New insights into cytosolic glucose levels during differentiation of 3T3-L1 fibroblasts into adipocytes

Cytosolic glucose concentration reflects the balance between glucose entry across the plasma membrane and cytosolic glucose utilization. In adipocytes, glucose utilization is considered very rapid, meaning that every glucose molecule entering the cytoplasm is quickly phosphorylated. Thus, the cytosolic free glucose concentration is considered to be negligible; however, it was never measured directly. In the present study, we monitored cytosolic glucose dynamics in 3T3-L1 fibroblasts and adipocytes by expressing a fluorescence resonance energy transfer (FRET)-based glucose nanosensor: fluorescent indicator protein FLIPglu-600μ. Specifically, we monitored cytosolic glucose responses by varying transmembrane glucose concentration gradient. The changes in cytosolic glucose concentration were detected in only 56% of 3T3-L1 fibroblasts and in 14% of 3T3-L1 adipocytes. In adipocytes, the resting cytosolic glucose concentration was reduced in comparison with the one recorded in fibroblasts. Membrane permeabilization increased cytosolic glucose concentration in adipocytes, and glycolytic inhibitor iodoacetate failed to increase cytosolic glucose concentration, indicating low adipocyte permeability for glucose at rest. We also examined the effects of insulin and adrenaline. Insulin significantly increased cytosolic glucose concentration in adipocytes by a factor of 3.6; however, we recorded no effect on delta ratio (ΔR) in fibroblasts. Adrenaline increased cytosolic glucose concentration in fibroblasts but not in adipocytes. However, in adipocytes in insulin-stimulated conditions, glucose clearance was significantly faster following adrenaline addition in comparison with controls (p < 0.001). Together, these results demonstrate that during differentiation, adipocytes develop more efficient mechanisms for maintaining low cytosolic glucose concentration, predominantly with reduced membrane permeability for glucose.     

在线推定和控制葡萄糖浓度改善谷氨酸发酵性能

谷氨酸发酵过程一般需要定时、人工分批式地添加葡萄糖。该流加操作方式会引起发酵罐内葡萄糖浓度的剧烈波动, 不利于高效、稳定的谷氨酸生产。谷氨酸发酵具有显著的非增殖耦联特征, 产酸期葡萄糖耗量与氨水耗量存在非常明显的关联性。通过在线计量氨水耗量推定糖耗以及葡萄糖浓度, 可比较准确地将谷氨酸发酵产酸期的糖浓度控制在预先设定的水平。当糖浓度控制在5 g/L~10 g/L的低水平时, 最终谷氨酸浓度可以达到80 g/L的较高水平, 高糖浓度下的渗透压效应有望得到缓解, 有利于发酵生产的稳定。     

Cerebrospinal Fluid Glucose: Turnover and Metabolism

The turnover of cerebrospinal fluid (CSF) glucose was studied in cats during steady-state perfusion. In all experiments, the perfusion fluid contained either tracer [¹⁴C]glucose alone or tracer glucose along with 4.45 mM unlabeled glucose. In some studies, serum glucose was lowered with insulin. The concentration of glucose and [¹⁴C]glucose in the effluent fluid was measured, and the distribution of ¹⁴C between glucose and lactate was determined by chromatography. From these values, the extraction of glucose and the metabolism of glucose to lactate were calculated. From the decrease in the specific activity of glucose in the perfusion fluid, the influx of glucose from serum was also determined. During steadystate perfusion, 71% of the radioactivity was recovered in the effluent fluid: 50% in the form of glucose, 6% in the form of lactate, and 15% in forms that were not identified. Thus, 50% of the perfusion fluid glucose was cleared, of which 29% was extracted and 21% metabolized. The influx of glucose was proportional to the serum glucose when the latter was about 2.5 mM or 10.0 mM. During perfusion with tracer glucose only, the concentration of glucose in the effluent fluid was 25% that of serum. The transport of glucose from serum was independent of the glucose concentration gradient between serum and perfusion fluid. However, when perfusion fluid glucose concentration was greater than that of serum, transport was inhibited. These studies suggest that in maintaining CSF glucose at a lower concentration than serum glucose, with equal amounts of glucose entering and leaving the CSF, 50% of CSF glucose concentration cleared is replaced by 25% of serum glucose concentration.     

溶液中木质素和葡萄糖的超滤分离

以木质素和葡萄糖的混合溶液为木质纤维素水解液模型,采用截留相对分子质量为5 000的卷式聚醚砜膜对葡萄糖和木质素进行全回流模式的分离,探讨了木质素和葡萄糖浓度、操作压力、错流速率对通量、木质素和葡萄糖截留率的影响。结果表明:在实验条件范围内,通量随葡萄糖浓度和木质素浓度的增加而降低,并随操作压力、错流速率的增加而增加。木质素截留率不受任何条件的影响,基本稳定在97%。葡萄糖截留率随木质素浓度的增加而增加,并随错流速率的增加而减小。在0.8 g/L的木质素质量浓度条件下,当错流速率从0.12 m/s增加到0.17 m/s时,葡萄糖截留率从14%减小到7.3%。由此可见,在混合溶液的超滤过程中,通过合理选择错流速率,能够改善木质素和葡萄糖的分离。     

Long-term effects of glucose on insulin release and glucose oxidation by mouse pancreatic islets maintained in tissue culture

Rates of glucose oxidation and insulin release in response to a wide range of glucose concentrations were studied in short-term experiments in isolated mouse pancreatic islets maintained in tissue culture for 6 days at either a physiological glucose concentration (6.7mm) or at a high glucose concentration (28mm). The curves relating glucose oxidation or insulin release to the extracellular glucose concentration obtained with islets cultured in 6.7mm-glucose displayed a sigmoid shape similar to that observed for freshly isolated non-cultured islets. By contrast islets that had been cultured in 28mm-glucose showed a linear relationship between the rate of glucose oxidation and the extracellular glucose concentration up to about 8mm-glucose. The maximal oxidative rate was twice that of the non-cultured islets and the glucose concentration associated with the half-maximal rate considerably decreased. In islets cultured at 28mm-glucose there was only a small increase in the insulin release in response to glucose, probably due to a depletion of stored insulin in those B cells that had been cultured in a high-glucose medium. It is concluded that exposure of B cells for 6 days to a glucose concentration comparable with that found in diabetic individuals causes adaptive metabolic alterations rather than degeneration of these cells.     

Glycaemic threshold for changes in electroencephalograms during hypoglycaemia in patients with insulin dependent diabetes

The relation between blood glucose concentration, the symptoms and signs of hypoglycaemia, and electroencephalographic changes in diabetic patients is not known. The effect of hypoglycaemia on brain function was studied in 13 patients with insulin dependent diabetes. During a gradual fall in blood glucose concentration induced by a bolus injection of insulin followed by an intravenous infusion of insulin, during 60 minutes of biochemical hypoglycaemia, and after restoration of normoglycaemia with intravenous glucose electroencephalograms were evaluated continuously by period-amplitude analysis; blood samples were taken every 10 minutes throughout. No changes were seen in electroencephalograms when the blood glucose concentration was above 3 mmol/l. At a median blood glucose concentration of 2·0 (95% confidence interval 1·7 to 2·3) mmol/l alpha activity decreased abruptly in the electroencephalograms concomitant with an increase in theta activity, reflecting neuronal dysfunction in the cortex. When the blood glucose concentration was further lowered changes were observed in the electroencephalograms indicating that deeper brain structures were affected. A normal electroencephalogram was re-established at a blood glucose concentration of 2·0 (1·8 to 2·1) mmol/l. There was no significant correlation between the blood glucose concentration at the onset of changes in the electroencephalograms and age, duration of diabetes, insulin dose, haemoglobin A_1c concentration, initial blood glucose concentration, rate of fall in blood glucose concentration, and appearance of symptoms and signs of hypoglycaemia.Changes in electroencephalograms during hypoglycaemia appear and disappear at such a narrow range of blood glucose concentrations that the term threshold blood glucose concentration for the onset of such changes seems justified.     

Effect of glucose concentrations on xanthan gum production by xanthomonas campestris

The effect of the glucose concentration on xantham gum production by Xanthomonas campestris ATCC 13951 was studied resulting that the glucose concentration between 30 and 40 g/kg broth was best for xanthan gum production. Controlling the glucose concentration at between 30 and 40 g/kg broth by intermittent addition of glucose prevented the inhibition of cell growth and the cessation of xanthan gum production, which were observation with a higher glucose concentration. By means of a glucose feeding strategy, the xanthan gum concentration reached 43 g/kg broth after 96-h cultivation.     

Kinetic characteristics of chloroplast glucose transport

Influx of labelled D-glucose into isolated spinach (Spinacia oleracea L. cv. Melody hybrid) chloroplasts was initially rapid followed by a period of slower influx. The stroma glucose concentration attained equilibrium rapidly with low external glucose concentrations and the two were linearly proportional. The period of slower influx resulted from conversion of glucose to acidic products that remained trapped in the chloroplast. As the external glucose concentration increased, the stroma glucose concentration increased less and less, attaining a maximal concentration of 72 mol m(-3). The maintenance of an equilibrium stroma glucose concentration lower than that in the external medium is evidence that plastid glucose efflux involves secondary active transport. The equilibrium stroma glucose concentration increased in response to light and protonophoric uncouplers. It is proposed that glucose efflux is coupled with a proton and the stroma glucose concentration equilibrates in response to the proton gradient across the membrane. To determine if glucose is a significant product of starch mobilization, chloroplasts were isolated from spinach leaves labelled with 14CO2 during the preceding light period. Chloroplasts degraded starch at the same rate as the intact leaf. Glucose, maltose, and isomaltose were the principal labelled products that appeared in the medium during starch mobilization. The glucose concentration in the chloroplast was 2 mol m(-3), which is similar to the measured Km for zero trans efflux. The data support the role of the glucose translocator as an important component in the pathway for sucrose synthesis at night.     

面向血液葡萄糖测量的生物组织阻抗模型

本文推导了细胞膜电容与血糖浓度之间的关系,并将这种关系应用到Pauly和Schwan提出的弛豫模型中,再将弛豫模型应用于多重Cole-Cole介电模型,最后通过介电模型得出阻抗模型,建立起人体血糖浓度与组织介电谱,阻抗谱之间的联系。通过分析发现,细胞膜电容随着血糖浓度增加而增大,且当血糖浓度趋于无穷大时,细胞膜电容等于血糖浓度为0时细胞膜电容的1.53倍。对介电谱的仿真得出,当外加信号频率小于1 MHz时,介电常数实部随着葡萄糖浓度改变而发生明显变化。阻抗谱的研究结果表明,随着葡萄糖浓度的升高,阻抗的实部减小,阻抗虚部模增大,且分别在频率为1.48 MHz和0.55 MHz时变化得最快。本文建立的模型为血糖无创检测提供了一种新的方法和手段。     

Kinetics of rat liver glucokinase. Co-operative interactions with glucose at physiologically significant concentrations.

The kinetics of glucokinase from rat liver were studied over wide ranges of glucose and MgATP2- concentrations. The initial rate shows a co-operative dependence on the glucose concentration, with Hill coefficients in the range 1.2-1.5. The degree of glucose co-operativity increases with the MgATP2- concentration, but no co-operativity was detected for the dependence of the rate on the MgATP2- concentration. The effects observed occur at physiologically reasonable concentrations of glucose and MgATP2- and are consistent with the presumed function of glucokinase in maintaining a constant concentration of glucose in the blood.     

体外循环超滤法影响小儿心内直视术患者血糖和血乳酸水平的研究

目的本研究旨在探讨体外循环超滤法对小儿心内直视术患者血糖和血乳酸的影响及临床意义.方法62例小儿先心病患者(男39例,女23例,体重9.5～20kg,年龄平均2.5岁)随机分成AB两组,A组用5%葡萄糖预充,B组无糖预充,各组再分成超滤组和对照组,两组病种基本相同.超滤组和对照组在复温前、停机时,分别采血和超滤液测定血糖和乳酸,进行对比.结果A组和B组超滤后血清中血糖相比(P＜0.01),两组统计学差异有显著性.超滤组与对照组血糖和乳酸浓度,无明显统计学差异.而超滤液中含有较高浓度糖和乳酸.结论无糖预充可以降低血糖和乳酸,用超滤法可直接排除心内直视术小儿患者高血糖和高乳酸,减少神经系统的受损,值得临床推广应用.     

谷氨酸发酵过程葡萄糖自动流加系统

补料（流加葡萄糖）操作是谷氨酸发酵过程最重要的操作之一,工业上有各种各样的补料操作方法。控制葡萄糖浓度于一个较为平稳且适中的水平有利于提高谷氨酸发酵的性能指标。通过在线计量谷氨酸发酵中的氨水耗量并据此在线推定发酵液中的葡萄糖浓度,构建了一个谷氨酸发酵自动在线补料系统。使用该控制系统,谷氨酸发酵过程的葡萄糖浓度可以控制在任意水平,平稳、无波动的谷氨酸发酵可以得到实现。     

The rate-limiting step for glucose transport into the hypothalamus is across the blood–hypothalamus interface

Specialized glucosensing neurons are present in the hypothalamus, some of which neighbor the median eminence, where the blood–brain barrier has been reported leaky. A leaky blood–brain barrier implies high tissue glucose levels and obviates a role for endothelial glucose transporters in the control of hypothalamic glucose concentration, important in understanding the mechanisms of glucose sensing We therefore addressed the question of blood–brain barrier integrity at the hypothalamus for glucose transport by examining the brain tissue-to-plasma glucose ratio in the hypothalamus relative to other brain regions. We also examined glycogenolysis in hypothalamus because its occurrence is unlikely in the potential absence of a hypothalamus–blood interface. Across all regions the concentration of glucose was comparable at a given plasma glucose concentration and was a near linear function of plasma glucose. At steady-state, hypothalamic glucose concentration was similar to the extracellular hypothalamic glucose concentration reported by others. Hypothalamic glycogen fell at a rate of ∼1.5 μmol/g/h and remained present in substantial amounts. We conclude for the hypothalamus, a putative primary site of brain glucose sensing that: the rate-limiting step for glucose transport into brain cells is at the blood–hypothalamus interface, and that glycogenolysis is consistent with a substantial blood -to- intracellular glucose concentration gradient.     

The effects of glucose and oxygen on the cytochromes and metabolic activity of yeast batch cultures. II. Candida utilis

Candida utilis was grown in batch culture with and without oxygen control. The concentrations of A-, B-, and C-type cytochromes were found to vary with the initial glucose concentration, with the dissolved oxygen concentration, and with time. A-type was the most sensitive. After glucose was essentially exhausted, the yeast catabolized ethanol, if it had been growing in a relatively low initial glucose concentration, or non-glucose carbohydrate, including some of that previously accumulated within the cell, if it had been growing in a high initial glucose concentration. This difference in metabolic pattern could explain why cytochrome derepression was initiated soon after glucose uptake ceased only if initial glucose had been relatively low. The effects of glucose and dissolved oxygen concentrations on yeast cytochromes and respiratory activity are discussed.     

Extracellular Glucose Concentrations in the Rat Hippocampus Measured by Zero-Net-Flux

Abstract : The concentration of glucose in the brain's extracellular fluid remains controversial, with recent estimates and measurements ranging from 0.35 to 3.3 m M . In the present experiments, we used the method of zero-net-flux microdialysis to determine glucose concentration in the hippocampal extracellular fluid of awake, freely moving rats. In addition, the point of zero-net-flux was measured across variations in flow rate to confirm that the results for glucose measurement were robust to such variations. In 3-month-old male Sprague-Dawley rats, the concentration of glucose in the hippocampal extracellular fluid was found to be 1.00 ± 0.05 m M , which did not vary with changes in flow rate. Three-month-old and 24-month-old Fischer-344 rats both showed a significantly higher hippocampal extracellular fluid glucose concentration, at 1.24 ± 0.07 and 1.21 ± 0.04 m M , respectively ; there was no significant difference between the two age groups. The present data demonstrate variation in extracellular brain glucose concentration between rat strains. When taken together with previous data showing a striatal extracellular glucose concentration on the order of 0.5 m M , the data also demonstrate variation in extracellular glucose between brain regions. Traditional models of brain glucose transport and distribution, in which extracellular concentration is assumed to be constant, may require revision.     

The correlation between glycogen–glycolysis metabolite pyruvate and vancomycin antibiotic productions of Amycolatopsis orientalis grown in glucose medium

The effect of glucose concentration in the growth medium on the relationship between glycolysis, glycogen accumulation and vancomycin production of Amycolatopsis orientalis was investigated depending on the incubation time. After a lag phase, bacterial growth of A. orientalis began and biomass concentration increased continuously up to 36th or 48th hours while glucose concentration in the culture medium was consumed rapidly in the same time of incubation. In addition, increase in glucose concentrations of the growth medium lead to increase intracellular glucose as well as glycerol levels. Intracellular pyruvate levels increased significantly up to 15 g/L while extracellular pyruvate levels with respect to increases in glucose concentration. A positive correlation between glucose kinase activities and glucose concentration was determined during the incubation period. Pyruvate kinase activity increased up to 15 g/L glucose and 48th hour of incubation. As a glycopeptide antibiotic, vancomycin production increased with the increases in glucose concentrations up to 15 g/L. These results indicated that glycogen accumulation with respect to glucose concentration of the growth medium was concomitant with the sporulation of A. orientalis. When the initial glucose concentration exceeded 15 g/L, pyruvate excretions as well as intracellular glycogen and glycerol productions were supported in spite of repression in vancomycin production of A. orientalis.     

血糖浓度检测技术的最新进展

本文阐述了血糖浓度检测技术的最新进展.从直接接触式的血糖浓度临床检测方法,以及市面上的各种微创式可随身携带的血糖浓度检测仪的测量技术,到近期正在研究的无创伤血糖浓度检测技术的描述,着重论述了目前国内外正在研究尚没应用于临床的无创伤血糖浓度检测的各种技术原理和方案,并对各种无创伤血糖浓度测量技术的工作原理和存在的临床应用问题进行分析,同时展望了血糖检测技术未来的发展之路.     

Hypoglycemia activates compensatory mechanism of glucose metabolism of brain

The effect of plasma glucose concentration on the cerebral uptake of [18F]-fluorodeoxy-D-glucose (FDG) was studied in a broad concentration range in a rabbit brain model using dynamic FDG PET measurements. Hypoglycemic and hyperglycemic conditions were maintained by manipulating plasma glucose applying i.v. glucose or insulin load. FDG utilization (K) and cerebral glucose metabolic rate (CGMR) were evaluated in a plasma glucose concentration range between 0.5 mM and 26 mM from the kinetic constant k1, k2, k3 obtained by the Sokoloff model of FDG accumulation. A decreasing set of standard FDG uptake values found with increasing blood glucose concentration was explained by competition between the plasma glucose and the radiopharmacon FDG. A similar trend was observed for the forward kinetic constants k1, and k3 in the entire concentration range studied. The same decreasing tendency of k2 was of a smaller magnitude and was reverted at the lowest glucose concentrations where a pronounced decrease of this backward transport rate constant was detected. Our kinetic data indicate a modulation of the kinetics of carbohydrate metabolism by the blood glucose concentration and report on a special mechanism compensating for the low glucose supply under conditions of extremely low blood glucose level.     

Regulation of glycogenolysis in the liver of the newborn rat in vivo inhibitory effect of glucose

Newborn rats were injected immediately after delivery with glucose or glucose plus mannoheptulose, and the time-courses of liver glycogen, plasma glucose, insulin and glucagon concentration were studied. The administration of glucose prevented both liver glycogenolysis and the increase in plasma glucagon concentration which normally occurs immediately after delivery. In addition, the administration of glucose prevented the decrease of plasma glucose and insulin concentration which normally occurs during the first hour of extrauterine life. Supplementation of glucose with mannoheptulose prevented the increase of plasma insulin concentrations caused by the administration of glucose; liver glycogenolysis, however, was not stimulated in these circumstances. The increase in the rate of glycogenolysis caused by the administration of glucagon was prevented in newborn rats previously treated with glucose. These results suggest that glucose exerts an inhibitory effect on the stimulation of neonatal liver glycogenolysis by glucagon.     

Glucose metabolism in mouse pancreatic islets

1. Rates of glucose oxidation, lactate output and the intracellular concentration of glucose 6-phosphate were measured in mouse pancreatic islets incubated in vitro. 2. Glucose oxidation rate, measured as the formation of (14)CO(2) from [U-(14)C]glucose, was markedly dependent on extracellular glucose concentration. It was especially sensitive to glucose concentrations between 1 and 2mg/ml. Glucose oxidation was inhibited by mannoheptulose and glucosamine but not by phlorrhizin, 2-deoxyglucose or N-acetylglucosamine. Glucose oxidation was slightly stimulated by tolbutamide but was not significantly affected by adrenaline, diazoxide or absence of Ca(2+) (all of which may inhibit glucose-stimulated insulin release), by arginine or glucagon (which may stimulate insulin release) or by cycloheximide (which may inhibit insulin synthesis). 3. Rates of lactate formation were dependent on the extracellular glucose concentration and were decreased by glucosamine though not by mannoheptulose; tolbutamide increased the rate of lactate output. 4. Islet glucose 6-phosphate concentration was also markedly dependent on extracellular glucose concentration and was diminished by mannoheptulose or glucosamine; tolbutamide and glucagon were without significant effect. Mannose increased islet fructose 6-phosphate concentration but had little effect on islet glucose 6-phosphate concentration. Fructose increased islet glucose 6-phosphate concentration but to a much smaller extent than did glucose. 5. [1-(14)C]Mannose and [U-(14)C]fructose were also oxidized by islets but less rapidly than glucose. Conversion of [1-(14)C]mannose into [1-(14)C]glucose 6-phosphate or [1-(14)C]glucose could not be detected. It is concluded that metabolism of mannose is associated with poor equilibration between fructose 6-phosphate and glucose 6-phosphate. 6. These results are consistent with the idea that glucose utilization in mouse islets may be limited by the rate of glucose phosphorylation, that mannoheptulose and glucosamine may inhibit glucose phosphorylation and that effects of glucose on insulin release may be mediated through metabolism of the sugar.