RESUMEN
Viruses hijack host metabolic pathways for their replicative advantage. In this study, using patient-derived multiomics data and in vitro infection assays, we aimed to understand the role of key metabolic pathways that can regulate severe acute respiratory syndrome coronavirus-2 reproduction and their association with disease severity. We used multiomics platforms (targeted and untargeted proteomics and untargeted metabolomics) on patient samples and cell-line models along with immune phenotyping of metabolite transporters in patient blood cells to understand viral-induced metabolic modulations. We also modulated key metabolic pathways that were identified using multiomics data to regulate the viral reproduction in vitro. Coronavirus disease 2019 disease severity was characterized by increased plasma glucose and mannose levels. Immune phenotyping identified altered expression patterns of carbohydrate transporter, glucose transporter 1, in CD8+ T cells, intermediate and nonclassical monocytes, and amino acid transporter, xCT, in classical, intermediate, and nonclassical monocytes. In in vitro lung epithelial cell (Calu-3) infection model, we found that glycolysis and glutaminolysis are essential for virus replication, and blocking these metabolic pathways caused significant reduction in virus production. Taken together, we therefore hypothesized that severe acute respiratory syndrome coronavirus-2 utilizes and rewires pathways governing central carbon metabolism leading to the efflux of toxic metabolites and associated with disease severity. Thus, the host metabolic perturbation could be an attractive strategy to limit the viral replication and disease severity.
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Proteínas Sanguíneas/metabolismo , COVID-19/etiología , SARS-CoV-2/fisiología , Adulto , Anciano , Sistema de Transporte de Aminoácidos y+/sangre , Aminoácidos/sangre , Biomarcadores/sangre , Proteínas Sanguíneas/análisis , COVID-19/metabolismo , COVID-19/virología , Carbohidratos/sangre , Estudios de Casos y Controles , Transportador de Glucosa de Tipo 1/sangre , Hospitalización , Humanos , Inmunofenotipificación , Manosa/sangre , Lectina de Unión a Manosa/sangre , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Replicación ViralRESUMEN
OBJECTIVE: Olaparib is a potent inhibitor of poly(ADP-ribose) polymerase (PARP)-1, 2, and 3 with potential activity in endometrial cancer (EC). METHODS: In this window-of-opportunity trial, women with operable type 1 EC received olaparib oral tablets (300mg) twice daily for 28days before surgery. The primary objective was to evaluate the effects of olaparib on EC in tissue samples taken at baseline and at treatment completion. Signal of activity was defined as significant changes in the expression of the cell cycle-related proteins cyclin D1, Ki67, and cleaved caspase-3. RESULTS: A total of 31 patients were included in the biomarker analysis. The median time of olaparib exposure was 24 days (1-39). Significant inhibition was found for cyclin D1 (p < 0.01), but not for Ki67 and active caspase 3 immunostaining. PARP-1 levels positively correlated with cyclin D1 levels (rho = 0.661, p = 0.0001). Both PARP-1 and cyclin D1 levels were significantly lower (p = 0.022 and p = 0.004, respectively) in patients with ARID1A[-] tumors than ARID1A[+] tumors. A significant relationship between plasma olaparib concentrations and decreased GLUT1 activity was observed (r = -0.5885; p < 0.05). Drug-related toxicity consisted mostly of gastrointestinal and grade 1 or 2 adverse events. CONCLUSIONS: Olaparib reduced expression of cyclin D1, which positively correlated with PARP-1 levels. This effect was more evident in ARID1A-deficient tumors. Olaparib further induced inhibition of GLUT1 plasma activity. Our findings could have noteworthy implications in predicting which patients with EC would benefit from olaparib-based strategies.
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Biomarcadores de Tumor/análisis , Neoplasias Endometriales/terapia , Terapia Neoadyuvante/métodos , Ftalazinas/administración & dosificación , Piperazinas/administración & dosificación , Inhibidores de Poli(ADP-Ribosa) Polimerasas/administración & dosificación , Administración Oral , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/genética , Quimioterapia Adyuvante/métodos , Ciclina D1/análisis , Ciclina D1/genética , Proteínas de Unión al ADN/genética , Relación Dosis-Respuesta a Droga , Neoplasias Endometriales/sangre , Neoplasias Endometriales/diagnóstico , Neoplasias Endometriales/genética , Endometrio/efectos de los fármacos , Endometrio/patología , Endometrio/cirugía , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Transportador de Glucosa de Tipo 1/antagonistas & inhibidores , Transportador de Glucosa de Tipo 1/sangre , Humanos , Histerectomía , Inmunohistoquímica , Persona de Mediana Edad , Estadificación de Neoplasias , Ftalazinas/efectos adversos , Piperazinas/efectos adversos , Poli(ADP-Ribosa) Polimerasa-1/antagonistas & inhibidores , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Inhibidores de Poli(ADP-Ribosa) Polimerasas/efectos adversos , Estudios Prospectivos , Comprimidos , Factores de Tiempo , Factores de Transcripción/genética , Resultado del TratamientoRESUMEN
Type 1 diabetic (T1D) adolescent children on insulin therapy suffer episodes of both hyper- and hypoglycemic episodes. Glucose transporter isoform GLUT1 expressed in blood-brain barrier (BBB) and red blood cells (RBC) compensates for perturbed circulating glucose toward protecting the supply to brain and RBCs. We hypothesized that RBC-GLUT1 concentration, as a surrogate for BBB-GLUT1, is altered in T1D children. To test this hypothesis, we measured RBC-GLUT1 by enzyme-linked immunosorbent assay (ELISA) in T1D children (n = 72; mean age 15.3 ± 0.2 yr) and control children (CON; n = 11; mean age 15.6 ± 0.9 yr) after 12 h of euglycemia and during a hyperinsulinemic-hypoglycemic clamp with a nadir blood glucose of ~3.3 mmol/L for 90 min (clamp I) or ~3 mmol/L for 45 min (clamp II). Reduced baseline RBC-GLUT1 was observed in T1D (2.4 ± 0.17 ng/ng membrane protein); vs. CON (4.2 ± 0.61 ng/ng protein) (p < 0.0001). Additionally, baseline RBC-GLUT1 in T1D negatively correlated with hemoglobin A1c (HbA1c) (R = -0.23, p < 0.05) but not in CON (R = 0.06, p < 0.9). Acute decline in serum glucose to 3.3 mmol/L (90 min) or 3 mmol/L (45 min) did not change baseline RBC-GLUT1 in T1D or CON children. We conclude that reduced RBC-GLUT1 encountered in T1D, with no ability to compensate by increasing during acute hypoglycemia over the durations examined, may demonstrate a vulnerability of impaired RBC glucose transport (serving as a surrogate for BBB), especially in those with the worst control. We speculate that this may contribute to the perturbed cognition seen in T1D adolescents.
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Barrera Hematoencefálica/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Regulación hacia Abajo , Eritrocitos/metabolismo , Transportador de Glucosa de Tipo 1/sangre , Modelos Biológicos , Adolescente , Biomarcadores/sangre , Glucemia/análisis , Barrera Hematoencefálica/efectos de los fármacos , Trastornos del Conocimiento/complicaciones , Diabetes Mellitus Tipo 1/sangre , Diabetes Mellitus Tipo 1/complicaciones , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Regulación hacia Abajo/efectos de los fármacos , Ensayo de Inmunoadsorción Enzimática , Eritrocitos/efectos de los fármacos , Técnica de Clampeo de la Glucosa , Hemoglobina Glucada/análisis , Humanos , Hiperglucemia/prevención & control , Hipoglucemia/inducido químicamente , Hipoglucemia/prevención & control , Hipoglucemiantes/farmacología , Hipoglucemiantes/uso terapéutico , Insulina/farmacología , Insulina/uso terapéutico , Estudios ProspectivosRESUMEN
BACKGROUND: Noninvoluting congenital hemangioma (NICH) is a distinct vascular tumor of infancy. OBJECTIVE: We describe the clinical characteristics, histopathology, imaging, and natural history of NICH and compare our findings with previous reports. METHODS: We conducted a retrospective review of charts and photographic databases from 2 vascular anomaly centers over a 15-year period. RESULTS: Thirty cases of NICH were identified. All patients had fully formed vascular lesions at birth that demonstrated a nonprogressive course. The trunk and lower extremities were preferred sites and there was a female predominance. Thirteen of 30 patients reported pain. Focal necrosis and scarring was seen in a minority. Doppler studies, when performed, confirmed high vascular flow. Microscopic evaluation of 4 excised lesions showed lobular areas of endothelial cell proliferation directly adjacent to ectatic malformed vessels. Immunohistochemical studies demonstrated absence of glucose transporter-1 protein expression in every case. Wilms tumor-1 positivity was observed in lobular areas. The larger vessels did not stain with Wilms tumor-1, but some displayed D2-40 positivity. LIMITATIONS: Patients were referred to university-based pediatric vascular anomaly centers, with potential bias toward more severe or extensive cases. CONCLUSIONS: This retrospective study highlights the unique clinical and histopathologic features of NICH.
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Hemangioma/diagnóstico , Neoplasias Cutáneas/diagnóstico , Niño , Preescolar , Femenino , Transportador de Glucosa de Tipo 1/sangre , Hemangioma/congénito , Hemangioma/diagnóstico por imagen , Hemangioma/patología , Humanos , Inmunohistoquímica , Masculino , Estudios Retrospectivos , Neoplasias Cutáneas/congénito , Neoplasias Cutáneas/diagnóstico por imagen , Neoplasias Cutáneas/patología , Ultrasonografía Doppler , Proteínas WT1/sangreRESUMEN
The hereditary stomatocytoses are a series of dominantly inherited hemolytic anemias in which the permeability of the erythrocyte membrane to monovalent cations is pathologically increased. The causative mutations for some forms of hereditary stomatocytosis have been found in the transporter protein genes, RHAG and SLC4A1. Glucose transporter 1 (glut1) deficiency syndromes (glut1DSs) result from mutations in SLC2A1, encoding glut1. Glut1 is the main glucose transporter in the mammalian blood-brain barrier, and glut1DSs are manifested by an array of neurologic symptoms. We have previously reported 2 cases of stomatin-deficient cryohydrocytosis (sdCHC), a rare form of stomatocytosis associated with a cold-induced cation leak, hemolytic anemia, and hepatosplenomegaly but also with cataracts, seizures, mental retardation, and movement disorder. We now show that sdCHC is associated with mutations in SLC2A1 that cause both loss of glucose transport and a cation leak, as shown by expression studies in Xenopus oocytes. On the basis of a 3-dimensional model of glut1, we propose potential mechanisms underlying the phenotypes of the 2 mutations found. We investigated the loss of stomatin during erythropoiesis and find this occurs during reticulocyte maturation and involves endocytosis. The molecular basis of the glut1DS, paroxysmal exercise-induced dyskinesia, and sdCHC phenotypes are compared and discussed.
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Transportador de Glucosa de Tipo 1/deficiencia , Transportador de Glucosa de Tipo 1/genética , Hiperpotasemia/congénito , Proteínas de la Membrana/deficiencia , Mutación , Secuencia de Aminoácidos , Animales , Catarata/sangre , Catarata/genética , Desoxiglucosa/metabolismo , Eritrocitos/metabolismo , Femenino , Transportador de Glucosa de Tipo 1/sangre , Transportador de Glucosa de Tipo 1/química , Humanos , Hiperpotasemia/sangre , Hiperpotasemia/genética , Hiperpotasemia/metabolismo , Técnicas In Vitro , Transporte Iónico , Proteínas de la Membrana/sangre , Modelos Moleculares , Datos de Secuencia Molecular , Proteínas Mutantes/sangre , Proteínas Mutantes/química , Proteínas Mutantes/genética , Oocitos/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homología Estructural de Proteína , Síndrome , Xenopus laevisRESUMEN
Water enables life and plays a critical role in biology. Considered as a versatile and adaptive component of the cell, water engages a wide range of biomolecular interactions. An organism can exist and function only if its self-assembled molecular structures are hydrated. It was shown recently that switching of AMP/ATP binding to the insulin-independent glucose transporter Human Erythrocyte Glucose Transport Protein (GLUT1) may greatly influence the ratio of bulk and bound water during regulation of glucose uptake by red blood cells. In this paper, we present the results on the hydration properties of AMP/ATP obtained by means of dielectric spectroscopy in aqueous solution and for fully ionized forms in solid amorphous films with the help of gravimetric studies.
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Adenosina Monofosfato/química , Adenosina Trifosfato/química , Transportador de Glucosa de Tipo 1/sangre , Agua/química , Adenosina Monofosfato/metabolismo , Adenosina Trifosfato/metabolismo , Espectroscopía Dieléctrica , Eritrocitos/química , Eritrocitos/metabolismo , Glucosa/metabolismo , Transportador de Glucosa de Tipo 1/química , Humanos , Insulina/química , Insulina/metabolismo , Soluciones/química , Agua/metabolismoRESUMEN
Little is known about the biological mechanisms underpinning the pathology of schizophrenia. We have analysed the proteome of stimulated and unstimulated peripheral blood mononuclear cells (PBMCs) from schizophrenia patients and controls as a potential model of altered cellular signaling using liquid-chromatography mass spectrometry proteomic profiling. PBMCs from patients and controls were stimulated for 72 h in vitro using staphylococcal enterotoxin B. In total, 18 differentially expressed proteins between first-onset, antipsychotic-naive patients and controls in the unstimulated and stimulated conditions were identified. Remarkably, eight of these proteins were associated with the glycolytic pathway and patient-control differences were more prominent in stimulated compared with unstimulated PBMCs. None of these proteins were altered in chronically ill antipsychotic-treated patients. Non-linear multivariate statistical analysis showed that small subsets of these proteins could be used as a signal for distinguishing first-onset patients from controls with high precision. Functional analysis of PBMCs did not reveal any difference in the glycolytic rate between patients and controls despite increased levels of lactate and the glucose transporter-1, and decreased levels of the insulin receptor in patients. In addition, subjects showed increased serum levels of insulin, consistent with the idea that some schizophrenia patients are insulin resistant. These results show that schizophrenia patients respond differently to PBMC activation and this is manifested at disease onset and may be modulated by antipsychotic treatment. The glycolytic protein signature associated with this effect could therefore be of diagnostic and prognostic value. Moreover, these results highlight the importance of using cells for functional discovery and show that it may not be sufficient to measure protein expression levels in static states.
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Antipsicóticos/administración & dosificación , Glucemia/metabolismo , Leucocitos Mononucleares/metabolismo , Esquizofrenia/metabolismo , Adulto , Antipsicóticos/uso terapéutico , Enterotoxinas/farmacología , Femenino , Transportador de Glucosa de Tipo 1/sangre , Hexoquinasa/metabolismo , Humanos , Insulina/sangre , Ácido Láctico/sangre , Leucocitos Mononucleares/efectos de los fármacos , Masculino , Proteoma/metabolismo , Proteómica/métodos , Receptor de Insulina/sangre , Esquizofrenia/sangre , Esquizofrenia/tratamiento farmacológicoAsunto(s)
Transportador de Glucosa de Tipo 1/sangre , Subunidad alfa del Factor 1 Inducible por Hipoxia/sangre , Talasemia/terapia , Adulto , Transfusión Sanguínea , Estudios de Casos y Controles , Femenino , Humanos , Hipoxia/sangre , Hipoxia/diagnóstico , Masculino , Persona de Mediana Edad , Talasemia/sangreRESUMEN
Anti-ischaemic drug mildronate suppresses fatty acid metabolism and increases glucose utilization in myocardium. It was proposed that it could produce a favourable effect on metabolic parameters and glucose transport in diabetic animals. Rats with streptozotocin diabetes mellitus were treated with mildronate (100 mg/kg daily, per os, 6 weeks). Therapeutic effect of mildronate was monitored by measuring animal weight, concentrations of blood glucose, insulin, blood triglycerides, free fatty acids, blood ketone bodies and cholesterol, glycated haemoglobin per cent (HbA1c%) and glucose tolerance. GLUT1 mRNA and protein expression in kidneys, heart, liver and muscles were studied by means of real time RT-PCR and immunohistochemistry correspondingly. In the streptozotocin + mildronate group, mildronate treatment caused a significant decrease in mean blood glucose, cholesterol, free fatty acid and HbA1c concentrations and improved glucose tolerance. Induction of streptozotocin diabetes mellitus provoked increase of both GLUT1 gene and protein expression in kidneys, heart and muscle, mildronate treatment produced normalization of the GLUT1 expression levels. In the liver a similar effect was observed for GLUT1 protein expression, while GLUT1 gene expression was increased by mildronate. Mildronate produces therapeutic effect in streptozotocin diabetes model. Mildronate normalizes the GLUT1 expression up-regulated by streptozotocin diabetes mellitus in kidneys, heart, muscle and liver. Copyright © 2011 John Wiley & Sons, Ltd.
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Glucemia/metabolismo , Diabetes Mellitus Experimental/tratamiento farmacológico , Transportador de Glucosa de Tipo 1/metabolismo , Hipoglucemiantes/farmacología , Metilhidrazinas/farmacología , ARN Mensajero/metabolismo , Estreptozocina/farmacología , Animales , Glucemia/efectos de los fármacos , Tamaño Corporal/efectos de los fármacos , Diabetes Mellitus/tratamiento farmacológico , Ácidos Grasos/sangre , Ácidos Grasos/metabolismo , Prueba de Tolerancia a la Glucosa , Transportador de Glucosa de Tipo 1/sangre , Transportador de Glucosa de Tipo 1/efectos de los fármacos , Hemoglobina Glucada/efectos de los fármacos , Hemoglobina Glucada/metabolismo , Insulina/sangre , Insulina/metabolismo , Metilhidrazinas/uso terapéutico , ARN Mensajero/efectos de los fármacos , Ratas , Ratas Wistar , Estreptozocina/efectos adversos , Triglicéridos/sangre , Triglicéridos/metabolismoRESUMEN
The interaction of ankyrin and spectrin yields the major anchor between the membrane skeleton and the lipid bilayer. It is critical for red cell deformability and stability, and it is also involved in the cellular localization of several proteins, in cell differentiation, and in neuron activity. Therefore, its nature is of great interest, and recently, several researchers have had varying degrees of success in elucidating the structural basis of ankyrin-spectrin recognition. In this short paper, we briefly summarize the data obtained and compare the resulting conclusions.
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Ancirinas/metabolismo , Espectrina/metabolismo , Ancirinas/sangre , Deformación Eritrocítica , Eritrocitos/fisiología , Transportador de Glucosa de Tipo 1/sangre , Transportador de Glucosa de Tipo 1/metabolismo , Glicoforinas/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular/sangre , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Cinética , Membrana Dobles de Lípidos/metabolismo , Modelos BiológicosRESUMEN
INTRODUCTION: Infantile hemangioma (IH) is a common vascular tumor in children. It is reported that IHs are associated with immunochemical markers such as vascular endothelial growth factor (VEGF)-A, glucose transporter isoform 1 (GLUT1), and insulin-like growth factor-2 (IGF-2). MATERIAL AND METHODS: This cross-sectional study focused on pediatric patients with IH. A total of 46 patients (mean age 14.2±21.9 months) with IH and 45 healthy controls (mean age 21.8±15.08 months) were enrolled. Demographic data, clinical findings, and laboratory parameters were recorded. Blood samples were collected. Serum GLUT1, IGF-2, VEGF-A, fibroblast growth factor 1 (FGF1), and angiopoietin 2 levels were assessed by enzyme-linked immunosorbent assay. RESULTS: Serum GLUT1, IGF-2, and VEGF-A levels were significantly higher in patients with IH than in healthy controls (8.80±4.07pg/mL vs. 5.66±4.34pg/mL, 281.10±84.12pg/mL vs. 234.19±75.38pg/mL, 1196.99±389.34pg/mL vs. 996.99±349.16pg/mL, respectively, p=0.026, p=0.030, and p=0.036). Serum GLUT1, IGF-2, and VEGF-A levels in patients with complicated hemangioma were significantly higher than in healthy controls (9.69±3.94pg/mL vs. 5.66±4.34pg/mL, 289.94±83.18pg/mL vs. 234.19±75.38pg/mL, 1276.22±388.24pg/mL vs. 996.99±349.16pg/mL, respectively, p=0.017, p=0.022, and p=0.011). Serum GLUT1, IGF-2, and VEGF-A levels in patients with hemangioma receiving propranolol treatment were significantly higher than in healthy controls. Serum FGF1 levels were higher in patients with IH, complicated hemangioma, and hemangioma receiving propranolol treatment than in healthy controls but the difference was not statistically significantly. CONCLUSION: Serum GLUT1, IGF-2, and VEGF-A levels were positively correlated with disease severity in patients with hemangioma, for example, in complicated hemangioma and hemangioma requiring propranolol treatment. However, further research on larger and different age subgroups is warranted to assess these markers.
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Angiopoyetina 2/sangre , Factor 1 de Crecimiento de Fibroblastos/sangre , Transportador de Glucosa de Tipo 1/sangre , Hemangioma/tratamiento farmacológico , Factor II del Crecimiento Similar a la Insulina/análisis , Propranolol/uso terapéutico , Factor A de Crecimiento Endotelial Vascular/sangre , Neoplasias Vasculares/tratamiento farmacológico , Angiopoyetina 2/uso terapéutico , Biomarcadores/sangre , Niño , Preescolar , Estudios Transversales , Ensayo de Inmunoadsorción Enzimática , Factor 1 de Crecimiento de Fibroblastos/uso terapéutico , Hemangioma/sangre , Hemangioma/patología , Humanos , Lactante , Masculino , Factor A de Crecimiento Endotelial Vascular/uso terapéutico , Neoplasias Vasculares/sangre , Neoplasias Vasculares/patologíaRESUMEN
We evaluated the effects of feeding high volumes of milk replacer on growth and reproductive performances in Japanese black heifers. Fifty-one heifers were fed milk replacer at 9 L/day for 60 days (9 L × 60 days; n = 18) or 41 days (9 L × 41 days; n = 15), or at 7 L/day for 40 days (7 L × 40 days; n = 18). Artificial insemination (AI) was performed on heifers with ≥270 kg body weight and ≥116 cm body height at 300 days of age. The age at the first AI was 0.35 month later for 7 L × 40 days than the other groups (p < .01). However, age at calving did not differ among treatments (22.1 months). The interval from the first AI to pregnancy tended to be ~2 months longer for the 9 L × 60 days than the other groups (p = .07). Our results showed that feeding high volumes of milk replacer may reduce the age at calving via an improved rate of growth. In addition, we propose that feeding a maximum of 7 L milk replacer for 40 days may be the most appropriate rearing regime because the success of pregnancy per AI may be reduced in calves fed a maximum of 9 L for 41 and 60 days.
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Alimentación Animal , Bovinos/sangre , Bovinos/fisiología , Leche , Reproducción , Factores de Edad , Animales , Glucemia/metabolismo , Bovinos/crecimiento & desarrollo , Dieta/veterinaria , Ácidos Grasos no Esterificados/sangre , Femenino , Transportador de Glucosa de Tipo 1/sangre , Inseminación Artificial/veterinaria , Insulina/sangre , Factor I del Crecimiento Similar a la Insulina/metabolismo , Hormona Luteinizante/sangre , Embarazo , Factores de TiempoRESUMEN
We investigated the impact of aerobic exercise (AE) on multiple organ dysfunction syndrome (MODS), aortic injury, pathoglycemia, and death during sepsis. ICR mice were randomized into four groups: Control (Con), Lipopolysaccharide (LPS), Exercise (Ex), and Exercise + LPS (Ex + LPS) groups. Mice were trained with low-intensity for 4 weeks. LPS and Ex + LPS mice received 5 mg/kg LPS intraperitoneally for induction of sepsis. Histopathological micrographs showed the organ morphology and damage. This study examined the effects of AE on LPS-induced changes in systemic inflammation, pulmonary inflammation, lung permeability, and bronchoalveolar lavage fluid (BALF) cell count, oxidative stress-related indicators in the lung, blood glucose levels, plasma lactate levels, serum insulin levels, plasma high-mobility group box 1 (HMGB1) levels, glucose transporter 1 (Glut1) and HMGB1, silent information regulator 1 (Sirt-1), and nuclear factor erythroid 2-related factor 2 (Nrf-2) mRNA expression levels in lung tissue. AE improved sepsis-associated multiple organ dysfunction syndrome (MODS), aortic injury, hypoglycemia, and death. AE prominently decreased pulmonary inflammation, pulmonary edema, and modulated redox balance during sepsis. AE prominently decreased neutrophil content in organ. AE prominently downregulated CXCL-1, CXCL-8, IL-6, TNF-α, Glu1, and HMGB1 mRNA expression but activated IL-1RN, IL-10, Sirt-1, and Nrf-2 mRNA expression in the lung during sepsis. AE decreased the serum levels of lactate and HMGB1 but increased blood glucose levels and serum insulin levels during sepsis. A 4-week AE improves sepsis-associated MODS, aortic injury, pathoglycemia, and death. AE impairs LPS-induced lactate and HMGB1 release partly because AE increases serum insulin levels and decreases the levels of Glut1. AE is a novel therapeutic strategy for sepsis targeting aerobic glycolysis.
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Endotoxemia/terapia , Ejercicio Físico , Glucólisis/fisiología , Insuficiencia Multiorgánica/prevención & control , Condicionamiento Físico Animal/fisiología , Animales , Glucemia/análisis , Líquido del Lavado Bronquioalveolar/citología , Citocinas/análisis , Endotoxemia/inducido químicamente , Endotoxemia/complicaciones , Transportador de Glucosa de Tipo 1/sangre , Proteína HMGB1/sangre , Humanos , Inmunidad Innata , Insulina/sangre , Lactatos/sangre , Lipopolisacáridos/toxicidad , Pulmón/patología , Masculino , Ratones , Ratones Endogámicos ICR , Insuficiencia Multiorgánica/etiología , Insuficiencia Multiorgánica/inmunología , Insuficiencia Multiorgánica/patología , Neutrófilos/patología , Estrés Oxidativo , Distribución Aleatoria , Vísceras/patologíaRESUMEN
AIMS: We aimed to assess whether expression of whole-blood RNA of sodium proton exchanger 1 (NHE1) and glucose transporter 1 (GLUT1) is associated with COVID-19 infection and outcome in patients presenting to the emergency department with respiratory infections. Furthermore, we investigated NHE1 and GLUT1 expression in the myocardium of deceased COVID-19 patients. METHODS AND RESULTS: Whole-blood quantitative assessment of NHE1 and GLUT1 RNA was performed using quantitative PCR in patients with respiratory infection upon first contact in the emergency department and subsequently stratified by SARS-CoV-2 infection status. Assessment of NHE1 and GLUT1 RNA using PCR was also performed in left ventricular myocardium of deceased COVID-19 patients. NHE1 expression is up-regulated in whole blood of patients with COVID-19 compared with other respiratory infections at first medical contact in the emergency department (control: 0.0021 ± 0.0002, COVID-19: 0.0031 ± 0.0003, P = 0.01). The ratio of GLUT1 to NHE1 is significantly decreased in the blood of COVID-19 patients who are subsequently intubated and/or die (severe disease) compared with patients with moderate disease (moderate disease: 0.497 ± 0.083 vs. severe disease: 0.294 ± 0.0336, P = 0.036). This ratio is even further decreased in the myocardium of patients who deceased from COVID-19 in comparison with the myocardium of non-infected donors. CONCLUSIONS: NHE1 and GLUT1 may be critically involved in the disease progression of SARS-CoV-2 infection. We show here that SARS-CoV-2 infection critically disturbs ion channel expression in the heart. A decreased ratio of GLUT1/NHE1 could potentially serve as a biomarker for disease severity in patients with COVID-19.
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COVID-19/metabolismo , Transportador de Glucosa de Tipo 1/sangre , Intercambiador 1 de Sodio-Hidrógeno/sangre , COVID-19/sangre , COVID-19/diagnóstico , Estudios de Casos y Controles , Servicio de Urgencia en Hospital , Femenino , Transportador de Glucosa de Tipo 1/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Valor Predictivo de las Pruebas , Estudios Prospectivos , ARN Mensajero/sangre , Índice de Severidad de la Enfermedad , Intercambiador 1 de Sodio-Hidrógeno/metabolismoRESUMEN
BACKGROUND: Hemoglobin A1c (HbA1c) is an accurate index of fluctuation in glycemia over the 2-3 months prior to quantitative assessment. During this time, hemoglobin (Hb) slowly glycates until it shows the properties of advanced glycation end-products. Glycation kinetics is intensified by prolonged glucose exposure. In subjects undergoing oral glucose tolerance testing (OGTT), immediately after ingestion, glucose is ostensibly transported by the glucose transporter 1 (GLUT1) to erythrocyte corpuscular hemoglobin. The earliest significant measurable level of hemoglobin glycation associated with this transportation is still not clear. SUBJECTS AND METHODS: We attempted to explore the early impact of short-term glucose load on HbA1c levels, because it is now known that transmembrane GLUT1-mediated glucose transport occurs immediately. A total of 88 participants (46 patients and 42 clinically healthy controls) underwent fasting plasma glucose quantitation during an OGTT. HbA1c, revealed by a monoclonal anti-glycation epitope antibody and adiponectin, was quantitated before (T0) and 2 hours (T120) after 80 g glucose ingestion. RESULTS: Wilcoxon test revealed that the HbA1c values did not significantly vary (P=0.15) during the OGTT, whereas glucose concentration varied strongly between T0 and T120. DISCUSSION: It is well known that quantitative estimation of HbA1c is informative for clinical care, independently of glucose level. The molecular mechanisms and dynamics by which glucose enters/exits red blood cells are incompletely known and may differ between individuals. We here show, for the first time, that HbA1c levels do not significantly increase during OGTT, supporting the view that non-enzymatic glycation of hemoglobin occurs slowly and that glycation during the 2 hours of an OGTT is insignificant.
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Glucemia/análisis , Prueba de Tolerancia a la Glucosa , Hemoglobina Glucada/análisis , Hemoglobinas/metabolismo , Adiponectina/sangre , Adolescente , Adulto , Glucemia/metabolismo , Niño , Diabetes Mellitus/sangre , Eritrocitos/metabolismo , Ayuno , Transportador de Glucosa de Tipo 1/sangre , Glicosilación , Humanos , Persona de Mediana Edad , Estado Prediabético/sangreRESUMEN
OBJECTIVE: White blood cells are essential in mediating immune and inflammatory responses. A prominent feature of these cells during activation of the immune function is increased glucose utilization, and this is dependent on the functioning of specific glucose transporter (GLUT) isoforms. The few data available on leukocyte glucose transporter expression are limited to type-2 diabetes mellitus, and nothing is known about its regulation. MATERIAL AND METHODS: Peripheral blood was drawn from 35 healthy controls and 35 diabetic subjects. Expression of GLUT1, GLUT3 and GLUT4 was determined in the leukocytes of healthy individuals and diabetic patients by flow cytometry, Western blot and semi-quantitative RT-PCR. RESULTS: GLUT 3 was decreased in granulocytes, lymphocytes and monocytes from diabetic patients. In monocytes, GLUT3 and GLUT4 were reduced in type-2 diabetic patients. In leukocytes of diabetic patients, GLUT1 and GLUT4, protein and mRNA were unchanged, but GLUT3 protein and mRNA levels were down-regulated compared to those of healthy controls. CONCLUSION: Elevated glucose concentration affects leukocyte GLUT expression. Decreased expression of GLUT isoforms in leukocytes may be responsible for diminished activation of diabetic leukocytes. These situations possibly contribute to a predisposition to infection and to a decreased immune response in diabetes.
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Diabetes Mellitus Tipo 2/metabolismo , Regulación hacia Abajo , Proteínas Facilitadoras del Transporte de la Glucosa/sangre , Adulto , Diabetes Mellitus Tipo 2/genética , Femenino , Citometría de Flujo , Proteínas Facilitadoras del Transporte de la Glucosa/genética , Transportador de Glucosa de Tipo 1/sangre , Transportador de Glucosa de Tipo 1/genética , Transportador de Glucosa de Tipo 3/sangre , Transportador de Glucosa de Tipo 3/genética , Transportador de Glucosa de Tipo 4/sangre , Transportador de Glucosa de Tipo 4/genética , Humanos , Leucocitos/metabolismo , Masculino , Persona de Mediana Edad , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Vitamin C is incorporated into the cerebrospinal fluid (CSF) through choroid plexus cells. While the transfer of vitamin C from the blood to the brain has been studied functionally, the vitamin C transporter, SVCT2, has not been detected in the basolateral membrane of choroid plexus cells. Furthermore, it is unknown how its expression is induced in the developing brain and modulated in scurvy conditions. We concluded that SVCT2 is intensely expressed in the second half of embryonic brain development and postnatal stages. In postnatal and adult brain, SVCT2 is highly expressed in all choroidal plexus epithelial cells, shown by colocalization with GLUT1 in the basolateral membranes and without MCT1 colocalization, which is expressed in the apical membrane. We confirmed that choroid plexus explant cells (in vitro) form a sealed epithelial structure, which polarized basolaterally, endogenous or overexpressed SVCT2. These results are reproduced in vivo by injecting hSVCT2wt-EYFP lentivirus into the CSF. Overexpressed SVCT2 incorporates AA (intraperitoneally injected) from the blood to the CSF. Finally, we observed in Guinea pig brain under scorbutic condition, that normal distribution of SVCT2 in choroid plexus may be regulated by peripheral concentrations of vitamin C. Additionally, we observed that SVCT2 polarization also depends on the metabolic stage of the choroid plexus cells.
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Ácido Ascórbico/metabolismo , Encéfalo/metabolismo , Transportador de Glucosa de Tipo 1/sangre , Transportadores de Sodio Acoplados a la Vitamina C/sangre , Animales , Barrera Hematoencefálica/crecimiento & desarrollo , Barrera Hematoencefálica/metabolismo , Encéfalo/crecimiento & desarrollo , Membrana Celular/metabolismo , Células Cultivadas , Plexo Coroideo/metabolismo , Desarrollo Embrionario/genética , Células Epiteliales/metabolismo , Células Epiteliales/patología , Regulación del Desarrollo de la Expresión Génica/genética , Cobayas , Ratones , Transportadores de Ácidos Monocarboxílicos/genética , Neuronas/metabolismo , Transportadores de Sodio Acoplados a la Vitamina C/líquido cefalorraquídeo , Porcinos , Simportadores/genéticaAsunto(s)
Transportador de Glucosa de Tipo 1/biosíntesis , Hemangioma/sangre , Hemangioma/diagnóstico , Malformaciones Vasculares/diagnóstico , Biomarcadores de Tumor/biosíntesis , Biomarcadores de Tumor/sangre , Angioma Venoso del Sistema Nervioso Central , Diagnóstico Diferencial , Transportador de Glucosa de Tipo 1/sangre , HumanosRESUMEN
We report the results of (18)F-fluorodeoxyglucose positron emission tomography (FDG PET) and immunohistochemical staining of glucose transporter 1 (Glut-1) and hexokinase II (HK-II) in patients with hepatocellular carcinoma (HCC) and cholangiocellular carcinoma (CCC) to observe the variation in (18)F-FDG uptake and variation in expression of Glut-1 and HK-II in these hepatic tumors. In the case of HCC, moderate (18)F-FDG uptake and strong expression of HK-II were detected, whereas Glut-1 was not expressed. Conversely, CCC showed high (18)F-FDG uptake and increased expression of Glut-1 but HK-II was not expressed. The variation in the (18)F-FDG uptake and expression of Glut 1 and HK-II in HCC and CCC might be owing to the difference in origin and the different mechanisms involved in glucose uptake, rate of glucose transporters, and hexokinase activity involved in the glycolytic pathway.