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Biomedical subjects

H Wiesinger

Publications and source records attributed to H Wiesinger.

At least 19 recordsLinked to original sources
PubMed1995

Stimulation of arginine transport and nitric oxide production by lipopolysaccharide is mediated by different signaling pathways in astrocytes.

Transport of L-arginine and generation of nitrite in microglia-free astroglial cultures derived from neonatal mouse brain were stimulated by bacterial lipopolysaccharide (LPS) in a time- and dose-dependent manner. LPS stimulated arginine transport between 1.3- and 2.5-fold; half-maximal stimulation was obtained with 0.3 micrograms/ml LPS. Acceleration of transport was detectable within 6 h of incubation with LPS. Cycloheximide or actinomycin D neutralized the effect of LPS. Stimulation of generation of nitrite was reduced when the cells were incubated simultaneously with LPS and either genistein or diethyldithiocarbamate, inhibitors of protein tyrosine kinase and nuclear transcription factor kappa, respectively. However, stimulation of arginine transport was not reduced in the presence of these compounds. Dexamethasone inhibited stimulation of nitric oxide (NO) production but not of arginine transport. Protein kinase C inhibitor staurosporine had no effect on either process. The results suggest that LPS-stimulated acceleration of arginine transport in astrocytes requires protein as well as RNA synthesis. Induction of synthesis of an astroglial cationic amino acid transport system appears to be mechanistically independent from stimulation of intracellular NO production.

Animals↗
PubMed1995

Lactate transport in cultured glial cells.

Uptake of L-lactate was investigated with a radioactive tracer method in cultured rat glioma cells and in astroglia-rich primary cultures derived from rat brain. In the glioma cells, a saturable component of uptake was identified with half-maximal uptake occurring at 1.0 +/- 0.4 mM lactate. In addition, a non-saturable component dominated the uptake at high concentrations of lactate. In astroglia-rich primary cultures, no saturable component of uptake could be detected in the concentration range studied (0.1-15 mM). In conclusion, lactate uptake at physiological concentrations is predominantly mediated in the glioma cells by a carrier-dependent process, whereas in astroglial cells, simple diffusion appears to be the dominant way of lactate transport.

Animals↗
PubMed1994

Transport of L-arginine in cultured glial cells.

Uptake of radiolabelled L-arginine was studied in four different kinds of glial cultures, in astroglia-rich primary cultures derived from neonatal rat and mouse brains, in pure murine astrocyte cultures, and in rat glioma cells C6-BU-1. A saturable component of uptake was found in all cases with KM values between 15 and 35 microM and Vmax values between 0.8 and 2.5 nmol.min-1.(mg protein)-1. In addition, in all cell types a non-saturable component dominated total uptake at high concentrations of extracellular arginine. Rates of uptake of arginine were not affected when Na+ or Cl- were absent from the incubation buffer. Carrier-mediated uptake of arginine was reduced by depolarizing concentrations of K+ and strongly inhibited by an excess of lysine or ornithine. Histidine, asparagine, glutamine, citrulline, creatine, NG-nitro-L-arginine, NG-monomethyl-L-arginine, or L-canavanine inhibited L-arginine transport to various degrees. Uptake of arginine was not reduced in the presence of serine or alanine cysteic acid, N-methyl-alpha-aminoisobutyric acid, or 2-aminobicyclo-(2.2.1)-heptane-2-carboxylic acid. Rates of uptake of arginine were increased when cells had been preloaded with lysine. Preincubation of primary cultures, but not glioma cells, with bacterial lipopolysaccharide stimulated transport of arginine by increasing the Vmax value of uptake. This stimulation was dependent on protein synthesis. The results suggest that, at physiological concentrations, arginine is taken up into the glial cells with the help of the transport system "y+" for basic amino acids. In glial primary cultures, uptake of arginine appears to be regulated by compounds which also exert influence on nitric oxide synthesis.

Amino Acids↗
PubMed1994

Utilization of mannose by astroglial cells.

Uptake and metabolism of mannose were studied in astroglia-rich primary cultures derived from neonatal rat brains. A saturable component of mannose uptake was found with half-maximal uptake at 6.7 +/- 1.0 mM mannose. In addition, a non-saturable component dominated the uptake at high concentrations of mannose. Glucose, cytochalasin B, or phloretin in the incubation buffer inhibited the carrier-mediated uptake of mannose. Within the astroglial cells mannose is phosphorylated to mannose-6-phosphate. In cell homogenates, the KM value of mannose-phosphorylating activity was determined to be 24 +/- 7 microM. The Vmax value of this activity is only 40% that of glucose-phosphorylating activity. Mannose-6-phosphate was converted to fructose-6-phosphate by mannose-6-phosphate isomerase. The specific activity of this enzyme in homogenates of astroglial cultures was higher than that of hexokinase. Two products of mannose utilization in astroglial cells are glycogen and lactate. The amounts of each of these products increased with increasing concentrations of mannose. In contrast to the generation of lactate, that of glycogen from mannose was enhanced in the presence of insulin. In conclusion, we suggest that mannose is taken up into the cells of astroglia-rich primary cultures by the glial glucose transporter and is metabolized to fructose-6-phosphate within the astroglial cells.

Animals↗
PubMed1993

Uptake of L-lactate by cultured rat brain neurons.

The uptake of L-lactate was investigated in neuronal primary cultures derived from embryonic rat brain with a radioactive tracer method. After preincubation of the cells in glucose-free buffer for 30 min, uptake increased with time for at least 10 min. A saturable component of uptake was found with half-maximal uptake at 10 mM lactate. This saturable component was abolished in the presence of 10 mM alpha-cyano-4-hydroxcinnamic acid. In addition, a non-saturable component dominated the uptake at high concentrations of lactate. Uptake was accelerated with decreasing pH, and was inhibited considerably by pyruvate. It is concluded that neurons are endowed with a lactate transport system which resembles in its properties the monocarboxylate carrier of peripheral tissues.

Animals↗
PubMed1993

Purification of cytosolic malic enzyme from bovine brain, generation of monoclonal antibodies, and immunocytochemical localization of the enzyme in glial cells of neural primary cultures.

Cytosolic malic enzyme (EC 1.1.1.40) was purified from bovine brain 5,600-fold to a specific activity of 47 U/mg. The enzyme is a homotetramer with a subunit molecular mass of 60 kDa and an isoelectric point of 6.2. Mouse monoclonal antibodies raised against this enzyme were purified and shown to be monospecific, as indicated by immunoblotting. Immunocytochemical examination of rat astroglia-rich primary cultures at the light microscopic level revealed colocalization of cytosolic malic enzyme with the astroglial marker glial fibrillary acidic protein. Also, a colocalization with the oligodendroglial marker myelin basic protein was found. Neurons in rat neuron-rich primary cultures did not show positive staining. The data suggest that cytosolic malic enzyme is a glial enzyme and is lacking in neurons.

Animals↗
PubMed1991

Replacement of glucose by sorbitol in growth medium causes selection of astroglial cells from heterogeneous primary cultures derived from newborn mouse brain.

Primary cultures derived from the brains of newborn mice are quantitatively dominated by astroglial cells, but contain also oligodendroglial, phagocytic and ependymal cells. When confluent cultures are fed with glucose-free growth medium containing 25 mM sorbitol for 14 days, oligodendroglial, phagocytic and ependymal cells are eliminated from the culture, as judged by morphological and immunocytochemical criteria. The remaining cells stain positively for vimentin and glial fibrillary acidic protein and, therefore, can be considered as astroglial cells. Inoculation of freshly dissociated mouse brain cells in the absence of glucose in a sorbitol-containing medium is not possible; however, feeding of the cultures from day 2 on with sorbitol instead of glucose results in a pure astroglial culture at confluency. Therefore glucose-free growth medium supplemented with sorbitol can be considered a selective medium for astroglial cells in primary mouse glial cultures.

Animals↗
PubMed1991

myo-inositol transport in mouse astroglia-rich primary cultures.

Uptake of radiolabeled myo-inositol was studied in astroglia-rich primary cultures derived from neonatal mouse brains. The uptake was saturable in the presence of Na+ with a Km of 25 microM and a Vmax of 60 pmol.min-1.(mg protein)-1, suggesting a high-affinity transport system for myo-inositol in astroglial cells. In addition, a Na(+)-independent, nonsaturable component was found. Carrier-mediated uptake was not inhibited by cytochalasin B (50 microM), but was reduced by depolarizing concentrations of K+ and, to different extents, in the presence of phloretin, ouabain, or amiloride (1 mM each). scyllo-Inositol, glucose, and galactose also reduced myo-inositol uptake; inhibition by the two hexoses was not reversed in the presence of 0.4 mM sorbinil. On the other hand, uptake of 2-deoxyglucose was not inhibited by high concentrations of myo-inositol. Preincubation of the cells with glucose-free or inositol-free medium stimulated uptake of myo-inositol and preincubation with 25 mM glucose in the presence of 0.4 mM sorbinil had no effect on the rate of uptake. The results suggest that myo-inositol is taken up into the astroglial cells by a transport mechanism that is distinct from that of glucose and probably is an active one. Sorbitol pathway activity does not interfere with myo-inositol uptake.

Animals↗
PubMed1990

Sorbitol pathway activity and utilization of polyols in astroglia-rich primary cultures.

Astroglia-rich rat primary cultures can be grown in a glucose-free medium containing 25 mM sorbitol. After 10 days under these conditions, the total number of cells and DNA content are reduced to 50-60% of those of control cultures, but remain constant thereafter. The specific activities of the sorbitol pathway enzymes, sorbitol dehydrogenase and aldose reductase, are increased 2.5-fold and unchanged, respectively, if the cells are grown in the presence of sorbitol instead of glucose. Treatment with the aldose reductase inhibitor sorbinil does not decrease the number of cells cultured in the glucose-free medium in the presence of sorbitol. Fructose is as good a substrate for the glial cells as sorbitol, whereas out of a number of other polyols tested only xylitol can support the primary cultures for more than 3 days. Neither neuron-rich rat brain primary cultures nor rat glioma cells can be cultured in the sorbitol-containing medium in the absence of glucose. With sorbitol substituting for glucose in the culture medium, effects of glucose deprivation on cellular functions like sugar transport and metabolism can be investigated in glial cultures for an extended period of time.

Aldehyde Reductase↗
PubMed1990

Increased nonshivering thermogenesis, brown fat cytochrome-c oxidase activity, GDP binding, and uncoupling protein mRNA levels after short daily cold exposure of Phodopus sungorus.

In their natural environment, burrowing rodents experience rather fluctuating ambient temperatures and are acutely cold exposed only for short periods outside their burrows. The effect of short daily cold exposure on basal metabolic rate, nonshivering thermogenesis, brown fat thermogenesis, and uncoupling protein mRNA was studied in the Djungarian hamster, Phodopus sungorus. They were kept at 23 degrees C and exposed to 5 degrees C daily either for one 4-h period or twice for 2 h (in 12-h intervals). At the same time control hamsters were kept continuously either at thermoneutrality (23 degrees C) or at 5 degrees C. Two 2-h cold exposures daily were sufficient to increase basal metabolic rate and nonshivering thermogenesis to the same level as continuous cold exposure, whereas one 4-h cold period per day did not result in a significant increase of both parameters. Brown fat thermogenesis (as measured by cytochrome-c oxidase activity and GDP binding to the mitochondrial uncoupling protein) increased to the same extent by both treatments with short daily cold exposure. However, this increase was less than in the chronically cold-exposed hamsters. A similar result was found for uncoupling protein mRNA: both short-term cold-exposed hamsters increased uncoupling protein mRNA levels to a similar extent, but less than after chronic cold treatment. It is concluded that short daily cold exposures are sufficient to cause adaptive increases of the capacity of metabolic heat production as well as brown fat thermogenic properties.

Adipose Tissue↗
PubMed1989

Estimation of free energy barriers in the cytoplasmic and mitochondrial aspartate aminotransferase reactions probed by hydrogen-exchange kinetics of C alpha-labeled amino acids with solvent.

The existence of the postulated quinonoid intermediate in the cytoplasmic aspartate amino-transferase catalyzed transamination of aspartate to oxaloacetate was probed by determining the extent of transfer of tritium from the C alpha position of tritiated L-aspartate to pyridoxamine 5'-phosphate in single turnover experiments in which washout from the back-reaction was obviated by product trapping. The maximum amount of transferred tritium observed was 0.7%, consistent either with a mechanism in which a fraction of the net transamination reaction proceeds through a quinonoid intermediate or with a mechanism in which this intermediate is formed off the main reaction pathway. It is shown that transfer of labeled hydrogen from the amino acid to cofactor cannot be used to differentiate a stepwise from a concerted transamination mechanism. The amount of tritium transferred is a function of the rate constant for torsional equilibration about the epsilon-amino group of Lys-258, the presumptive abstractor of the C alpha proton; the relative rate constants for hydrogen exchange with solvent versus cofactor protonation; and the tritium isotope effect on this ratio. The free energy barriers facing the covalent intermediate between aldimine and keto acid product (i.e., ketimine and possibly quinonoid) were evaluated relatively by comparing the rates of C alpha-hydrogen exchange in starting amino acid with the rates of keto acid formation. The value of theta (= kexge/kprod) was found to be 2.6 for the reaction of cytoplasmic isozyme with aspartate and ca. 0.5 for that of the mitochondrial form with glutamate.

Animals↗
PubMed1989

Effect of photoperiod and acclimation temperature on nonshivering thermogenesis and GDP-binding of brown fat mitochondria in the Djungarian hamster Phodopus s. sungorus.

Acclimation to short photoperiod at 23 degrees C constant Ta caused P. sungorus to improve their NST capacity from 752 to 1,082 mW. Chronic cold exposure in short photoperiod further enhanced the NST capacity, reaching a maximum level of 1,573 mW at -5 degrees C acclimation temperature. Improvements in NST capacity were always accompanied by an increase in brown fat mitochondrial mass and GDP-binding of brown fat mitochondria, in proportion with the cold load applied during temperature acclimation (23 degrees, 15 degrees, 5 degrees, -5 degrees C). Brown fat mitochondrial protein increased from 7.41 mg (23 degrees C Ta, long photoperiod) through 21.6 mg (23 degrees C Ta, short photoperiod) and 81.6 mg (-5 degrees C Ta, short photoperiod). This approximately 10-fold increase was accompanied by a approximately 35-fold increase in GDP-binding (2.0, 7.3 and 71.6 nmol GDP bound, respectively), demonstrating that the increase in capacity for uncoupled respiration in brown fat is of primary significance for thermogenic acclimation to cold as well as to short photoperiod.

Acclimatization↗
PubMed1989

Purification and characterization of sorbitol dehydrogenase from bovine brain.

Sorbitol dehydrogenase (EC 1.1.1.14) was isolated from bovine brain and purified 3,000-fold to apparent homogeneity, as judged by polyacrylamide gel electrophoresis. The purified enzyme had a specific activity of 36 units/mg of protein; a molecular weight of 39,000 for each of the four identical subunits and 155,000 for the intact enzyme were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel exclusion chromatography, respectively. The presence of one Zn2+ per subunit was confirmed by atom absorption spectroscopy; inactivation of the enzyme by metal-chelating agents points to the essential role that Zn2+ plays in the catalytically competent enzyme. The enzyme is also inactivated by thiol-blocking reagents; with respect to inactivation by sodium pyrophosphate, sorbitol dehydrogenase is different from closely related alcohol dehydrogenase.

Aldehyde Reductase↗
PubMed1989

Characteristics of sorbitol uptake in rat glial primary cultures.

Uptake of [U-14C]sorbitol was studied in astrogliarich rat primary cultures. Initial rate of sorbitol uptake is proportional to sorbitol concentration between 20 microM and 400 mM. Sorbitol transport is not inhibited by glucose, fructose, and a variety of structurally related polyols, or by cytochalasin B, an inhibitor of glucose transport. Phloretin, phlorizin, filipin, and n-hexanol, all compounds that alter the properties of biological membranes, and the sulfhydryl reagent p-chloromercuribenzoate inhibit sorbitol uptake to various degrees. Variation in the concentrations of extracellular Na+ and K+ does not affect transfer of sorbitol across the cell membrane. It is concluded that sorbitol is taken up into glial cells by a diffusion process, not involving a carrier and probably not through the lipid bilayer, but through a proteinaceous channel-like structure.

Animals↗
PubMed1989

Short photoperiod and cold activate brown fat lipoprotein lipase in the Djungarian hamster.

Activation of lipoprotein lipase (LPL) in brown fat of the Djungarian hamster ensures substrate supply for nonshivering thermogenesis. Cold acclimation [5 degrees C ambient temperature (Ta)] in long photoperiod [light-dark (LD) 16:8] increased brown fat LPL activity from 4.7 to 22.7 nmol oleate.mg-1.min-1 within 1 day. Slight reduction of this high LPL activity was observed during prolonged cold exposure for up to 4 wk. Deacclimation (transfer from 5 to 23 degrees C Ta) caused total inactivation of brown fat LPL within 4 days. Short photoperiod (LD 8:16) also stimulates brown fat LPL activity. Its effect can be summarized by three different observations. At thermoneutrality short photoperiod elevated LPL activity to 7.1 instead of 4.7 nmol.mg-1.min-1 observed in long photoperiod. Second, at low ambient temperature short photoperiod reinforces the effect of cold acclimation. Maximum LPL activity of cold-exposed Djungarian hamsters averaged 23.7 nmol.mg-1.min-1 in long photoperiod and was elevated to 35.5 nmol.mg-1.min-1 in short photoperiod. Third, inactivation of LPL during deacclimation was delayed in short photoperiod. These results demonstrate that photoperiod as well as ambient temperature may be cooperatively used as environmental cues for seasonal acclimation of brown fat substrate supply for nonshivering thermogenesis.

Acclimatization↗
PubMed1989

Photoperiod and thermoregulation in vertebrates: body temperature rhythms and thermogenic acclimation.

Evidence has recently begun to accumulate that photoperiodic responses of mammals and birds may affect the control of energy balance and thermoregulation. Exposure to short photoperiod can lower the set point for body temperature regulation in birds and mammals, as well as the voluntarily selected body temperature in ectothermic lizards. This decrease is accompanied by a reorganization of circadian or ultradian rhythms of body temperature, particularly an increase in periods spent at rest with minimum body temperatures. Short photoperiod is also used as an environmental cue for induction of seasonal torpor or facilitation of hibernation. During winter, cold tolerance of small mammals is improved by an increase of nonshivering thermogenesis in brown fat. Thermogenic capacity of brown fat (respiratory enzymes, mitochondria, uncoupling protein) is enhanced in response to short photoperiod. This response is mediated via an increase in the activity of sympathetic innervation in brown fat. Moreover, an exposure to short photoperiod prior to low temperatures may act in preparing brown fat for facilitated thermogenesis during acclimation to cold. This shows that photoperiodic control not only affects energy balance indirectly via the control of reproduction or body mass, but may directly interact with central control of thermoregulation and may influence the process of acclimatization.

Activity Cycles↗
PubMed1988

[Spontaneous course of Wegener's granulomatosis].

Two cases of Wegener's granulomatosis are reported. During the 5 and 9 years course of the disease complete remissions and the disappearance of pulmonary opacities were found without specific treatment. Since 1970 cytotoxic drugs, e.g. cyclophosphamide, are frequently used for the treatment of Wegener's granulomatosis. The evaluation of the follow-up showed in these patients that in most cases the side-effects of the therapy, and not the primary disease, limited the life-expectancy. This especially applies for patients with mild forms of Wegener's granulomatosis, in which the initiation of a cytotoxic therapy should carefully be evaluated.

Aged↗