RESUMEN
AIMS/HYPOTHESIS: In adipocytes, triacylglycerol synthesis depends on the formation of glycerol 3-phosphate, which originates either from glucose, through glycolysis, or from lactate, through glyceroneogenesis. However, glucose is traditionally viewed as the main precursor of the glycerol backbone and thus, enhanced glucose uptake would be expected to result in increased triacylglycerol synthesis and contribute to obesity. METHODS: To further explore this issue, we generated a mouse model with chronically increased glucose uptake in adipose tissue by expressing Gck, which encodes the glucokinase enzyme. RESULTS: Here we show that the production of high levels of glucokinase led to increased adipose tissue glucose uptake and lactate production, improved glucose tolerance and higher whole-body and skeletal muscle insulin sensitivity. There was no parallel increase in glycerol 3-phosphate synthesis in vivo, fat accumulation or obesity. Moreover, at high glucose concentrations, in cultured fat cells overproducing glucokinase, glycerol 3-phosphate synthesis from pyruvate decreased, while glyceroneogenesis increased in fat cells overproducing hexokinase II. CONCLUSIONS/INTERPRETATIONS: These findings indicate that the absence of glucokinase inhibition by glucose 6-phosphate probably led to increased glycolysis and blocked glyceroneogenesis in the mouse model. Furthermore, this study suggests that under physiological conditions, when blood glucose increases, glyceroneogenesis may prevail over glycolysis for triacylglycerol formation because of the inhibition of hexokinase II by glucose 6-phosphate. Together these results point to the indirect pathway (glucose to lactate to glycerol 3-phosphate) being key for fat deposition in adipose tissue.
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Tejido Adiposo/enzimología , Glucosa/metabolismo , Insulina/farmacología , Ácido Láctico/metabolismo , Obesidad/prevención & control , Células 3T3-L1 , Tejido Adiposo/efectos de los fármacos , Animales , Northern Blotting , Células Cultivadas , Glucoquinasa/genética , Glucoquinasa/metabolismo , Glicerofosfatos/metabolismo , Hipoglucemiantes/farmacología , Masculino , Ratones , Ratones Transgénicos , Obesidad/genética , Obesidad/metabolismo , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
AIM: To provide a summary of the different experimental models of traumatic brain injury (TBI) designed under both in vivo and in vitro conditions. A comprehensible review of the specific types of brain lesions induced, as well as the technical details to reproduce each model at the laboratory is given. DEVELOPMENT: Outcome of patients suffering from a TBI has significantly improved with the rapid application of vital supporting measures in addition to a strict control of blood and intracranial pressure at the intensive care units. However no specific treatment for post-traumatic brain lesions has proven as efficacious in the clinical settings. A deeper knowledge of the physiopathological events associated with TBI is necessary for the development of new specific therapies. Due to the heterogeneity of the human TBI, each experimental model has been designed to reproduce a different type of brain lesion. Experimental TBI models allow the study of the dynamic evolution of brain injuries under controlled conditions. Usefulness of experimental models is limited by their reliability and reproducibility among different researchers. Small rodents have been the preferred animals to reproduce TBI injuries, mainly due to the similar cerebral physiology shared by these animals and the human beings. CONCLUSION: The use of experimental models of TBI is the most appropriate tool to study the mechanisms underlying this type of injury. However their simplicity precludes an exact reproduction of the heterogeneous cerebral damage observed in clinical settings. This could be the main reason for the discrepancies observed in the therapeutic effects of treatments between experimental and clinical studies.
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Lesiones Encefálicas , Modelos Animales , Animales , Lesiones Encefálicas/patología , Lesiones Encefálicas/terapia , Humanos , Reproducibilidad de los Resultados , Resultado del TratamientoRESUMEN
The value of extracellular pH (pH(e)) in tumors is an important factor in prognosisand choice of therapy. We demonstrate here that pH(e) can be mappedin vivo in a rat brain glioma by (1)H magnetic resonance spectroscopic imaging (SI) of the pH buffer (+/-)2-imidazole-1-yl-3-ethoxycarbonylpropionic acid (IEPA). (1)H SI also allowed us to map metabolites, and, to better understand the determinants of pH(e), we compared maps of pH(e), metabolites, and the distribution of the contrast agent gadolinium1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraaceticacid (Gd-DOTA). C6 cells injected in caudate nuclei of four Wistar rats gave rise to gliomas of approximately 10 mm in diameter. Three mmols of IEPA were injected in the right jugular vein from t = 0 to t = 60 min. From t = 50 min to t = 90 min, spin-echo (1)H SI was performed with an echo time of 40 ms in a 2.5-mm slice including the glioma (nominal voxel size, 2.2 microl). IEPA resonances were detected only within the glioma and were intense enough for pH(e) to be calculated from the chemical shift of the H2 resonance in almost all voxels of the glioma. (1)H spectroscopic images with an echo time of 136 ms were then acquired to map metabolites: lactate, choline-containing compounds (tCho), phosphocreatine/creatine, and N-acetylaspartate. Finally, T(1)-weighted imaging after injection of a bolus of Gd-DOTA gave a map indicative of extravasation. On average, the gradient of pH(e) (measured where sufficient IEPA was present) from the center to the periphery was not statistically significant. Mean pH(e) was calculated for each of the four gliomas, and the average was 7.084 +/- 0.017 (+/- SE; n = 4 rats), which is acid with respect to pH(e) of normal tissue. After normalization of spectra to their water peak, voxel-by-voxel comparisons of peak areas showed that N-acetylaspartate, a marker of neurons, correlated negatively with IEPA (P < 0.0001) and lactate (P < 0.05), as expected of a glioma surrounded by normal tissue. tCho (which may indicate proliferation) correlated positively with pH(e) (P < 0.0001). Lactate correlated positively with tCho (P < 0.0001), phosphocreatine/creatine (P < 0.001), and Gd-DOTA (P < 0.0001). Although lactate is exported from cells in association with protons, within the gliomas, no evidence was observed that pH(e) was significantly lower where lactate concentration was higher. These results suggest that lactate is produced mainly in viable, well-perfused, tumoral tissue from which proton equivalents are rapidly cleared.
Asunto(s)
Neoplasias Encefálicas/metabolismo , Glioma/metabolismo , Concentración de Iones de Hidrógeno , Animales , Ácido Aspártico/análogos & derivados , Ácido Aspártico/metabolismo , Tampones (Química) , Colina/metabolismo , Medios de Contraste , Creatina/metabolismo , Espacio Extracelular/metabolismo , Femenino , Compuestos Heterocíclicos , Imidazoles/metabolismo , Ácido Láctico/metabolismo , Espectroscopía de Resonancia Magnética/métodos , Masculino , Compuestos Organometálicos , Fosfocreatina/metabolismo , Propionatos/metabolismo , Protones , Ratas , Ratas WistarRESUMEN
Triggering of the macrophage cell line RAW 264.7 with lipopolysaccharide and interferon-gamma promoted apoptosis that was prevented by inhibitors of type 2 nitric oxide synthase or caspase. Using (1)H NMR analysis, we have investigated the changes of the intracellular transverse relaxation time (T(2)) and apparent diffusion coefficient (ADC) as parameters reflecting the rotational and translational motions of water in apoptotic macrophages. T(2) values decreased significantly from 287 to 182 ms in cells treated for 18 h with NO-donors. These changes of T(2) were prevented by caspase inhibitors and were not due to mitochondrial depolarization or microtubule depolymerization. The decrease of the intracellular values of T(2) and ADC in apoptotic macrophages was observed after caspase activation, but preceded phosphatidylserine exposure and nucleosomal DNA cleavage. The changes of water motion were accompanied by an enhancement of the hydrophobic properties of the intracellular milieu, as detected by fluorescent probes. These results indicate the occurrence of an alteration in the physicochemical properties of intracellular water during the course of apoptosis.
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Apoptosis , Agua Corporal/química , Caspasas/metabolismo , Macrófagos/citología , Clorometilcetonas de Aminoácidos/farmacología , Animales , Línea Celular , Inhibidores de Cisteína Proteinasa/farmacología , Citoplasma/química , Difusión , Activación Enzimática , Humanos , Células Jurkat , Cinética , Espectroscopía de Resonancia Magnética , Movimiento , Óxido Nítrico/fisiología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo IIRESUMEN
We describe a multivariate analysis procedure to classify human cerebral tumors nonhistologically in vitro, combining the use of 1H magnetic resonance spectroscopy (MRS) with automatic amino acid analysis of biopsy extracts. Eighty-one biopsies were obtained surgically and classified histologically in eight classes: high-grade astrocytomas (class 1, n = 19), low-grade astrocytomas (class 2, n = 10), normal brain (class 3, n = 9), medulloblastomas (class 4, n = 4), meningiomas (class 5, n = 18), metastases (class 6, n = 8), neurinomas (class 7, n = 9), and oligodendrogliomas (class 8, n = 4). Perchloric acid extracts were prepared from every biopsy and analyzed by high resolution 1H MRS and automatic amino acid analysis by ionic exchange chromatography. Intensities of 27 resonances and ratios of resonances were measured in the 1H MRS spectra, and 17 amino acid concentrations were determined in the chromatograms. Linear discriminant analysis provided the most adequate combination of these variables for binary classifications of a biopsy between any two possible classes and in multiple choice comparisons, involving the eight possible classes considered. Correct diagnosis was obtained when the class selected by the computer matched the histological diagnosis. In binary comparisons, consideration of the amino acid profile increased the percentage of correct classifications, being always higher than 75% and reaching 100% in many cases. In multilateral comparisons, scores were: high-grade astrocytomas, 80%; low-grade astrocytomas, 74%; normal brain, 100%; medulloblastomas, 100%; meningiomas, 94.5%; metastases, 86%; neurinomas, 100%; and oligodendrogliomas, 75%. These results indicate that statistical multivariate procedures, combining 1H MRS and amino acid analysis of tissue extracts, provide a valuable classifier for the nonhistological diagnosis of biopsies from brain tumors in vitro.
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Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/diagnóstico , Espectroscopía de Resonancia Magnética/métodos , Análisis de Secuencia de Proteína/métodos , Algoritmos , Astrocitoma/diagnóstico , Astrocitoma/metabolismo , Biopsia , Encéfalo/diagnóstico por imagen , Encéfalo/metabolismo , Neoplasias Encefálicas/metabolismo , Cromatografía por Intercambio Iónico , Humanos , Meduloblastoma/diagnóstico , Meduloblastoma/metabolismo , Meningioma/diagnóstico , Meningioma/metabolismo , Modelos Estadísticos , Neurilemoma/diagnóstico , Neurilemoma/metabolismo , Oligodendroglioma/diagnóstico , Oligodendroglioma/metabolismo , Radiografía , Factores de TiempoRESUMEN
The effects of adult-onset hypothyroidism on the metabolic compartmentation of the cerebral tricarboxylic acid cycle and the gamma-aminobutyric acid (GABA) shunt have been investigated by 13C nuclear magnetic resonance spectroscopy. Rats thyroidectomized as adults and age-matched controls were infused in the right jugular vein with unlabeled or (1,2-13C2) acetate solutions for 60 min. At the end of the infusion, the brains were frozen in situ and perchloric acid extracts were prepared and analyzed by 13C nuclear magnetic resonance and reverse-phase HPLC. Thyroidectomized animals showed a decrease in the incorporation of 13C from (1,2-13C2) acetate in cerebral metabolites and an increase in the concentrations of unlabeled glutamate and GABA. Computer-assisted interpretation of the 13C multiplets observed for the carbons of glutamate, glutamine, and GABA indicated that adult-onset hypothyroidism produced 1) a decrease in the contribution of infused (1,2-13C2) acetate to the glial tricarboxylic acid cycle; 2) an increase in the contribution of unlabeled acetyl-CoA to the neuronal tricarboxylic acid cycle; and 3) impairments in the exchange of glutamate, glutamine, and GABA between the neuronal and glial compartments. Despite the fact that the adult brain has often been considered metabolically unresponsive to thyroid hormone status, present results show metabolic alterations in the neuronal and glial compartments that are reversible with substitution therapy.
Asunto(s)
Acetatos/metabolismo , Encéfalo/metabolismo , Hipotiroidismo/metabolismo , Edad de Inicio , Animales , Química Encefálica , Isótopos de Carbono , Procesamiento Automatizado de Datos , Femenino , Ácido Glutámico/metabolismo , Glutamina/metabolismo , Espectroscopía de Resonancia Magnética , Modelos Neurológicos , Ratas , Ratas Wistar , Tiroidectomía , Tiroxina/farmacología , Extractos de Tejidos/metabolismo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
A previously unassigned group of prominent multiplets of the 360 MHz 1H NMR spectrum of acid stable metabolite extracts from rat brain is shown to arise from free myo-inositol. This conclusion is derived from a systematic analysis of the high-resolution 1H NMR spectra of brain acid extracts, in which appropriate conditions and optimal proton signals have been selected for the quantitative analysis of up to 15 metabolites. Developmental variations in the cerebral content of myo-inositol could be readily detected using this approach, which provides a novel alternative to study myo-inositol metabolism under physiological or pathological conditions.
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Química Encefálica , Inositol/análisis , Animales , Encéfalo/crecimiento & desarrollo , Espectroscopía de Resonancia Magnética , Masculino , Ratas , Ratas EndogámicasRESUMEN
The sources of ammonia used by isolated, intact rat liver mitochondria in the production of citrulline have been investigated in situ using a novel methodology based on the analysis of 13C-15N heteronuclear couplings observed by 13C NMR. Isolated mitochondria from rat liver were incubated with ornithine, 13CO3H- and 15NH4Cl, using unlabeled glutamate or glutamine as alternative, intramitochondrial nitrogen donors. The production of (7-13C, 8-15N) or (7-13C, 8-14N) citrulline was determined in situ by 13C NMR and the relative proportions of 15N- and 14N-citrullines confirmed by high resolution 13C NMR analysis of the C-7 citrulline resonance observed in perchloric acid extracts prepared at the end of the incubations. The 15N fractional enrichment of the intramitochondrial NH3 pool was manipulated either by modifying the 15N enrichment of added 15NH4Cl, or by altering the concentration of the unlabeled nitrogen donors in the incubation medium. Fractional 15N enrichments measured in the N-8 nitrogen of the resulting (7-13C) citrulline closely paralleled those of the external 15NH4Cl with minor dilutions derived from the unlabeled nitrogen contribution from the alternative substrates. In the presence of 10 mM 15NH4Cl, 10 mM glutamate contributed 4% of the citrulline N-8 nitrogen. Under similar conditions, the contribution of nitrogen from 10 mM glutamine to N-8 citrulline was 6%. These results indicate that the primary source of ammonia used for citrulline synthesis by isolated, intact rat liver mitochondria is extramitochondrial, providing also an illustration of the use of 13C-15N spin coupling patterns observed by 13C NMR, as a new tool in the study of ammonia metabolism.
Asunto(s)
Amoníaco/metabolismo , Citrulina/biosíntesis , Espectroscopía de Resonancia Magnética/métodos , Mitocondrias Hepáticas/metabolismo , Animales , Carbono , Glutamatos/metabolismo , Ácido Glutámico , Glutamina/metabolismo , Técnicas In Vitro , Masculino , Isótopos de Nitrógeno , Ratas , Ratas WistarRESUMEN
Ex vivo ¿(13)C, (2)H¿ NMR spectroscopy allowed to estimate the relative sizes of neuronal and glial glutamate pools and the relative contributions of (1-(13)C) glucose and (2-(13)C, 2-(2)H(3)) acetate to the neuronal and glial tricarboxylic acid cycles of the adult rat brain. Rats were infused during 60 min in the right jugular vein with solutions containing (2-(13)C, 2-(2)H(3)) acetate and (1-(13)C) glucose or (2-(13)C, 2-(2)H(3)) acetate only. At the end of the infusion the brains were frozen in situ and perchloric acid extracts were prepared and analyzed by high resolution (13)C NMR spectroscopy (90.5 MHz). The relative sizes of the neuronal and glial glutamate pools and the contributions of acetyl-CoA molecules derived from (2-(13)C, (2)H(3)) acetate or (1-(13)C) glucose entering the tricarboxylic acid cycles of both compartments, could be determined by the analysis of (2)H-(13)C multiplets and (2)H induced isotopic shifts observed in the C4 carbon resonances of glutamate and glutamine. During the infusions with (2-(13)C, 2-(2)H(3)) acetate and (1-(13)C) glucose, the glial glutamate pool contributed 9% of total cerebral glutamate being derived from (2-(13)C, 2-(2)H(3)) acetyl-CoA (4%), (2-(13)C) acetyl-CoA (3%) and recycled (2-(13)C, 2-(2)H) acetyl-CoA (2%). The neuronal glutamate pool accounted for 91% of the total cerebral glutamate being mainly originated from (2-(13)C) acetyl-CoA (86%) and (2-(13)C, 2-(2)H) acetyl-CoA (5%). During the infusions of (2-(13)C, 2-(2)H(3)) acetate only, the glial glutamate pool contributed 73% of the cerebral glutamate, being derived from (2-(13)C, 2-(2)H(3)) acetyl-CoA (36%), (2-(13)C, 2-(2)H) acetyl-CoA (27%) and (2-(13)C) acetyl-CoA (10%). The neuronal pool contributed 27% of cerebral glutamate being formed from (2-(13)C) acetyl-CoA (11%) and recycled (2-(13)C, 2-(2)H) acetyl-CoA (16%). These results illustrate the potential of ¿(13)C, (2)H¿ NMR spectroscopy as a novel approach to investigate substrate selection and metabolic compartmentation in the adult mammalian brain.
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Acetatos/metabolismo , Química Encefálica/fisiología , Encéfalo/citología , Glucosa/metabolismo , Neuroglía/metabolismo , Neuronas/metabolismo , Animales , Ácido Glutámico/metabolismo , Glutamina/metabolismo , Espectroscopía de Resonancia Magnética , Masculino , Ratas , Ratas WistarRESUMEN
Pyruvate and malate (P/M) increase the contribution of mitochondria to neuronal calcium homeostasis. We have now found that cortical neuronal cultures utilize pyruvate preferentially over glucose. The supply of pyruvate and malate protects hippocampal and cortical neurons against delayed cell death occurring 24 h after glutamate exposure. High [Ca2+]i levels attained during and after glutamate exposure were reduced when neurons were incubated in the presence of P/M. At the single cell level, this was reflected in a decrease in the number of neurons that respond to glutamate with high rises in [Ca2+]i. The results suggest that the ability to prevent large increases in [Ca2+]i may underlie the beneficial effects of pyruvate and malate during glutamate excitotoxicity.
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Corteza Cerebral/citología , Ácido Glutámico/toxicidad , Hipocampo/citología , Malatos/farmacología , Neuronas/efectos de los fármacos , Neurotoxinas/toxicidad , Ácido Pirúvico/farmacología , Animales , Calcio/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Embrión de Mamíferos , Cinética , Malatos/metabolismo , Neuronas/citología , Neuronas/metabolismo , Ácido Pirúvico/metabolismo , RatasRESUMEN
AIMS: To analyze the functional reasons justifying the existence of the blood brain barrier with an emphasis on its fundamental role supporting neuroglial coupling. DEVELOPMENT: We review in an integrated manner the contributions of different research areas in physiology and metabolism of the central nervous system which allow to understand the functional need for the existence of the blood brain barrier. In particular, we describe the physiological basis of the metabolic functional coupling and the metabolic interactions between neurons and glial cells, two properties directly derived from the presence of the blood brain barrier. Likewise the blood brain barrier is presented as an important determinant of the heterogeneous activation of cerebral tissue as detected by neuroimaging technologies as positron emission tomography and functional magnetic resonance imaging. CONCLUSIONS: The main function of the blood brain barrier is to maintain a stable composition of the extracellular milieu in nervous tissue. This allows the changes in ionic composition and neurotransmitter concentration in the extracellular milieu, to reflect indirectly the generation of action potentials and the state of neurotransmission of neuronal circuits. Glial cells induce the development of the blood brain barrier and are the main sensors of neuronal function, due to their important take up capacity for extracellular potassium and neurotransmitters. Glial homeostasis of the extracellular milieu is circuit specific, limiting the functional metabolic coupling to discrete regions of the brain and generating the classical pattern of heterogeneous activity in the different modules of the nervous tissue.
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Barrera Hematoencefálica/fisiología , Sistema Nervioso Central/fisiología , Barrera Hematoencefálica/ultraestructura , Sistema Nervioso Central/anatomía & histología , Líquido Extracelular/química , Glucosa/metabolismo , Ácido Glutámico/metabolismo , Glutamina/metabolismo , Homeostasis , Humanos , Red Nerviosa , Neuroglía/metabolismo , Neuroglía/ultraestructura , Neuronas/metabolismo , Neuronas/ultraestructura , Potasio/metabolismo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Murine models are the most commonly used and best investigated among the animal models of HGG. They constitute an important weapon in the development and testing of new anticancer drugs and have long been used in preclinical trials. Neuroimaging methods, particularly MR imaging, offer important advantages for the evaluation of treatment response: shorter and more reliable treatment end points and insight on tumor biology and physiology through the use of functional imaging DWI, PWI, BOLD, and MR spectroscopy. This functional information has been progressively consolidated as a surrogate marker of tumor biology and genetics and may play a pivotal role in the assessment of specifically targeted drugs, both in clinical and preclinical trials. The purpose of this Research Perspectives was to compile, summarize, and critically assess the available information on the neuroimaging features of different murine models of HGGs, and explain how these correlate with human disease and reflect tumor biology.
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Neoplasias Encefálicas/patología , Encéfalo/patología , Modelos Animales de Enfermedad , Glioma/patología , Imagen por Resonancia Magnética/métodos , Animales , Humanos , Ratones , Especificidad de la EspecieAsunto(s)
Coenzimas/farmacología , Grasas de la Dieta/farmacología , Etanol/farmacología , Hígado/efectos de los fármacos , Animales , Coenzima A/farmacología , Citidina Difosfato Colina/farmacología , Hígado/enzimología , Masculino , Ratas , S-Adenosilmetionina/farmacología , Uridina Difosfato Glucosa/farmacologíaRESUMEN
An overview of 13C nuclear magnetic resonance (NMR) spectroscopy methods and their applications in the study of the metabolism of brain cells in vitro and in the in vivo brain is presented as well as their implications for modern molecular imaging techniques. Various topics will be discussed, such as general properties of the 13C NMR spectrum, 13C NMR spectroscopy acquisition protocols, determination of fractional 13C enrichment, 13C(2H) NMR methodologies, and the use of 13C hyperpolarized substrates for NMR spectroscopy and imaging. Some illustrative applications are described, both in vitro and in vivo.
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Encéfalo/patología , Isótopos de Carbono/farmacología , Espectroscopía de Resonancia Magnética/métodos , Imagen Molecular/métodos , Neuroquímica/métodos , Animales , Ácido Glutámico/química , Glutamina/metabolismo , Humanos , Hidrógeno/química , Concentración de Iones de Hidrógeno , Litio/química , Angiografía por Resonancia Magnética/métodos , Modelos Biológicos , RatasRESUMEN
Hepatic triglyceride (HTG) accumulation from peripheral dietary sources and from endogenous de novo lipogenesis (DNL) was quantified in adult Sprague-Dawley rats by combining in vivo localized (1)H MRS measurement of total hepatic lipid with a novel ex vivo (2)H NMR analysis of HTG (2)H enrichment from (2)H-enriched body water. The methodology for DNL determination needs further validation against standard methodologies. To examine the effect of a high-fat diet on HTG concentrations and sources, animals (n = 5) were given high-fat chow for 35 days. HTG accumulation, measured by in vivo (1)H MRS, increased significantly after 1 week (3.85 +/- 0.60% vs 2.13 +/- 0.34% for animals fed on a standard chow diet, P < 0.05) and was maintained until week 5 (3.30 +/- 0.60% vs 1.12 +/- 0.30%, P < 0.05). Animals fed on a high-fat diet were glucose intolerant (13.3 +/- 1.3 vs 9.4 +/- 0.8 mM in animals fed on a standard chow diet, for 60 min glycemia after glucose challenge, P < 0.05). In control animals, DNL accounted for 10.9 +/- 1.0% of HTG, whereas in animals given the high-fat diet, the DNL contribution was significantly reduced to 1.0 +/- 0.2% (P < 0.01 relative to controls). In a separate study to determine the response of HTG to weaning from a high-fat diet, animals with raised HTG (3.33 +/- 0.51%) after 7 days of a high-fat diet reverted to basal HTG concentrations (0.76 +/- 0.06%) after an additional 7 days of weaning on a standard chow diet. These studies show that, in healthy rats, HTG concentrations are acutely influenced by dietary lipid concentrations. Although the DNL contribution to HTG content is suppressed by a high-fat diet in adult Sprague-Dawley rats, this effect is insufficient to prevent overall increases in HTG concentrations.
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Grasas de la Dieta/administración & dosificación , Grasas de la Dieta/farmacología , Conducta Alimentaria/efectos de los fármacos , Salud , Hígado/efectos de los fármacos , Hígado/metabolismo , Triglicéridos/metabolismo , Animales , Glucosa/administración & dosificación , Glucosa/farmacología , Lipogénesis/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Masculino , Ratas , Ratas Sprague-Dawley , Agua/metabolismoRESUMEN
(1)H MRS is evolving into an invaluable tool for brain tumor classification in humans based on pattern recognition analysis, but there is still room for improvement. Here we propose a new approach: to challenge tumor metabolism in vivo by a defined perturbation, and study the induced changes in MRS pattern. For this we recorded single voxel (1)H MR spectra from mice bearing a stereotactically induced GL261 grade IV brain glioma during a period of induced acute hyperglycemia. A total of 29 C57BL/6 mice were used. Single voxel spectra were acquired at 7 T with point resolved spectroscopy and TE of 12, 30 and 136 ms. Tumors were induced by stereotactic injection of 10(5) GL261cells in 17 mice. Hyperglycemia (up to 338 +/- 36 mg/dL glucose in the blood) was induced by intraperitoneal bolus injection. Maximal increases in glucose resonances of up to 2.4-fold were recorded from tumors in vivo. Our observations are in agreement with extracellular accumulation of glucose, which may suggest that glucose transport and/or metabolism are working close to their maximum capacity in GL261 tumors. The significant and specific MRS pattern changes observed when comparing euglycemia and hyperglycemia may be of use for future pattern-recognition studies of animal and human brain tumors by enhancing MRS-based discrimination between tumor types and grades.
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Glioma , Hiperglucemia , Interpretación de Imagen Asistida por Computador , Animales , Glucemia/metabolismo , Encéfalo/metabolismo , Encéfalo/patología , Femenino , Glioma/metabolismo , Glioma/patología , Glucosa/química , Glucosa/metabolismo , Humanos , Espectroscopía de Resonancia Magnética , Masculino , Ratones , Ratones Endogámicos C57BL , Estadificación de NeoplasiasRESUMEN
We review the different methods for measuring pH by magnetic resonance imaging and magnetic resonance spectroscopy and discuss their potential diagnostic repercussions. We begin with a brief description of intra- and extra-cellular pH regulation in physiological and pathological conditions. Then we present the main 31P or 1H magnetic resonance spectroscopy procedures, which are based on the dependence of the pH on the chemical displacements of the intrinsic intracellular inorganic phosphate or of the H2 proton of imidazole in extrinsic indicators. Finally, we describe the procedures that use magnetic resonance imaging, whose main tool is the dependence of the pH (i) on the relaxivity of certain paramagnetic contrast agents, or (ii) on the processes of magnetic transference between diamagnetic molecules (DIACEST) or paramagnetic molecules (PARACEST) and the free water in the tissues. We briefly illustrate the potential clinical applications of these new procedures.
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Líquido Extracelular/química , Líquido Intracelular/química , Imagen por Resonancia Magnética , Espectroscopía de Resonancia Magnética , Humanos , Concentración de Iones de HidrógenoRESUMEN
PURPOSE: Chemotherapy increases survival in breast cancer patients. Consequently, cerebral metastases have recently become a significant clinical problem, with an incidence of 30-40% among breast carcinoma patients. As this phenomenon cannot be studied longitudinally in humans, models which mimic brain metastasis are needed to investigate its pathogenesis. Such models may later be used in experimental therapeutic approaches. MATERIAL AND METHODS/RESULTS: We report a model in which 69% of the animals (9/13 BALB/c nude mice) developed MR-detectable abnormal masses in the brain parenchyma within a 20 to 62-day time window post intra-carotid injection of 435-Br1 human cells. The masses detected in vivo were either single (7 animals) or multiple (2 animals). Longitudinal MR (MRI/MRS) studies and post-mortem histological data were correlated, revealing a total incidence of experimental brain metastases of 85% in the cases studied (11/13 animals). ADC maps perfectly differentiated edema and/or CSF areas from metastasis. Preliminary MRS data also revealed additional features: decrease in N-acetyl aspartate (NAA) was the first MRS-based marker of metastasis growth in the brain (micrometastasis); choline-containing compounds (Cho) rose and creatine (Cr) levels decreased as these lesions evolved, with mobile lipids and lactate also becoming visible. Furthermore, MRS pattern recognition-based analysis suggested that this approach may help to discriminate different growth stages. CONCLUSIONS: This study paves the way for further in vivo studies oriented towards detection of different tumor progression states and for improving treatment efficiency.
Asunto(s)
Neoplasias Encefálicas/patología , Neoplasias Mamarias Animales/patología , Animales , Ácido Aspártico/análogos & derivados , Ácido Aspártico/metabolismo , Neoplasias Encefálicas/secundario , Línea Celular Tumoral , Colina/metabolismo , Creatina/metabolismo , Femenino , Humanos , Imagen por Resonancia Magnética/métodos , Espectroscopía de Resonancia Magnética/métodos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de NeoplasiasRESUMEN
The characterization of a new class of hydrophilic liver-targeted agents for gamma-scintigraphy and MRI, consisting, respectively, of [(153)Sm](3+) or Gd(3+) complexes of DOTA monoamide or bisamide linked glycoconjugates (DOTA = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), is reported. In vitro studies show high uptake of radiolabeled [(153)Sm]-DOTAGal(2) by the human hepatocyte carcinoma cell line Hep G2 containing the asialoglycoprotein receptor (ASGP-R), which is decreased to less than 50% by the presence of its high-affinity ligand asialofetuin (ASF). In vivo biodistribution, gamma-imaging and pharmacokinetic studies on Wistar rats using the [(153)Sm](3+)-labeled glycoconjugates show a high uptake in the receptor-rich organ liver of the radiolabeled compounds containing terminal galactosyl groups, but very little uptake for those compounds with terminal glycosyl groups. Blocking the receptor in vivo reduced liver uptake by 90%, strongly suggesting that the liver uptake of these compounds is mediated by their binding to the asyaloglycoprotein receptor (ASGP-R). This study also demonstrated that the valency increase improves the targeting capability of the glycoconjugates, which is also affected by their topology. However despite the specific liver uptake of the radiolabeled galactose-bearing multivalent compounds, the animal MRI assessment of the corresponding Gd(3+) chelates shows liver-to-kidney contrast effects which are not significantly better than those shown by GdDTPA. This probably results from the quick wash-out from the liver of these highly hydrophilic complexes, before they can be sufficiently concentrated within the hepatocytes via receptor-mediated endocytosis.
Asunto(s)
Receptor de Asialoglicoproteína/metabolismo , Endocitosis , Gadolinio/metabolismo , Glicoconjugados/metabolismo , Compuestos Heterocíclicos con 1 Anillo/metabolismo , Hígado/metabolismo , Samario/metabolismo , Animales , Quelantes/metabolismo , Gadolinio/farmacocinética , Glicoconjugados/química , Glicoconjugados/farmacocinética , Compuestos Heterocíclicos con 1 Anillo/química , Compuestos Heterocíclicos con 1 Anillo/farmacocinética , Humanos , Hígado/citología , Hígado/diagnóstico por imagen , Imagen por Resonancia Magnética , Radiografía , Cintigrafía , Ratas , Ratas Wistar , Samario/farmacocinética , Factores de TiempoRESUMEN
The potential use of 13C multilabeled substrates has been studied in biological applications using in vivo and in vitro proton and 13C NMR spectroscopy. In 13C NMR spectroscopy, multilabeled compounds allow the simultaneous observation of several nuclei or increase distinctly the signal to noise ratio due to a higher degree of enrichment. Contiguous labeling of substrates leads to homonuclear 13C-13C spin couplings and provides a simple means to distinguish between endogenous stores of metabolites and metabolites derived from added substrates.