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1.
Ann Palliat Med ; 10(8): 8746-8752, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34488364

RESUMO

BACKGROUND: The early diagnosis and treatment of diabetes mellitus has significant clinical benefits. However, the current diagnostic tools available for community-based populations are limited. This study sought to evaluate the clinical benefits of combining serum 1,5-anhydroglucitol (1,5-AG) with fasting plasma glucose (FPG) to detect diabetes in a community-based population with hypertension. METHODS: A total of 359 subjects were enrolled in this diagnostic study, all of whom underwent a 75-g oral glucose tolerance test (OGTT). Venous blood samples were collected to measure FPG, 2 h postprandial plasma glucose (2h-PG), and hemoglobin A1c (HbA1c). Serum 1,5-AG levels were tested using the Glycomark assay, and a receiver operating characteristic (ROC) curve was used to assess the diagnostic value of this tool for diabetes and determine the optimal cut-point value to provide the maximum Youden's index. A Spearman correlation analysis was performed to analyze the relationship between 1,5-AG and other indexes. RESULTS: A total of 102 participants were diagnosed with diabetes, indicating a prevalence of 28.4% in the community-based population. The Spearman correlation analysis showed that 1,5-AG was negatively correlated with FPG and 2h-PG (r=-0.367 and -0.487, respectively; both P<0.05). For the estimation of 2h-PG ≥11.1 mmol/L using 1,5-AG, the area under the curve (AUC) for the ROC analysis was 0.850 (95% confidence interval: 0.809-0.891). The corresponding optimal cut-off for 1,5-AG was 13.23 µg/mL, which yielded a sensitivity of 89.7% and a specificity of 73.5%. Compared with FPG alone, FPG combined with 1,5-AG had a higher sensitivity for detecting diabetes (97.1% vs. 47.1%; P<0.001). CONCLUSIONS: FPG combined with 1,5-AG substantially improved the sensitivity in detecting diabetes relative to FPG alone in a community-based population with hypertension, and may be a simple and efficient tool for screening diabetes.


Assuntos
Diabetes Mellitus , Hipertensão , Glicemia , Desoxiglucose , Diabetes Mellitus/diagnóstico , Jejum , Humanos , Hipertensão/diagnóstico
2.
Molecules ; 26(16)2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-34443330

RESUMO

5-Hydroxymethylfurfural (5-HMF) is a harmful substance generated during the processing of black garlic. Our previous research demonstrated that impregnation of black garlic with epigallocatechin gallate (EGCG) could reduce the formation of 5-HMF. However, there is still a lack of relevant research on the mechanism and structural identification of EGCG inhibiting the production of 5-HMF. In this study, an intermediate product of 5-HMF, 3-deoxyglucosone (3-DG), was found to be decreased in black garlic during the aging process, and impregnation with EGCG for 24 h further reduced the formation of 3-DG by approximately 60% in black garlic compared with that in the untreated control. The aging-mimicking reaction system of 3-DG + EGCG was employed to determine whether the reduction of 3-DG was the underlying mechanism of decreased 5-HMF formation in EGCG-treated black garlic. The results showed that EGCG accelerated the decrease of 3-DG and further attenuated 5-HMF formation, which may be caused by an additional reaction with 3-DG, as evidenced by LC-MS/MS analysis. In conclusion, this study provides new insights regarding the role of EGCG in blocking 5-HMF formation.


Assuntos
Catequina/análogos & derivados , Desoxiglucose/análogos & derivados , Furaldeído/análogos & derivados , Alho/efeitos dos fármacos , Alho/metabolismo , Catequina/farmacologia , Desoxiglucose/biossíntese , Desoxiglucose/metabolismo , Relação Dose-Resposta a Droga , Furaldeído/metabolismo
3.
IUBMB Life ; 73(10): 1198-1204, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34418270

RESUMO

Mechanisms discovered to drive increased glucose metabolism in cancer cells are found to be similar to those in viral-infected cells. In this mini review, we summarize the major pathways by which the sugar analog, 2-Deoxy-d-glucose, has been shown to exploit increased glucose metabolism in cancer and how this information applies to viral-infected cells. Moreover, we highlight the relevance of these findings to the emergency approval of 2-Deoxy-d-glucose in India to be used against SARS-CoV-2, the virus responsible for COVID-19.


Assuntos
COVID-19/metabolismo , Desoxiglucose/farmacologia , Glucose/metabolismo , Neoplasias/metabolismo , SARS-CoV-2/efeitos dos fármacos , COVID-19/virologia , Humanos , Índia , SARS-CoV-2/isolamento & purificação , SARS-CoV-2/patogenicidade
4.
J Vet Sci ; 22(4): e55, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34313040

RESUMO

BACKGROUND: Naringenin and its glycoside naringin are well known citrus flavonoids with several therapeutic benefits. Although the anti-adipogenic effects of naringenin and naringin have been reported previously, the detailed mechanism underlying their anti-adipogenesis effects is poorly understood. OBJECTIVES: This study examined the anti-adipogenic effects of naringenin and naringin by determining differential gene expression patterns in these flavonoids-treated 3T3-L1 adipocytes. METHODS: Lipid accumulation and triglyceride (TG) content were determined by Oil red O staining and TG assay. Glucose uptake was measured using a 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose fluorescent d-glucose analog. The phosphorylation levels of AMP-activated protein kinase (AMPK) and acetyl Co-A carboxylase (ACC) were observed via Western blot analysis. Differential gene expressions in 3T3-L1 adipocytes were evaluated via RNA sequencing analysis. RESULTS: Naringenin and naringin inhibited both lipid accumulation and TG content, increased phosphorylation levels of both AMPK and ACC and decreased the expression level of 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) in 3T3-L1 adipocytes. RNA sequencing analysis revealed that 32 up-regulated (> 2-fold) and 17 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Scd1, Mogat1, Dgat, Lipin1, Cpt1a, and Lepr, were normalized to the control level in naringenin-treated adipocytes. In addition, 25 up-regulated (> 2-fold) and 25 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Fabp5, Scd1, Srebf1, Hmgcs1, Cpt1c, Lepr, and Lrp1, were normalized to the control level by naringin. CONCLUSIONS: The results indicate that naringenin and naringin have anti-adipogenic potentials that are achieved by normalizing the expression levels of lipid metabolism-related genes that were perturbed in differentiated 3T3-L1 cells.


Assuntos
Adipócitos/efeitos dos fármacos , Flavanonas/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células 3T3-L1 , 4-Cloro-7-nitrobenzofurazano/análogos & derivados , 4-Cloro-7-nitrobenzofurazano/metabolismo , Adipócitos/metabolismo , Adipogenia , Animais , Transporte Biológico , Desoxiglucose/análogos & derivados , Desoxiglucose/metabolismo , Camundongos
5.
Molecules ; 26(12)2021 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-34207699

RESUMO

Pancreatic cancer is an aggressive disease that progresses in a relatively symptom-free manner; thus, is difficult to detect and treat. Essential oil is reported to exhibit pharmacological properties, besides its common and well-known function as aromatherapy. Therefore, this study herein aimed to investigate the anti-proliferative effect of essential oil extracted from leaves of Garcinia atroviridis (EO-L) against PANC-1 human pancreatic cancer cell line. The cell growth inhibitory concentration at 50% (IC50) and selective index (SI) values of EO-L analyses were determined as 78 µg/mL and 1.23, respectively. Combination index (CI) analysis revealed moderate synergism (CI values of 0.36 to 0.75) between EO-L and 2 deoxy-d-glucose (2-DG) treatments. The treatments of PANC-1 cells with EO-L, 2-DG and EOL+2DG showed evidence of depolarization of mitochondrial membrane potential, cell growth arrest and apoptosis. The molecular mechanism causing the anti-proliferative effect between EO-L and 2-DG is potentially through pronounced up-regulation of P53 (4.40-fold), HIF1α (1.92-fold), HK2 (2.88-fold) and down-regulation of CYP3A5 (0.11-fold), as supported by quantitative mRNA expression analysis. Collectively, the current data suggest that the combination of two anti-proliferative agents, EO-L and 2-DG, can potentially be explored as therapeutic treatments and as potentiating agents to conventional therapy against human pancreatic cancer.


Assuntos
Desoxiglucose/farmacologia , Garcinia/química , Óleos Voláteis/farmacologia , Neoplasias Pancreáticas/tratamento farmacológico , Apoptose/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sinergismo Farmacológico , Humanos , Potencial da Membrana Mitocondrial , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Folhas de Planta/química
7.
Molecules ; 26(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201708

RESUMO

Caffeine is a plant alkaloid present in food and beverages consumed worldwide. It has high lipid solubility with recognized actions in the central nervous system and in peripheral tissues, notably the adipose depots. However, the literature is scant regarding caffeine's influence on adipocyte functions other than lipolysis, such as glucose incorporation into lipids (lipogenesis) and amine oxidation. The objective of this study was to explore the direct effects of caffeine and of isobutylmethylxanthine (IBMX) on these adipocyte functions. Glucose transport into fat cells freshly isolated from mice, rats, or humans was monitored by determining [3H]-2-deoxyglucose (2-DG) uptake, while the incorporation of radiolabeled glucose into cell lipids was used as an index of lipogenic activity. Oxidation of benzylamine by primary amine oxidase (PrAO) was inhibited by increasing doses of caffeine in human adipose tissue preparations with an inhibition constant (Ki) in the millimolar range. Caffeine inhibited basal and insulin-stimulated glucose transport as well as lipogenesis in rodent adipose cells. The antilipogenic action of caffeine was also observed in adipocytes from mice genetically invalidated for PrAO activity, indicating that PrAO activity was not required for lipogenesis inhibition. These caffeine inhibitory properties were extended to human adipocytes: relative to basal 2-DG uptake, set at 1.0 ± 0.2 for 6 individuals, 0.1 mM caffeine tended to reduce uptake to 0.83 ± 0.08. Insulin increased uptake by 3.86 ± 1.11 fold when tested alone at 100 nM, and by 3.21 ± 0.80 when combined with caffeine. Our results reinforce the recommendation of caffeine's potential in the treatment or prevention of obesity complications.


Assuntos
Adipócitos/efeitos dos fármacos , Aminas Biogênicas/metabolismo , Cafeína/farmacologia , Glucose/metabolismo , Lipogênese/efeitos dos fármacos , Monoaminoxidase/metabolismo , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Animais , Benzilaminas/metabolismo , Transporte Biológico/efeitos dos fármacos , Desoxiglucose/metabolismo , Humanos , Insulina/metabolismo , Lipólise/efeitos dos fármacos , Camundongos , Ratos , Xantinas/farmacologia
8.
Arch Biochem Biophys ; 708: 108962, 2021 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-34116007

RESUMO

The involvement of metabolic reprogramming has been suggested to contribute to the pathophysiology of rheumatoid arthritis (RA). Glycolysis is enhanced in synovial cell metabolism in RA patients. Inhibitors of glycolysis are known to have anti-inflammatory effects. But, changes in the metabolism of normal synovial membranes or synovial cells during the early stages of inflammation remains unknown. Moreover, there are still many aspects of inflammatory signaling pathways altered by glycolysis inhibitors, that remain unclear. In this study we found that, in normal, non-pathological bovine synovial cells, most of ATP synthesis was generated by mitochondrial respiration. However, during the early of stages inflammation, initiated by lipopolysaccharide (LPS) exposure, synovial cells shifted to glycolysis for ATP production. The glycolysis inhibitor 2-deoxyglucose (2DG) reversed LPS induced increases in glycolysis for ATP production and suppressed the expression of inflammatory cytokines and proteolytic enzymes. 2DG suppressed the phosphorylation of the transcription factor cAMP response element binding protein (CREB) enhanced by LPS. Treatment with a CREB inhibitor reversed the expression of LPS-stimulated inflammatory cytokines and proteolytic enzymes. This study showed that changes in metabolism occur during the early stages of inflammation of synovial cells and can be reversed by 2DG and signaling pathways associated with CREB phosphorylation.


Assuntos
Anti-Inflamatórios/farmacologia , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Desoxiglucose/farmacologia , Membrana Sinovial/metabolismo , Membrana Sinovial/patologia , Animais , Bovinos , Citocinas/metabolismo , Glicólise/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Membrana Sinovial/efeitos dos fármacos
9.
J Agric Food Chem ; 69(23): 6650-6664, 2021 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-34080840

RESUMO

ß-Carbolines are naturally occurring bioactive alkaloids. In this work, carbohydrate-derived ß-carbolines (ßCs), 1-(1,3,4,5-tetrahydroxypent-1-yl)-ß-carboline isomers (1a/b), 1-(1,4,5-trihydroxypent-1-yl)-ß-carboline (2), 1-(1,5-dihydroxypent-3-en-1-yl)-ß-carboline (3), and 1-(1,2,3,4,5-pentahydroxypent-1-yl)-ß-carboline (4) were identified and analyzed in commercial foods. The concentrations of ßCs 1-4 in foods ranged from undetectable to 11.4 µg/g levels, suggesting their intake in the diet. Processed foods contained higher amounts than fresh or unprocessed foods, and the highest content was found in processed tomato and fruit products, sauces, and baked foods. ßCs 1-3 were formed in foods during heating, and 1a/b were the main compounds. The formation of carbohydrate-derived ßCs was studied in model reactions of tryptophan and carbohydrates. They formed in reactions of tryptophan with glucose under acidic conditions at temperatures higher than 80 °C. The formation of 1a/b was favored, but 2-3 increased at high temperatures. Noticeably, the ßCs 1-3 formed in the reactions of tryptophan with fructose or sucrose, and the formation from fructose was much higher than from glucose. Thus, fructose was the main carbohydrate involved in the formation of 1-3, whereas sucrose gave these ßCs after acid hydrolysis. It is shown for the first time that the mechanism of formation of ßCs 1-3 occurs from the sugar intermediate 3-deoxyglucosone that reacts with tryptophan affording these carbohydrate-derived ßCs. A mechanism of reaction to give ßCs 1-3 is proposed that relies on the tautomerism (keto-enediol or enamine-imine) of intermediates involved in the reaction. Carbohydrate ßCs 1-4 were assessed as inhibitors of monoamine oxidase (MAO), as antioxidants, and for their interaction with DNA. They were not good inhibitors of MAO-A or -B, were poor antioxidants, and did not appreciably interact with DNA.


Assuntos
Alcaloides , Frutose , Carboidratos , Carbolinas , Desoxiglucose/análogos & derivados
10.
Methods Mol Biol ; 2276: 271-283, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34060049

RESUMO

Several methods are available to measure ATP production by isolated mitochondria or permeabilized cells but have several limitations, depending upon the particular assay employed. These limitations may include poor sensitivity or specificity, complexity of the method, poor throughput, changes in mitochondrial inner membrane potential as ATP is consumed, and/or inability to simultaneously assess other mitochondrial functional parameters. Here we describe a novel nuclear magnetic resonance (NMR)-based assay that can be carried out with high efficiency in a manner that alleviates the above problems.


Assuntos
Trifosfato de Adenosina/metabolismo , Peróxido de Hidrogênio/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Animais , Desoxiglucose/metabolismo , Metabolismo Energético , Hexoquinase/metabolismo , Potencial da Membrana Mitocondrial , Camundongos , Mitocôndrias Cardíacas/metabolismo , Mitocôndrias Hepáticas/metabolismo , ATPases Mitocondriais Próton-Translocadoras/metabolismo
11.
Artigo em Inglês | MEDLINE | ID: mdl-34167955

RESUMO

INTRODUCTION: Saliva collection is a non-invasive test and is convenient. 1,5-anhydroglucitol (1,5-AG) is a new indicator reflecting short-term blood glucose levels. This study aimed to explore the relationship between saliva 1,5-AG and insulin secretion function and insulin sensitivity. RESEARCH DESIGN AND METHODS: Adult patients with type 2 diabetes who were hospitalized were enrolled. Based on blood glucose and C-peptide, homeostasis model assessment 2 for ß cell secretion function, C-peptidogenic index (CGI), △2-hour C-peptide (2hCP)/△2-hour postprandial glucose (2hPG), ratio of 0-30 min area under the curve for C-peptide and area under the curve for glucose (AUCCP30/AUCPG30), and AUC2hCP/AUC2hPG were calculated to evaluate insulin secretion function, while indicators such as homeostasis model assessment 2 for insulin resistance were used to assess insulin sensitivity. RESULTS: We included 284 subjects (178 men and 106 women) with type 2 diabetes aged 20-70 years. The saliva 1,5-AG level was 0.133 (0.089-0.204) µg/mL. Spearman's correlation analysis revealed a significantly negative correlation between saliva 1,5-AG and 0, 30, and 120 min blood glucose, glycated hemoglobin A1c, and glycated albumin (all p<0.05), and a significantly positive association between saliva 1,5-AG and CGI (r=0.171, p=0.004) and AUC CP30 /AUC PG30 (r=0.174, p=0.003). The above correlations still existed after adjusting for age, sex, body mass index, and diabetes duration. In multiple linear regression, saliva 1,5-AG was an independent factor of CGI (standardized ß=0.135, p=0.015) and AUC CP30 /AUC PG30 (standardized ß=0.110, p=0.020). CONCLUSIONS: Saliva 1,5-AG was related to CGI and AUCCP30/AUCPG30 in patients with type 2 diabetes. TRIAL REGISTRATION NUMBER: ChiCTR-SOC-17011356.


Assuntos
Diabetes Mellitus Tipo 2 , Adulto , China , Desoxiglucose , Diabetes Mellitus Tipo 2/diagnóstico , Feminino , Teste de Tolerância a Glucose , Humanos , Insulina/metabolismo , Secreção de Insulina , Masculino , Saliva/metabolismo
12.
Chem Commun (Camb) ; 57(45): 5530-5533, 2021 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-33959731

RESUMO

We report a non-antibody GLUT1 inhibitor probe NBDQ that is 30 times more sensitive than the traditional GLUT1 transportable tracer for cancer cell imaging and Warburg effect-based tumor detection. NBDQ reveals significant advantages in terms of tumor selectivity, fluorescence stability and in vivo biocompatibility in xenograft tumor imaging, including triple-negative breast cancer.


Assuntos
4-Cloro-7-nitrobenzofurazano/análogos & derivados , Biomarcadores Tumorais/análise , Desoxiglucose/análogos & derivados , Corantes Fluorescentes/química , Transportador de Glucose Tipo 1/antagonistas & inibidores , Neoplasias de Mama Triplo Negativas/diagnóstico por imagem , 4-Cloro-7-nitrobenzofurazano/química , Animais , Materiais Biocompatíveis/química , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular , Desoxiglucose/química , Transportador de Glucose Tipo 1/genética , Humanos , Camundongos , Imagem Multimodal , Neoplasias Experimentais , Imagem Óptica , Tomografia por Emissão de Pósitrons
13.
Curr Protoc ; 1(5): e139, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34033261

RESUMO

The liver is central in maintaining glucose homeostasis. Indeed, impaired hepatic glucose uptake has been implicated in the development of hyperglycemia in type II diabetes (T2D) and non-alcoholic fatty liver disease (NAFLD). However, current approaches to evaluate glucose mobilization rely on indirect measurements that do not provide spatial and temporal information. Here, we describe confocal-based intravital microscopy (IVM) of the liver that allows the identification of hepatocyte spatial organization and glucose transport. Specifically, we describe a method to fluorescently label hepatic landmarks to identify different compartments within the liver. In addition, we outline an in vivo fluorescent glucose uptake assay to quantitatively measure glucose mobilization in space and time. These protocols allow direct investigation of hepatic glycemic control and can be further applied to murine models of liver disease. © Published 2021. This article is a U.S. Government work and is in the public domain in the USA. Basic Protocol 1: Mouse surgical procedure and positioning for liver intravital imaging Basic Protocol 2: Fluorescent labeling and intravital imaging of mouse hepatic compartments Basic Protocol 3: Mouse hepatic glucose uptake assay and intravital imaging analysis.


Assuntos
Glucose/metabolismo , Microscopia Intravital , Fígado/metabolismo , 4-Cloro-7-nitrobenzofurazano/análogos & derivados , 4-Cloro-7-nitrobenzofurazano/metabolismo , Animais , Desoxiglucose/análogos & derivados , Desoxiglucose/metabolismo , Corantes Fluorescentes/química , Hepatócitos/metabolismo , Imageamento Tridimensional , Camundongos
14.
Int J Mol Sci ; 22(7)2021 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-33918425

RESUMO

The results of structural studies on a series of halogen-substituted derivatives of 2-deoxy-D-glucose (2-DG) are reported. 2-DG is an inhibitor of glycolysis, a metabolic pathway crucial for cancer cell proliferation and viral replication in host cells, and interferes with D-glucose and D-mannose metabolism. Thus, 2-DG and its derivatives are considered as potential anticancer and antiviral drugs. X-ray crystallography shows that a halogen atom present at the C2 position in the pyranose ring does not significantly affect its conformation. However, it has a noticeable effect on the crystal structure. Fluorine derivatives exist as a dense 3D framework isostructural with the parent compound, while Cl- and I-derivatives form layered structures. Analysis of the Hirshfeld surface shows formation of hydrogen bonds involving the halogen, yet no indication for the existence of halogen bonds. Density functional theory (DFT) periodic calculations of cohesive and interaction energies (at the B3LYP level of theory) have supported these findings. NMR studies in the solution show that most of the compounds do not display significant differences in their anomeric equilibria, and that pyranose ring puckering is similar to the crystalline state. For 2-deoxy-2-fluoro-D-glucose (2-FG), electrostatic interaction energies between the ligand and protein for several existing structures of pyranose 2-oxidase were also computed. These interactions mostly involve acidic residues of the protein; single amino-acid substitutions have only a minor impact on binding. These studies provide a better understanding of the structural chemistry of halogen-substituted carbohydrates as well as their intermolecular interactions with proteins determining their distinct biological activity.


Assuntos
Desoxiglucose/análogos & derivados , Halogênios/química , Desoxiglucose/metabolismo , Ligantes , Espectroscopia de Ressonância Magnética , Conformação Molecular , Proteínas/metabolismo , Difração de Raios X
15.
Molecules ; 26(6)2021 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-33802864

RESUMO

The aim and novelty of this paper are found in assessing the influence of inhibitors and antibiotics on intact cell MALDI-TOF mass spectra of the cyanobacterium Synechococcus sp. UPOC S4 and to check the impact on reliability of identification. Defining the limits of this method is important for its use in biology and applied science. The compounds included inhibitors of respiration, glycolysis, citrate cycle, and proteosynthesis. They were used at 1-10 µM concentrations and different periods of up to 3 weeks. Cells were also grown without inhibitors in a microgravity because of expected strong effects. Mass spectra were evaluated using controls and interpreted in terms of differential peaks and their assignment to protein sequences by mass. Antibiotics, azide, and bromopyruvate had the greatest impact. The spectral patterns were markedly altered after a prolonged incubation at higher concentrations, which precluded identification in the database of reference spectra. The incubation in microgravity showed a similar effect. These differences were evident in dendrograms constructed from the spectral data. Enzyme inhibitors affected the spectra to a smaller extent. This study shows that only a long-term presence of antibiotics and strong metabolic inhibitors in the medium at 10-5 M concentrations hinders the correct identification of cyanobacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF).


Assuntos
Antibacterianos/toxicidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Synechococcus/química , Synechococcus/efeitos dos fármacos , Antimicina A/análogos & derivados , Antimicina A/toxicidade , Azidas/toxicidade , Respiração Celular/efeitos dos fármacos , Cloranfenicol/toxicidade , Ciclo do Ácido Cítrico/efeitos dos fármacos , Desoxiglucose/toxicidade , Fluoracetatos/toxicidade , Glicólise/efeitos dos fármacos , Malonatos/toxicidade , Biossíntese de Proteínas/efeitos dos fármacos , Piruvatos/toxicidade , Reprodutibilidade dos Testes , Estreptomicina/toxicidade , Synechococcus/isolamento & purificação , Synechococcus/metabolismo , Ausência de Peso
16.
Front Immunol ; 12: 632512, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33815385

RESUMO

Visceral leishmaniasis (VL) is a fatal parasitic disease if untreated. Treatment options of VL diminish due to emerging drug resistance. Although the principal host cells for the multiplication of Leishmania are macrophages, neutrophils are the first cells infected with the parasites rapidly after parasite inoculation. Leishmania can survive in neutrophils despite the potent antimicrobial effector functions of neutrophils that can eliminate the parasites. Recently, the growing field of immunometabolism provided strong evidence for the therapeutic potential in targeting metabolic processes as a means of controlling immune effector functions. Therefore, the understanding of the immunometabolic profile of neutrophils during Leishmania infection could provide new promising targets for host-directed therapies against VL. To our knowledge, this is the first study addressing the bioenergetics profile of L. donovani-infected primary human neutrophils. Transcriptome analysis of L. donovani-infected neutrophils revealed an early significant upregulation of several glycolytic enzymes. Extracellular flux analysis showed that glycolysis and glycolytic capacity were upregulated in L. donovani-infected neutrophils at 6 h post infection. An increased glucose uptake and accumulation of glycolytic end products were further signs for an elevated glycolytic metabolism in L. donovani-infected neutrophils. At the same time point, oxidative phosphorylation provided NADPH for the oxidative burst but did not contribute to ATP production. Inhibition of glycolysis with 2-DG significantly reduced the survival of L. donovani promastigotes in neutrophils and in culture. However, this reduction was due to a direct antileishmanial effect of 2-DG and not a consequence of enhanced antileishmanial activity of neutrophils. To further address the impact of glucose metabolism during the first days of infection in vivo, we treated C57BL/6 mice with 2-DG prior to infection with L. donovani and assessed the parasite load one day and seven days post infection. Our results show, that seven days post-infection the parasite load of 2-DG treated animals was significantly higher than in mock treated animals. This data indicates that glycolysis serves as major energy source for antimicrobial effector functions against L. donovani. Inhibition of glycolysis abrogates important neutrophil effector functions that are necessary the initial control of Leishmania infection.


Assuntos
Glucose/metabolismo , Leishmania donovani/fisiologia , Leishmaniose Visceral/imunologia , Neutrófilos/imunologia , Animais , Células Cultivadas , Desoxiglucose/efeitos adversos , Desoxiglucose/farmacologia , Glicólise/efeitos dos fármacos , Humanos , Leishmania donovani/efeitos dos fármacos , Leishmaniose Visceral/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Neutrófilos/parasitologia , Fosforilação Oxidativa , Carga Parasitária , Espécies Reativas de Oxigênio/metabolismo , Explosão Respiratória
17.
Cells ; 10(2)2021 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-33672651

RESUMO

Hepatocellular carcinoma (HCC) that is triggered by metabolic defects is one of the most malignant liver cancers. A much higher incidence of HCC among men than women suggests the protective roles of estrogen in HCC development and progression. To begin to understand the mechanisms involving estrogenic metabolic effects, we compared cell number, viability, cytotoxicity, and apoptosis among HCC-derived HepG2 cells that were treated with different concentrations of 2-deoxy-d-glucose (2-DG) that blocks glucose metabolism, oxamate that inhibits lactate dehydrogenase and glycolysis, or oligomycin that blocks ATP synthesis and mitochondrial oxidative phosphorylation. We confirmed that HepG2 cells primarily utilized glycolysis followed by lactate fermentation, instead of mitochondrial oxidative phosphorylation, for cell growth. We hypothesized that estrogen altered energy metabolism via its receptors to carry out its anticancer effects in HepG2 cells. We treated cells with 17ß-estradiol (E2), 1,3,5-tris(4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT) an estrogen receptor (ER) α (ERα) agonist, or 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN), an ERß agonist. We then used transcriptomic and metabolomic analyses and identified differentially expressed genes and unique metabolite fingerprints that are produced by each treatment. We further performed integrated multi-omics analysis, and identified key genes and metabolites in the gene-metabolite interaction contributed by E2 and ER agonists. This integrated transcriptomic and metabolomic study suggested that estrogen acts on estrogen receptors to suppress liver cancer cell growth via altering metabolism. This is the first exploratory study that comprehensively investigated estrogen and its receptors, and their roles in regulating gene expression, metabolites, metabolic pathways, and gene-metabolite interaction in HCC cells using bioinformatic tools. Overall, this study provides potential therapeutic targets for future HCC treatment.


Assuntos
Estrogênios/metabolismo , Neoplasias Hepáticas/metabolismo , Metabolômica , Contagem de Células , Proliferação de Células/efeitos dos fármacos , Desoxiglucose/farmacologia , Estradiol/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Redes e Vias Metabólicas/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Nitrilas/farmacologia , Oligomicinas/farmacologia , Pirazóis/farmacologia , Receptores de Estrogênio/metabolismo , Transcriptoma/genética
18.
Helicobacter ; 26(3): e12797, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33682972

RESUMO

PURPOSE: Helicobacter pylori (HP) infection is reported to increase 18 F-fluoro-2-deoxyglucose (FDG) accumulation in the stomach. The accumulation of FDG by positron-emission tomography (FDG-PET) in the stomach for the voluntary health examinees of cancer checkup was examined before and after the HP eradication. SUBJECTS AND METHODS: From March 2013 to October 2015, eighty-one subjects were performed FDG-PET to detect cancer at the health checkup. All of them were also surveyed by esophagogastroduodenoscopy. Subjects were classified as the 33 cases of HP positive (group A), 38 cases of originally negative (group B), and the 10 negative cases by HP eradication therapy (group C). Group A was treated by combination of amoxicillin, clarithromycin, and proton pump inhibitor for a week, and all of them eradicated HP. A part of group A (n = 7) was serially performed FDG-PET one to five years after the treatment and compared the maximum standard uptake value of FDG (SUV) around the fundic gland region. RESULTS: SUV of group A (3.55 ± 0.69) was significantly higher than those of both group B (2.96 ± 0.72) and group C (2.89 ± 0.51) (p < 0.01, respectively). Groups B and C are almost comparable and showed no significant difference during the course. In group A, HP eradication significantly decreased the SUV to 3.1 ± 0.43 (P < .01). SUV after the eradication was significantly reduced (P < .01) in the mild to moderate atrophy (C1-C3) group according to Kimura and Takemoto classification of chronic gastritis of group A. Although SUV in the advanced atrophy group (O1-O3) tended to decline after the eradication, the change was not significant. CONCLUSION: HP-infected stomach showed higher FDG uptake in the fundic gland region and HP eradication decreased the uptake in the mild to moderate atrophic gastritis but not in the severe atrophic gastritis.


Assuntos
Fluordesoxiglucose F18/farmacocinética , Mucosa Gástrica/metabolismo , Gastrite/diagnóstico , Infecções por Helicobacter , Tomografia por Emissão de Pósitrons , Amoxicilina/uso terapêutico , Antibacterianos/uso terapêutico , Claritromicina/uso terapêutico , Desoxiglucose/uso terapêutico , Quimioterapia Combinada , Gastrite/microbiologia , Infecções por Helicobacter/tratamento farmacológico , Helicobacter pylori , Humanos , Estômago , Neoplasias Gástricas , Tomografia
19.
Methods Mol Biol ; 2265: 73-80, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33704706

RESUMO

Melanoma cells have high glycolytic capacity. Glucose uptake is a key rate-limiting step in glucose utilization. Here we describe a simple protocol for measuring direct glucose uptake in living melanoma cells by flow cytometry.


Assuntos
Citometria de Fluxo/métodos , Glucose/metabolismo , Melanoma/metabolismo , 4-Cloro-7-nitrobenzofurazano/análogos & derivados , 4-Cloro-7-nitrobenzofurazano/química , Transporte Biológico , Técnicas de Cultura de Células/métodos , Linhagem Celular Tumoral , Desoxiglucose/análogos & derivados , Desoxiglucose/química , Fluorescência , Corantes Fluorescentes/química , Humanos
20.
Biochem Biophys Res Commun ; 546: 90-96, 2021 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-33578294

RESUMO

Although, numerous in vitro studies showed that cancer cells are killed after exposure to pharmacological doses of ascorbic acid (AA), significant clinical data proving the efficacy of AA is still absent. A hallmark of most tumor cells is an altered glucose metabolism characterized by an upregulation of glycolysis despite normoxic conditions (Warburg effect). Since pyruvate is capable of detoxifying hydrogen peroxide (H2O2), the alleged mediator of AA-induced toxicity, it seems likely that enhanced glycolysis and subsequent higher pyruvate formation might be an explanation for the attenuated effect of pharmacological AA in vivo. Therefore, inhibition of glycolysis might be a promising approach to enhance the anticancer effect of AA by diminishing the generation of pyruvate. Considering the altered metabolism of cancer cells, we examined the cytotoxic potential of 2-DG and/or AA using SRB assay in two different cell lines: a glycolytic human melanoma (451Lu) and a non-glycolytic breast cancer (MCF-7) cell line. Inhibition of glycolysis increased AA cytotoxicity in 451Lu cells, whereas same treatment had a marginal effect on MCF-7 cells. We also investigated the influence of glycolysis inhibition on H2O2 generation. H2O2 concentrations were higher in presence of 451Lu cells when pretreated with 2-DG, but not in MCF-7 cells. Treatment with 10 mM 2-DG decreased pyruvate and lactate concentrations in both cell lines in a concentration-dependent manner. In summary, 2-DG enhances the cytotoxic effect of AA in glycolytic 451Lu cells by increasing AA-induced H2O2 concentration. This result indicates that lower pyruvate levels, as a result of glycolysis inhibition, may be responsible for the enhanced effect of 2-DG on AA toxicity. Further experiments are needed to clarify the underlying mechanism and the potential use in cancer therapy.


Assuntos
Ácido Ascórbico/farmacologia , Desoxiglucose/farmacologia , Glucose/metabolismo , Glicólise , Melanoma/metabolismo , Melanoma/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Peróxido de Hidrogênio/metabolismo , Ácido Láctico/metabolismo , Ácido Pirúvico/metabolismo
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