Effect Of Different Glucose Concentrations Research Articles

Oral stimulation with glucose and fructose, but not sucrose, accelerates gastric emptying in humans.

Postprandial regulation of the gastric emptying (GE) rate plays an important role in food intake. Although oral sweetening with glucose may accelerate GE, the effects of different sweetness intensities of glucose (10% and 20%, w/v) and other energy sweeteners (e.g. fructose and sucrose) remain uncertain. The purpose of this study was to determine the effects of different glucose concentrations (Experiment 1) and different sugars with the same sweet taste intensity (Experiment 2) on postprandial GE. In both experiments, after ingesting a 200 kcal carbohydrate solution containing 50g of maltodextrin, participants repeatedly sipped, but did not swallow, one of three (water, 10% and 20%, w/v glucose) or four (water and equally sweet 20%, w/v glucose, 12%, w/v fructose, and 14%, w/v sucrose) solutions for 1min every 5min over a 30min period. GE was evaluated by measuring the temporal change in the cross-sectional area of the gastric antrum using ultrasound. In Experiment 1, oral stimulation with 20% (w/v) glucose resulted in greater GE than the control stimulus (i.e. water), but the effect of stimulation with 10% (w/v) glucose on GE was not different from that of the control stimulus. In Experiment 2, stimulation with 20% (w/v) glucose or 12% (w/v) fructose resulted in greater GE than the control stimulus. However, the effect of stimulation with 14% (w/v) sucrose on GE did not differ from that of the control stimulus. Consequently, oral stimulation with glucose or fructose solutions of moderate to high sweetness following a meal facilitates postprandial GE.

Understanding the Effect of Different Glucose Concentrations in the Oligotrophic Bacterium Bacillus subtilis BS-G1 through Transcriptomics Analysis.

Glucose is an important carbon source for microbial growth, and its content in infertile soils is essential for the growth of bacteria. Since the mechanism of oligotrophic bacterium adaptation in barren soils is unclear, this research employed RNA-seq technology to examine the impact of glucose concentration on the oligotrophic bacterium B. subtilis BS-G1 in soil affected by desertification. A global transcriptome analysis (RNA-Seq) revealed that the significantly differentially expressed genes (DEGs) histidine metabolism, glutamate synthesis, the HIF-1 signaling pathway, sporulation, and the TCA cycle pathway of B. subtilis BS-G1 were significantly enriched with a 0.015 g/L glucose concentration (L group), compared to a 10 g/L glucose concentration (H group). The DEGs amino acid system, two-component system, metal ion transport, and nitrogen metabolism system of B. subtilis BS-G1 were significantly enriched in the 5 g/L glucose concentration (M group), compared with the H group. In addition, the present study identified the regulation pattern and key genes under a low-glucose environment (7 mRNAs and 16 sRNAs). This study primarily investigates the variances in the regulatory pathways of the oligotrophic B. subtilis BS-G1, which holds substantial importance in comprehending the mechanism underlying the limited sugar tolerance of oligotrophic bacteria.

The Effect of Different Glucose Concentrations on the Antiproliferative Activity of Metformin in MCF-7 Breast Cancer Cells.

The glucose-lowering drug metformin has been reported to have anticancer properties through unknown mechanisms. Other unknown factors that may influence its anticancer potential include the glycemic status of the patient. Therefore, the objective of this study is to determine the effect of different glucose environments on the antiproliferative potency and the cellular mechanism of action of metformin. Human breast cancer cells, MCF-7, were incubated in low, normal, elevated, and high glucose environments and treated with metformin. The antiproliferative potential of metformin and its effect on protein expression as well as its ability to induce cellular apoptosis and autophagy under different glucose environments, were determined using different molecular techniques. Metformin significantly inhibited cellular proliferation in a time- and glucose-concentration-dependent manner. In comparison to elevated glucose, low normal glucose alone induced a significant level of autophagy that was further increased in the presence of metformin. While glucose concentration did not appear to have an effect on the antiproliferative potency of metformin, the cellular basis of action was shown to be glucose-dependent. The antiproliferative mechanism of action of metformin in elevated and low normal glucose environments is mTOR-dependent, whereas, in the high glucose environment, the antiproliferative mechanism is independent of mTOR. This is the first study to report that both the antiproliferative potency and the cellular mechanism of action aredependent on the concentration of glucose.

In vitro effect of high glucose concentrations on erythrocyte morphology assessed by scanning electron microscopy

AimsTo evaluate the in vitro effect of different glucose concentrations and the time series in erythrocyte cell viability and morphology parameters. MethodsDifferent glucose concentrations were assessed, from a normal physiological glucose concentration (5 mM) to a diabetes scenario (100 mM). Cell viability was obtained by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The erythrocyte morphology (diameter, height, and axial ratio) was assessed by scanning electron microscopy (SEM). ResultsA significant decrease in cell viability was observed in erythrocytes with higher glucose concentration (100 mM). There was also a significant increase in the cells’ diameter and height exposed to 100 mM, vs. those cultured with 5, 20, and 45 mM glucose, but only at 24 and 48 h. When comparing the same glucose concentration by a time series, it was observed that erythrocytes from the culture with 45 and 100 mM of glucose had a significant continuous decrease (approximately -0.40 μm) in diameter and height (24, 48, and 72 h), but there was no difference in the axial ratio observed. ConclusionContinuous exposure to high glucose concentrations in diabetes models produced less cell viability and changes in erythrocyte morphology (larger and scrambled cells), which in vivo might impact in microvascular complications.

SUMO1 modification of IGF-1R combining with SNAI2 inhibited osteogenic differentiation of PDLSCs stimulated by high glucose

BackgroundPeriodontal disease, an oral disease characterized by loss of alveolar bone and progressive teeth loss, is the sixth major complication of diabetes. It is spreading worldwide as it is difficult to be cured. The insulin-like growth factor 1 receptor (IGF-1R) plays an important role in regulating functional impairment associated with diabetes. However, it is unclear whether IGF-1R expression in periodontal tissue is related to alveolar bone destruction in diabetic patients. SUMO modification has been reported in various diseases and is associated with an increasing number of biological processes, but previous studies have not focused on diabetic periodontitis. This study aimed to explore the role of IGF-1R in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) in high glucose and control the multiple downstream damage signal factors.MethodsPDLSCs were isolated and cultured after extraction of impacted teeth from healthy donors or subtractive orthodontic extraction in adolescents. PDLSCs were cultured in the osteogenic medium with different glucose concentrations prepared by medical 5% sterile glucose solution. The effects of different glucose concentrations on the osteogenic differentiation ability of PDLSCs were studied at the genetic and cellular levels by staining assay, Western Blot, RT-PCR, Co-IP and cytofluorescence.ResultsWe found that SNAI2, RUNX2 expression decreased in PDLSCs cultured in high glucose osteogenic medium compared with that in normal glucose osteogenic medium, which were osteogenesis-related marker. In addition, the IGF-1R expression, sumoylation of IGF-1R and osteogenic differentiation in PDLSCs cultured in high glucose osteogenic medium were not consistent with those cultured in normal glucose osteogenic medium. However, osteogenic differentiation of PDLCSs enhanced after adding IGF-1R inhibitors to high glucose osteogenic medium.ConclusionOur data demonstrated that SUMO1 modification of IGF-1R inhibited osteogenic differentiation of PDLSCs by binding to SNAI2 in high glucose environment, a key factor leading to alveolar bone loss in diabetic patients. Thus we could maximize the control of multiple downstream damage signaling factors and bring new hope for alveolar bone regeneration in diabetic patients.

A high glucose concentration during early stages of in vitro equine embryo development alters expression of genes involved in glucose metabolism.

Equineembryos exhibit an unusual pattern of glucose tolerance in vitro and are currently cultured in hyperglycaemic conditions. Our main objective was to analyse the effect of different glucose concentrations on in vitro-produced equine embryo development and quality. Experiments comparing in vitro and in vivo produced embryos. Oocytes (n=641) were collected from post-mortem ovaries, matured in vitro and fertilised by intracytoplasmic sperm injection (ICSI). Embryo culture was divided from Day 0 to Day 4 and from Day 4 to Day 9 in three groups: 5-10 (5 and 10mmol/L glucose respectively; n=87); 5-17 (5 and 17.5mmol/L; n=66); and 10-17 (10 and 17.5mmol/L; n=117). A control group of 20 in vivo produced blastocysts was included. Cleavage and blastocyst rates were evaluated and embryos were snap-frozen for analysis of the relative mRNA expression of genes related to mitochondrial function, DNA methylation, apoptosis, glucose transport and metabolism. No differences were observed in the cleavage or blastocyst rates among in vitro groups. Under high glucose conditions in vitro (10-17 group), BAX/BCL2 was higher, and PFKP, LDHA and COX2 were overexpressed compared to all other groups. The two groups with 5mmol/L glucose concentration during the first culture stage (5-10 and 5-17) displayed similar patterns which differed to the 10-17 group. Conclusions related to embryo quality are based on gene expression patterns. Transfer of in vitro-produced embryos would reveal whether the observed differences improve embryo developmental competence. Five mM glucose during the first days of culture seems to be preferable to avoid over-activation of embryonic glycolytic pathways. Further studies are necessary to determine whether this improves embryo developmental competence.

The effects of different concentrations of glucose on glucose sensors and GLP-1 secretion in the enteroendocrine cell line STC-1

Glucose triggers glucagon-like peptide (GLP)-1 secretion from L cells involving several glucose sensors including sodium-glucose transporter (SGLT)1, glucose transporter (GLUT)2, and sweet taste receptors (STRs). This study investigated the effects of different glucose concentrations on GLP-1 secretion, intracellular concentrations of Ca2+ and cAMP, glucose uptake, and protein levels of SGLT1, GLUT2, and STRs in STC-1 cells. Low glucose (5.6 mM) increased GLP-1 secretion, intracellular Ca2+ concentration, and SGLT1 protein level compared with glucose-free group. GLP-1 secretion and intracellular Ca2+ concentration triggered by low glucose were inhibited by the SGLT1 inhibitor. GLP-1 secretion or intracellular Ca2+ concentration in high-glucose (25, 100, 200 mM) groups was significantly higher than that of low-glucose group. Elevation of cAMP level was observed in concentration-dependent manner, and decreased glucose uptake was observed in 100 or 200 mM glucose group. High glucose increased protein levels of STRs and GLUT2 in comparison to low-glucose group. GLP-1 secretion and intracellular levels of Ca2+ and cAMP triggered by high glucose were inhibited in the presence of the GLUT2 or STR inhibitor. These results suggest that SGLT1 is dominantly responsible for GLP-1 secretion triggered by low glucose, and that STRs and GLUT2 are involved in GLP-1 secretion induced by high glucose.

Senescence-associated miR-34a and miR-126 in middle-aged Indians with type 2 diabetes.

Rapid urbanization and unhealthy dietary patterns critically increase the risk of type 2 diabetes(T2D) in middle-aged Indians. However, despite recent evidence of senescence-associated microRNAs (SA-miRNAs) in regulating complex pathways of ageing, their expressions in middle-aged Indians with T2D remain unexplored. Hence we aimed to investigate the changes in expressions of SA-miRNAs miR-34a and miR-126 in middle-aged T2D patients. A total of 30 T2D patients and 30 controls were recruited of age 31-50years. The expressions of plasma miR-34a and miR-126 were determined by quantitative PCR. Oxidized LDL (OxLDL) and malondialdehyde (MDA) levels were quantified using enzyme-linked immunosorbent assay (ELISA). The effect of different glucose concentrations on miR-34a, miR-126, senescence-associated, and oxidative stress-responsive genes were also studied in an in vitro model of mice pancreatic β-cells. MiR-34a was significantly upregulated, whereas miR-126 was nonsignificantly reducedin T2D patients as compared to controls. T2D patients showed elevated levels of oxidative stress markers than controls. Analysis of cultured mice pancreatic β-cells exposed to high glucose showed significant upregulation of miR-34a, miR-126, p53, and superoxide dismutase 2 (SOD2). We found that circulating miR-34a levels and oxidative stress markers levels were elevated in the middle-aged Indians with T2D as compared to controls. The presence of diabetes may aggravate the normal ageing process in the middle-aged Indians. These SA-miRNAs can also be used to check the cellular dysfunctions and ageing of pancreatic β-cells.

Adult Goat Retinal Neuronal Culture: Applications in Modeling Hyperglycemia.

Culture of adult neurons of the central nervous system (CNS) can provide a unique model system to explore neurodegenerative diseases. The CNS includes neurons and glia of the brain, spinal cord and retina. Neurons in the retina have the advantage of being the most accessible cells of the CNS, and can serve as a reliable mirror to the brain. Typically, primary cultures utilize fetal rodent neurons, but very rarely adult neurons from larger mammals. Here, we cultured primary retinal neurons isolated from adult goat up to 10 days, and established an in vitro model of hyperglycemia for performing morphological and molecular characterization studies. Immunofluorescence staining revealed that approximately 30–40% of cultured cells expressed neuronal markers. Next, we examined the relative expression of cell adhesion molecules (CAMs) in adult goat brain and retina. We also studied the effect of different glucose concentrations and media composition on the growth and expression of CAMs in cultured retinal neurons. Hyperglycemia significantly enhances neurite outgrowth in adult retinal neurons in culture. Expression of CAMs such as Caspr1, Contactin1 and Prion is downregulated in the presence of high glucose. Hyperglycemia downregulates the expression of the transcription factor CCAAT/enhancer binding protein (C/EBP α), predicted to bind CAM gene promoters. Collectively, our study demonstrates that metabolic environment markedly affects transcriptional regulation of CAMs in adult retinal neurons in culture. The effect of hyperglycemia on CAM interactions, as well as related changes in intracellular signaling pathways in adult retinal neurons warrants further investigation.

Effect of Different Glucose Concentrations on Small RNA Levels and Adherence of Streptococcus mutans.

Streptococcus mutans (S. mutans) adheres to the tooth surface, metabolizes carbohydrates, and produces acid products, leading to enamel demineralization-the onset of dental caries. Rapid acidification by S. mutans has been observed in the presence of glucose. However, little is known about the role of small RNAs (sRNAs) in S. mutans in the presence of glucose and their relationship to tooth adherence. The objective of this study was to evaluate the role of sRNAs in S. mutans (18-50 nucleotides) regarding adherence capacity under 1% and 5% glucose concentrations. The pH drop and adherence capacity in the 1% glucose condition were similar to these parameters under conditions of 5% sucrose that were published in our previous study. A total of 2149 candidate sRNA with at least 100 average reads in the 5% and 1% glucose libraries were obtained. Between the two libraries, 581 sRNAs were differentially expressed and 43 sRNAs were verified. However, the expression levels of the predicted target genes gtfC and spaP were similar between the 1% and 5% glucose conditions. The bioinformatic analysis suggested that differentially expressed sRNAs may be involved in several pathways. These findings indicate that sRNAs were induced under these glucose concentrations and a series of sRNAs were specifically induced, respectively. sRNAs that are induced under glucose stress may be involved in regulating adherence of S. mutans.

The Effect of Different Glucose Concentrations on E2F1 Level and Cell Proliferation

The Effect of Different Glucose Concentrations on E2F1 Level and Cell Proliferation

Effects of different glucose concentrations on the leptin signaling pathway in MCF-7 and T47D breast cancer cells.

Leptin activates multiple intracellular signaling pathways, including JAK/STAT, by binding to its receptor. Leptin is also an important regulator of glucose homeostasis. Although both glucose and leptin increase breast cancer cell proliferation in vitro, whether the enhancing effect of glucose on the proliferation of breast cancer cells is mediated by the leptin signaling pathway is not known. The aim of this study was to investigate the effect of different glucose concentrations on the leptin signaling pathway in MCF-7 and T47D breast cancer cells. MCF-7 and T47D cell proliferation in different glucose concentrations (2.5 mM, 5 mM, 25 mM, or 50 mM) were assayed using CCK-8 assay. Leptin, leptin receptors (ObR, ObRb) as well as STAT3 mRNA and protein levels in both cell lines in different glucose concentrations were examined by RT-PCR and western blot, respectively. Incubation in 2.5 mM, 5 mM, 25 mM, or 50 mM glucose for 72h significantly increased the proliferation of both MCF-7 and T47D cells compared to 0 mM glucose incubated cells (P<0.001). mRNA levels of leptin, ObR, ObRb or STAT3 in 2.5 mM, 5 mM, 25 mM, or 50 mM glucose incubated cells were not significantly different in both cell lines compared to 0 mM (p>0.05). However, ObR protein levels in MCF-7 cells incubated in 25 mM glucose was significantly lower compared to 0 mM glucose by western blot (p<0.05). Our data suggest that the enhancing effect of glucose on breast cancer cell proliferation is not mediated by the JAK/STAT pathway.

Tetramethylpyrazine protects against high glucose-induced vascular smooth muscle cell injury through inhibiting the phosphorylation of JNK, p38MAPK, and ERK.

ObjectivesHigh glucose-induced alterations in vascular smooth muscle cell behavior have not been fully characterized. We explored the protective mechanism of tetramethylpyrazine (TMP) on rat smooth muscle cell injury induced by high glucose via the mitogen-activated protein kinase (MAPK) signaling pathway.MethodsVascular smooth muscle cells (VSMCs) isolated from rat thoracic aortas were divided into control, high glucose (HG), and pre-hatching TMP groups. The effect of different glucose concentrations on cell viability and on the migration activity of VSMC cells was examined using MTT analysis and the wound scratch assay, respectively. Superoxide dismutase (SOD) and malondialdehyde (MDA) levels were measured using enzyme-linked immunoassays. The levels of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38MAPK, and MAPK phosphorylation were assessed by western blotting.ResultsCell proliferation was remarkably increased by increased glucose concentrations. Compared with the HG group, the migratory ability of VSMC cells was reduced in the presence of TMP. TMP also decreased the MDA content in the supernatant, but significantly increased the SOD activity. Western blotting showed that TMP inhibited the phosphorylation of JNK, p38MAPK, and ERK.ConclusionsTMP appears to protect against HG-induced VSMC injury through inhibiting reactive oxygen species overproduction, and p38MAPK/JNK/ERK phosphorylation.

Mass Spectrometric Analysis of Proteins of L6 Skeletal Muscle Cells Under Different Glucose Conditions and Vitamin D Supplementation.

Type 2 Diabetes (T2D) is a metabolic disease which affects glucose homeostasis caused due to inability of the target cells to respond to insulin. Role of vitamin D in the pathogenesis and prevention of T2D has sparked widespread interest. Vitamin D plays a classical role in Ca++ homeostasis as well as regulates insulin secretion from β-cells and its action on various target cells. Proteins are the vital components of all cellular processes and their expression alters in response to various external or internal stimuli. Alteration in protein structure, function may contribute to the pathogenesis of many diseases including diabetes. Protein expression during the exposure of the cells to different glucose concentrations may alter and can give vital information about the pathogenesis of T2D. To study the effect of different glucose concentrations and supplementation of vitamin D on proteomic profile of L6 cell lines. L6 skeletal muscle cells were exposed to different Glucose (G) concentrations (0mM, 8mM, 16mM and 25mM) supplemented with Vitamin D (VD) for 48 hours. Total cell protein was extracted and protein profile was studied using SDS-PAGE. Three distinct bands observed in SDSPAGE in samples obtained from cells which were exposed to 8mM (G), 8mM (G) + VD and 16mM (G). The distinct bands were excised, in gel digestion were performed and MALDI-TOF analysis of the samples were done. MALDI-TOF analysis revealed these bands as mitochondrial uncoupling protein 3 (UCP3 MOUSE), Insulin gene enhancer protein 2 (ISL2 MOUSE) and Tubulin polyglutamylase complex1 (TPGS1 MOUSE) respectively. UCP3 protein is primarily expressed in the skeletal muscle cells and is involved in energy homeostasis and modulates insulin sensitivity. ISL2 protein plays an important role in differentiation and maintenance of the tissues. TPGS1 helps in microtubule polymerization might be helping in glucose transport and also play crucial role in the cellular movement, organization of intracellular structure, and intracellular transport. These identified proteins may provide information about disease pathophysiology and can serve as potential targets for therapeutic intervention of T2D. Further studies on the changes of protein expression under high glucose concentration and supplementation with vitamin D will lead to better understanding of the molecular mechanisms of T2D.

Effects of glucose concentration on osteogenic differentiation of type II diabetes mellitus rat bone marrow-derived mesenchymal stromal cells on a nano-scale modified titanium.

Diabetes mellitus (DM) is a common disease worldwide. Patients with DM have an increased risk of losing their teeth compared with other individuals. Dental implants are a standard of care for treating partial or full edentulism, and various implant surface treatments have recently been developed to increase dental implant stability. However, some studies have reported that DM reduces osseointegration and the success rate of dental implants. The purpose of this study was to determine the effects of high glucose levels for hard tissue formation on a nano-scale modified titanium surface. Titanium disks were heated at 600°C for 1 h after treatment with or without 10 m NaOH solution. All disks were incubated with type II DM rat bone marrow-derived mesenchymal stromal cells before exposure to one of four concentrations of glucose (5.5, 8.0, 12.0 or 24.0 mm). The effect of different glucose concentrations on bone marrow-derived mesenchymal stromal cell osteogenesis and inflammatory cytokines on the nano-scale modified titanium surface was evaluated. Alkaline phosphatase activity decreased with increasing glucose concentration. In contrast, osteocalcin production and calcium deposition were significantly decreased at 8.0 mm glucose, but increased with glucose concentrations over 8.0 mm. Differences in calcium/phosphate ratio associated with the various glucose concentrations were similar to osteocalcin production and calcium deposition. Inflammatory cytokines were expressed at high glucose concentrations, but the nano-scale modified titanium surface inhibited the effect of high glucose concentrations. High glucose concentration increased hard tissue formation, but the quality of the mineralized tissue decreased. Furthermore, the nano-scale modified titanium surface increased mineralized tissue formation and anti-inflammation, but the quality of hard tissue was dependent on glucose concentration.

Enhancement of Surfactin yield by improving the medium composition and fermentation process.

Surfactin is one of the most promising biosurfactants due to its extraordinary surface activity. Commonly, the well-established Cooper medium, a glucose-based mineral salt medium, is utilized for the microbial production of Surfactin. The current study investigated the enhancement of Surfactin yields by analyzing the effects of different glucose concentrations, next to the introduction of an alternative chelating agent and nitrogen source. The utilization of 8 g/L glucose, 0.008 mM Na3citrate and 50 mM (NH4)2SO4 increased Surfactin yields from 0.7 to 1.1 g/L during shake flask experiments applying Bacillus subtilis DSM10T. Consequentially conducted shake flask experiments, employing five other Surfactin producer strains during cultivation in the former and enhanced version of the Cooper medium, suggest a general enhancement of Surfactin yields during application of the enhanced version of the Cooper medium. The enhancement of the medium composition is therefore most likely independent from the employed producer strain. The following utilization of the enhanced medium composition during fed-batch fermentation with integrated foam fractionation yielded 30 % more Surfactin in comparison to batch fermentations with integrated foam fractionation employing the former version of the Cooper medium.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-015-0145-0) contains supplementary material, which is available to authorized users.

EFFECTS OF ACUTE HYPERGLYCEMIA AND INSULIN INTERVENTION ON THE SPONTANEOUS FIELD POTENTIAL OF SINOATRIAL NODE TISSUE BY USING MICROELECTRODE ARRAYS

Traditional studies on the relationship between hyperglycemia and heart diseases generally focused on the impact of chronic and long-term effect of diabetes on cardiac functions. Most of the methods were culturing myocardial cells and giving outside stimulations. However, recent studies show that acute hyperglycemia might play a significant role in spontaneous cardiac electrophysiology. In this research we applied microelectrode arrays (MEA) to record the spontaneous sinoatrial node field potentials of C57/BL6J mice and analyzed the effects of different glucose concentrations in time domain and frequency domain by using statistical method, vector maps and fast Fourier transform (FFT). Meanwhile, we studied the effects of insulin interference in the experimental process. When the concentration of the glucose solution was greater than 40 mmol/L, the spontaneous sinoatrial node field potential changed markedly. In the time domain, the amplitude decreased rapidly and the conductive characteristics were disordered. In the frequency domain, the two spectrum peaks decreased rapidly. These changes were irreversible. However, insulin preconditioning could inhibit the impact of high glucose.

Effect of different glucose concentrations on cAMP generation and insulin secretion in INS-1 cell

Objective To analyze the effect of glucose on cAMP generation and insulin secretion in INS-1 cell.Method We detected the changes of cAMP and insulin in INS-1 cell at 30 min stimulation by glucose of different concentrations (2.8,7.5,15 mmol/L).Result cAMP generation and insulin secretion in INS-1 cell were obviously stimulated as glucose concentration increased (P ＜ 0.05) and were relative to the concentration of glucose.Conclusion The increasing concentration of glucose could induce the cAMP production and insulin secretion in INS-1 cell. Key words: Glucose; cAMP; ELISA; Second messengers; Insulin secretion

Metabolome Response to Glucose in the β-Cell Line INS-1 832/13

Glucose-stimulated insulin secretion (GSIS) from pancreatic β-cells is triggered by metabolism of the sugar to increase ATP/ADP ratio that blocks the KATP channel leading to membrane depolarization and insulin exocytosis. Other metabolic pathways believed to augment insulin secretion have yet to be fully elucidated. To study metabolic changes during GSIS, liquid chromatography with mass spectrometry was used to determine levels of 87 metabolites temporally following a change in glucose from 3 to 10 mM glucose and in response to increasing concentrations of glucose in the INS-1 832/13 β-cell line. U-[(13)C]Glucose was used to probe flux in specific metabolic pathways. Results include a rapid increase in ATP/ADP, anaplerotic tricarboxylic acid cycle flux, and increases in the malonyl CoA pathway, support prevailing theories of GSIS. Novel findings include that aspartate used for anaplerosis does not derive from the glucose fuel added to stimulate insulin secretion, glucose flux into glycerol-3-phosphate, and esterification of long chain CoAs resulting in rapid consumption of long chain CoAs and de novo generation of phosphatidic acid and diacylglycerol. Further, novel metabolites with potential roles in GSIS such as 5-aminoimidazole-4-carboxamide ribotide (ZMP), GDP-mannose, and farnesyl pyrophosphate were found to be rapidly altered following glucose exposure.

Medium formulation and impeller design on the biosynthesis of high molecular weight hyaluronic acid by Streptococcus zooepidemicus ATCC 39920

The effects of medium formulation and impeller design (Rushton turbine and helical ribbon) on hyaluronic acid (HA) biosynthesis by Streptococcus zooepidemicus ATCC 39920 were investigated using a 2 L stirred-tank bioreactor. The effect of different glucose concentrations (20, 30, 50 and 60 g/L), nitrogen sources ((NH4)2S2O8, (NH4)2PO4, yeast extract, and tryptone) and carbon/nitrogen ratios on the growth of the strain and on HA biosynthesis were initially investigated. Organic nitrogen sources (yeast extract and tryptone) were proven to be favourable in media for HA biosynthesis compared to inorganic nitrogen sources. About 2.442 g/L of HA with a high molecular weight (4.36 x 106Da) was synthesised at an optimal C/N of 2.5:1 (using a mixture of yeast extract and tryptone) in a 2 L stirred-tank bioreactor equipped with a Rushton turbine impeller. When using an optimal medium formulation at equal HA production levels, the helical ribbon impeller resulted in a higher molecular weight of HA (5.20 x 106 Da) compared to the Rushton turbine impeller. Key words: Streptococcus spp., hyaluronic acid, helical ribbon impeller, carbon/nitrogen.