L-Aspartate,L-Ornithine and L-Ornithine-L-Aspartate (LOLA) and Their Impact on Brain Energy Metabolism

L-Ornithine-L-aspartate (LOLA), a crystalline salt, is used primarily in the management of hepatic encephalopathy. The degree to which it might penetrate the brain, and the effects it might have on metabolism in brain are poorly understood. Here, to investigate the effects of LOLA on brain energy metabolism we incubated brain cortical tissue slices from guinea pig (Cavea porcellus) with the constituent amino acids of LOLA, L-ornithine or L-aspartate, as well as LOLA, in the presence of [1-¹³C]D-glucose and [1,2-¹³C]acetate; these labelled substrates are useful indicators of brain metabolic activity. L-Ornithine produced significant “sedative” effects on brain slice metabolism, most likely via conversion of ornithine to GABA via the ornithine aminotransferase pathway, while L-aspartate showed concentration-dependent excitatory effects. The metabolic effects of LOLA reflected a mix of these two different processes and were concentration-dependent. We also investigated the effect of an intraperitoneal bolus injection of L-ornithine, L-aspartate or LOLA on levels of metabolites in kidney, liver and brain cortex and brain stem in mice (C57Bl6J) 1 h later. No significant changes in metabolite levels were seen following the bolus injection of L-aspartate, most likely due to rapid metabolism of aspartate before reaching the target tissue. Brain cortex glutamate was decreased by L-ornithine but no other brain effects were observed with any other compound. Kidney levels of aspartate were increased after injection of L-ornithine and LOLA which may be due to interference by ornithine with the kidney urea cycle. It is likely that without optimising chronic intravenous infusion, LOLA has minimal impact on healthy brain energy metabolism due to systemic clearance and the blood - brain barrier.

     

Ghrelin: Impact on Muscle Energy Metabolism in Sepsis

We investigated the effects of exogenous ghrelin on energy levels and tissue histology in skeletal muscle in experimentally lipopolysaccharide (LPS) induced septic rats. Male Wistar albino rats 200–250 g were separated into four groups; Control, LPS (5 mg/kg), Ghrelin (10 nmol/kg i.v.), and ghrelin+LPS. Gastrocnemius muscle tissue was taken and stained using modified Gomori trichrome (MGT), succinic dehydrogenase (SDH), and cytochrome oxidase (COX) and hematoxylin and eosin. In stained sections, histological score value was calculated according to the intensity and the distribution for MGT, SDH and COX stainings. Creatine, creatine phosphate, adenosine triphosphate (ATP), adenosine monophosphate (AMP) levels, and the ratios of AMP/ATP and CreaP/ATP were investigated using high performance liquid chromatography (HPLC) in muscle tissue. Significances between experimental groups were calculated with an analysis of variance (ANOVA) followed by Tukey’s tests. Myopathic changes were seen in the 50% of rats in the LPS group as rounding of muscle fibers and fiber size variation. In the ghrelin+LPS group, ghrelin treatment was reduced damage in skeletal muscle structure. There was no change in creatine or AMP levels between the groups. Ghrelin treatment significantly increased ATP values (P?<?0.01) and improved tissue histology in septic rats. Ratios of both AMP/ATP and CreaP/ATP were found increased in the septic group, but there were decreaments in both the ghrelin and ghrelin-treated septic groups. Ghrelin could play an important role in energy balance and muscle morphology in skeletal muscle during sepsis.     

Modeling Brain Energy Metabolism and Function: A Multiparametric Monitoring Approach

Mathematical modeling of brain function is an important tool needed for a better understanding of experimental results and clinical situations. In the present study, we are constructing and testing a mathematical model capable of simulating changes in brain energy metabolism that develop in real time under various pathophysiological conditions. The model incorporates the following parameters: cerebral blood flow, partial oxygen pressure, mitochondrial NADH redox state, and extracellular potassium. Accordingly, all the model variables are only time dependent (`point-model' approach). Numerical runs demonstrate the ability of the model to mimic pathological conditions, such as complete and partial ischemia, cortical spreading depression under normoxic and partial ischemic conditions. They also show that, when properly tuned, a model of this type permits the monitoring of only one or two crucial variables and the computation of the remaining variables in real time during clinical or experimental procedures.     

Effects of 3-Nitropropionic Acid on Synaptosomal Energy and Transmitter Metabolism: Relevance to Neurodegenerative Brain Diseases

Abstract: 3-Nitropropionic acid (3-NPA) inhibited synaptosomal respiration in a dose-dependent manner; the degree of inhibition by the same concentration of the compound was greater, however, when respiration was stimulated by concomitant increase in ATP usage. The most rapid event after addition of 3-NPA was a decrease in [creatine phosphate]/[creatine] ([CrP]/[Cr]) and an increase in [lactate]/[pyruvate]. A fall in [ATP]/[ADP] and [GTP]/[GDP] was initially less pronounced but closely followed that in [CrP]/[Cr]. In the absence of glutamine, 3-NPA caused a pronounced decrease in internal aspartate level and a small reduction in glutamate concentration, whereas [GABA] rose; the sum of these three amino acids inside synaptosomes fell, but there were no increases in their external levels. With glutamine in the medium, the reduction in intrasynaptosomal aspartate was accompanied by increases in intrasynaptosomal glutamate and GABA. The external concentration of glutamate rose substantially in the presence of the inhibitor. 3-NPA had no effect on basal release of either glutamate (and GABA) or biogenic amines but increased efflux occurring upon addition of nonsaturating concentrations of the depolarizing agents veratridine and KCI. The results allow the following predictions with respect to the behavior of brain metabolism in neurodegenerative diseases that involve restrictions of mitochondrial function: (1) The extent of inhibition of mitochondrial ATP generation is expected to be greater in cells with high energy demand. The earliest signs of impairment of the respiratory chain function are a fall in [PCr]/[Cr] (or a rise in [Pi]/[CrP]) and an increase in [lactate]/[pyruvate]. (2) A fall in [GTP]/[GDP] can limit protein synthesis. This may be one of the factors that contributes to cell death. (3) An increase in the concentration of inorganic phosphate stimulates neuronal glutaminase activity and leads to a release of glutamate into the external environment; the latter could activate excitatory amino acid receptors. (4) A lowered energy level limits the cell's ability to restore ion gradients. Stimulated release of transmitters from neurons may, therefore, be enhanced and their reuptake delayed.     

Clonidine and Brain Mitochondrial Energy Metabolism: Pharmacodynamic Insights Beyond Receptorial Effects

Neurochemical Research - Clonidine is an anti-hypertensive drug that inhibits the release of norepinephrine from pre-synaptic terminals binding to pre-synaptic α2-adrenoreceptors. Some studies...     

Influence of Severe Hypoglycemia on Brain Extracellular Calcium and Potassium Activities, Energy, and Phospholipid Metabolism

Abstract: In the cerebral cortices of rats, during insulininduced hypoglycemia, changes in the concentrations of labile phosphate compounds [ATP, ADP, AMP, and phosphocreatine (PCr)] and glycolytic metabolites (lactate, pyruvate, and glucose) as well as phospholipids and free fatty acids (FFAs) were studied in relation to extracellular potassium and calcium activities. Changes in extracellular calcium and potassium activities occurred at approximately the onset of isoelectricity. The extracellular calcium activity dropped from 1.17 ± 0.14 mM to 0.18 ± 0.28 mM and the potassium activity rose from 3.4 ± 0.94 mM to 48 ± 12 mM (means ± SD). Minutes prior to this ionic change the levels of ATP, PCr, and phospholipids were unchanged while the levels of FFAs remained unchanged or slightly elevated. Following the first ionic change the steady-state levels of ATP decreased by 40%, from 2.42 to 1.56 μmol/g. PCr levels decreased by 75%, from 4.58 to 1.26 μmol/g. Simultaneously, the levels of FFAs increased from 338 to 642 nmol/g, arachidonic acid displaying the largest relative increase, 33 to 130 nmol/g. The first ionic change was followed by a short period of normalization of ionic concentrations followed by a sustained ionic change. This was accompanied by a small additional decrease in ATP (to 1.26 μmol/g). The FEA levels increased to 704 nmol/g. There was a highly sig nificant negative correlation between the levels of FFAs and the energy charge of the tissue. The formation of FFAs was accompanied by a decrease in the phospholipid pool. The largest relative decrease was observed in the inositol phosphoglycerides, followed by serine and ethanolamine phosphoglycerides. After 10 min of isoelectricity the levels of phospholipids had decreased by 5.12 μmol/g while the levels of FFAs had increased by 0.46 μmol/g, indicating oxidative metabolism or washout of the released FFAs. The attenuation of the rapid initial changes in the levels of the energy metabolites and FFAs as well as the correlation between the energy charge and the levels of FFAs suggests that a new steady state is established following the first ionic change. The importance of these reactions for the development of hypogiycemic neuronal damage is discussed.     

LOLA ZRREGULARIS (CHLOROPHYTA-CLADOPHORACEAE): A NEW SPECIES FROM THE ROSS SEA,ANTARCTICA1

A small, loose lying, unbranched, filamentous alga from the Ross Sea, Antarctica, is described as a new species, Lola irregularis. The genus Lola is closely related to the genera Hormiscia, Rhizoclonium, and Chaetomorpha in the family Cladophoraceae. A key to all known species of Lola is included. This is the first record of a species of Lola south of the Antarctic convergence.     

The Development of Enzymes of Energy Metabolism in the Brain of a Precocial (Guinea Pig) and Non-Precocial (Rat) Species

Abstract: Key enzymes of ketone body metabolism (3-hydroxybutyrate de-hydrogenase, 3-oxo-acid: CoA transferase, acetoacetyl-CoA thiolase) and glucose metabolism (hexokinase, lactate dehydrogenase, pyruvate dehydrogenase, citrate synthase) have been measured in the brains of foetal, neonatal and adult guinea pigs and compared to those in the brains of neonatal and adult rats. The activities of the guinea pig brain ketone-body-metabolising enzymes remain relatively low in activity throughout the foetal and neonatal periods, with only slight increases occurring at birth. This contrasts with the rat brain, where three- to fourfold increases in activity occur during the suckling period (0–21 days post partum), followed by a corresponding decrease in the adult. The activities of the hexokinase (mitochondrial and cytosolic), pyruvate dehydrogenase, lactate dehydrogenase and citrate synthase of guinea pig brain show marked increases in the last 10–15 days before birth, so that at birth the guinea pig possesses activities of these enzymes similar to the adult state. This contrasts with the rat brain where these enzymes develop during the late suckling period (10–15 days after birth). The development of the enzymes of aerobic glycolytic metabolism correlate with the onset of neurological competence in the two species, the guinea pig being a "precocial" species born neurologically competent and the rat being a "non-precocial" species born neurologically immature. The results are discussed with respect to the enzymatic activities required for the energy metabolism of a fully developed, neurologically competent mammalian brain and its relative sensitivity to hypoxia.     

LOLA IRREGULARIS (CHLOROPHYTA-CLADOPHORACEAE) : A NEW SPECIES FROM THE ROSS SEA,ANTARCTICA1

A small, loose lying, unbranched, filamentous alga from the Ross Sea, Antarctica, is described as a new species, Lola irregularis. The genus Lola is closely related to the genera Hormiscia, Rhizoclonium, and Chaetomorpha in the family Cladophoraceae. A key to all known species of Lola is included. This is the first record of a species of Lola south of the Antarctic convergence.     

Effects of Amphetamine on Protein Synthesis and Energy Metabolism in Mouse Brain: Role of Drug-Induced Hyperthermia

Abstract: Changes in brain protein synthesis activity, and in brain levels of glucose, glycogen, and several high-energy phosphate metabolites, were evaluated under conditions of amphetamine-induced hyperthermia in mice. Protein synthesis showed a striking dependence on rectal temperature ( T _R), falling abruptly at T _R above 40°C. A similar result was obtained following direct heating of the animals. Protein synthesis activity in liver showed the same temperature dependence observed for brain. Increased synthesis of a protein with characteristics of the major mammalian stress protein, hsp 70, was demonstrated in both brain and liver following amphetamine administration. Brain protein synthesis showed significant recovery within 2 h after amphetamine administration whereas that of liver remained below 30% of control activity, suggesting significant temporal and quantitative differences in the response of individual tissues to elevated temperatures. Brain glycogen levels after amphetamine administration were significantly lower under conditions of ambient temperature which resulted in more severe drug-induced hyperthermia but did not correlate as strikingly as protein synthesis with the temperatures of individual animals. Brain glycogen also fell in animals whose temperatures were increased by brief exposure at high ambient temperature. Brain glucose levels did not consistently change with hyperthermia. Slight decreases in high-energy phosphates with increasing T _R were likely the result of fixation artifact. These results demonstrate the fundamental role of hyperthermia in the reduction of protein synthesis in brain and other tissues by amphetamine, and suggest that temperature also constitutes a significant source of variability in the effects of this drug on brain energy metabolism, in particular glycogenolysis.     

Interactions of Triethyltin-Chloride (TET) with the Energy Metabolism of Cultured Rat Brain Astrocytes: Studies by Multinuclear Magnetic Resonance Spectroscopy

The effect of triethyltin-chloride (TET), a highly neurotoxic compound, on the cellular metabolism of rat brain astrocytes in vitro was examined by nuclear magnetic resonance (NMR) spectroscopy. 5-week-old cultures were exposed to TET (0.2–40 M) either for (1) acute (3h), (2) 24 h, or (3) chronic treatment (8 d). Cells were labeled with 1-¹³C-glucose, cell extracts were prepared and ³¹P, ¹H, and ¹³C spectra were analyzed. Cytotoxic effects of TET were assessed by vital dye uptake assay using neutral red (NR) and by exclusion of trypan blue (TB). Cells were examined ultrastructurally by electron microscopy. The data show that the major target of TET at concentrations already causing morphological effects on cultured astrocytes is not the energy metabolism, but that TET rather alters the intracellular concentrations of organic osmolytes, such as myo-inositol, taurine and hypotaurine, which are part of the control of ion and volume regulation and osmotic balance in astrocytes.     

Energy Metabolism in Postlarval Ontogenesis of Planorbarius corneus (Planorbidae,Gastropoda)

Russian Journal of Developmental Biology - The change in the rate and mass-specific rate of oxygen consumption was studied during individual ontogenesis of nine Planorbarius corneus individuals...     

Impact of Silver(I) on the Metabolism of Shewanella oneidensis

Anaerobic cultures of Shewanella oneidensis MR-1 reduced toxic Ag(I), forming nanoparticles of elemental Ag(0), as confirmed by X-ray diffraction analyses. The addition of 1 to 50 μM Ag(I) had a limited impact on growth, while 100 μM Ag(I) reduced both the doubling time and cell yields. At this higher Ag(I) concentration transmission electron microscopy showed the accumulation of elemental silver particles within the cell, while at lower concentrations the metal was exclusively reduced and precipitated outside the cell wall. Whole organism metabolite fingerprinting, using the method of Fourier transform infrared spectroscopy analysis of cells grown in a range of silver concentrations, confirmed that there were significant physiological changes at 100 μM silver. Principal component-discriminant function analysis scores and loading plots highlighted changes in certain functional groups, notably, lipids, amides I and II, and nucleic acids, as being discriminatory. Molecular analyses confirmed a dramatic drop in cellular yields of both the phospholipid fatty acids and their precursor molecules at high concentrations of silver, suggesting that the structural integrity of the cellular membrane was compromised at high silver concentrations, which was a result of intracellular accumulation of the toxic metal.Silver is an element that has been used widely in industrial processes as diverse as photographic processing, catalysis, mirror production, electroplating, alkaline battery production, and jewelry making (18). It has been known for some time that silver ions and silver-based compounds can be highly toxic to microorganisms, and with increasing concern about pathogenic “superbugs” with high resistance to conventional antibiotics, silver is attracting much interest as a potential biocide (11, 18, 36, 42). Silver has no known physiological functions and can exist in several oxidation states, although it is most commonly encountered in its elemental [Ag(0)] and monovalent [Ag(I)] forms. Although use of nanoscale elemental Ag(0) as a biocide has been increasing, for example, in wound dressings and as an antimicrobial coating on consumer products, little is known about its mode of toxicity. This is despite the surprising ability of actively growing Fe(III)-reducing bacteria such as Geobacter sulfurreducens to precipitate nanoscale Ag(0) particles within and around the cell surface via reduction of Ag(I) (18). Ionic Ag(I), in contrast, has been the focus of more studies on the mode of metal toxicity. Previous research showed that silver ions have antimicrobial activities against a wide diversity of bacteria (19). They have been shown to disrupt the respiratory chain of Escherichia coli (3) and inhibit the exchange of phosphate and its uptake (34). Ag(I) has also been linked to copper metabolism in E. coli, potentially competing with copper binding sites on the cell surface and subsequent copper transport into the cell (8). However, the toxicity of silver is not limited to prokaryotes, as long-term exposure in humans can cause argyria, impaired night vision, and abdominal pain (31, 32, 36). The detailed mechanism of toxicity in prokaryotes or eukaryotes remains to be identified, although it has been proposed that silver ions react with cellular proteins via SH groups (16), leading to the disruption of cellular metabolism.Microbial cells have evolved an extremely diverse range of mechanisms to survive high concentrations of toxic metals. The mechanisms invoked include biosorption, bioaccumulation, special efflux systems, alteration of solubility and toxicity via reduction or oxidation, extracellular complexation or precipitation of metals, and lack of specific metal transport systems (1). For example, for silver ions the bacterial cell wall can be an efficient permeability barrier to block the uptake of metal (21), with additional complexation in the periplasm by specific silver-binding proteins (35). Redox transformations also offer the potential to detoxify Ag(I) ions, e.g., through the reduction to insoluble elemental Ag(0) (30). In addition, the energy-dependent efflux of toxic Ag(I) is perhaps the best-studied resistance mechanism for silver, mediated via ATPases and chemiosmotic cation/protons antiporters (9).Shewanella spp., Gram-negative, dissimilatory metal-reducing bacteria, can use a wide variety of terminal electron acceptors for growth (23, 39), including high oxidation state metals such as Fe(III), Mn(IV), Cr(VI), U(VI), and Au(III) (5, 17, 26, 28, 41). Shewanella species also have the potential to reduce Ag(I), given their similar activities against Au(III), and the reduction of Ag(I) to form nanoscale deposits of Ag(0) within the cell has been documented for other Fe(III)-reducing bacteria (18). This metabolic versatility offers considerable potential for bioremediation applications, for example, via reduction of U(VI) to insoluble U(IV) (5, 17, 26, 28, 41), and the recovery of precious metals such as silver and gold via reductive precipitation. It also offers an interesting model organism to study the metabolism of toxic metals such as silver, including the physiological impact of ionic Ag(I) and nanoscale Ag(0).This paper describes interactions of Shewanella oneidensis MR-1 with various concentrations of Ag(I), including demonstrations of the reduction and deposition of silver nanoparticles under anaerobic conditions. A range of techniques, including X-ray diffraction (XRD) and analytical transmission electron microscopy (TEM), were used to investigate the nature and cellular localization of the precipitates, while Fourier transform infrared (FT-IR) spectroscopy metabolic profiling techniques were used to identify the impact of toxic metal accumulation on the cell. The disruption of membrane integrity was implied by these investigations and confirmed by fatty acid methyl ester (FAME) analysis, which showed a dramatic decrease in the quantities of membrane lipid components.     

The Relation of Energy Metabolism and Body Weight in Bivalves (Mollusca: Bivalvia)

On the basis of experimental and published data, the interspecific and intraspecific (ontogenetic) dependence of energy metabolism on body weight in bivalves was calculated. Changes in the parameters of intraspecific allometric dependence under the effect of environmental factors were analyzed. The rate of comparable standard metabolism (coefficient a at k = 0.76) was shown to vary in different taxonomic and zoogeographic groups of bivalves.