in rat heart.

The behavior of several enzymes was studied during rat heart development (4 days before birth to adult stage). Hexokinase has its highest activity during the fetal period; it decreases at birth and remains with low activity in the adult. The alpha-glycerophosphate dehydrogenase and alpha-glycerophosphate oxidase profiles are similar up to the 15th day of development. From there onwards, both profiles diverge, the cytoplasmic activity increasing 3-fold, while the mitochondrial activity remains unchanged. The developmental profiles of the malate dehydrogenases are almost parallel. The development of citrate synthase and succinate dehydrogenase results in a 2- to 4-fold increase in their activities. However, ATPase increases dramatically (20-fold) over the same period. With respect to the enzymes of the adenine nucleotide metabolism, adenylate kinase is fully expressed throughout all ages examined, showing no variation during development. AMP deaminase and creatine kinase increase during development, the cytoplasmic creatine kinase reaching a high level at birth whereas the increases of the mitochondrial enzymes take place gradually during development.

AMP Deaminase↗

The further metabolism of sorbitol-3-phosphate and fructose-3-phosphate in the mature rat lens.

Glucose and phosphorus metabolism in mature (8-month-old) rat lenses were examined with NMR spectroscopy. Nondiabetic mature lenses contained sorbitol-3-phosphate (S3P) and fructose-3-phosphate (F3P) which were absent from young (1- to 2-month-old) normal rat lenses. The concentrations of these two phosphates can be changed through (1) diabetes induction with streptozotocin - this results in a dramatic increase in both compounds; and (2) oral dosing with a drug known to prevent sorbitol production - both metabolites disappeared. When normal mature lenses were incubated in 35.5 mM 13C1-glucose, both 13C1-lactate and 13C3-lactate were produced. Preservation of the 13C label at C1 is likely through the formation of 13C1-S3P and -F3P, which were then split through an aldolase-like mechanism into two 3-carbon compounds, one an unlabeled glycerol and the other 13C1-alpha-glycerophosphate (from S3P) and 13C1-dihydroxyacetone phosphate (from F3P). These reactions can contribute to the increase in alpha-glycerophosphate observed in both the streptozotocin-induced diabetic lenses and lenses incubated in high glucose.

Aging↗

Iron deficiency in the rat. Physiological and biochemical studies of muscle dysfunction.

Work performance on a treadmill has been evaluated in normal and iron-deficient rats. Anemia was removed as a variable by adjusting the hemoglobin of all animals to the same concentration. At a hemoglobin compatible with normal work performance, iron-deficient animals showed a marked impairment of running ability as compared to control animals. Iron therapy corrected the disability within 4 days. Concentrations of the cytochrome pigments and myoglobin, and rates of oxidative phosphorylation with pyruvate-malate, succinate, and alpha-glycerophosphate as substrates were all reduced in mitochondrial preparations from skeletal muscle of iron-deficient rats, but only the rate of phosphorylation with alpha-glycerophosphate as substrate increased significantly and in parallel with the recovery in work performance of the iron-deficient rats treated with iron dextran.

Anemia, Hypochromic↗

Lactic acidosis as a result of iron deficiency.

Iron-deficient rats have an impaired work performance, even when their anemia is corrected by exchange transfusion. Muscle activity is associated with a higher blood lactate concentration than is observed in iron-replete animals. The accumulation of lactate is a result of excessive production as lactate clearance from the blood was shown to be unaffected. By adjusting the work load to a lower level, it was possible to divide iron-deficient animals into two groups, one capable of continued treadmill running and another in which animals stopped before 20 min. In the former, blood lactate concentration reached a plateau at moderate levels, whereas it continued to increase in the latter until the animal stopped running. Levels of alpha-glycerophosphate oxidase in skeletal muscle mitochondria were found to be much lower in the second group (P < 0.001). Lactate infusion into normal animals was shown to interfere with work performance, and maintenance of a normal pH in iron-deficient and iron-replete animals did not prevent the impairment in work associated with high blood lactate concentrations. Additional evidence was obtained that energy substrate (blood glucose and free fatty acids, muscle glycogen) was adequate in irondeficient animals. Oxygen tension in their vena caval blood was higher than in controls. Furthermore, the in situ behavior of electrically stimulated gastroenemius and soleus muscles appeared similar to that of control animals. Because the stimulation of the single muscle in the iron-deficient animal did not result in appreciable elevation of blood lactate and did not show impaired contractility further supported the hypothesis that the elevation of blood lactate caused the decreased work performance. It is concluded that iron deficiency by a depletion in the iron-containing mitochondrial enzyme, alpha-glycerophosphate oxidase, impairs glycolysis, resulting in excess lactate formation, which at high levels leads to cessation of physical activity.

Acidosis↗

Are iodine-deficient rats euthyroid?

Inhabitants of many severe endemic goiter areas have low serum T4 and high circulating TSH, despite normal levels of T3. This situation may be produced experimentally chronically feeding rats a low iodine diet (LID). We fed rats a Remington-type LID and gave them 1% NaClO4 in their drinking water for 2 days. After this, the animals were divided into three groups. One group was fed LID, supplemented with 5 micrograms I/rat.day and was used as the control group. Another group was fed LID alone. The third group was fed LID and given 1% NaClO4 to drink. The latter treatment was used to induce severe hypothyroidism. Animals were killed 1, 2, 3, and 5 weeks after the onset of these treatment schedules. The following measurements were made on some or all groups of animals: body and thyroid weights; thyroidal I content; soluble labeled iodoprotein profile; thyroidal labeled iodoamino acid distribution pattern; plasma T4, T3, and TSH; pituitary GH content; and liver intramitochondrial alpha-glycerophosphate dehydrogenase and cytosolic malic enzyme activities. T4 and T3 concentrations were also measured in liver nuclei of the animals killed 5 weeks after the onset of treatment. As assessed from various indices of thyroid function, the LID rats became iodine deficient, although not as markedly as those given LID and ClO4-, The plasma T4 decreased to undetectable levels, and plasma TSH increased, whereas circulating T3 remained normal throughout in the LID rats. In rats given LID and ClO4-, plasma T4 decreased sooner than in rats given LID alone; plasma T3 levels also became undetectable, and TSH increased more markedly and sooner than in rats given LID alone. As measured at the end of 5 weeks of treatment, pituitary GH content, and liver alpha-glycerophosphate dehydrogenase and malic enzyme activities were lower in rats given LID than in the euthyroid LID- and I--treated controls. They were not, however, as markedly reduced as in the severely hypothyroid LID- and ClO4--treated rats. In spite of normal plasma T3 levels, the concentration of T3 in liver nuclei of the rats given LID alone was significantly lower than that of the LID- and I--treated controls. The results show that the thyrotrophs, somatotrophs, and livers of rats given LID alone are not like those of euthyroid rats despite normal circulating T3 levels. In iodine-deficient rats, there is a discrepancy between the measured indices of thyroid hormone action in the liver and the circulating T3 level, but not between biological activity and liver nuclear T3 concentration. It remains to be seen whether the same is true in the anterior pituitary.

Animals↗

Diabetes decreases liver and kidney nuclear 3,5,3'-triiodothyronine receptors in rats.

The liver and kidney nuclear T3 content and the maximal nuclear T3-binding capacity (MBC) were measured 1 month after streptozotocin administration and compared with values in controls either fed ad libitum (C) or offered a restricted diet (FR). A group of insulin-treated diabetic (D+I) rats was also included. Plasma T4 and T3 concentrations decreased to low levels in diabetic (D) rats. Plasma T3 levels were decreased in FR rats, whereas circulating T4 was in the normal range for C animals. The MBC (nanograms of T3 per mg DNA) for liver and kidney nuclear T3 was determined by an in vivo saturation technique. The respective results for all groups were as follows (asterisks denote values differing from C with P values less than 0.05): C, 0.601 and 0.414; FR, 0.583 and 0.369; D, 0.310 and 0.220; D+I, 0.630 and 0.394. Nuclear T4 and T3 concentrations were determined by an isotopic equilibrium technique. Nuclear T3 (nanograms per mg DNA) for liver and kidney were, respectively, 0.298 and 0.176 for C, 0.208 and 0.135 for FR, 0.109 and 0.070 for D, and 0.270 and 0.168 for D+I rats. The decreased liver and kidney nuclear T3 content in D rats appears to be due to a marked reduction of their available intracellular T4 pool, from which T3 could be generated, but most likely represents a decreased T3 uptake into liver and kidney nuclei, as the nuclear to plasma ratios of labeled T3 were decreased in D rats. The low levels of T3 in nuclei of FR rats could be attributed to an inhibition of T4 to T3 conversion, since the intracellular pool of T4 appears to be normal. The possibility that diabetes and food restriction might affect the thyroid activity was examined by measurement of the activities of alpha-glycerophosphate dehydrogenase and cytosol malic enzyme, two liver and kidney enzymes regulated by thyroid hormone. Furthermore, although the measurements made in FR rats excluded the possibility that the alterations in MBC found in D animals were nutrition dependent, the reduced nuclear T3 content concomitant with food restriction may account for some of the quantitative changes in the alpha-glycerophosphate dehydrogenase and cytosol malic enzyme activity found in D rat tissues. In conclusion, the present findings suggest that the observed changes in indices of thyroid hormone action in liver and kidney of D rats could be related to alterations in nuclear T3 receptor concentrations and the concentration of T3 bound to the receptor.

Animals↗

1,25-dihydroxyvitamin D3 stimulates adipocyte differentiation in cultures of fetal rat calvaria cells: comparison with the effects of dexamethasone.

Progenitor cells for several mesenchymally derived cell types exist within freshly isolated fetal rat calvaria (RC) cell populations. We have characterized the effects of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] on the differentiation of adipocytes from primary RC cells and compared these effects with those of dexamethasone (Dex). RC cells were plated at 3 x 10(4)/35-mm dish, and cultures were maintained for 14-19 days in alpha-Minimum Essential Medium containing 10% fetal bovine serum, 50 micrograms/ml ascorbic acid, 10 mM Na beta-glycerophosphate, and 0.1-100 nM 1,25-(OH)2D3 or 1-1000 nM Dex. Morphological (quantitation of adipocyte foci number and area after staining cultures with Sudan IV) and biochemical (glycerol-3-phosphate dehydrogenase activity) methods of assessing adipogenesis were used. In the presence of 1,25-(OH)2D3, adipocyte foci developed about 3 days after confluency as clusters of rounded or stellate cells. Stimulation of adipocyte foci development was dose dependent from 0.1-100 nM and was maximal with 10 nM 1,25-(OH)2D3; half-maximal stimulation occurred at about 1 nM. The presence of ascorbic acid and beta-glycerophosphate was not required for 1,25-(OH)2D3-induced stimulation of adipocytes, but both significantly increased the number of adipocyte foci in the presence of 1,25-(OH)2D3. The critical period for initiation of adipocyte differentiation with 1,25-(OH)2D3 was between 1-9 days, and once committed along the adipogenic pathway, adipocytes maintained their differentiated state in the absence of 1,25-(OH)2D3. Short term (48-h) pulses of 1,25-(OH)2D3 resulted in slight, but significant, increases in adipocyte formation. Other vitamin D3 metabolites were less effective than 1,25-(OH)2D3 in stimulating adipocyte differentiation. Dex (1-100 nM) also caused a dose-dependent increase in the differentiation of adipocyte foci in RC cell cultures. The adipocyte foci that developed in the presence of Dex frequently appeared earlier in culture, i.e. when cells reached confluency on days 6-7, and were more diffuse than those forming with 1,25-(OH)2D3. The stimulation of adipocyte differentiation by 1,25-(OH)2D3, however, was greater than that by Dex in mixed RC II-V cells. The combined effects of 1,25-(OH)2D3 and Dex were additive at low concentrations and synergistic at higher concentrations of either 1,25-(OH)2D3 or Dex. The data show that bone cell populations isolated from fetal RC contain adipocyte progenitors and that 1,25-(OH)2D3 as well as Dex are potent regulators of adipocyte differentiation within these bone cell populations.

Adipocytes↗

Effect of chlorpromazine administration on enzyme activity concerning phosphatidic acid metabolism in rat liver.

The effect of chlorpromazine on phospholipid metabolism was studied in rat liver endoplasmic reticulum. Administration of chlorpromazine per os caused a marked increase in microsomal phospholipid content without affecting its composition. The rate of incorporation of both [(32)P]orthophosphate and [2-(3)H)glycerol into phosphatidic acid and phosphatidylinositol increased by the administration of chlorpromazine. Enhanced labeling of phosphatidylinositol was also observed when myo-[2-(3)H]inositol was used as a precursor. The activity of glycerophosphate acyltransferase significantly increased and that of phosphatidate phosphohydrolase markedly decreased by the administration of chlorpromazine. The activity of phosphatidate cytidylyltransferase was not affected by the drug treatment. These findings suggested that the administration of chlorpromazine increases the availability of phosphatidic acid by activating glycerophosphate acyltransferase and inhibiting phosphatidate phosphohydrolase, leading to enhanced synthesis of phosphatidylinositol.

Animals↗

Thyroid hormone inhibits growth and stimulates terminal differentiation of epiphyseal growth plate chondrocytes.

As a continuation of our studies on mineralization in epiphyseal growth plate (GP) chondrocyte cultures, the effects of tri-iodothyronine (T3) in both beta-glycerophosphate-containing, serum-free (HL-1) and beta-glycerophosphate-free, serum-containing medium (DATP5) were studied. The GP cells responded to T3 in a serum-, stage-, and dosage-dependent manner. Added at graded levels (0.1-10.0 nM) to preconfluent cultures (from day 7) in both HL-1 and DATP5, T3 caused progressive decreases in protein, collagen, and DNA synthesis but increased mineral deposition. In postconfluent cultures, these effects of T3 were generally muted. In preconfluent cultures, proteoglycan (PG) levels were not significantly affected in DATP5, although in HL-1 they were decreased by approximately 50%. In postconfluent cultures, T3 increased PG levels in DATP5 but had no effect in HL-1. In HL-1, alkaline phosphatase (ALP) activity was progressively increased by 200-500% in both pre- and postconfluent cultures. In DATP5 in preconfluent cultures, T3 initially stimulated but later suppressed ALP; in postconfluent cultures, T3 also transiently increased ALP but did not suppress activity upon longer exposure. The inhibitory effects of T3 on protein, PG, and DNA levels of GP chondrocytes suggest that in vivo its effects on bone growth must occur primarily after cellular proliferation. Apparently by binding to the 50 kDa thyroxine-binding globulin, which cannot penetrate the PG barrier, accessibility of T3 to GP chondrocytes is limited until the time of vascular penetration when its stimulatory effects on ALP and mineral deposition become critical for continued bone development.

Alkaline Phosphatase↗

Influence of 2-deoxy-D-glucose and energy substrates on guinea-pig sperm capacitation and acrosome reaction.

When guinea-pig spermatozoa were suspended in a minimal culture medium (MCM-PL), 2-deoxy-D-glucose (100 microM) and 2-amino-2-deoxy-D-glucose (100 microM) were potent inhibitors of the acrosome reaction without affecting the sperm motility, whereas the N-acetyl derivative 2-acetamido-2-deoxy-D-glucose (2 mM) had no inhibitory effect. The addition of D-glucose (2 mM) partly inhibited the percentage acrosome reaction of spermatozoa suspended in Medium MCM-PL, but DL-alpha-glycerophosphate (2 mM) and myo-inositol (2 mM) had no effect. In addition, DL-alpha-glycerophosphate (2 mM) did not overcome the inhibitory effect of 2-deoxy-D-glucose on the sperm acrosome reaction. The inhibitory action of 2-deoxy-D-glucose (100 microM) on the sperm acrosome reaction assessed after a 3-h incubation was irreversible and was only completely effective if the sugar was added within 30 min of the start of incubation. When spermatozoa suspended in Medium MCM-PL were treated with 2-deoxy-D-glucose (100-200 microM) for an extended incubation up to 6 h, the inhibitory effect of 2-deoxy-D-glucose was partly overcome. Spermatozoa treated with 2-deoxy-D-glucose had significantly reduced concentrations of ATP after incubation for 2 h in Ca2+-free media, compared with the ATP concentrations of spermatozoa preincubated for 2 h in Ca2+-free media that supported acrosome reactions. The addition of Ca2+ (5 mM) caused a rapid decrease in ATP concentrations of spermatozoa suspended in Medium MCM-PL, while the addition of the monovalent ionophore monensin (50 microM) and Ca2+ stimulated sperm acrosome reactions as well as an additional decline in the sperm ATP concentrations. However, monensin (50 microM) in the absence of Ca2+ caused only a slight decline in the sperm ATP concentrations over the 15-min incubation period. The depletion of the sperm ATP concentrations by 2-deoxy-D-glucose may retard completion of the capacitation process and the resultant acrosome reaction.

Acrosome↗

Expression of extracellular matrix proteins in human periodontal ligament cells during mineralization in vitro.

Periodontal regeneration is a complex process that requires coordinated responses from several cell types within the periodontium. It is generally accepted that the periodontal ligament (PDL) has a heterogeneous cell population, where some of the cells may be capable of differentiating into either cementoblasts or osteoblasts. Thus, it has been hypothesized that PDL cells play a role in promoting periodontal regeneration. However, definitive evidence to support this concept is lacking. Previously, we reported that PDL cells induce biomineralization as determined by Von Kossa histochemistry and transmission electron microscopy. To further determine the osteoblast-like properties of PDL cells, human PDL cells were exposed to dexamethasone (DEX) in order to promote an osteoblast phenotype, and then cell activity monitored during mineral nodule formation in vitro. For mineralization studies, cells were cultured in DMEM containing 10% FBS and a) vehicle only; b) ascorbic acid (50 micrograms/ml) and beta-glycerophosphate (10 mM); or c) ascorbic acid, beta-glycerophosphate and DEX (100 nM) for 30 days. In addition, the effects of DEX on PDL cells in non-mineralizing media were determined. Cells were stained weekly to evaluate mineral-like nodules, using the Von Kossa method. Northern blot analyses for mRNA steady state levels for several bone-associated proteins, i.e., osteopontin (OPN), bone sialoprotein (BSP), alkaline phosphatase (ALP), osteocalcin (OCN), alpha 2(1)(type 1) collagen and osteonectin (ON), were performed. DNA levels were also determined during the 30-day mineralization period. Under phase contrast microscopy, PDL cells in non-mineralizing media treated with DEX exhibited a more spindle-shaped morphology when compared with similar cells not exposed to DEX. Mineralizing conditions were required to induce mineral nodule formation. However, in this situation, mineral induction was independent of DEX; and furthermore, DEX-treated cells did not exhibit a different morphological pattern when compared with non-DEX treated cells. Mineral-like nodules were first seen at day 15, in concert with an increase followed by a decrease in expression of type I collagen and ON mRNA in both DEX-treated and non-treated cultures. Using Northern blot analysis for detection of specific proteins, we found that PDL cells did not express OPN, BSP, OCN, or ALP under any of the conditions used in this study. DEX did not alter DNA content in the cultures during the mineralization period. These results confirm that human periodontal ligament cells can be induced to mineralize in vitro and indicate that dexamethasone does not significantly alter the extent and pattern of mineralization.

Alkaline Phosphatase↗

Hydrologic influence on stability of organic phosphorus in wetland detritus.

Accretion of organic matter in wetlands provides long-term storage for nutrients and other contaminants. Water-table fluctuations and resulting alternate flooded and drained conditions may substantially alter the stability of stored materials including phosphorus (P). To study the effects of hydrologic fluctuation on P mobilization in wetlands, recently accreted detrial material (derived primarily from Typha spp.) was collected from the Everglades Nutrient Removal Project (ENRP), a constructed wetland used to treat agricultural drainage water in the northern Everglades. The detrital material was subjected to different periods of drawdown and consecutive reflooding under laboratory conditions. The 31P nuclear magnetic resonance (31P NMR) spectroscopy analysis revealed that sugar phosphate, glycerophosphate, polynucleotides, and phospholipids (glycerophosphoethanolamine and glycerophosphocholine) were the major forms of P in the detrital material. After 30 d of drawdown, polynucleotides were reduced to trace levels, whereas sugar phosphate, glycerophosphate, and phospholipids remained the major fractions of organic P. Microorganisms seemed to preferentially utilize nucleic acid P, perhaps to obtain associated nutrients including carbon and nitrogen. At the end of the 30-d reflooding period, cumulative P flux from detritus to water column accounted for 3% of the total P (< or = 15 d of drawdown) and further decreased to 2% at 30 d of drawdown, but increased to 8% at 60 d of drawdown. The drawdown (< or = 30 d) not only reduced P flux to the water column, but also increased the humification and microbial immobilization of P. Excessive drawdown (60 d), however, triggered the release of P into the water column as the water content of detritus decreased from 95 to 11%.

Ecosystem↗

Glycerolipid biosynthesis in porcine adipose tissue in vitro: effect of adiposity and depot site.

To compare genetic differences in glycerolipid biosynthesis, rates were determined in s.c. adipose tissue of lean and obese pigs at 28, 60 and 110 d of age. To compare depot-specific differences, glycerolipid biosynthetic rates were determined in outer s.c., middle s.c., perirenal and omental adipose tissues obtained from 105-kg contemporary pigs. Rates were determined with a 700 x g infranatant fraction of an adipose tissue homogenate by measuring glycerophosphate incorporation into total lipids (mostly phosphatidic acid) during 4 min. This assay represents entrance of substrates into the glycerolipid synthesis pathway or glycerophosphate acyltransferase (GPAT) activity. Rates measured for 60 min represent maximal synthesis of glycerolipid (more triacylglycerol than phosphatidic acid) or lipid synthesis capacity (LSC). Adipocyte diameter and volume were greater for adipose tissue of obese than of lean pigs both at 60 and 110 d. When expressed per cell, activity of GPAT and LSC were similar for lean and obese pigs at 28 d. At 60 d and 110 d, LSC was greater for obese than for lean pigs; GPAT activity was greater at 60 but not at 110 d in obese than in lean pigs. Expressed on a cell basis, GPAT activity was highest in omental and outer s.c., intermediate in perirenal and lowest in middle s.c. adipose tissue depots. Lipid synthesis capacity was highest in perirenal and lowest in outer and middle s.c. depots. Our results indicate that the LSC assay was more closely related to the accretion of fat in vivo than to GPAT activity.

Adipose Tissue↗

Effects of nicotinic acid on glucose tolerance and glucose incorporation into adipose tissue in hypertriglyceridaemia.

Nicotinic acid 4 g daily was given to 28 weight-stable hypertriglyceridaemic patients. The aim was to study its effects on serum lipoprotein lipid levels, intravenous glucose tolerance (k-values) and glucose incorporation into subcutaneous adipose tissue (GLIAT) in vitro. The investigations were performed prior to the nicotinic acid therapy, after 6 weeks, and 6 months of drug treatment. Fasting blood glucose levels increased by 13%, whereas k-values fell by 26% after the nicotinic acid treatment. Decrease in k-values was predicted from the initial k-values (R2-value = 0.76). GLIAT increased by 76%, while in the subgroup of eight patients, treated for 6 months with nicotinic acid, GLIAT increased by 331%. The changes in k-values and GLIAT were not significantly interrelated. Serum triglyceride levels were strongly decreased. The most likely explanation for the decrease in intravenous glucose tolerance is that nicotinic acid stimulates glucose output from the liver and that this effect outweighs the stimulating effects of the drug on glucose utilization in extrahepatic tissues. The latter is reflected by the increased uptake of glucose in adipose tissue. A stimulated GLIAT, reflecting formation of alpha-glycerophosphate in adipose tissue, might contribute to the reduction of serum triglyceride levels induced by nicotinic acid, since alpha-glycerophosphate is the acceptor of fatty acids assumed to be liberated from circulating triglycerides by lipoprotein lipase.

Adipose Tissue↗