Glucose Transport Activity Research Articles

MicroRNA-6077 enhances the sensitivity of patients-derived lung adenocarcinoma cells to anlotinib by repressing the activation of glucose transporter 1 pathway

Anlotinib is a novel molecular targeted agent targeting the vascular endothelial growth factor receptor, which differs from the other currently available non-small cell lung cancer (NSCLC) molecular targeted drugs targeting this receptor. Although the application of anlotinib may bring new hope for patients with advanced NSCLC, the cost of treatment is high. The results of this study showed that microRNA-6077 (miR-6077) represses the expression of GLUT1 (glucose transporter 1) and enhances the sensitivity of patient-derived lung adenocarcinoma (AC) cells to anlotinib. The miR-6077, which potentially binds to the 3’untranslated region of GLUT1, was identified and screened by miRDB, an online tool; sequences of miR-6077 were prepared as lentivirus particles. A549 cells (a lung adenocarcinoma cell line) and five patient-derived AC cell lines were infected with control miRNA or miR-6077, and subsequently treated with the indicated concentration of anlotinib. The expression of proteins, such as GLUT1, was determined by western blotting. The antitumor effect of anlotinib was identified through in-vitro (e.g., MTT) or in-vivo methods (e.g., subcutaneous tumor model). Overexpression of miR-6077 repressed the expression of GLUT1 and decreased the glucose uptake, lactate production, or ATP generation in AC cells. In addition, MiR-6077 may enhance the antitumor effect of anlotinib on A549 or patient-derived AC cell lines. Therefore, our results indicated that miR-6077 represses the expression of GLUT1 and enhances the sensitivity of patients-derived lung AC cells to anlotinib.

Methylamine Activates Glucose Uptake in Human Adipocytes Without Overpassing Action of Insulin or Stimulating its Secretion in Pancreatic Islets.

Background: Methylamine, a natural soluble amine present in foods, is known to be a substrate of primary amine oxidase (PrAO) widely expressed in animal tissues. Methylamine has been reported to activate glucose transport in fat cells and to facilitate glucose disposal in rabbits but the interests and limits of such insulin-mimicking actions have not been further explored. This work aimed to perform a preclinical study of the inter-individual variations of these biological properties to study the putative link between PrAO activity and insulin resistance. Methods: Methylamine was tested on human adipocyte preparations and in rabbit pancreatic islets to determine its influence on glucose uptake and insulin release, respectively. PrAO activity and related responses were determined in adipose tissues obtained from two cohorts of non-obese and obese women. Results: Adipose tissue PrAO activity was negatively correlated with insulin resistance in high-risk obese women. PrAO-dependent activation of glucose uptake was negatively correlated with body mass index and reflected the decrease of insulin responsiveness of human fat cells with increasing obesity. Methylamine exhibited antilipolytic properties in adipocytes but was unable to directly activate insulin secretion in isolated pancreatic islets. Conclusions: PrAO activation by its substrates, e.g., methylamine, increases glucose utilization in human adipocytes in a manner that is linked to insulin responsiveness. Methylamine/PrAO interaction can therefore contribute to adipose tissue enlargement but should be considered as potentially useful for diabetes prevention since it could limit lipotoxicity and facilitate glucose handling, at the expense of favoring healthy fat accumulation.

Ginsenoside compound K ameliorates Alzheimer's disease in HT22 cells by adjusting energy metabolism.

Energy metabolism disorders have been shown to exert detrimental effects on the pathology of Alzheimer's disease (AD). The ginsenoside compound K (CK), a major intestinal metabolite underlying the pharmacological actions of orally administered ginseng, has an ameliorating effect against AD, but the relevant molecular mechanism remains unclear. We hypothesized that the improvement of AD by CK is mediated by the energy metabolism signaling pathway induced by amyloid β peptide (Aβ) and tested this hypothesis in HT22 cells. HT22 cells were incubated with CK and exposed to Aβ. Cell viability was analyzed using the MTT assay. Cell growth curves were derived from real-time cell analysis. Apoptosis was determined by flow cytometry, Aβ localization and expression by immunofluorescence, and ATP content by a specific assay kit. The expression of proteins related to the energy metabolism signaling pathway was analyzed using Western blotting. CK treatment improved cell viability, cell growth, and apoptosis induced by Aβ, and the cellular localization and expression of Aβ. Moreover, CK increased ATP content by promoting the activity of glucose transporters (GLUTs). Therefore, the neuroprotective effect of CK against Aβ injury was mainly realized through the activation of the energy metabolism signaling pathway. CK treatment inhibits neuronal damage caused by Aβ through the activation of the energy metabolism signaling pathway, revealing that CK might be one of the key bioactive ingredients of ginseng in the treatment of Alzheimer's disease and may serve as a preventive or therapeutic agent for Alzheimer's disease.

364 Impact of porcine reproductive and respiratory syndrome virus challenge and deoxynivalenol on intestinal integrity and nutrient transport

Abstract Porcine reproductive and respiratory syndrome (PRRS) virus infections and mycotoxin contaminated feedstuffs are two common issues facing the pork industry as both act, independently, to antagonize pig intestinal function, growth performance, and health. However, the combined impact of PRRS and mycotoxin exposure on pig intestinal health and function has not been investigated. Therefore, the objective of this experiment was to examine the impact of the mycotoxin deoxynivalenol (DON) on the intestinal integrity and function of PRRS infected pigs, employing an ex vivo model. Jejunum samples were collected from PRRS naïve (control; n = 6) and 21 day post inoculated PRRS virus infected (n = 8) gilts (10 weeks of age). In duplicate, explants were mounted into modified Ussing Chambers and incubated with or without 25 mM DON for 45 minutes prior to assessment of transepithelial resistance (TER), mucosal to serosal macromolecule (FD4) flux, and active glucose and glutamine transport. Data were analyzed using the mixed model procedure of SAS and a split plot design to examine the fixed effects of PRRS, DON, and their interaction. There were no PRRS by DON interactions for any ex vivo parameters assessed. Irrespective of DON, PRRS jejunum segments had decreased TERs (57%, P < 0.001), increased FD4 flux (P = 0.005), and tended to have increased active glucose transport (P = 0.093) compared with uninfected controls. Irrespective of PRRS status, DON increased FD4 flux (113%, P = 0.013), and tended to increase active glucose transport (P = 0.088) compared with jejunal explants not treated with DON. However, TERs and active glutamine transport did not differ due to DON treatment. Taken together, these data highlight the impacts that viral health challenge and mycotoxin contamination have on aspects of intestinal integrity and function. However, it does not appear that PRRS challenged pigs are more sensitive to the effects of DON than their healthy counterparts.

PSIV-18 Effect of sub-therapeutic antibiotics and pharmacological zinc oxide on nursery pig performance and intestinal integrity and function

Abstract The objective of this study was to evaluate the impact of sub-therapeutic chlortetracycline (CTC) and pharmacological zinc oxide on nursery pig performance and intestinal integrity and function. Twenty-four newly weaned pigs (4.51 ± 0.21 kg BW) were randomly assigned to one of three dietary treatments for 21 d (n = 8/treatment): 1) Control diet (NC), 2) NC + Zinc oxide at 3000 ppm for 7 d followed by 2000 ppm for 14 d (ZnO), and 3) NC + 40 ppm CTC for 21 d (sCTC). Pig ADG, ADFI, and G:F were determined for the overall 21 d test period. At d 21, all pigs were euthanized for tissue collection. Jejunum and ileum ex vivo transepithelial resistance (TER), macromolecule (FD4) flux, and active glucose and glutamine transport were assessed in modified Ussing Chambers. Additionally, jejunum brush border enzyme and Na+/K+ ATPase activities were determined. Morphology and T-cell abundance were determined in all ileum sections by H&E staining and immunohistochemistry, respectively. Compared to the NC, sCTC and ZnO increased ADG (0.14, 0.20 and 0.23 kg/d, respectively, P = 0.020). Overall ADFI was increased by 150% due to sCTC compared to the NC and ZnO pigs (P < 0.05). G:F was increased due to ZnO and sCTC compared to the NC (0.82, 0.63 and 0.47, respectively, P < 0.01). Jejunum and ileum ex vivo function and integrity did not differ. Ileal villus height was increased in the NC compared to sCTC treatment (P < 0.01), but not different from the ZnO treatment. In the ileum, total T-cell number per unit area was significantly lower in the ZnO segments compared to sCTC and NC (0.95, 2.29 and 2.59 count per μm2, respectively, P = 0.05). These data indicate that ZnO and sCTC improved pig performance but did not directly alter small intestinal integrity and function. However, mucosal T cell abundance was altered.

SGLT1 is required for the survival of triple‐negative breast cancer cells via potentiation of EGFR activity

Sodium/glucose cotransporter 1 (SGLT1), an essential active glucose transport protein that helps maintain high intracellular glucose levels, was previously shown to interact with epidermal growth factor receptor (EGFR); the SGLT1–EGFR interaction maintains intracellular glucose levels to promote survival of cancer cells. Here, we explore the role of SGLT1 in triple‐negative breast cancer (TNBC), which is the most aggressive type of breast cancer. We performed TCGA analysis coupled to in vitro experiments in TNBC cell lines as well as in vivo xenografts established in the mammary fat pad of female nude mice. Tissue microarrays of TNBC patients with information of clinical–pathological parameters were also used to investigate the expression and function of SGLT1 in TNBC. We show that high levels of SGLT1 are associated with greater tumour size in TNBC. Knockdown of SGLT1 compromises cell growth in vitro and in vivo. We further demonstrate that SGLT1 depletion results in decreased levels of phospho‐EGFR, and as a result, the activity of downstream signalling pathways (such as AKT and ERK) is inhibited. Hence, targeting SGLT1 itself or the EGFR–SGLT1 interaction may provide novel therapeutics against TNBC.

Rosmarinic acid improves hypertension and skeletal muscle glucose transport in angiotensin II-treated rats

BackgroundRosmarinic acid (RA) is a natural pure compound from herbs belonging to the Lamiaceae family, such as rosemary, sage, basil, and mint. The antioxidant, angiotensin-converting enzyme inhibitory, and vasodilatory effects of RA have been revealed. Angiotensin II (ANG II) is a potent agent that generates hypertension and oxidative stress. Hypertension and skeletal muscle insulin resistance are strongly related. The aim of this study was to evaluate the effects of acute and chronic RA treatment on blood pressure and skeletal muscle glucose transport in ANG II-induced hypertensive rats.MethodsEight-week-old male Sprague Dawley rats were separated into SHAM and ANG II-infused (250 ng/kg/min) groups. ANG II rats were treated with or without acute or chronic RA at 10, 20, or 40 mg/kg. At the end of the experiment, body weight, liver and heart weights, oral glucose tolerance, skeletal muscle glucose transport activity, and signaling proteins were evaluated.ResultsBoth acute and chronic RA treatment decreased systolic, diastolic, and mean arterial blood pressure. Only acute RA at 40 mg/kg resulted in a reduction of fasting plasma glucose levels and an induction of skeletal muscle glucose transport activity. These effects might involve increased ERK activity in skeletal muscle. Meanwhile, chronic RA treatment with 10, 20, and 40 mg/kg prevented ANG II-induced hyperglycemia.ConclusionsBoth acute and chronic RA treatment attenuated ANG II-induced cardiometabolic abnormalities in rats. Therefore, RA would be an alternative strategy for improving skeletal muscle glucose transport and protecting against ANG II-induced hypertension and hyperglycemia.

Human placental GLUT1 glucose transporter expression and the fetal insulin-like growth factor axis in pregnancies complicated by diabetes

We have previously described regulation of syncytial GLUT1 glucose transporters by IGF-I. Despite this, it is not clear what signal regulates transplacental glucose transport. In this report we asked whether changes in GLUT1 expression and glucose transport activity in diabetic pregnancies were associated with alterations in the fetal IGF axis. Cord blood samples and paired syncytial microvillous and basal membranes were isolated from normal term pregnancies and pregnancies characterized by gestational diabetes type A2 (GDM A2) and pre-existing insulin-dependent diabetes mellitus (IDDM). Circulating IGF-I, basal membrane GLUT1 expression and glucose transporter activity were correlated with birth weight, but only in control, not diabetic groups. Basal membrane GLUT1 and transporter activity were correlated with IGF-I concentrations in control, but not diabetic groups. IGF binding protein (IGFBP) binding capacity showed a ≥50% reduction in the diabetic groups compared to control; both showed a higher level of free IGF-I. The absence of a correlation between birth weight and factors such as fetal IGF-I or GLUT1 expression in the diabetic groups suggests that IGF-I-stimulated effects may have reached a limiting threshold, such that further increases in IGF-I (or GLUT1) are without effect. These data support that fetal IGF-I acts as a fetal nutritional signal, modulating placental GLUT1 expression and birth weight via altered levels of fetal circulating IGFBPs. Diabetes appears to exert its effects on fetal and placental factors prior to the third trimester and, despite good glycemic control immediately prior to, and in the third trimester, these effects persist to term.

Active transport of glucose across the jejunal epithelium decreases with age in broiler chickens

Intestinal glucose absorption varies with growth; however, the dynamics of these variations has not been yet fully elucidated in broiler chickens. The present study aimed to compare jejunal glucose uptake and maltose digestion in broilers of 2 different ages, i.e., 1- vs. 5 wk old. Oral D-maltose gavage, everted sac, and Ussing chamber experiments were carried out to investigate intestinal glucose absorption and mRNA expression of glucose-transport-related genes as well as jejunal maltase activity. Upon gavage, glucose concentrations peaked at 10min post-administration in 1-wk-old chicks, while they peaked at 40min in 5-wk-old chickens. Glucose concentrations at 10min were significantly higher in the 1-wk-old chicks (P = 0.010). Using the everted sacs experimental setup, 5 intestinal regions i.e., duodenum, proximal jejunum, distal jejunum, proximal ileum, and distal ileum, were targeted to examine D-maltose digestion and glucose transport across the intestinal mucosa. In the distal and proximal ileum, glucose concentrations were found to be significantly higher in the serosal compartment of the 1-wk-old chicks upon incubation with D-maltose (25mM) (P < 0.05), while in the mucosal compartment the levels were significantly higher in the 5-wk-old chickens (P < 0.05). An Ussing chamber setup was employed to measure glucose-induced short-circuit current (ΔIsc) in the mucosal epithelium of the jejunum. In response to the addition of D-maltose (10mM) into the mucosal compartment, ΔIsc was significantly higher in the 1-wk-old chicks (P = 0.018). Furthermore, no variations in jejunal maltase activity were observed between the 2 age groups. While jejunal glucose absorption was lower in the 5-wk-old chickens, the mRNA expression levels of jejunal SGLT1, GLUT2, and Na+/K+-ATPase did not show any significant differences between the 2 age groups. Our results suggest that the active transport of glucose across the jejunal epithelium decreases upon growth in broiler chickens but is not accompanied by any variations in maltase activity or in the expression of glucose-absorption-related genes.

Andrographis Paniculata Ameliorates Insulin Resistance in High Fat Diet-fed Mice and 3T3-L1 Adipocytes (P21-074-19)

ObjectivesPrevious studies have shown that Andrographis paniculata (AP) possesses antiinflammatory activity and is able to reduce blood glucose level in diabetes. The results suggested that AP reduced blood glucose level via activation of glucose transporter 4 (GLUT4) membrane translocation; however, the mechanism in adipocytes remained unclear. Proinflammatory cytokines have been shown to play a role in blood glucose dysregulation, which leads to hyperglycemia in vivo. The aims of the present study were to investigate the effects of bioactive compounds of AP on GLUT4 membrane translocation in 3T3-L1 cells and hyperglycemia in high fat diet-fed mice. MethodsThe ethanolic extract of AP (APE) was prepared and 14-deoxy-11,12-didehydroandrographolide (deAND) was purified from APE. The purity of deAND was identified by LC/MS, which showed the purity for deAND was 98%. The 6-weeks old C57BL/6JNarl mice were fed normal fat diet (ND, 12.1% kcal from fat), high fat diet (HFD, 60% kcal from fat), HFD plus 0.1% AND, 0.05% deAND, or 0.4% APE for 16 weeks. 3T3-L1 cells were treated with 10 ng/mL TNFα in the presence or absence of 20 μM AND, 20 μM deAND, or 100 μg/mL APE for 24 hours. The blood glucose and insulin levels were determined by commercial assay kits and the protein expression was assayed by Western blotting. ResultsHFD significantly induced hyperglycemia as compared with ND. Treatment with deAND and APE significantly decreased the fasting blood glucose and HOMA-IR as compared with HFD. TNFα, a proinflammatory cytokine, significantly inhibited glucose uptake in 3T3-L1 cells because impairment of IRS1-AKT-AS160 signaling pathway and GLUT4 membrane translocation. In contrast, treatment with AND, deAND, and APE attenuated the lower glucose uptake induced by TNFα. The protective effects of deAND and APE were associated with improvement of IRS1-AKT-AS160 signaling pathway, which resulted in more GLUT4 membrane translocation. ConclusionsHFD induced hyperglycemia and elevated blood TNFα level in vivo. TNFα dysregulated insulin signaling pathway in vitro. Treatment with deAND and APE reversed defects in IRS1-AKT-AS160-GLUT4 signaling pathway induced by TNFα in 3T3-L1 cells, which accounted for the anti-diabetic effect of AP. Funding SourcesThis study was supported by Ministry of Science and Technology, Taiwan.

Impairments in Protein Digestion and Absorption Attenuate the Anabolic Response to Feeding in Patients with Congestive Heart Failure (FS09-03-19)

ObjectivesGastrointestinal symptoms are prevalent extracardiac systemic manifestations of Congestive Heart Failure (CHF). We developed a comprehensive panel of methods to unravel gut dysfunction in CHF and its impact on the anabolic response to feeding. MethodsWe recruited 14 clinically stable CHF patients (ejection fraction: 33.9 ± 2.1, NYHA class: 2.3 ± 0.2) and 17 healthy controls matched for age and gender. Stable tracers of L-phenylalanine (PHE)-[ring-2H5] and L-tyrosine (TYR)-[13C9,15 N] were administered intravenously for 5 hours via primed constant and continuous infusion. After 2 hours, participants ingested a complete high protein meal containing L-PHE-[1–13C] and spirulina-[U-15 N]. We sampled blood throughout the study to analyze enrichments by LC-MS/MS. We calculated the anabolic response to feeding before and after correction for changes in protein digestion and absorption, assessed by spirulina degradation ratio (L-PHE-[15 N]/[1–13C]). Moreover, we measured small intestinal membrane integrity and active carrier-mediated glucose transport by urinary recovery of the orally ingested inert sugars lactulose, rhamnose, and 3-O-methyl-glucose. Disease severity was assessed by medical chart and history. Statistical analysis was performed by unpaired t-tests. Data are expressed as mean ± SEM. ResultsIn CHF patients, protein digestion and absorption were reduced (0.66 ± 0.04 vs. 0.82 ± 0.04, P < 0.01), which further attenuated the anabolic response to feeding (28.3 ± 3.8 vs. 54.0 ±5.5 μmol/kg FFM/meal, P < 0.001). Disturbances in protein digestion and absorption as well as anabolic response in CHF were independent of disease severity. Small intestinal permeability and active carrier-mediated glucose transport did not differ between the groups indicating a preserved enterocyte function in CHF patients. ConclusionsWe hypothesize that enhancing protein digestion and absorption in patients with CHF can improve the availability of nutrients and protein anabolism. Funding SourcesNational Institutes of Health

Glucose transporter inhibitor-conjugated insulin mitigates hypoglycemia

Insulin therapy in the setting of type 1 and advanced type 2 diabetes is complicated by increased risk of hypoglycemia. This potentially fatal complication could be mitigated by a glucose-responsive insulin analog. We report an insulin-facilitated glucose transporter (Glut) inhibitor conjugate, in which the insulin molecule is rendered glucose-responsive via conjugation to an inhibitor of Glut. The binding affinity of this insulin analog to endogenous Glut is modulated by plasma and tissue glucose levels. In hyperglycemic conditions (e.g., uncontrolled diabetes or the postprandial state), the in situ-generated insulin analog-Glut complex is driven to dissociate, freeing the insulin analog and glucose-accessible Glut to restore normoglycemia. Upon overdose, enhanced binding of insulin analog to Glut suppresses the glucose transport activity of Glut to attenuate further uptake of glucose. We demonstrate the ability of this insulin conjugate to regulate blood glucose levels within a normal range while mitigating the risk of hypoglycemia in a type 1 diabetic mouse model.

A Subsidiary Cell-Localized Glucose Transporter Promotes Stomatal Conductance and Photosynthesis.

It has long been recognized that stomatal movement modulates CO2 availability and as a consequence the photosynthetic rate of plants, and that this process is feedback-regulated by photoassimilates. However, the genetic components and mechanisms underlying this regulatory loop remain poorly understood, especially in monocot crop species. Here, we report the cloning and functional characterization of a maize (Zea mays) mutant named closed stomata1 (cst1). Map-based cloning of cst1 followed by confirmation with the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR associated protein 9 system identified the causal mutation in a Clade I Sugars Will Eventually be Exported Transporters (SWEET) family gene, which leads to the E81K mutation in the CST1 protein. CST1 encodes a functional glucose transporter expressed in subsidiary cells, and the E81K mutation strongly impairs the oligomerization and glucose transporter activity of CST1. Mutation of CST1 results in reduced stomatal opening, carbon starvation, and early senescence in leaves, suggesting that CST1 functions as a positive regulator of stomatal opening. Moreover, CST1 expression is induced by carbon starvation and suppressed by photoassimilate accumulation. Our study thus defines CST1 as a missing link in the feedback-regulation of stomatal movement and photosynthesis by photoassimilates in maize.

New insight into the classification and evolution of glucose transporters in the Metazoa.

Because glucose is an essential energy source for living organisms, glucose transporters (GLUTs) are present in all species worldwide. Encoded by the solute carrier family 2 gene family, the GLUT proteins generally have 12 transmembrane helices (TMHs). In total, 14 GLUT proteins have been identified in humans (hGLUTs), and they are divided into 3 classes on the basis of their transport characteristics and sequence similarities. Herein, we report the use of protein sequence similarity networks (SSNs) to visualize the sequence trends of 4101 GLUT proteins across the Metazoa. The SSNs separated the metazoan proteins into 3 new classes that were different from the traditional classification system. In the new system, 9 of the 14 hGLUTs (hGLUT1-5, 7, 9, 11, and 14) were grouped into class I, 3 (hGLUT10, 12, and 13) were grouped into class II, and 2 (hGLUT6 and 8) were grouped into class III, as also supported by the phylogenetic tree. Multiple sequence alignments further showed that the conserved residues in each class were different. Furthermore, the hGLUTs in each class showed unique evolutionary characteristics, with similar nonsynonymous-to-synonymous divergence ratios and similar regions under conservative selection pressure. Of note, GLUTs with 3, 6, 18, 24, and 36 TMHs were identified among the metazoan genomes, and 1 Chinese hamster protein with 6 TMHs showed GLUT activity. In summary, this large-scale sequence analysis provided new insights into the classification and evolution of GLUTs and further showed that gene duplication and fusion could have been important drivers during the evolution of these transporter molecules.-Jia, B., Yuan, D. P., Lan, W. J., Xuan, Y. H., Jeon, C. O. New insight into the classification and evolution of glucose transporters in the Metazoa.

Modeling studies of heterogeneities in glycolytic oscillations in HeLa cervical cancer cells.

Previous experiments demonstrated that a population of HeLa cells starved of glucose or both glucose and serum exhibited a strong heterogeneity in the glycolytic oscillations in terms of the number of oscillatory cells, periods of oscillations, and duration of oscillations. Here, we report numerical simulations of this heterogeneous oscillatory behavior in HeLa cells by using a newly developed mathematical model. It is simple enough that we can apply a mathematical analysis, but capture the core of the glycolytic pathway and the activity of the glucose transporter (GLUT). Lognormal distributions of the values of the four rate constants in the model were obtained from the experimental distributions in the periods of oscillations. Thus, the heterogeneity in the periods of oscillations can be attributed to the difference in the rate constants of the enzymatic reactions. The activity of GLUT is found to determine whether the HeLa cells were oscillatory or non-oscillatory under the same experimental conditions. Simulation with the log-normal distribution of the maximum uptake velocity of glucose and the four randomized rate constants based on the log-normal distributions successfully reproduced the time-dependent number of oscillatory cells (oscillatory ratios) under the two starving conditions. The difference in the initial values of the metabolites has little effect on the simulated results.

Antidiabetic Activity of Zingiber officinale Roscoe Rhizome Extract: an In Vitro Study

The potential roles of Zingiber officinale Roscoe (ginger) for treating and preventing diabetes have been investigated in both humans and experimental animals. However, the mode of its action has not yet been elucidated. This study aimed to investigate the effects of ginger extract on glucose uptake activity and its activation pathway in L6 myotubes. Cells were co-cultured for 24 h with a variable concentration of either ginger extract or 2 mM metformin or 200 nM insulin or 20 μM Troglitazone (TGZ), followed by a 10-min 2-[3H]-deoxy-D-glucose (2-DG) uptake. The levels of glucose transporters 1 (GLUT1) and GLUT4 protein and mRNA expression were determined. Ginger extract at 400 μg/ml significantly enhanced glucose uptake in L6 myotubes (208.03 ± 10.65% above basal value, p&lt;0.05) after co-culture for 24 h. The ginger-enhancement of glucose uptake was inhibited by 3.5 μM cycloheximide, a protein synthesis inhibitor, 1 μM wortmannin (Phosphatidylinositol 3-Kinase (PI3 kinase) inhibitor) and 15 nM rapamycin (mammalian target of rapamycin (mTOR) inhibitor). The enhancement of glucose transport by ginger extract at 400 μg/ml was accompanied with the increased expression of GLUT1 protein (1.60 ± 0.20, 2.03 ± 0.19, and 2.25 ± 0.35 folds of basal at 4, 8, and 24 h, respectively p&lt;0.05) and mRNA (1.22 ± 0.96, 1.45 ± 0.93, 1.91 ± 0.75, 2.32±0.92, and 2.20 ± 0.64 folds of basal at 1, 2, 4, 8, and 24 h, respectively p&lt;0.05) in a time-dependent manner. Z. officinale Roscoe rhizome extract increase glucose transport activity of L6 myotubes by enhancing GLUT1 expression, the results of PI3-Kinase and 5’-AMP-activated kinase (AMPK) stimulation.

Analysis of chemical constituents from Polygonatum cyrtonema after “Nine-Steam-Nine-Bask” processing

Seven new compounds, known as polygonatine N1‒N7 (1‒7), and a known compound (8) were isolated from the ‘Nine-Steam-Nine-Bask’ processing product of Polygonatum cyrtonema Hua. The compounds’ structures were determined by spectroscopic analysis. All compounds were tested for dipeptidyl peptidase-4 (DPP-4) inhibition, glucose transport, and anti-inflammatory activities. Compound 8 suppressed NO production with an IC50 of 35.4 μM.

An Evaluation of Boar Spermatozoa as a Biosensor for the Detection of Sublethal and Lethal Toxicity.

A novel, objective, and rapid computed motility inhibition (CMI) assay was developed to identify and assess sublethal injury in toxin-exposed boar spermatozoa and compared with a subjective visual motility inhibition (VMI) assay. The CMI values were calculated from digital micrographic videos using a custom MATLAB® script by contrasting the motility index values of each experiment with those of the background and control experiments. Following a comparison of the CMI and VMI assays results, it was determined that their agreement depended on the shape of the dose-response curve. Toxins that exhibited a steep slope were indicative of good agreement between the assays. Those depicted by a gentle decline in the slope of the dose-response curve, the CMI assay were shown to be two times more sensitive than the VMI assay. The CMI assay was highly sensitive to the inhibition of mitochondrial function and glucose transport activity by sublethal doses of toxins and to disruption of cellular cation homeostasis by carrier ionophoric toxins, when compared to the cytotoxicity and lethal toxicity assays (i.e., that evaluated the inhibition of cell proliferation in somatic cell lines (FL, PK-15, and MNA cells)) and disruption to spermatozoa membrane integrity. The CMI assay recognized subtle sublethal toxicity changes in metabolism, manifested as a decrease in boar spermatozoa motility. Thus, it was feasible to effectively compare the objectively-measured numerical values for motility inhibition using the CMI assay against those reflecting lethal damage in the spermatozoa cells and somatic cell lines using a cytotoxicity assay.

The Brown Alga Ascophyllum nodosum As A Nutraceutical Useful for the Control of Type II Diabetes

In the last few decades, diabetes has become a global health concern, due to its increasing prevalence in developed and developing countries. The most common form of diabetes is type 2 diabetes (T2D), characterized by an impairment of insulin in activating glucose transport system into the cells and an inappropriate hyper-insulinemia due to insulin resistance. The maintenance of normal blood glucose levels is one of the major goals in diabetes management and could be achieved by reducing the glycemic index (GI) of foods. The brown seaweed Ascophyllum nodosum is an easily available food traditionally used by coastal communities.

Molecular and functional characterization of glucose transporter genes of the fox tapeworm Echinococcus multilocularis

Alveolar echinococcosis (AE) is a zoonotic parasitosis caused by larvae of the fox tapeworm, Echinococcus multilocularis. E. multilocularis is distributed widely in the Northern hemisphere, causing serious health problems in various animals and humans. E. multilocularis, like other cestodes, lacks a digestive tract and absorbs essential nutrients, including glucose, across the syncytial tegument on its external surface. Therefore, it is hypothesized that E. multilocularis uses glucose transporters on its surface similar to a closely-related species, Taenia solium. Based on this hypothesis, we cloned and characterized glucose transporter homologues from E. multilocularis. As a result, we obtained full-length sequences of 2 putative glucose transporter genes (EmGLUT1 and EmGLUT2) from E. multilocularis. In silico analysis predicted that these were classified in the solute carrier family 2 group. Functional expression analysis using Xenopus oocytes demonstrated clear uptake of 2-deoxy-D-glucose (2-DG) by EmGLUT1, but not by EmGLUT2 in this experimental system. EmGLUT1 was shown to have relatively high glucose transport activity. Further analyses using the Xenopus oocyte system revealed that 2-DG uptake of EmGLUT1 did not depend on the presence or concentration of Na+ nor H+, respectively. Immunoblot analyses using cultured metacestode, ex vivo protoscolex, and adult worm samples demonstrated that both EmGLUTs were stably expressed during each developmental stage of the parasite. Based on the above-mentioned findings, we conclude that EmGLUT1 is a simple facilitated glucose transporter and possibly plays an important role in glucose uptake by E. multilocularis throughout its life cycle.