Glucose Respiration in Zygorhyncus moelleri; the Entry of Glucose into the Cells

When glucose was added to carbohydrate-starved cells of Zygorhyncus,moelleri the rate of oxygen uptake did not immediately riseto a constant value, but there was a lag period of 2 or 3 hoursbefore it reached its maximum level. The length of this lagperiod increased from a few minutes for short periods of starvationto 2–3 hours after 12 hours in a carbohydrate-free medium.Factors believed to affect cellular permeability (a cationicdetergent, adjustments of the pH, and of the potassium/calciumratio) reduced the length of the lag period by not more than40 per cent. of the original value without affecting the finalrate of oxygen uptake. Investigation of the entry of glucoseinto the cells showed that the rate of oxygen uptake was notlimited by the concentration of intra-cellular glucose for morethat about 11 per cent. of the lag period in starved cells.The reasons for this difference in the percentage of the lagperiod apparently due to a permeability barrier are tentativelydiscussed in connexion with the route by which glucose entersthe cells.     

Clinical Use of the Glucose Disappearance Rate

An intravenous glucose tolerance test, coupled with a graphical or mathematical analysis, has been used in normal, diabetic, and glycosuric subjects, and patients suspected of having hypoglycemia. From the analysis, a single value, K, is derived which integrates the disappearance of the injected glucose. A significant difference was found between normals and known diabetics: this difference becomes even more distinct when the disappearance of the glucose load is measured as K1 or Increment Index, as opposed to decrease of the total blood glucose, K2 or Total Index. A 300-g. carbohydrate diet was shown to be important to the success of this test. More than 400 patients have been tested in the investigation of different disorders of glucose metabolism. Renal glycosuria was diagnosed in 35 patients. Rapid disappearance of the glucose load has been observed in 18 of 40 patients suspected of having hypoglycemia. Of 64 patients diagnosed in the hospital as previously unknown diabetics, 59 presented K1 values below the normal range.     

The Proteomic Response of Bacillus pumilus Cells to Glucose Starvation

Since starvation for carbon sources is a common condition for bacteria in nature and it can also occur in industrial fermentation processes due to mixing zones, knowledge about the response of cells to carbon starvation is beneficial. The preferred carbon source for bacilli is glucose. The response of Bacillus pumilus cells to glucose starvation using metabolic labeling and quantitative proteomics was analyzed. Glucose starvation led to an extensive reprogramming of the protein expression pattern in B. pumilus. The amounts of proteins of the central carbon metabolic pathways (glycolysis and TCC) remained stable in starving cells. Proteins for gluconeogenesis were found in higher amounts during starvation. Furthermore, many proteins involved in acquisition and usage of alternative carbon sources were present in elevated amounts in starving cells. Enzymes for fatty acid degradation and proteases and peptidases were also found in higher abundance when cells entered stationary phase. Among the proteins found in lower amounts were many enzymes involved in amino acid and nucleotide synthesis and several NRPS and PKS proteins.     

Glucose Oxidation during Shoot Initiation in Tobacco Callus Cultures

Involvement of the Embden-Meyerhof Parnas and the pentose phosphatepathways in glucose oxidation in glucose oxidation in tobaccocallus was examined. Marked changes in the activities of glucokinase,aldolase, glucose-6-phosphate dehydrogenase, and phosphogluconatedehydrogenase were observed during culture of tobacco callusunder shoot-forming and non-shoot-forming conditions. Activitiesof these enzymes were higher in shoot-forming tissue than innon-shoot-forming tissue. Furthermore, the activities of thepentose phosphate pathway enzymes showed greater differencesthan those of the Embden-Meyerhof-Parnas pathway. Confirmationof these findings was obtained by investigating the contributionsof ¹⁴C from [14C-1]- and [¹⁴C–6]-glucose to CO2 released.The significance of these findings on glucose oxidation in relationto the shoot-initiation process are discussed.     

Metabolic Studies on Intermediates in the myo-Inositol Oxidation Pathway in Lilium longiflorum Pollen: III. Polysaccharidic Origin of Labeled Glucose

myo-Inositol-linked glucogenesis in germinated lily (Lilium longiflorum Thunb., cv. Ace) pollen was investigated by studying the effects of added l-arabinose or d-xylose on metabolism of myo-[2-³H]inositol and by determining the distribution of radioisotope in pentosyl and hexosyl residues of polysaccharides from pollen labeled with myo-[2-¹⁴C]inositol, myo-[2-³H]inositol, l-[5-¹⁴C]arabinose, and d-[5R,5S-³H]xylose.     

Effect of Glucose Starvation on Glucose Transport in Neuronal Cells in Primary Culture from Rat Brain

The regulation of glucose transport into cultured brain cells during glucose starvation was studied. On glucose deprivation for 40 h, 2-deoxy-D-glucose (2-DG) uptake was stimulated twofold in neuronal cells but was not changed significantly in astrocytes. On refeeding, the increased activity of neuronal cells rapidly returned to the basal level, an observation indicating that the effect of glucose starvation was reversible. The increase was due solely to change in the Vmax, a finding suggesting that the number of glucose transporters on the plasma membrane is increased in starved cells. Cycloheximide inhibited this increase. In the presence of cycloheximide, the activity of 2-DG uptake of starved cells remained constant for 12 h and then slowly decreased, whereas that of fed cells decreased rapidly. These findings suggest that glucose starvation regulates glucose transport by changing the rate of net synthesis of the transporter in neuronal cells in culture.     

Increased Fatty Acid beta-Oxidation after Glucose Starvation in Maize Root Tips

The effects of glucose starvation on the oxidation of fatty acids were studied in excised maize (Zea mays L.) root tips. After 24 hours of glucose starvation, the rate of oxidation of palmitic acid to CO₂ by the root tips was increased 2.5-fold. Different enzyme activities were tested in a crude particulate fraction from nonstarved root tips and those starved for 24 hours. The activities of the β-oxidation enzymes crotonase, hydroxyacyl-coenzyme A (CoA) dehydrogenase, and thiolase and those of catalase, malate synthase, and peroxisomal citrate synthase were higher after starvation. However, no isocitrate lyase activity was detected, thus suggesting that the glyoxylate cycle does not operate. The overall β-oxidation activity was assayed as the formation of [¹⁴C]acetyl-CoA from [¹⁴C]palmitic acid after high-performance liquid chromatography analysis of the CoA derivatives. An activity was detected in sugar-fed root tips, and it was increased by two-to fivefold in starved roots. Because the recovery of enzyme activities is only marginally better in starved roots compared with nonstarved roots, these results indicate that the β-oxidation activity in the tissues is increased during sugar starvation. This increase is probably an essential part of the response to a situation in which lipids and proteins replace carbohydrates as the major respiratory substrates. These results are discussed in relation to the metabolic changes observed in senescing plant tissues.     

Biogenesis of Autophagosome in Trichomonas vaginalis during Macroautophagy Induced by Rapamycin‐treatment and Iron or Glucose Starvation Conditions

Autophagy is an adaptive response for cell survival in which cytoplasmic components and organelles are degraded in bulk under normal and stress conditions. Trichomonas vaginalis is a parasite highly adaptable to stress conditions such as iron (IR) and glucose restriction (GR). Autophagy can be traced by detecting a key autophagy protein (Atg8) anchored to the autophagosome membrane by a lipid moiety. Our goal was to perform a morphological and cellular study of autophagy in T. vaginalis under GR, IR, and Rapamycin (Rapa) treatment using TvAtg8 as a putative autophagy marker. We cloned tvatg8a and tvatg8b and expressed and purified rTvAtg8a and rTvAtg8b to produce specific polyclonal antibodies. Autophagy vesicles were detected by indirect immunofluorescence assays and confirmed by ultrastructural analysis. The biogenesis of autophagosomes was detected, showing intact cytosolic cargo. TvAtg8 was detected as puncta signal with the anti‐rTvAtg8b antibody that recognized soluble and lipid‐associated TvAtg8b by Western blot assays in lysates from stress‐inducing conditions. The TvAtg8b signal co‐localized with the CytoID and lysotracker labeling (autolysosomes) that accumulated after E‐64d treatment in GR parasites. Our data suggest that autophagy induced by starvation in T. vaginalis results in the formation of autophagosomes for which TvAtg8b could be a putative autophagy marker.     

Polysulfides as Intermediates in the Oxidation of Sulfide to Sulfate by Beggiatoa spp.

Zero-valent sulfur is a key intermediate in the microbial oxidation of sulfide to sulfate. Many sulfide-oxidizing bacteria produce and store large amounts of sulfur intra- or extracellularly. It is still not understood how the stored sulfur is metabolized, as the most stable form of S⁰ under standard biological conditions, orthorhombic α-sulfur, is most likely inaccessible to bacterial enzymes. Here we analyzed the speciation of sulfur in single cells of living sulfide-oxidizing bacteria via Raman spectroscopy. Our results showed that under various ecological and physiological conditions, all three investigated Beggiatoa strains stored sulfur as a combination of cyclooctasulfur (S₈) and inorganic polysulfides (S_n²⁻). Linear sulfur chains were detected during both the oxidation and reduction of stored sulfur, suggesting that S_n²⁻ species represent a universal pool of bioavailable sulfur. Formation of polysulfides due to the cleavage of sulfur rings could occur biologically by thiol-containing enzymes or chemically by the strong nucleophile HS⁻ as Beggiatoa migrates vertically between oxic and sulfidic zones in the environment. Most Beggiatoa spp. thus far studied can oxidize sulfur further to sulfate. Our results suggest that the ratio of produced sulfur and sulfate varies depending on the sulfide flux. Almost all of the sulfide was oxidized directly to sulfate under low-sulfide-flux conditions, whereas only 50% was oxidized to sulfate under high-sulfide-flux conditions leading to S⁰ deposition. With Raman spectroscopy we could show that sulfate accumulated in Beggiatoa filaments, reaching intracellular concentrations of 0.72 to 1.73 M.     

Oxidation of Two Pregnane Steroids Catalyzed by the Fungus Cephalosporium aphidicola

Two pregnane steroids, pregnane (1) and 3β-hydroxypregnane (2), were oxidized by fermentation with the fungus Cephalosporium aphidicola. The fermentation of pregnane (1) yielded 3β-hydroxypregnane (2) and 3β, 6β,11α-trihydroxypregnane (3), while that of 3β-hydroxypregnane (2) afforded 6β,11α-dihydroxypregn-3,20-dione (4), 3β,6β,15α-trihydroxypregn-20-one (5) and 3β,5α,11α-trihydroxypregn-20-one (6). The metabolites are characterized by detailed physical and spectroscopic studies.     

Effects of Starvation and Refeeding on Glucose Metabolism and Immune Responses in Macrobrachium rosenbergii

Marine Biotechnology - Starvation is a common challenge for aquatic animals in both natural and cultured environments. To investigate the effects of starvation and refeeding on glucose metabolism...     

Changes in Enzyme Activity of Fungi during Nitrogen Starvation

The intracellular proteinase enzymes of Penicillium griseofulvumand other fungi increase to levels several times higher thanthat in growing mycelium in the first few hours of nitrogenstarvation, and remain at the higher level unless more assimilablenitrogen is supplied. The rise in proteinase does not take placein absence of oxygen or in the presence of p-fluorophenylalanine,and is not accompanied by net protein breakdown in the myceliumunless synthetic processes are inhibited by lack of oxygen orexogenous carbohydrate. The proteinase activity changes during the life cycle of thefungus, being absent in the spores, very low in the early stagesof germination, and tending to rise steadily during growth.In some conditions the induction of sporulation is associatedwith a rise in proteinase, but there is no simple relation betweenthe two processes. The results are considered to support theview that intracellular proteinase plays a part in protein turnoverin the fungal cell.