Glucose Concentration Gradient Research Articles

Bridging the gap: glucose transporters, Alzheimer's, and future therapeutic prospects.

Glucose is the major source of chemical energy for cell functions in living organisms. The aim of this mini-review is to provide a clearer and simpler picture of the fundamentals of glucose transporters as well as the relationship of these transporters to Alzheimer's disease. This study was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Electronic databases (PubMed and ScienceDirect) were used to search for relevant studies mainly published during the period 2018-2023. This mini-review covers the two main types of glucose transporters, facilitated glucose transporters (GLUTs) and sodium-glucose linked transporters (SGLTs). The main difference between these two types is that the first type works through passive transport across the glucose concentration gradient. The second type works through active co-transportation to transport glucose against its chemical gradient. Fluctuation in glucose transporters translates into a disturbance of normal functioning, such as Alzheimer's disease, which may be caused by a significant downregulation of GLUTs most closely associated with insulin resistance in the brain. The first sign of Alzheimer's is a lack of GLUT4 translocation. The second sign is tau hyperphosphorylation, which is caused by GLUT1 and 3 being strongly upregulated. The current study focuses on the use of glucose transporters in treating diseases because of their proven therapeutic potential. Despite this, studies remain insufficient and inconclusive due to the complex and intertwined nature of glucose transport processes. This study recommends further understanding of the mechanisms related to these vectors for promising future therapies.

PH-Sensitive Glucose-Powered Nanomotors for Enhanced Intracellular Drug Delivery and Ferroptosis Efficiency.

We propose a glucose-powered Janus nanomotor where two faces are functionalized with glucose oxidase (GOx) and polydopamine-Fe3+ chelates (PDF), respectively. In the glucose fuel solution, the GOx on the one side of these Janus nanomotors catalytically decomposes glucose fuels into gluconic acid and hydrogen peroxide (H2 O2 ) to drive them at a speed of 2.67 μm/s. The underlying propulsion mechanism is the glucose-based self-diffusiophoresis owing to the generated local glucose concentration gradient by the enzymatic reaction. Based on the enhanced diffusion motion, such nanomotors with catalytic activity increase the uptake towards cells and subsequently exhibit excellent capabilities for Fe3+ ions delivery and H2 O2 generation for enhancing ferroptosis efficiency for inducing cancer cell death. In particular, the Fe3+ ions are released from nanomotors in a slightly acidic environment, and subsequently generate toxic hydroxyl radicals via Fenton reactions, which accumulation reactive oxygen species (ROS) level (~300 %) and further lipid peroxidation (LPO) strengthened ferroptosis therapy for cancer treatment. The as-developed glucose-powered Janus nanomotor with efficient diffusion and Fe ions delivery capabilities show great promise as a potential in biomedical applications.

Metabolic-fluid dynamics model construction and scale-down design for an industrial penicillin chrysogenum fermentation with combined dissolved oxygen and glucose concentration dynamics

This study focuses on the metabolic impacts of simultaneous glucose and oxygen concentration gradients on penicillin production in an industrial-scale fermentor, using the computational fluid dynamics-cellular reaction dynamics approach. Inclusion of oxygen-coupling considerably impacts the glucose uptake and resulting penicillin productivity. This is characterised by six metabolic regimes; lifeline data reconstructed from experimental results, recorded from the cellular perspective, indicates rapid dynamics in glucose and dissolved oxygen uptake by the microorganisms. The results are highly sensitive to variations in the oxygen-related model parameters, requiring accurate insight into the multiphase hydrodynamics and metabolic processes. Hypothetical scenarios with stronger glucose-oxygen limitations than tested experimentally were further explored. A precision scale-down (SD) simulator was designed based on the lifeline data, requiring considerable operational dynamics, with increasing system complexity and implementation difficulty. These insights may inspire further research into alternative SD configurations better suited to mimic the rapid dynamics of large-scale fermentation processes.

Performing in spite of starvation: How Saccharomyces cerevisiae maintains robust growth when facing famine zones in industrial bioreactors.

In fed-batch operated industrial bioreactors, glucose-limited feeding is commonly applied for optimal control of cell growth and product formation. Still, microbial cells such as yeasts and bacteria are frequently exposed to glucose starvation conditions in poorly mixed zones or far away from the feedstock inlet point. Despite its commonness, studies mimicking related stimuli are still underrepresented in scale-up/scale-down considerations. This may surprise as the transition from glucose limitation to starvation has the potential to provoke regulatory responses with negative consequences for production performance. In order to shed more light, we performed gene-expression analysis of Saccharomyces cerevisiae grown in intermittently fed chemostat cultures to study the effect of limitation-starvation transitions. The resulting glucose concentration gradient was representative for the commercial scale and compelled cells to tolerate about 76 s with sub-optimal substrate supply. Special attention was paid to the adaptation status of the population by discriminating between first time and repeated entry into the starvation regime. Unprepared cells reacted with a transiently reduced growth rate governed by the general stress response. Yeasts adapted to the dynamic environment by increasing internal growth capacities at the cost of rising maintenance demands by 2.7%. Evidence was found that multiple protein kinase A (PKA) and Snf1-mediated regulatory circuits were initiated and ramped down still keeping the cells in an adapted trade-off between growth optimization and down-regulation of stress response. From this finding, primary engineering guidelines are deduced to optimize both the production host's genetic background and the design of scale-down experiments.

Estimation of Oxygen and Glucose Concentration Distribution in the Rat Brain Arterial System

Glucose and oxygen concentration gradients are the key indicators that form the trophic tissue supply in mammalian brain. To describe them in detail it is essential to combine the solution of both hemodynamics and the convection-diffusion-reaction problems in the tissue. Visualization of spatio-temporal distributions of the metabolites noted above can be carried out both using the gradients themselves and the corresponding probability density functions. In the case of considering large parts of the brain, as well as the entire organ as a whole, the second method for metabolite heterogeneity description is of greater interest for practical purposes. This paper presents an approach to obtain a probability density functions based on structural segmentation of the diffusion region using Delaunay triangulation and the spherical source diffusion field method. It is shown that the average values of the estimated distributions deviate by 8 % from the experimentally obtained results and it corresponds to the best match during the validation by the finite element method in the triangulation simplices of basic topology. Given the relatively low computational complexity of both the segmentation process and the estimation of concentration in a single segment, the proposed method to obtain integral distributions of various compounds, in particular glucose and oxygen, can be used as an affordable alternative to precise calculation of the concentration gradients in the whole brain and its distinct anatomical structures.

Application of a multicomponent model of convectional reaction-diffusion to description of glucose gradients in a neurovascular unit.

A supply of glucose to a nervous tissue is fulfilled by a cerebrovascular network, and further diffusion is known to occur at both an arteriolar and a microvascular level. Despite a direct relation, a blood flow dynamic and reaction-diffusion of metabolites are usually considered separately in the mathematical models. In the present study they are coupled in a multiphysical approach which allows to evaluate the effects of capillary blood flow changes on near-vessels nutrient concentration gradients evidently. Cerebral blood flow (CBF) was described by the non-steady-state Navier-Stokes equations for a non-Newtonian fluid whose constitutive law is given by the Carreau model. A three-level organization of blood–brain barrier (BBB) is modelled by the flux dysconnectivity functions including densities and kinetic properties of glucose transporters. The velocity of a fluid flow in brain extracellular space (ECS) was estimated using Darcy’s law. The equations of reaction-diffusion with convection based on a generated flow field for continues and porous media were used to describe spatial-time gradients of glucose in the capillary lumen and brain parenchyma of a neurovascular unit (NVU), respectively. Changes in CBF were directly simulated using smoothing step-like functions altering the difference of intracapillary pressure in time. The changes of CBF cover both the decrease (on 70%) and the increase (on 50%) in a capillary flow velocity. Analyzing the dynamics of glucose gradients, it was shown that a rapid decrease of a capillary blood flow yields an enhanced level of glucose in a near-capillary nervous tissue if the contacts between astrocytes end-feet are not tight. Under the increased CBF velocities the amplitude of glucose concentration gradients is always enhanced. The introduced approach can be used for estimation of blood flow changes influence not only on glucose but also on other nutrients concentration gradients and for the modelling of distributions of their concentrations near blood vessels in other tissues as well.

MicroRNA-148a-3p alleviates high glucose-induced diabetic retinopathy by targeting TGFB2 and FGF2.

Diabetic retinopathy (DR), a common complication of type 1 or type 2 diabetes mellitus, has become the leading cause of blindness among adults in working age. The dysregulation of microRNA has been reported to be strongly related to the initiation or progression of DR. However, neither the biological role nor the molecular mechanism of miR-148a-3p has been researched in DR. This study is designed to investigate the function and mechanism of miR-148a-3p in DR. The bioinformatics analysis (Targetscan: https://www.targetscan.org/vert_72/ ) and numerous experiments including real-time quantitative polymerase chain reaction, terminal deoxynucleotidyltransferase dUTP nick end labeling, CCK-8, western blot, vasculogenesis and luciferase reporter assays were used to research the function and mechanism of miR-148a-3p in DR. We constructed DR cell model by treating human retinal microvascular endothelial cells (HRECs) with different concentration gradients of high glucose (HG). Additionally, HG treatment reduced miR-148a-3p level in HRECs. In function, overexpression of miR-148a-3p caused an increase in cell viability and a decrease in cell apoptosis. Besides, miR-148a-3p overexpression led to a damage on blood-retinal barrier (BRB) and suppressed angiogenesis. In mechanism, miR-148a-3p specifically bound to 3' untranslated region of TGFB2 and FGF2. At least, rescue assays demonstrated that the inhibitive influence of miR-148a-3p mimics on BRB injury was offset by overexpression of TGFB2 and the attenuation of angiogenesis resulting from miR-148a-3p mimics was abrogated by overexpression of FGF2 CONCLUSIONS: In a word, we discovered that miR-148a-3p alleviated HG-induced DR by targeting TGFB2 and FGF2. This novel discovery indicated miR-148a-3p as a potential target for DR diagnosis or treatment.

The circadian clock regulates RPE-mediated lactate transport via SLC16A1 (MCT1)

Multiple retinal cells harbor a circadian oscillator, including retinal pigment epithelial cells (RPE). However, little is known about the functions that are regulated by the RPE clock. The aim of this study was to investigate whether the circadian clock in the RPE regulates the transport of glucose and its glycolytic metabolic by-product - lactate. To that end, we first characterized the mRNA expression profile of glucose and monocarboxylate transporters in ARPE-19 cells. We found that SLC2A1 and SLC16A1 were, respectively, the most abundantly expressed glucose and lactate (monocarboxylate) transporters. We further observed that the protein products of SLC2A1 (encoding GLUT1) and SLC16A1 (encoding MCT1) localize on the apical membrane of ARPE-19 monolayers. In a subsequent time-course experiment, we found that SLC2A1 and SLC16A1 mRNA oscillated in ARPE-19 monolayers, but not in dispersed cells, suggesting that monolayer cellular organization is necessary for rhythmic regulation of these transporters. In these monolayers, we found that MCT1 proteins varied over time, in contrast to GLUT1 proteins which did not vary over time. Spectrophotometric measurements of supernatants sampled from ARPE-19 monolayer cultures revealed that glucose concentrations did not significantly differ between apical (Api) supernatants and basolateral (BL) ones. In addition, we did not find rhythms in Api or BL glucose concentrations. Conversely, we found higher lactate concentrations in Api supernatants than BL ones. Further, we found that Api lactate concentrations were rhythmic. Pearson's r revealed that the concentration gradients (Api - BL) of glucose and lactate correlated with the gene expression of respective SLC2A1 and SLC16A1 transporters. Incubation with photoreceptor outer segments (POS) affected the mRNA expression of SLC16A1 and SLC2A1 in ARPE-19 monolayers in a time-dependent manner, thus suggesting that the retina might modulate the RPE clock-controlled expression of transporters via interactions with POS. In conclusion, this work provides evidence that the transport of lactate is regulated by the circadian clock in the RPE.

Biomarker Guided Diagnosis of Septic Peritonitis in Dogs

Septic peritonitis (SP) is common in dogs and is associated with high mortality. Early recognition is essential to maximizing survival and may be aided by biomarker measurement. The present study aimed to evaluate the ability of biomarkers to discriminate septic peritonitis from non-septic ascites (NSA). Eighteen dogs with SP and 19 age-matched controls with NSA were enrolled. Contemporaneous blood and peritoneal effusion samples were obtained. Concentrations of cell-free DNA (cfDNA), cytokines, glucose, lactate, N-terminal pro-C-type natriuretic peptide (NT-proCNP), nucleosomes, and procalcitonin (PCT) were measured using commercial reagents and assays. Paired biomarker concentrations were compared with the Wilcoxon matched-pairs signed rank test, and biomarker concentrations between groups were compared with the Mann-Whitney U-test. P-values were adjusted for multiple comparisons using the Bonferroni correction. Receiver operating characteristic curves were generated to assess the ability of the above biomarkers to discriminate SP from NSA. Dogs with SP had significantly greater blood CCL2 concentrations than dogs with NSA (P = 0.032). Dogs with SP had significantly greater effusion CCL2, IL-6, IL-10, and lactate concentrations than dogs with NSA (P ≤ 0.0121). Blood-effusion concentration gradients of CCL2, glucose, IL-6, IL-10, and lactate were significantly different in dogs with SP compared to dogs with NSA (P ≤ 0.0165). Effusion lactate concentration had the highest AUROC value (0.866, 95% CI 0.751–0.980, P = 0.0001), although other biomarkers performed similarly. An effusion lactate concentration of 4.2 mmol/L was 72.2% (95% CI 46.5–90.3%) sensitive and 84.2% (95% CI 60.4–96.6%) specific for the diagnosis of SP.

YY1: A novel therapeutic target for diabetic nephropathy orchestrated renal fibrosis

BackgroundRenal fibrosis promotes the development of diabetic nephropathy (DN). A growing number of studies have reported that Yin Yang 1 (YY1), which is involved in cellular proliferation and differentiation, plays a crucial role in the pathogenesis of many diseases, such as pulmonary fibrosis, hepatic steatosis and cancer. MethodsWe detected the expression of YY1 under various glucose concentration and time gradient conditions. Rapamycin was used to verify the mTORC1/p70S6K/YY1 signaling pathway in HK-2 cells. We used db/db mice to examine the connection between renal fibrosis and YY1. A luciferase assay and chromatin immunoprecipitation (ChIP) assay were used to identify whether YY1 directly regulated α-SMA by binding to the α-SMA promoter. RNA silencing and overexpression were performed by using a YY1 expression/knockdown plasmid to investigate the function of YY1 in renal fibrosis of DN. ResultsYY1 expression and subsequent nuclear translocation were upregulated in a glucose- and time-dependent manner via the mTORC1/p70S6K signaling pathway in HK-2 cells. YY1 expression and nuclear translocation was significantly upregulated in db/db mice. Furthermore, YY1 upregulated α-SMA expression and activity in high-glucose-cultured HK-2 cells. Overexpression of YY1 promoted renal fibrosis in db/m mice mainly by upregulating α-SMA expression and inducing epithelial-mesenchymal transition (EMT) in vitro and in vivo. Finally, downregulation of YY1 reversed renal fibrosis by improving EMT in vivo and in vitro. ConclusionsThese results reveal that upregulation of YY1 plays a critical role in HG-induced deregulation of EMT-associated protein expression, which finally results in renal fibrosis of DN. Therefore, decreasing YY1 expression might represent a new therapeutic target for diabetic nephropathy-induced renal fibrosis.

Dynamic mathematical modeling of heat and mass transfer incorporating with the local nutrient and biomass limitation of growth in a packed-bed solid-state bioreactor

This research develops on our previous semi-mechanistic model that describes the dynamic physical and biochemical processes taking place in a packed-bed bioreactor to analyze the relationship of nutrient limitation, biomass accumulation, metabolic heat generation, and mathematical description of packed-bed porous media. The experimental and simulation data proved that glucose concentration gradients in the biofilm could be neglected due to small biofilm thickness and high diffusivity of glucose in the biofilm. The prediction results also showed that an increase in the initial substrate concentration leads to a rise in the temperature gradient in the bed. The model proposes that if the diameter of substrate particle is too large (r > 0.1 cm), the growth rate will decrease significantly due to the high biomass accumulation in the biofilm, and temperature gradients decrease in the column. This can be used as a strategy to control the overheating problem in the bed.

An agent-based model for drug-radiation interactions in the tumour microenvironment: Hypoxia-activated prodrug SN30000 in multicellular tumour spheroids.

Multicellular tumour spheroids capture many characteristics of human tumour microenvironments, including hypoxia, and represent an experimentally tractable in vitro model for studying interactions between radiotherapy and anticancer drugs. However, interpreting spheroid data is challenging because of limited ability to observe cell fate within spheroids dynamically. To overcome this limitation, we have developed a hybrid continuum/agent-based model (ABM) for HCT116 tumour spheroids, parameterised using experimental models (monolayers and multilayers) in which reaction and diffusion can be measured directly. In the ABM, cell fate is simulated as a function of local oxygen, glucose and drug concentrations, determined by solving diffusion equations and intracellular reactions. The model is lattice-based, with cells occupying discrete locations on a 3D grid embedded within a coarser grid that encompasses the culture medium; separate solvers are employed for each grid. The generated concentration fields account for depletion in the medium and specify concentration-time profiles within the spheroid. Cell growth and survival are determined by intracellular oxygen and glucose concentrations, the latter based on direct measurement of glucose diffusion/reaction (in multilayers) for the first time. The ABM reproduces known features of spheroids including overall growth rate, its oxygen and glucose dependence, peripheral cell proliferation, central hypoxia and necrosis. We extended the ABM to describe in detail the hypoxia-dependent interaction between ionising radiation and a hypoxia-activated prodrug (SN30000), again using experimentally determined parameters; the model accurately simulated clonogenic cell killing in spheroids, while inclusion of reversible cell cycle delay was required to account for the marked spheroid growth delay after combined radiation and SN30000. This ABM of spheroid growth and response exemplifies the utility of integrating computational and experimental tools for investigating radiation/drug interactions, and highlights the critical importance of understanding oxygen, glucose and drug concentration gradients in interpreting activity of therapeutic agents in spheroid models.

Glucose promotes the formation of tunneling nanotubes between bladder cancer cells by up-regulating Fyn/Rho-associated kinase signaling pathway

Objective To investigate the effects of glucose in formation of tunneling nanotubes (TNTs) between bladder cancer cells and the potential mechanism. Methods The highly invasive bladder cancer cells T24 were labeled with Mito-Tracker Deep Red, and then co-cultured with the less invasive bladder cancer cells RT4 for 24 hours. Actin-Tracker Green was used to label the cytoskeletal protein to detect the TNTs between T24 and RT4 cells. T24 and RT4 cells were cultured separately for 12 hours and 24 hours respectively, and the medium was collected and analyzed by liquid chromatography and mass spectrometry to screened out the difference factors between the micro-environment. T24 cells were cultured with the factors to confirm whether they could induce and enhance TNTs formation. The expression level of Fyn, Rho-associated kinase (ROCK) and p-paxillin proteins were detected and compared by Western blotting analysis. Results The formation of TNTs was found between T24 and RT4 cells. And the main component of TNTs was F-actin. Metabolomics analysis revealed that glucose was a differential factor between the micro-environments of T24 and RT4 cells, which could induce the formation of TNTs between T24 and RT4 cells (P=0.005). Concentration gradient of glucose between T24 and RT4 cells can up-regulate the expression of Fyn/ROCK and p-paxillin proteins (P=0.001, P=0.004, P=0.010). Conclusion Concentration gradient of glucose in the micro-environment of bladder cancer cells can promote the formation of TNTs between highly invasive and less invasive bladder cancer cells. The potential mechanism of the formation of TNTs induced by glucose might be explained by concentration gradient of glucose in the micro-environment of bladder cancer cells which can up-regulate the activity of Fyn/ROCK signaling pathway. Key words: Tunneling nanotubes; Bladder cancer; Cell conjunction; Micro-environment

Stearoyl-CoA desaturase-1 promotes colorectal cancer metastasis in response to glucose by suppressing PTEN

BackgroundDiabetic patients have a higher risk factor for colorectal cancer (CRC) metastasis. Stearoyl-CoA desaturase 1 (SCD1), the main enzyme responsible for producing monounsaturated fatty acids(MUFA) from saturated fatty acids, is frequently deregulated in both diabetes and CRC. The function and mechanism of SCD1 in metastasis of CRC and its relevance to glucose remains largely unknown.MethodsSCD1 expression levels were analyzed in human CRC tissues and the Cancer Browser database (https://genome-cancer.ucsc.edu/). CRC cell lines stably transfected with SCD1 shRNAs or vector were established to investigate the role of SCD1 in modulating migration and invasion of CRC cells. A glucose concentration gradient was set to investigate regulation of SCD1 in CRC relevant to diabetic conditions.ResultsThe clinical data analysis showed high expression of SCD1 in CRC tissues with a negative correlation with the prognosis of CRC. In vitro experiments revealed that SCD1 increased CRC progression through promoting epithelial–mesenchymal transition (EMT). Lipidomic analysis demonstrated that SCD1 increased MUFA levels and MUFA administration could rescue migration and invasion defect of CRC cells induced by SCD1 knockdown. Furthermore, SCD1-mediated progression of CRC was promoted by carbohydrate response-element binding protein (ChREBP) in response to high glucose. Mechanistically, hyperglycemia-SCD1-MUFA induced CRC cell migration and invasion by regulating PTEN.ConclusionsOur findings show that SCD1 promotes metastasis of CRC cells through MUFA production and suppressing PTEN in response to glucose, which may be a novel mechanism for diabetes-induced CRC metastasis.

Computational fluid dynamics simulation of an industrial P. chrysogenum fermentation with a coupled 9-pool metabolic model: Towards rational scale-down and design optimization

We assess the effect of substrate heterogeneity on the metabolic response of P. chrysogenum in industrial bioreactors via the coupling of a 9-pool metabolic model with Euler-Lagrange CFD simulations. In this work, we outline how this coupled hydrodynamic-metabolic modeling can be utilized in 5 steps. (1) A model response study with a fixed spatial extra-cellular glucose concentration gradient, which reveals a drop in penicillin production rate qp of 18–50% for the simulated reactor, depending on model setup. (2) CFD-based scale-down design, where we design a 1-vessel scale down simulator based on the organism lifelines. (3) Scale-down verification, numerically comparing the model response in the proposed scale-down simulator with large-scale CFD response. (4) Reactor design optimization, reducing the drop in penicillin production by a change of feed location. (5) Long-term fed-batch simulation, where we verify model predictions against experimental data, and discuss population heterogeneity. Overall, these steps present a coupled hydrodynamic-metabolic approach towards bioreactor evaluation, scale-down and optimization.

Investigation of the Influence of Glucose Concentration on Cancer Cells by Using a Microfluidic Gradient Generator without the Induction of Large Shear Stress.

A microfluidic device capable of precise chemical control is helpful to mimic tumor microenvironments in vitro, which are closely associated with malignant progression, including metastasis. Cancer cells under a concentration gradient of oxygen and other sustenance materials inside a tumor in vivo have recently been reported to increase the probability of metastasis. The influence of glucose concentration on cancer cells has not been measured well, whereas that of oxygen concentration has been thoroughly examined using microfluidic devices. This is because glucose concentrations can be controlled using microfluidic concentration gradient generators, which trade off temporal stability of the glucose concentration and shear stress on the cells; by contrast, oxygen concentration can be easily controlled without microfluidic device-induced shear stresses. To study cell division and migration responses as a function of glucose concentration, we developed a microfluidic device to observe cell behaviors under various chemical conditions. The device has small-cross-section microchannels for generating a concentration gradient and a large-cross-section chamber for cell culture. With this design, the device can achieve both a cell culture with sufficiently low shear stress on cell activity and a stable glucose concentration gradient. Experiments revealed that a low glucose concentration increased the total migration length of HeLa cells and that HeLa cells under a glucose concentration gradient exhibit random motion rather than chemotaxis.

The fetal glucose steal: an underappreciated phenomenon in diabetic pregnancy.

Adverse neonatal outcomes continue to be high for mothers with type 1 and type 2 diabetes, and are far from eliminated in mothers with gestational diabetes mellitus. This is often despite seemingly satisfactory glycaemic control in the latter half of pregnancy. Here we argue that this could be a consequence of the early establishment of fetal hyperinsulinaemia, a driver that exaggerates the fetal glucose steal. Essentially, fetal hyperinsulinaemia, through its effect on lowering fetal glycaemia, will increase the glucose concentration gradient across the placenta and consequently the glucose flux to the fetus. While the steepness of this gradient and glucose flux will be greatest at times when maternal hyperglycaemia and fetal hyperinsulinaemia coexist, fetal hyperinsulinaemia will favour a persistently high glucose flux even at times when maternal blood glucose is normal. The obvious implication is that glycaemic control needs to be optimised very early in pregnancy to prevent the establishment of fetal hyperinsulinaemia, further supporting the need for pre-pregnancy planning and early establishment of maternal glycaemic control. An exaggerated glucose steal by a hyperinsulinaemic fetus could also attenuate maternal glucose levels during an OGTT, providing an explanation for why some mothers with fetuses with all the characteristics of diabetic fetopathy have ‘normal’ glucose tolerance.

Scale-down of continuous protein producing Saccharomyces cerevisiae cultivations using a two-compartment system.

In the biotechnological industry, economic decisions in investment are typically based on laboratory-scale experiments. Scale-down as a tool is therefore of high industrial importance to transfer the processes into larger production scale without loss in performance. In this study, large-scale prolonged continuous cultivations with a heterologous protein producing Saccharomyces cerevisiae strain have been scaled-down to a two-compartment scale-down reactor system. The effects of glucose, pH, and oxygen concentration gradients have been investigated by comparison with corresponding 300 mL standard continuous cultivations. It was found that substrate gradients within a limited range result in increased productivity of the heterologous protein under regulation of glycolytic TPI promoter and delay the decrease of protein and trehalose production during continuous cultivation. Based on these results, it is argued that introduction of variations in substrate concentration can be beneficial for industrial continuous cultivations.

Cultivation of yeast in diffusion-based microfluidic device

The capacity to create a diffusive chemical concentration gradient in microfluidic systems has the potential to improve the study of microbial processes. These tools allow the evaluation of microbial cell performance under different and controlled conditions. Diffusion-based gradient generators, in particular, have the capacity to maintain spatiotemporally constant gradient concentrations necessary to evaluate cell behavior in a precise environment. This work uses a known microfluidic device capable of generating a diffusive glucose concentration gradient to evaluate for the first time the behavior of Saccharomyces cerevisiae ATCC 7754 inside a microchannel. The cell growth along the microfluidic microchambers was observed and the kinetic parameters determined, with values statistically similar to those of conventional batch cultivation. Monod kinetic parameters could also be determined in the microfluidic device using small substrate concentrations. These results show the potential of this microbioreactor to investigate yeast growth with microliter samples and to evaluate experiments in triplicate performed and in parallel. The diffusive concentration gradient in a microfluidic device allowed the acquisition of results in a more practical way when compared to conventional techniques.

Pre-treatment with glucagon-like Peptide-1 protects against ischemic left ventricular dysfunction and stunning without a detected difference in myocardial substrate utilization.

This study sought to determine whether pre-treatment with intravenous glucagon-like peptide-1 (GLP-1)(7-36) amide could alter myocardial glucose use and protect the heart against ischemic left ventricular (LV) dysfunction during percutaneous coronary intervention. GLP-1 has been shown to have favorable cardioprotective effects, but its mechanisms of action remain unclear. Twenty patients with preserved LV function and single-vessel left anterior descending coronary artery disease undergoing elective percutaneous coronary intervention were studied. A conductance catheter was placed into the LV, and pressure-volume loops were recorded at baseline, during 1-min low-pressure balloon occlusion (BO), and at 30-min recovery. Patients were randomized to receive an infusion of either GLP-1(7-36) amide at 1.2 pmol/kg/min or saline immediately after baseline measurements. Simultaneous coronary artery and coronary sinus blood sampling was performed at baseline and after BO to assess transmyocardial glucose concentration gradients. BO caused both ischemic LV dysfunction and stunning in the control group but not in the GLP-1 group. Compared with control subjects, the GLP-1 group had a smaller reduction in LV performance during BO (delta dP/dTmax, -4.3 vs. -19.0%, p = 0.02; delta stroke volume, -7.8 vs. -26.4%, p = 0.05), and improved LV performance at 30-min recovery. There was no difference in transmyocardial glucose concentration gradients between the 2 groups. Pre-treatment with GLP-1(7-36) amide protects the heart against ischemic LV dysfunction and improves the recovery of function during reperfusion. This occurs without a detected change in myocardial glucose extraction and may indicate a mechanism of action independent of an effect on cardiac substrate use. (Effect of Glucgon-Like-Peptide-1 [GLP-1] on Left Ventricular Function During Percutaneous Coronary Intervention [PCI]; ISRCTN77442023).