Glycogen Consumption Research Articles

The nutritional status of Methanosarcina acetivorans regulates glycogen metabolism and gluconeogenesis and glycolysis fluxes.

Gluconeogenesis is an essential pathway in methanogens because they are unable to use exogenous hexoses as carbon source for cell growth. With the aim of understanding the regulatory mechanisms of central carbon metabolism in Methanosarcina acetivorans, the present study investigated gene expression, the activities and metabolic regulation of key enzymes, metabolite contents and fluxes of gluconeogenesis, as well as glycolysis and glycogen synthesis/degradation pathways. Cells were grown with methanol as a carbon source. Key enzymes were kinetically characterized at physiological pH/temperature. Active consumption of methanol during exponential cell growth correlated with significant methanogenesis, gluconeogenic flux and steady glycogen synthesis. After methanol exhaustion, cells reached the stationary growth phase, which correlated with the rise in glycogen consumption and glycolytic flux, decreased methanogenesis, negligible acetate production and an absence of gluconeogenesis. Elevated activities of carbon monoxide dehydrogenase/acetyl-CoA synthetase complex and pyruvate: ferredoxin oxidoreductase suggested the generation of acetyl-CoA and pyruvate for glycogen synthesis. In the early stationary growth phase, the transcript contents and activities of pyruvate phosphate dikinase, fructose 1,6-bisphosphatase and glycogen synthase decreased, whereas those of glycogen phosphorylase, ADP-phosphofructokinase and pyruvate kinase increased. Therefore, glycogen and gluconeogenic metabolites were synthesized when an external carbon source was provided. Once such a carbon source became depleted, glycolysis and methanogenesis fed by glycogen degradation provided the ATP supply. Weak inhibition of key enzymes by metabolites suggested that the pathways evaluated were mainly transcriptionally regulated. Because glycogen metabolism and glycolysis/gluconeogenesis are not present in all methanogens, the overall data suggest that glycogen storage might represent an environmental advantage for methanosarcinales when carbon sources are scarce. Also, the understanding of the central carbohydrate metabolism in methanosarcinales may help to optimize methane production.

Effects of sulphide on anoxia-driven mortality and anaerobic metabolism in the ark shellAnadara kagoshimensis

Reduction in survival under hypoxic conditions in the presence of sulphide has been repeatedly demonstrated in various benthic invertebrates. However, the reason for this reduction has not yet been clearly elucidated. In this study, the effects of sulphide accumulation on anoxic survival and anaerobic metabolism were investigated in the ark shell Anadara kagoshimensis. Ark shells from western Japan were experimentally exposed to 3 sulphide-accumulation levels under sustained anoxic conditions: accumulated H2S treatment (static incubation), decreased H2S treatment (semi-static incubation with daily replacement of incubation media), and inhibited H2S treatment (static incubation with the addition of antibiotics). Moreover, the effect of antibiotics on anoxic survival was examined under sulphide exposure. The decreased H2S and inhibited H2S treatments resulted in 1.5- and 3-fold increase in the anoxic survival time, respectively, when compared with the accumulated H2S treatment. Under anoxic sulphide exposure, the antibiotics addition did not affect survival time, suggesting the shorter survival time in the accumulated H2S incubation was probably due to sulphide toxicity. Glycogen consumption and propionate accumulation, which indicate activation of anaerobic metabolism, were observed in both accumulated and inhibited H2S treatments. However, glycogen consumption was significantly higher in the accumulated H2S treatment after a significant sulphide accumulation was detected in the incubation media. In addition, survival in the accumulated H2S treatment decreased rapidly, whereas no significant mortality was observed in the inhibited H2S throughout the experiment. These results likely suggest that the accelerated anoxic-driven mortality in sulphide-rich environments was partly due to the faster breakdown of glycogen.

Lipid-Liver Crosstalk in Health and Disease

The liver is a major metabolic organ, essentially controlling glucose and fat metabolism. Because lipids are water-insoluble, they need to be transported in the circulation in association with proteins in the form of lipoproteins. These lipoproteins play a key role in the absorption and transport of dietary lipids by the small intestine, in the transport of lipids from the liver to peripheral tissues, and in the transport of lipids from peripheral tissues to the liver and intestine. Upon feeding, energy is provided by glycolysis and the unused glucose is stored as glycogen in the liver. Excess glucose is used to synthesize fatty acids through de novo lipogenesis. Fatty acids are incorporated into triacylglycerol, phospholipids, or cholesterol esters in hepatocytes. These complex lipids are stored in lipid droplets or secreted into the circulation as very low-density lipoprotein particles. Upon fasting, after consumption of stored glycogen, the liver secretes glucose through gluconeogenesis. Fasting also promotes lipolysis in adipose tissue, resulting in release of free fatty acids which are metabolized in hepatic mitochondria though beta-oxidation for energy production, while excess fatty acids are stored by the liver leading to hepatic steatosis. Disturbances in lipid metabolism, as in alcoholic and non-alcoholic steatohepatitis, obesity and diabetes, will affect liver performance and function. Likewise, disturbed liver functions by acute or chronic liver disease, as in viral hepatitis, will affect lipid homeostasis. This may carry an increased risk of atherosclerosis and ischemic heart disease which may endanger life. Dietary restriction or fasting was found to have a positive impact on restoring lipid homeostasis leading to improved quality of life.

Metabolic and hormonal response to intermittent high-intensity and continuous moderate intensity exercise in individuals with type 1 diabetes: a randomised crossover study.

To investigate exercise-related fuel metabolism in intermittent high-intensity (IHE) and continuous moderate intensity (CONT) exercise in individuals with type 1 diabetes mellitus. In a prospective randomised open-label cross-over trial twelve male individuals with well-controlled type 1 diabetes underwent a 90 min iso-energetic cycling session at 50% maximal oxygen consumption ([Formula: see text]), with (IHE) or without (CONT) interspersed 10 s sprints every 10 min without insulin adaptation. Euglycaemia was maintained using oral (13)C-labelled glucose. (13)C Magnetic resonance spectroscopy (MRS) served to quantify hepatocellular and intramyocellular glycogen. Measurements of glucose kinetics (stable isotopes), hormones and metabolites complemented the investigation. Glucose and insulin levels were comparable between interventions. Exogenous glucose requirements during the last 30 min of exercise were significantly lower in IHE (p = 0.02). Hepatic glucose output did not differ significantly between interventions, but glucose disposal was significantly lower in IHE (p < 0.05). There was no significant difference in glycogen consumption. Growth hormone, catecholamine and lactate levels were significantly higher in IHE (p < 0.05). IHE in individuals with type 1 diabetes without insulin adaptation reduced exogenous glucose requirements compared with CONT. The difference was not related to increased hepatic glucose output, nor to enhanced muscle glycogen utilisation, but to decreased glucose uptake. The lower glucose disposal in IHE implies a shift towards consumption of alternative substrates. These findings indicate a high flexibility of exercise-related fuel metabolism in type 1 diabetes, and point towards a novel and potentially beneficial role of IHE in these individuals. ClinicalTrials.gov NCT02068638 FUNDING: Swiss National Science Foundation (grant number 320030_149321/) and R&A Scherbarth Foundation (Switzerland).

Effects of 6 weeks of aerobic exercise combined with conjugated linoleic acid on the physical working capacity at fatigue threshold.

The purpose of this study was to examine the effects of conjugated linoleic acid (CLA) supplementation in conjunction with 6 weeks of aerobic exercise training on the physical working capacity at the fatigue threshold (PWCFT), timed sit-ups, and the standing long jump. Thirty-three untrained to moderately trained men (mean ± SD; age = 21.6 ± 2.8 years) participated in this double-blind, placebo-controlled study and were randomly assigned to either a CLA (Clarinol A-80; n = 17) or placebo (PLA; sunflower oil; n = 16) group. Before and after 6 weeks of aerobic training (50% VO2peak for 30 minutes, twice per week) and supplementation (8 ml CLA or PLA per day), each subject completed an incremental cycle ergometer test to exhaustion to determine the PWCFT, maximal number of sit-ups in 1 minute, and the standing long jump. There were no differences (all p ≥ 0.23) between the CLA and PLA groups for the analysis of covariance-adjusted posttest mean values for PWCFT, sit-ups, or standing long jump. The PWCFT increased from pre- to posttraining in the CLA (p = 0.003) and PLA (p = 0.003) groups. There were no differences (p > 0.05) from pre- to posttraining for sit-ups and standing long jump in either the CLA or PLA groups. There was no effect of CLA on the training-induced increases in PWCFT, nor were there any effects of CLA or aerobic training on the maximum number of sit-ups or standing long jump. Thus, CLA had no ergogenic benefits on this model of aerobic training-induced improvements in neuromuscular fatigue, or on field tests of muscle endurance and power.

The effect of substrate competition on the metabolism of polyphosphate accumulating organisms (PAOs)

The type of carbon source present in the wastewater is one factor that affects the competition between polyphosphate accumulating organisms (PAO) and glycogen accumulating organisms (GAO) and therefore, the efficiency of the enhanced biological phosphorus removal (EBPR) process. This study investigated the impact of the carbon source composition on the anaerobic and aerobic kinetics of PAOs and the EBPR performance of an 85% PAO enrichment. When both acetate (HAc) and propionate (HPr) were present, propionate was depleted more quickly, with a constant uptake rate of 0.18 ± 0.02 C-mol/(C-mol biomass·h), while the acetate uptake rate decreased with an increase in propionate concentration, due to the substrate competition between acetate and propionate. The metabolic model for PAOs was modified to incorporate the anaerobic substrate competition effect. The aerobic rates for phosphorus (P) uptake, glycogen production and polyhydroxyalkanoates (PHA) degradation were within the same range for all tests, indicating that these rates are essentially independent of the acetate and propionate concentration, simplifying the calibration procedure for metabolic models. The metabolic model applied to describe the anaerobic and aerobic activity agreed well with the experimental data of HAc, HPr, P, PHA and biomass growth. The low glycogen consumption observed suggest that some reducing equivalents were generated anaerobically through the TCA cycle. The results of this work suggest that the propionate uptake kinetics by PAOs can provide them an advantage over GAOs in EBPR systems, even when the propionate fraction of the influent is relatively low.

Nitrogen removal from medium-age landfill leachate via post-denitrification driven by PHAs and glycogen in a single sequencing batch reactor

An anaerobic/aerobic/anoxic (AOA) process in a sequencing batch reactor (SBR) was proposed to treat typical medium-age landfill leachate without extra carbon addition. In a steady-state, the average removal efficiencies of NH4+-N, total nitrogen (TN) and COD were 99.7±0.1%, 98.3±0.3% and 89.8±1.4%, when influent NH4+-N, TN and COD were 1025–1327mg/L, 1346–1854mg/L and 6430–9372mg/L, respectively. In the anaerobic stage, dissolved organic matter was taken up partially and stored as polyhydroxyalkanoates (PHAs) with concomitant consumption of glycogen. In the aerobic stage, PHAs was oxidized and glycogen was replenished in the bacterial cells, when TN of 75.4mg/L was removed via simultaneous nitrification and denitrification (SND). The residual nitrate and nitrite were denitrified completely by utilizing residual PHAs and glycogen as electron donors in the anoxic phase. Denitrifying glycogen accumulating organisms (GAOs) were considered to be playing the major role in the process.

When shape matters: Strategies of different Antarctic ascidians morphotypes to deal with sedimentation

Climate change leads to increased melting of tidewater glaciers in the Western Antarctic Peninsula region and sediment bearing glacial melt waters negatively affects filter feeding species as solitary ascidians. In previous work the erect-forms Molgula pedunculata and Cnemidocarpa verrucosa (Order Stolidobranchiata) appeared more sensitive than the flat form Ascidia challengeri (Order Phlebobranchiata). Sedimentation exposure is expected to induce up-regulation of anaerobic metabolism by obstructing the organs of gas exchange (environmental hypoxia) or causes enhanced squirting activity (functional hypoxia). In this study we evaluated the possible relationship between ascidian morphotype and their physiological response to sedimentation. Together with some behavioural observations, we analysed the response of anaerobic metabolic parameters (lactate formation and glycogen consumption) in different tissues of three Antarctic ascidians, exposed to high sediment concentrations (200 mgL−1). The results were compared to experimental hypoxia (10% pO2) and exercise (induced muscular contraction) effects, in order to discriminate the effect of sediment on each species and morpho-type (erect vs. flat forms). Our results suggest that the styled (erect) C. verrucosa increases muscular squirting activity in order to expulse excessive material, while the flat-form A. challengeri reacts more passively by down-regulating its aerobic metabolism under sediment exposure. Contrary, the erect ascidian M. pedunculata did not show any measurable response to the treatments, indicating that filtration and ingestion activities were not reduced or altered even under high sedimentation (low energetic material) which could be disadvantageous on the long-term and could explain why M. pedunculata densities decline in the study area.

Contributions of glycogen to astrocytic energetics during brain activation.

Glycogen is the major store of glucose in brain and is mainly in astrocytes. Brain glycogen levels in unstimulated, carefully-handled rats are 10-12 μmol/g, and assuming that astrocytes account for half the brain mass, astrocytic glycogen content is twice as high. Glycogen turnover is slow under basal conditions, but it is mobilized during activation. There is no net increase in incorporation of label from glucose during activation, whereas label release from pre-labeled glycogen exceeds net glycogen consumption, which increases during stronger stimuli. Because glycogen level is restored by non-oxidative metabolism, astrocytes can influence the global ratio of oxygen to glucose utilization. Compensatory increases in utilization of blood glucose during inhibition of glycogen phosphorylase are large and approximate glycogenolysis rates during sensory stimulation. In contrast, glycogenolysis rates during hypoglycemia are low due to continued glucose delivery and oxidation of endogenous substrates; rates that preserve neuronal function in the absence of glucose are also low, probably due to metabolite oxidation. Modeling studies predict that glycogenolysis maintains a high level of glucose-6-phosphate in astrocytes to maintain feedback inhibition of hexokinase, thereby diverting glucose for use by neurons. The fate of glycogen carbon in vivo is not known, but lactate efflux from brain best accounts for the major metabolic characteristics during activation of living brain. Substantial shuttling coupled with oxidation of glycogen-derived lactate is inconsistent with available evidence. Glycogen has important roles in astrocytic energetics, including glucose sparing, control of extracellular K(+) level, oxidative stress management, and memory consolidation; it is a multi-functional compound.

Shifts in the Distribution of Mass Densities Is a Signature of Caloric Restriction in Caenorhabditis elegans

Although the starvation response of the model multicellular organism Caenorhabditis elegans is a subject of much research, there is no convenient phenotypic readout of caloric restriction that can be applicable to large numbers of worms. This paper describes the distribution of mass densities of populations of C. elegans, from larval stages up to day one of adulthood, using isopycnic centrifugation, and finds that density is a convenient, if complex, phenotypic readout in C. elegans. The density of worms in synchronized populations of wildtype N2 C. elegans grown under standard solid-phase culture conditions was normally distributed, with distributions peaked sharply at a mean of 1.091 g/cm3 for L1, L2 and L3 larvae, 1.087 g/cm3 for L4 larvae, 1.081 g/cm3 for newly molted adults, and 1.074 g/cm3 at 24 hours of adulthood. The density of adult worms under starvation stress fell well outside this range, falling to a mean value of 1.054 g/cm3 after eight hours of starvation. This decrease in density correlated with the consumption of stored glycogen in the food-deprived worms. The density of the worms increased when deprived of food for longer durations, corresponding to a shift in the response of the worms: worms sacrifice their bodies by retaining larvae, which consume the adults from within. Density-based screens with the drug Ivermectin on worms cultured on single plates resulted in a clear bimodal (double-peaked) distribution of densities corresponding to drug exposed and non-exposed worms. Thus, measurements of changes in density could be used to conduct screens on the effects of drugs on several populations of worms cultured on single plates.

Direct conversion of Spirulina to ethanol without pretreatment or enzymatic hydrolysis processes

Oxygenic photosynthetic microorganisms such as cyanobacteria and microalgae have attracted attention as feedstocks for next-generation biofuels. To date, however, there have been no reports on efficient bioethanol production from cyanobacterial glycogen by yeast fermentation. Additionally, multiple pretreatment and enzymatic hydrolysis steps of polysaccharides are required for conventional ethanol production from agricultural crops and microalgae. Here, we investigate direct ethanol production from Arthrospira (Spirulina) platensis, a fast-growing halophilic cyanobacterium that accumulates large amounts of glycogen, using lysozyme and a recombinant amylase-expressing yeast strain to eliminate the need for biomass pretreatment and amylase hydrolysis. In the direct conversion process from A. platensis to ethanol, 6.5 g L−1 (ethanol productivity of 1.08 g per L per day) of ethanol was produced. The total ethanol yield based on glycogen consumption was 86% of theoretical yield, which to our knowledge, is the highest yield of bioethanol from an oxygenic photosynthetic microorganism. The present findings indicate that A. platensis is a remarkable carbohydrate feedstock in the form of glycogen, which is a promising material for the production of bioethanol and various other commercially valuable chemicals.

The Effects of Ara-C, Simvastatin and combo therapy on energy metabolism of HL-60 promyolocytic leukemia cell lines

1 Biochemistry, 4 Surgery, 5 Internal ABSTRACT Objective: Cancer cells choose their metabolic pathway depending on the oxygen content and substrate concentration of the medium. A wide spectrum of therapeutic agents regulating the energy metabolism of cancer cells are in still in use. Cytosine arabinoside (Ara-C) is a pyrimidine analogue used in the treatment of acute myeloid leukemia (AML) and simvastatin is an inhibitor of HMG-CoA (3-hydroxy-3-methyl-glutaryl-CoA) reductase, which regulates cholesterol biosynthesis. Thus, this study aimed to assess the energy metabolism of HL-60 promyelocytic leukemia cells and healthy white blood cells, additionally to determine the effects of simvastatin and Ara-C, alone or in combination on the energy metabolism of these cells. Materials and Methods: Healthy white blood cells, untreated and treated HL-60 promyo- locytic leukemia cell lines were incubated for 4 hours with radiolabelled glucose. Following incubation, lactate, which is one of the end products of the carbohydrate catabolism, and radiolabelled CO 2 produced by cells were collected and measured by the liquid scintillation device. In addition, glycogen consumption per hour was determined in each group. Results and Conclusion: We found that untreated HL-60 promyolocytic cells use anaero- bic glycolytic pathway whereas healthy white blood cells use aerobic glycolysis for energy gain. It was concluded that combined use of Ara-C and Simvastatin might lead to significant increase in the rate of aerobic glycolysis of HL-60 promyelocytic cells and the metabolism of these leukemia cells become more similar to the metabolism of healthy white blood cells which they originate from.

Akt Activation Protects Liver Cells from Apoptosis in Rats during Acute Cold Exposure

Accidental deaths due to exposure to extremely low natural temperature happen every winter. Exposure to extreme cold causes injury of multiple organs. However, early responses of the bodies to acute extreme cold exposure remain incompletely understood. In this study, we found that hepatic glycogen was rapidly reduced in rats exposed to -15°C, and the key enzymes required for glycogenesis were upregulated in the livers of the cold-exposed rats. In line with the rapid consumption of glycogen, acute cold exposure induced a transient elevation of cellular ATP level, which lasted about one hour. The ATP level went back to basal level after two hours of cold exposure. Four hours of cold exposure resulted in cellular ATP depletion and cell apoptosis. The dynamic change of cellular ATP levels was well associated with Akt activation in cold-exposed liver cells. The activation of Akt was required for cold exposure-induced ATP elevation. Blockade of Akt activation diminished the transient increase of intracellular ATP content and exacerbated cell apoptosis during acute cold exposure. These results suggest that Akt activation plays a pivotal role in maintaining cellular bioenergy balance and promoting liver cell survival during acute cold exposure.

Effects of celiac superior mesenteric ganglionectomy on glucose homeostasis and hormonal changes during oral glucose tolerance testing in rats

The liver plays an important role in maintaining glucose homeostasis in the body. In the prandial state, some of the glucose which is absorbed by the gastrointestinal tract is converted into glycogen and stored in the liver. In contrast, the liver produces glucose by glycogenolysis and gluconeogenesis while fasting. Thus, the liver contributes to maintaining blood glucose level within normoglycemic range. Glycogenesis and glycogenolysis are regulated by various mechanisms including hormones, the sympathetic and parasympathetic nervous systems and the hepatic glucose content. In this study, we examined a rat model in which the celiac superior mesenteric ganglion (CSMG) was resected. We attempted to elucidate how the celiac sympathetic nervous system is involved in regulating glucose homeostasis by assessing the effects of CSMG resection on glucose excursion during an oral glucose tolerance test, and by examining hepatic glycogen content and hepatic glycogen phosphorylase (GP) activity. On the oral glucose tolerance test, CSMG-resected rats demonstrated improved glucose tolerance and significantly increased GP activity compared with sham-operated rats, whereas there were no significant differences in insulin, glucagon or catecholamine levels between the 2 groups. These results suggest that the celiac sympathetic nervous system is involved in regulating the rate of glycogen consumption through GP activity. In conclusion, the examined rat model showed that the celiac sympathetic nervous system regulates hepatic glucose metabolism in conjunction with vagal nerve innervations and is a critical component in the maintenance of blood glucose homeostasis.

High phosphorus shock loading induced glycogen accumulating organisms in anaerobic/oxic sequencing batch reactor

This study investigated the long-term adaptability of an acetate-fed anaerobic–oxic sequencing batch reactor, operated at 15 days of solid retention time (SRT), to the influent P/COD loading raised from 15/400 to 40/400 mg/mg. Experiment results showed that after four SRTs operation, the anaerobic P release, aerobic P uptake and sludge P content decreased apparently, implying the metabolism of phosphate-accumulating organisms (PAOs) was inhibited. In contrast, anaerobic glycogen consumption and polyhydroxyalkanoates (PHAs) accumulation per acetate uptake increased observably, and 3HV/PHAs ratio elevated gradually to 21.8%. These results indicated that the proliferation of glycogen-accumulating organisms (GAOs) was encouraged. When influent phosphorus was reduced back to 15 mg/L, sludge P content diminished and a low P release/acetate uptake ratio was achieved, suggesting that PAOs’ metabolism was not restored. The above findings concluded that long-term operation of the high phosphorus influent inhibited the proliferation of PAOs, and more importantly provided GAOs a competitive advantage over PAOs.

Glycogen is the primary source of glucose during the lag phase of E. coli proliferation

In the studies of Escherichia coli (E. coli), metabolomics analyses have mainly been performed using steady state culture. However, to analyze the dynamic changes in cellular metabolism, we performed a profiling of concentration of metabolites by using batch culture. As a first step, we focused on glucose uptake and the behavior of the first metabolite, G6P (glucose-6-phosphate). A computational formula was derived to express the glucose uptake rate by a single cell from two kinds of experimental data, extracellular glucose concentration and cell growth, being simulated by Cell Illustrator. In addition, average concentration of G6P has been measured by CE-MS. The existence of another carbon source was suggested from the computational result. After careful comparison between cell growth, G6P concentration, and the computationally obtained curve of glucose uptake rate, we predicted the consumption of glycogen in lag phase and its accumulation as an energy source in an E. coli cell for the next proliferation. We confirmed our prediction experimentally. This behavior indicates the importance of glycogen participation in the lag phase for the growth of E. coli. This article is part of a Special Issue entitled: Computational Methods for Protein Interaction and Structural Prediction.

Metastasis-associated in colon cancer-1 is associated with poor prognosis in hepatocellular carcinoma, partly by promoting proliferation through enhanced glucose metabolism

Metastasis-associated in colon cancer-1 (MACC1) is a newly identified gene that is involved in the development and progression of hepatocellular carcinoma (HCC), however its investigation has not been comprehensive. In the present study, in vitro techniques, including immunohistochemistry, western blotting, reverse transcription quantitative polymerase chain reaction, metabolic assay, MTT assay, colony formation assay and prognostic analysis were used to confirm the involvement of MACC1 in HCC. Histological examination confirmed that the protein expression of MACC1 was upregulated in HCC and was associated with the hexokinase-2 (HK2) protein, which also indicates a poor prognosis. Knockdown of MACC1 induced the reduction of glycogen consumption and lactate production, which then lead to a marked reduction of proliferation in the MHCC-97H cells. However, the overexpression of MACC1 produced the opposite results in the HepG2 cells. These results suggested that MACC1 leads to a poor prognosis in HCC, partly by promoting proliferation via enhancement in glucose metabolism by HK2. Therefore, this pathway has the potential to become an important therapeutic target in HCC.

Hydroxypropylated distarch phosphate versus unmodified tapioca starch: fat oxidation and endurance in C57BL/6J mice

An RS4-type resistant starch is a chemically modified starch that shows reduced availability in comparison to the corresponding unmodified starch. Hydroxypropylated distarch phosphate (HDP) is an RS4-type resistant starch that increases energy expenditure and prevents high-fat diet-induced obesity through increased hepatic fatty acid oxidation. The aim of this study was to clarify the acute effects of HDP from tapioca starch (HPdTSP) on physical performance in mice. Male C57BL/6J mice were used to examine the effects of a single administration of 2 mg/g body weight HPdTSP or unmodified tapioca starch (TS) on postprandial responses in serum metabolic parameters, running endurance capacity on a treadmill, whole-body energy metabolism during exercise, activity of enzymes involved in fatty acid oxidation, liver and gastrocnemius muscle glycogen content, and serum glucose, insulin, non-esterified fatty acid, lactate, and triglyceride levels after exercise. Running time to fatigue was significantly greater in HPdTSP mice than in TS mice. Furthermore, HPdTSP maintained higher fat oxidation and this was associated with a greater activity of enzymes in fatty acid oxidation in the muscle during exercise. The blood lactate and serum insulin levels after exercise was significantly lower in HPdTSP mice than in TS mice. Liver glycogen was significantly higher in HPdTSP mice than in TS mice. These results suggest that acute oral administration of the RS4-type resistant starch, HPdTSP, maintained higher fat oxidation and reduced liver glycogen consumption during exercise and increased running endurance capacity in mice.

Genomic and Physiological Analysis of Carbon Storage in the Verrucomicrobial Methanotroph “Ca. Methylacidiphilum Fumariolicum” SolV

“Candidatus Methylacidiphilum fumariolicum” SolV is a verrucomicrobial methanotroph that can grow in extremely acidic environments at high temperature. Strain SolV fixes carbon dioxide (CO2) via the Calvin–Benson–Bassham cycle with methane as energy source, a trait so far very unusual in methanotrophs. In this study, the ability of “Ca. M. fumariolicum” to store carbon was explored by genome analysis, physiological studies, and electron microscopy. When cell cultures were depleted for nitrogen, glycogen storage was clearly observed in cytoplasmic storage vesicles by electron microscopy. After cessation of growth, the dry weight kept increasing and the bacteria were filled up almost entirely by glycogen. This was confirmed by biochemical analysis, which showed that glycogen accumulated to 36% of the total dry weight of the cells. When methane was removed from the culture, this glycogen was consumed within 47 days. During the period of glycogen consumption, the bacteria kept their viability high when compared to bacteria without glycogen (from cultures growing exponentially). The latter bacteria lost viability already after a few days when starved for methane. Analysis of the draft genome of “Ca. M. fumariolicum” SolV demonstrated that all known genes for glycogen storage and degradation were present and also transcribed. Phylogenetic analysis of these genes showed that they form a separate cluster with “Ca. M. infernorum” V4, and the most closely related other sequences only have an identity of 40%. This study presents the first physiological evidence of glycogen storage in the phylum Verrucomicrobia and indicates that carbon storage is important for survival at times of methane starvation.

Effects Of Thalidomide And rhGM-CSF on Carbohydrate Metabolism In HL-60 Acute Promyelocytic Leukemia Cell Line

The purpose of this study is to investigate the in vitro effect of thalidomide and rhGM-CSF on energy metabolism rate in HL-60 cell lines. HL-60 cells were incubated with radiolabelled glucose for 4 hours. Following incubation, radiolabelled CO2 was isolated and collected in specially designed scintillation vials. The other end-products of carbohydrate catabolism collected via anion-exchange chromatography were analyzed using liquid scintillation. Protein and glycogen levels for each group were also determined. The study results indicated that the use of drugs for 2-days had no significant effect when compared with aerobic and anaerobic controls. However, groups treated with the drugs for 5 days showed significant differences. It was obvious that rhGM-CSF caused HL-60 leukemia cells to use aerobic glycolytic pathway for production of energy. When thalidomide and rhGM-CSF combination therapy results were compared with untreated control cells, it was observed that glycogen consumption was decreased by 50.37%, and CO2 production was increased by 94.03%. When compared with those of anaerobic controls, glycogen consumption and CO2 production rates were found to be decreased by 54.01% and increased by 96.59%, respectively. As a consequence, we found that those cell lines treated with combined use of Thalidomide and rhGM-CSF caused HL-60 leukemia cells to predominantly prefer aerobic glycolytic pathways for energy production.