Glucose Transporter Protein Research Articles

The potential application of gold-apoferritin nanocages conjugated with 2-amino-2-deoxy-glucose for imaging of breast cancer cells.

Development of biocompatible and multifunctional nanoprobes for tumor targeting, imaging, and therapy still remains a great challenge. Herein, gold nanoparticles (AuNPs) were synthesized in the cavity of horse spleen apoferritin protein (HoSAF) and protein surface was labeled with 2-amino-2-deoxy-glucose (2DG) as a cell surface glucose transport protein specific targeting probe to study the feasibility of its usage as a computer tomography (CT) contrast agent with tumor targeting capability through in vitro experiments. 2DG conjugated and gold-loaded apoferritin (Au-HoSAF-2DG) nanoparticles (NPs) showed selective targeting for human breast adenocarcinoma (MCF-7) cells when compared to normal breast (MCF-10A) cells. This AuNP-based imaging agent was found to be non-cytotoxic in a given concentration range with an apoptotic effect upon longer exposure times towards MCF-7 cells, while MCF-10A cells were affected less. This selective cell death would also be useful for further cancer treatments with the ability of X-ray attenuation in in vitro X-ray and computed tomography (CT) imaging.

Targeting Cancer Metabolism to Enhance Tumor Radiation Response

A key emerging hallmark of cancer includes a deregulated cellular metabolism that can be reprogrammed to preferentially exhibit dependence on glycolysis over oxidative phosphorylation (OXPHOS) even in the presence of oxygen (Warburg effect). Pre-clinical data suggests that targeting lactate dehydrogenase (LDH) enzyme and components of mitochondrial OXPHOS metabolism in tumor cells can have a potent anti-tumor effects, yet the interactions of these anti-tumor approaches with standard therapies, such as RT is less clear. clonogenic assay, immunoblotting, tumor regrowth delay, Seahorse studies, LC/MS Mass spectrometry We characterized novel and specific inhibitors targeting LDH enzyme A and B isoform (LDHi) and novel OXPHOS inhibitor (IACS-010759) targeting mitochondrial complex I in combination with ionizing radiation (IR) for their anti-cancer and radio-sensitization effects across various tumor types. Targeting LDH enzyme in vitro leads to significant (p<0.001) reduction in extracellular lactate levels and radiosensitization of variety of tumor cell lines with a dose modification factor (DMF) ranging between 1.5-2.2. Clonogenic survival studies of variety of pancreatic, prostate and lung cancer cells indicated that inhibiting LDH in conjunction with IR, can also enhance radiosensitivity under both hypoxic (1% oxygen) (DMF = 2.0) and normoxic (DMF = 1.65) conditions while not affecting non-glycolytic/normal cells (1522, skin fibroblasts, DMF = 1.0) in vitro. Further, we established that this enhanced radiosensitivity upon LDH inhibition could be partly attributed to reduced DNA repair as seen by enhanced expression of gamma-H2AX (4 fold), reduced ATP generation (80% reduction compared to control) as well as altered expression of glucose and lactate transporter proteins (Glut-1, MCT-1, MCT-4, PFKP and PKM2) as seen by immunoblotting. Tumor regrowth delay studies with MiaPaCa and H460 xenografts demonstrated differential LDH response with no significant tumor radiosensitization. We attributed this failure to tumor metabolic reprogramming confirmed by increased oxygen consumption rate of tumor cells upon LDH inhibition in vitro. Targeting OXPHOS pathway by IACS-010759 in conjunction with LDHi promoted metabolic synthetic lethality in vitro (survival reduced to 10-5). Similar synthetic lethality was also observed in the H460 xenograft; additive enhancement in radiation response. However, inhibition of OXPHOS by complex I inhibitor and fractionated IR led to additive tumor regrowth delay in both MiaPaCa and H460 xenograft models. Furthermore, steady state global metabolomics study of inhibiting LDH and complex I of mitochondria either alone or in combination lead to significant decrease in glycolytic and TCA cycle intermediates leading to severe energy stress. In summary, our results presently indicate that targeting mitochondrial metabolism may be a more effective means of enhancing tumor radiosensitivity.

An insight into anti-adipogenic properties of an Oroxylum indicum (L.) Kurz extract

BackgroundOroxylum indicum fruit extract (OIE) has been reported to inhibit the development of adipocytes. However, the exact mechanism of its metabolic activity is not clearly defined. This study attempted to investigate whether OIE was involved in disrupting the cell cycle, glucose metabolism, and mitochondrial function in 3 T3-L1 cells.MethodsThe effect of the OIE on cell cycle progression was measured by flow cytometry along with observing the expression of the cycle regulator by immunoblotting. The effect of the OIE on glucose metabolism was investigated. The amount of glucose uptake (2-NBDG) influenced by insulin was determined as well as the protein tyrosine phosphorylation (PY20), and glucose transporter4 (GLUT4) expression was determined by immunoblotting assay. Mitochondria are also essential to metabolic processes. This study investigated mitochondrial activity using fluorescent lipophilic carbocyanine dye (JC-1) and mitochondria mass by MitoTracker Green (MTG) staining fluorescence dyes. Finally, cellular ATP concentration was measured using an ATP chemiluminescence assay.ResultsTreatment with OIE plus adipogenic stimulators for 24 h arrested cell cycle progression in the G2/M phase. Moreover, 200 μg/mL of OIE significantly diminished the expression of the insulin receptor (IR) and GLUT4 protein compared to the untreated-adipocytes (P < 0.05). The mitochondrial membrane potential (MMP) was significantly reduced (24 h) and increased (day 12) by OIE compared to untreated-adipocytes (P < 0.05). However, OIE maintained MMP and ATP at a similar level compared to the pre-adipocytes (day 12). Transmission electron microscope (TEM) results demonstrated that OIE could protect mitochondria deformation compared to the untreated-adipocytes.ConclusionThese results suggest that the inhibitory effect of the OIE on adipogenesis may potentially inhibit the cell cycle and phosphorylation of IR, leading to a decrease in glucose uptake to the cells. The OIE also slows down the mitochondrial activity of the early phase of cell differentiation, which can also inhibit the development of fat cells.

Abundance of the skeletal muscle Glut-4 glucose transport protein in Standardbred foals during development and exercise

The objective of this study was to investigate the changes in Glut-4 expression in gluteus muscle, and in the plasma concentrations of hormones and metabolites during the suckling period, after weaning, and after exercise training in foals. Our hypotheses were Glut-4 abundance will decrease following metabolite and hormonal concentration between neonatal period and late development but will increase with exercise training. Eight clinically normal Standardbred foals were used in this study, and they stayed with their mothers from birth until weaning at 6 months. After weaning, the foals were randomly divided in two groups: an exercise group (EX) (n=4; ~282 kg) which trained 3 d/wk for 12 wks and a control group (n=4; ~271 kg) without structured exercise. Venous blood samples were obtained immediately after birth (&lt;30 min), at 24 h, at day 7 and day 14, and at 1, 3, 6, 9 and 12 months after parturition. Results were analysed using one-way ANOVA for repeated measures. Post-hoc differences were identified using the Tukey test and correlations were obtained using Pearson product moment analysis. The abundance of Glut-4 in skeletal muscle was highest immediately after birth and with growth and development (P&lt;0.05). Exercise training did not change Glut-4 abundance (P&gt;0.05). Plasma lactate, insulin and cortisol concentrations decreased during the first three months of lactation while glucose and leptin concentrations increased (P&lt;0.05). There were no effects of exercise training on any of the above plasma parameters (P&gt;0.05). In conclusion, Glut-4 abundance in gluteus muscle was highest at the birth and decreased during development and exercise training for 12 weeks did not change its’ abundance.

MicroRNA-1291-5p Sensitizes Pancreatic Carcinoma Cells to Arginine Deprivation and Chemotherapy through the Regulation of Arginolysis and Glycolysis.

Cancer cells are dysregulated and addicted to continuous supply and metabolism of nutritional glucose and amino acids (e.g., arginine) to drive the synthesis of critical macromolecules for uncontrolled growth. Recent studies have revealed that genome-derived microRNA (miRNA or miR)-1291-5p (miR-1291-5p or miR-1291) may modulate the expression of argininosuccinate synthase (ASS1) and glucose transporter protein type 1 (GLUT1). We also developed a novel approach to produce recombinant miR-1291 agents for research, which are distinguished from conventional chemo-engineered miRNA mimics. Herein, we firstly demonstrated that bioengineered miR-1291 agent was selectively processed to high levels of target miR-1291-5p in human pancreatic cancer (PC) cells. After the suppression of ASS1 protein levels, miR-1291 perturbed arginine homeostasis and preferably sensitized ASS1-abundant L3.3 cells to arginine deprivation therapy. In addition, miR-1291 treatment reduced the protein levels of GLUT1 in both AsPC-1 and PANC-1 cells, leading to a lower glucose uptake (deceased > 40%) and glycolysis capacity (reduced approximately 50%). As a result, miR-1291 largely improved cisplatin efficacy in the inhibition of PC cell viability. Our results demonstrated that miR-1291 was effective to sensitize PC cells to arginine deprivation treatment and chemotherapy through targeting ASS1- and GLUT1-mediated arginolysis and glycolysis, respectively, which may provide insights into understanding miRNA signaling underlying cancer cell metabolism and development of new strategies for the treatment of lethal PC.SIGNIFICANCE STATEMENTMany anticancer drugs in clinical use and under investigation exert pharmacological effects or improve efficacy of coadministered medications by targeting cancer cell metabolism. Using new recombinant miR-1291 agent, we revealed that miR-1291 acts as a metabolism modulator in pancreatic carcinoma cells through the regulation of argininosuccinate synthase– and glucose transporter protein type 1–mediated arginolysis and glycolysis. Consequently, miR-1291 effectively enhanced the efficacy of arginine deprivation (pegylated arginine deiminase) and chemotherapy (cisplatin), offering new insights into development of rational combination therapies.

TUG is a calpain-10 substrate involved in the translocation of GLUT4 in adipocytes.

The calpain-10 (CAPN10) protease is implicated in the translocation of the glucose transporter 4 (GLUT4), which is retained in the Golgi matrix via the Tether containing a UBX domain for GLUT4 (TUG) protein. Insulin stimulation induces the proteolytic processing of TUG, which leads to the translocation of GLUT4 to the cell membrane. We tested whether TUG is a CAPN10 substrate. Proteolysis of TUG by calpains was assessed using a cell-free system containing calpain-1 and TUG. In situ proteolysis of TUG by calpains was demonstrated in 3T3-L1 adipocytes in the presence of insulin or calpain inhibitors to modulate calpain activity. Proteolysis of TUG by CAPN10 was confirmed using transient or stable silencing of CAPN10 in 3T3-L1 adipocytes. Calpains proteolyzed the C-terminus of TUG in vitro. In adipocytes, insulin-induced cleavage of TUG was correlated with the activation of calpains. Treatment with calpain inhibitors reduced TUG cleavage, resulting in impaired GLUT4 translocation without altering Akt phosphorylation. Furthermore, CAPN10 but not calpain-1 or calpain-2 colocalized with GLUT4 in the absence of insulin, and their colocalization was reduced after stimulation with insulin. Finally, we demonstrated that CAPN10 knockdown reduced the proteolysis of TUG without altering the phosphorylation of Akt or the expression of the Usp25m protease. Thus, our results provide evidence that the TUG protein is cleaved by CAPN10 to regulate GLUT4 translocation.

MiR-150 regulates glucose utilization through targeting GLUT4 in insulin-resistant cardiomyocytes.

MicroRNAs (miRNAs) play an important role in cardiac function and metabolism. However, whether they regulate insulin resistance (IR) of cardiomyocytes remains unclear. The aim of the present study was to shed light on this issue with a focus on miR-150. We found here that miR-150 level was elevated in myocardium of type 2 diabetes mellitus (T2DM) rat model and in insulin-resistant cardiomyocytes induced by high glucose (25 mM) and high insulin (1 μM). Deregulation of miR-150 downregulated the protein and mRNA levels of glucose transporter 4 (GLUT4) as assessed by western blot, real-time polymerase chain reaction (qPCR), and immunofluorescence assays. Overexpression of miR-150 inhibited glucose utilization in cardiomyocytes as detected by 2-deoxyglucose transport and glucose consumption assays. In contrast, knockdown of miR-150 significantly increased glucose uptake in cardiomyocytes. Moreover, GLUT4 translocation was increased after transfection of miR-150 inhibitor (AMO-150). Collectively, miR-150 reduced glucose utilization by directly decreasing the expression and translocation of GLUT4 in the cardiomyocytes with IR and therefore might be a new therapeutic target for metabolic diseases such as T2DM.

18F]FDG Uptake in Adipose Tissue Is Not Related to Inflammation in Type 2 Diabetes Mellitus

Purpose2-deoxy-2-[18F]fluoro-d-glucose ([18F]FDG) uptake is a marker of metabolic activity and is therefore used to measure the inflammatory state of several tissues. This radionuclide marker is transported through the cell membrane via glucose transport proteins (GLUTs). The aim of this study is to investigate whether insulin resistance (IR) or inflammation plays a role in [18F]FDG uptake in adipose tissue (AT).ProceduresThis study consisted of an in vivo clinical part and an ex vivo mechanistic part. In the clinical part, [18F]FDG uptake in abdominal visceral AT (VAT) and subcutaneous AT (SAT) was determined using PET/CT imaging in 44 patients with early type 2 diabetes mellitus (T2DM) (age 63 [54–66] years, HbA1c [6.3 ± 0.4 %], HOMA-IR 5.1[3.1–8.5]). Plasma levels were measured with ELISA. In the mechanistic part, AT biopsies obtained from 8 patients were ex vivo incubated with [18F]FDG followed by autoradiography. Next, a qRT-PCR analysis was performed to determine GLUT and cytokine mRNA expression levels. Immunohistochemistry was performed to determine CD68+ macrophage infiltration and GLUT4 protein expression in AT.ResultsIn vivo VAT [18F]FDG uptake in patients with T2DM was inversely correlated with HOMA-IR (r = − 0.32, p = 0.034), and positively related to adiponectin plasma levels (r = 0.43, p = 0.003). Ex vivo [18F]FDG uptake in VAT was not related to CD68+ macrophage infiltration, and IL-1ß and IL-6 mRNA expression levels. Ex vivo VAT [18F]FDG uptake was positively related to GLUT4 (r = 0.83, p = 0.042), inversely to GLUT3 (r = − 0.83, p = 0.042) and not related to GLUT1 mRNA expression levels.ConclusionsIn vivo [18F]FDG uptake in VAT from patients with T2DM is positively correlated with adiponectin levels and inversely with IR. Ex vivo [18F]FDG uptake in AT is associated with GLUT4 expression but not with pro-inflammatory markers. The effect of IR should be taken into account when interpreting data of [18F]FDG uptake as a marker for AT inflammation.

Propofol disrupts cell carcinogenesis and aerobic glycolysis by regulating circTADA2A/miR-455-3p/FOXM1 axis in lung cancer

ABSTRACT The involvement of propofol and circular RNAs (circRNAs) in lung cancer progression has been identified. However, the relationship between propofol and circRNAs as well as the underlying molecular mechanisms on lung cancer development remain unclear. Cell viability, migration and invasion were measured by cell counting kit-8 assay, 5-bromo-2-deoxyuridine (BrdU) and transwell assay. Glycolytic metabolism was calculated by measuring the glucose consumption, lactate production and extracellular acidification. Western blot was used to detect the protein of glucose transporter 1 (GLUT1), glycolysis enzymes, and forkhead box M1 (FOXM1). The expression of circRNA transcriptional adaptor 2A (circTADA2A), microRNA (miR)-455-3p and FOXM1 mRNA was detected by quantitative real-time polymerase chain reaction. The interaction between miR-455-3p and circTADA2A or FOXM1 was analyzed using the dual-luciferase reporter assay. Murine xenograft model was established to perform in vivo experiments. We found propofol treatment alleviated lung cancer cell proliferation, migration, invasion and aerobic glycolysis in vitro as well as inhibited tumor growth in vivo. Propofol decreased the level of circTADA2A and exerted anti-tumor effects by regulating circTADA2A. MiR-455-3p directly interacted with circTADA2A and FOXM1 in lung cancer cells, and circTADA2A could regulate FOXM1 expression by binding to miR-455-3p. Subsequently, rescue assay showed that propofol inhibited cell proliferation, migration, invasion and aerobic glycolysis by regulating circTADA2A/miR-455-3p/FOXM1 axis in lung cancer. Collectively, propofol suppressed cell carcinogenesis and aerobic glycolysis by regulating circTADA2A/miR-455-3p/FOXM1 axis in lung cancer, providing an effective clinical implication for propofol to prevent the development of lung cancer.

Sex hormone-binding globulin regulates glucose metabolism in human placental trophoblasts via cAMP/PKA/CREB1.

AimThis study was conducted to investigate the effects of sex hormone‐binding globulin (SHBG) on glucose metabolism and insulin resistance in human placental trophoblasts and involvement of the cAMP/PKA/CREB1 signaling pathway in these effects.MethodsAn insulin resistance cell model of human trophoblasts was established. An SHBG‐overexpression plasmid was transfected into these cells, and the expression of glucose transporter 1 (GLUT1), CREB and p‐CREB was detected and analyzed in normal cells, model cells and all groups of transfected cells by real‐time PCR and western blotting; cAMP, PKA, glucose consumption and pyruvic acid levels were also detected.ResultsAmong the four groups, there was no significant difference in the expression of CREB mRNA or GLUT1 mRNA (P > 0.05); however, CREB, p‐CREB, GLUT1 protein, cAMP and PKA showed low expression (P < 0.05) and cell glucose consumption and pyruvate production were decreased (P < 0.05) in the model group, compared to the normal group. SHBG overexpression in insulin‐resistant cells partially increased the levels of p‐CREB, GLUT1, cAMP and PKA (P < 0.05). Intracellular glucose consumption and pyruvate production were nearly restored to the levels observed in cells from the normal group.ConclusionSex hormone‐binding globulin regulates GLUT1 expression via the cAMP/PKA/CREB1 pathway and affects glucose transport in the placenta, which can induce insulin resistance and gestational diabetes.

Sex-dimorphic moderate hypoglycemia preconditioning effects on Hippocampal CA1 neuron bio-energetic and anti-oxidant function.

Hypoglycemia is a detrimental complication of rigorous management of type 1 diabetes mellitus. Moderate hypoglycemia (MH) preconditioning of male rats partially affords protection from loss of vulnerable brain neurons to severe hypoglycemia (SH). Current research investigated whether MH preconditioning exerts sex-dimorphic effects on hippocampal CA1 neuron bio-energetic and anti-oxidant responses to SH. SH up-regulated CA1 glucose or monocarboxylate transporter proteins in corresponding hypoglycemia-naïve male versus female rats; precedent MH amplified glucose transporter expression in SH irrespective of sex. Sex-differentiating SH effects on glycolytic and tricarboxylic pathway markers correlated with elevated tissue ATP content and diminished CA1 5'-AMP-activated protein kinase (AMPK) activation in females. MH-preconditioned suppression of mitochondrial energy pathway enzyme profiles and tissue ATP in SH rats coincided with amplified CA1 AMPK activity in both sexes. Anti-oxidative stress enzyme protein responses to SH were primarily sex-contingent; preconditioning amplified most of these profiles, yet exacerbated expression of lipid and protein oxidation markers in SH male and female rats, respectively. Results show that MH preconditioning abolishes female CA1 neuron neuroprotection of positive energy balance through SH, resulting in augmented CA1 AMPK activity and oxidative injury and diminished tissue ATP in hypoglycemia-conditioned versus naïve rats in each sex. It is unclear if SH elicits differential rates of CA1 neuronal destruction in the two sexes, or how MH may impact sex-specific cell loss. Further research is needed to determine if molecular mechanism(s) that maintain female CA1 neuron metabolic stability in the absence of MH preconditioning can be leveraged for therapeutic prevention of hypoglycemic nerve cell damage.

Evaluating the effect of arachidonic acid and eicosapentaenoic acid on induction of adipogenesis in human adipose-derived stem cells.

Objective(s):Adipose tissue is one of the most important endocrine organs that liberates many metabolic mediators such as hormones, cytokines, and chemokines. Different types of fatty acids have key roles in adipogenesis. The aim of this study was to evaluate the effects of two essential fatty acids, including Arachidonic acid (AA) and Eicosapentaenoic acid (EPA), on the process of adipogenicity in human Adipose-Derived Stem Cells (hADSCs). Materials and Methods:After immunophenotyping of hADSCs by flowcytometry, they were differentiated into adipocytes and simultaneously exposed to 30 μM and 60 μM of AA and 25 μM and 50 μM of EPA. Further, along with the MTS assay, the activity of glycalaldehyde-3-phosphate dehydrogenase (GAPDH) was also measured. In addition, expression of lipid markers including peroxisome proliferator-activated receptor γ2 (PPARγ2) and glucose transporter 4 (GLUT4) was evaluated, and the neutral lipid contents were determined using Oil red O staining.Results:MTS evaluation showed a significant decrease in proliferation in all treatment groups compared to the control group. Based on oil red O staining, fat droplets in the AA treatment groups were higher than in controls. The expression of PPARγ2 and GLUT4 genes and proteins increased in almost all AA and EPA groups compared to control. In addition, GAPDH activity was higher in AA groups than in the control group. In general, while different concentrations of EPA did not increase the adipogenic process compared to the control group, stimulation of differentiation to adipocytes was largely determined by the AA. Conclusion:The result indicates a positive effect of omega-6 versus omega-3 in stimulating the pathways of adipogenesis.

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Host tissue response to floating microelectrode arrays chronically implanted in the feline spinal nerve

Objective. Neural interfacing technologies could significantly improve quality of life for people living with the loss of a limb. Both motor commands and sensory feedback must be considered; these complementary systems are segregated from one another in the spinal nerve. Approach. The dorsal root ganglion-ventral root (DRG-VR) complex was targeted chronically with floating microelectrode arrays designed to record from motor neuron axons in the VR or stimulate sensory neurons in the DRG. Hematoxylin and eosin and Nissl/Luxol fast blue staining were performed. Characterization of the tissue response in regions of interest and pixel-based image analyses were used to quantify MAC387 (monocytes/macrophages), NF200 (axons), S100 (Schwann cells), vimentin (fibroblasts, endothelial cells, astrocytes), and GLUT1 (glucose transport proteins) reactivity. Implanted roots were compared to non-implanted roots and differences between the VR and DRG examined. Main results. The tissue response associated with chronic array implantation in this peripheral location is similar to that observed in central nervous system locations. Markers of inflammation were increased in implanted roots relative to control roots with MAC387 positive cells distributed throughout the region corresponding to the device footprint. Significant decreases in neuronal density and myelination were observed in both the VR, which contains only neuronal axons, and the DRG, which contains both neuronal axons and cell bodies. Notably, decreases in NF200 in the VR were observed only at implant times less than ten weeks. Observations related to the blood-nerve barrier and tissue integrity suggest that tissue remodeling occurs, particularly in the VR. Significance. This study was designed to assess the viability of the DRG-VR complex as a site for neural interfacing applications and suggests that continued efforts to mitigate the tissue response will be critical to achieve the overall goal of a long-term, reliable neural interface.

PdpaMn inhibits fatty acid synthase-mediated glycolysis by down-regulating PI3K/Akt signaling pathway in breast cancer.

Novel manganese complex, PdpaMn ([(Pdpa)MnCl2]), was developed to induce apoptosis in breast cancer cells. The impact of phosphoinositide-(3)-kinase pathway onto fatty acid synthase (FASN) has an effect on cellular metabolism in breast cancer. However, reverse actions from FASN towards PI3K/Akt are still indefinable. Perhaps, loss of FASN could regulate glycolysis. Previously we established that PdpaMn inhibits FASN and involve in mitochondrial function. This study investigated the activity of PdpaMn on glycolysis and its mechanism. PdpaMn was used to suppress FASN expression in tumor. Expression of ATP and lactic acid level was measured to investigate the glycolysis variance in cells and animals. MCF-7 and 4T1 cells were treated with G28UCM, an inhibitor of FASN and PdpaMn, western blotting to detect PI3K/Akt signaling pathway. The capacity of proliferation was investigated by western blotting and immunohistochemistry. PdpaMn selectively inhibits cancer cells and tumor growth but also block FASN expression and suppresses the content of free fatty acid. Lactate dehydrogenase (LDHA) protein level was down-regulated as G28UCM and PdpaMn inhibited FASN, glucose transporter (Glut1), and pyruvate kinase (PKM2) proteins level were not affected. PI3K, p-Akt in the experimental group evidently declined compared to the control group. Proliferation was suppressed in FASN-arbitrated glycolysis. Our study supports the hypothesis that loss of FASN by PdpaMn suppressed glycolysis via down-regulating PI3K/Akt signaling pathway revealing the direct link between FASN and glycolysis. The results have paved the way to unravel the mechanisms of FASN and mitochondrial will be useful for designing novel co-targeting strategies for breast cancer.

Glycosylated-imidazole aldoximes as reactivators of pesticides inhibited AChE: Synthesis and in-vitro reactivation study

The present armamentarium of commercially available antidotes provides limited protection against the neurological effects of organophosphate exposure. Hence, there is an urgent need to design and develop molecules that can protect and reactivate inhibited-AChE in the central nervous system. Some natural compounds like glucose and certain amino acids (glutamate, the anion of glutamic acid) can easily cross the blood brain barrier although they are highly polar. Glucose is mainly transported by systems like glucose transporter protein type 1 (GLUT1). For this reason, a series of non-quaternary and quaternary glycosylated imidazolium oximes with different alkane linkers have been designed and synthesized. These compounds were evaluated for their in-vitro reactivation ability against pesticide (paraoxon-ethyl and paraoxon-methyl) inhibited-AChE and compared with standards antidote AChE reactivators pralidoxime and obidoxime. Several physicochemical properties including acid dissociation constant (pKa), logP, logD, HBD and HBA, have also been assessed for reported compounds. Out of the synthesized compounds, three have exhibited comparable potency with a standard antidote (pralidoxime).

Effects of acute and chronic exercise on fat PI3K/AKT/GLUT4 signal pathway in type 2 diabetic rats

To study the effects of acute and chronic exercise on phosphatidylinositol 3-hydroxy kinase(PI3K)/protein kinase B(AKT)/glucose transporter 4(GLUT4)signaling pathway in adipose tissue of rats with type 2 diabetes mellitus (T2DM) induced by high-fat diet and low-dose streptozotocin (STZ). A total of 52 SD male rats aged 15 months were randomly divided into normal control group (13) and high-fat group (39), and fed normal and high fat diets. After 8 weeks, the body weight of the high-fat group was higher 20% than that of the normal control group. After a small dose of STZ, the blood glucose level was ＞16.7 mmol/l, and the model was successfully established. The diabetic model group was randomly divided into a diabetic control group (DC, n=13), a diabetic chronic exercise group (DCE, n=13), and a diabetic acute exercise group (DAE, n=13). The DCE group underwent an 8-week swimming exercise and the DAE group performed a one-time swimming exercise. Blood lipids, blood glucose and serum insulin levels were measured, and the contents of fat PI3K, AKT and GLUT4 proteins were determined by Western blot method. The levels of body weight, blood lipids, blood glucose and insulin in the diabetic group were significantly higher than those in the normal control group (P＜0.01); high density liptein cholesterol(HDL-C) levels were decreased (P＜0.05), and the expressions of PI3K, AKT and GLUT4 protein in adipose tissue were decreased (P＜0.01). After 8 weeks of swimming training, the levels of body weight, blood lipids, blood glucose and insulin all were decreased significantly (P＜0.01); while the level of HDL-C was increased (P＜0.05), and the expressions of PI3K, AKT and GLUT4 protein were increased (P＜0.01). After acute exercise, the levels of blood lipids, blood glucose and insulin were decreased (P＜0.05); the level of HDL-C was increased (P＜0.05), and the expression levels of fat PI3K, AKT and GLUT4 were increased significantly (P＜0.05). ①High fat diet combined with low-dose STZ induced damage to the PI3K/AKT pathway in adipose tissue of T2DM rats reduced insulin sensitivity. ②Acute and chronic aerobic exercise can improve the disorder of glucose and lipid metabolism through PI3K/AKT pathway, and the chronic exercise is better than acute exercise.

MiR-200c inhibits proliferation and promotes apoptosis of Wilms tumor cells by regulating akt signaling pathway.

The aim of this study was to explore the effect of micro ribonucleic acid (miR)-200c on the proliferation and apoptosis of Wilms tumor cells, and to further elucidate its potential mechanisms. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was used to detect the expression level of miR-200c in cancer tissues and adjacent normal tissues of 20 patients with Wilms tumor. Human primary Wilms tumor cells were taken as research objects, and were divided into Control group and miR-200c mimic group. In miR-200c mimic group, miR-200c was overexpressed in Wilms tumor cells using liposome transfection technology. Subsequently, the proliferation and apoptosis of cells in each group were observed by functional assays. The expression levels of phosphorylated protein kinase B (p-Akt), total Akt (T-Akt) and glucose transporter protein type 1 (GLUT1) in each group of cells were finally detected by Western blotting. In Wilms tumor patients, the expression level of miR-200c in cancer tissues was notably lower than that in adjacent normal tissues (p<0.05). Wilms tumor cells were cultured in vitro, and were transfected with miR-200c mimic. Subsequent cell counting kit-8 (CCK-8) assay results showed that the proliferation ability of cells in miR-200c mimic group was remarkably weakened (p<0.05). Colony formation assay indicated the number of formed colonies in miR-200c mimic group was remarkably less than that in Control group (p<0.05). Western blotting results manifested that overexpression of miR-200c markedly increased the ratio of B-cell lymphoma protein 2 (Bcl-2) to Bcl-2-associated X protein (Bax) (p<0.05). Flow cytometry results revealed that miR-200c overexpression significantly elevated the apoptosis rate of Wilms tumor cells (p<0.05). In addition, it was discovered that the overexpression of miR-200c could prominently reduce the phosphorylation level of intracellular Akt and the expression of its downstream protein GLUT1. Finally, immunohistochemical staining results verified that the expression levels of p-Akt and GLUT1 in cancer tissues of Wilms tumor patients were significantly higher than those in adjacent normal tissues (p<0.05). MiR-200c was lowly expressed in cancer tissues of Wilms tumor patients. Besides, overexpression of miR-200c inhibited the proliferation and promoted the apoptosis of cells through targeted inhibition of the Akt/GLUT1 signaling pathway.

Fundamentals about onset and progressive disease character of type 2 diabetes mellitus.

ResearchGate is a world wide web for scientists and researchers to share papers, ask and answer questions, and find collaborators. As one of the more than 15 million members, the author uploads research output and reads and responds to some of the questions raised, which are related to type 2 diabetes. In that way, he noticed a serious gap of knowledge of this disease among medical professionals over recent decades. The main aim of the current study is to remedy this situation through providing a comprehensive review on recent developments in biochemistry and molecular biology, which can be helpful for the scientific understanding of the molecular nature of type 2 diabetes. To fill up the shortcomings in the curricula of medical education, and to familiarize the medical community with a new concept of the onset of type 2 diabetes, items are discussed like: Insulin resistance, glucose effectiveness, insulin sensitivity, cell membranes, membrane flexibility, unsaturation index (UI; number of carbon-carbon double bonds per 100 acyl chains of membrane phospholipids), slow-down principle, effects of temperature acclimation on phospholipid membrane composition, free fatty acids, energy transport, onset of type 2 diabetes, metformin, and exercise. Based on the reviewed data, a new model is presented with proposed steps in the development of type 2 diabetes, a disease arising as a result of a hypothetical hereditary anomaly, which causes hyperthermia in and around the mitochondria. Hyperthermia is counterbalanced by the slow-down principle, which lowers the amount of carbon-carbon double bonds of membrane phospholipid acyl chains. The accompanying reduction in the UI lowers membrane flexibility, promotes a redistribution of the lateral pressure in cell membranes, and thereby reduces the glucose transporter protein pore diameter of the transmembrane glucose transport channel of all Class I GLUT proteins. These events will set up a reduction in transmembrane glucose transport. So, a new blood glucose regulation system, effective in type 2 diabetes and its prediabetic phase, is based on variations in the acyl composition of phospholipids and operates independent of changes in insulin and glucose concentration. UI assessment is currently arising as a promising analytical technology for a membrane flexibility analysis. An increase in mitochondrial heat production plays a pivotal role in the existence of this regulation system.

Non-insulin therapies in addition to insulin in Type 1 DM treatment.

Complications of Type 1 diabetes (T1DM) remain prevalent due to suboptimal glycaemic control despite advances in analogue insulin, its delivery and technological advances in glucose monitoring. Intensive insulin therapy is associated with hypoglycaemia and weight gain. Non-insulin-dependent glucose lowering strategies may provide a strategy in improving glycaemic control without hypoglycaemia and weight gain. Research papers and reviews about adjunctive treatment with insulin in T1DM in the published literature. Non-insulin-dependent strategies may be beneficial inT1DM particularly when there is insulin resistance, but the evidence for benefit at the current time is limited. Although there have been trials with various drugs as adjunctive therapy to insulin in T1DM currently in the UK, there is only one sodium glucose transport protein 2 (SGLT2) inhibitor with a marketing authorization for use in this indication. Potential for harm with SGLT2 inhibitors in T1DM is a potential issue, particularly euglycaemic diabetic ketoacidosis. Clinical trials confirm that there is a risk albeit small, but emerging safety data have led to questions as to whether the risk of euglycaemic diabetic ketoacidosis is higher with the use of SGLT2 inhibitors in clinical practice. Patient education is paramount-the work being done in T1DM to ensure safe use of SGLT2 inhibitors may help improve safety in the prescribing of SGLT2 inhibitors in Type 2 diabetes. There is a need for larger clinical trials with SGLT2 inhibitors in T1DM and real world studies to clarify safety.