RESUMEN
Tetrodotoxin (3 x 10(-8) molar) promoted the release of thiamine from perfused rat and frog nerve preparations in a manner similar to other neuroactive drugs. When the rats were injected with thiamine labeled with sulfur-35, analyses of brain, spinal cord, and sciatic nerve homogenates revealed labeled thiamine in membrane, synaptosomes, and mitochondrial subfractions. However, on incubation of these fractions with tetrodotoxin, thiamine was released only from the membrane fragments.
Asunto(s)
Membranas/efectos de los fármacos , Neurosecreción/efectos de los fármacos , Tetrodotoxina/farmacología , Tiamina/metabolismo , Animales , Anuros , Encéfalo/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Perfusión , Ratas , Nervio Ciático/efectos de los fármacos , Médula Espinal/efectos de los fármacosRESUMEN
Extracts of tissue fluids from a patient with subacute necrotizing encephalomyelopathy inhibit thiamine pyrophosphate-adenosine triphosphate phosphotransferase of rat brain. Brain tissue from the patient, in contrast to normal brain tissue, contained essentially no thiamine triphosphate, although thiamine and its other phosphate esters were present in normal concentrations. These findings suggest a relation between this disease and thiamine triphosphate.
Asunto(s)
Encefalopatías , Fosfatos , Fosfotransferasas , Enfermedades de la Médula Espinal , Deficiencia de Tiamina , Animales , Encéfalo/enzimología , Encefalopatías/sangre , Encefalopatías/líquido cefalorraquídeo , Encefalopatías/genética , Encefalopatías/orina , Cerebelo/análisis , Niño , Preescolar , Lóbulo Frontal/análisis , Humanos , Riñón/análisis , Hígado/análisis , Fosfatos/análisis , Ratas , Enfermedades de la Médula Espinal/sangre , Enfermedades de la Médula Espinal/líquido cefalorraquídeo , Enfermedades de la Médula Espinal/genética , Enfermedades de la Médula Espinal/orina , Tiamina/análisis , Tiamina Pirofosfato/análisisRESUMEN
Transketolase (sedoheptulose-7-phosphate: D-glyceraldehyde-3-phosphate glycolaldehydetransferase, EC 2.2.1.1) was purified 16 000-fold from human red blood cells, using DEAE-Sephadex A-50, Sephadex G-150, FPLC on Mono P, and Sephadex G-100. The purified enzyme migrated as a single protein band on SDS-polyacrylamide gel electrophoresis. The FPLC step resolved transketolase into three peaks, designated I, II and III. From results of re-FPLC on Mono P, SDS-polyacrylamide gel electrophoresis, gel filtration, catalytic studies, amino acid analysis and immunological studies, it was concluded that I, II and III were originally the same protein, modified during storage and purification. Transketolase had a subunit (Mr 70 000) and appeared to be composed of two identical subunits. 1 mol of subunit contained 0.9 mol of thiamine pyrophosphate. The pH optimum of the reaction lay within the range 7.6-8.0, and the Km values were determined to be 1.5 X 10(-4) M for xylulose 5-phosphate and 4.0 X 10(-4) M for ribose 5-phosphate. Hg2+ and p-chloromercuribenzoate inhibited the enzyme reaction, and the inhibition of the latter disappeared upon the addition of cysteine. Thiamine and its phosphate esters did not, but cysteine (1 X 10(-2) M) and ethanol (10% and 1% v/v) did activate the enzyme reaction. Antibody prepared to II bound all forms of transketolase in the hemolysate, but inhibited the reaction only about 20%.
Asunto(s)
Anticuerpos , Eritrocitos/enzimología , Transcetolasa/aislamiento & purificación , Aminoácidos/análisis , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Electroforesis en Gel de Poliacrilamida , Hemólisis , Humanos , Inmunodifusión , Inmunoglobulina G/inmunología , Punto Isoeléctrico , Metales/farmacología , Peso Molecular , Transcetolasa/inmunologíaRESUMEN
Male Wistar rats were separated into 4 groups: group 1, thiamine sufficient diet (control); group 2, thiamine sufficient diet with intraperitoneal administration of furosemide (20 mg . kg-1 of body weight); group 3, thiamine deficient diet; group 4, thiamine deficient diet within tra-peritoneal administration of furosemide. After 4 weeks, the rats were killed and the thiamine levels and activity of transketolase were assayed. Thiamine concentration and transketolase activity were significantly decreased and thiamine pyrophosphate effect was significantly increased in the blood, and various tissues in group 2 and 4 compared with group 1 and 3, respectively. The intraperitoneal administration of various concentrations of furosemide (20 mg, 10 mg, and 2 mg . kg-1 of body weight) resulted in a significant increase in urinary thiamine excretion. Thus, it is assumed that long-term administration of furosemide could induce a thiamine deficiency.
Asunto(s)
Furosemida/toxicidad , Deficiencia de Tiamina/inducido químicamente , Animales , Activación Enzimática/efectos de los fármacos , Masculino , Ratas , Tiamina , Tiamina Pirofosfato/farmacología , Transcetolasa/metabolismoRESUMEN
Male Wistar rats (15 weeks old), the ankle joints of one hindlimb of which were immobilized in the extended position for 12 days, were injected with salicylate as a trapping agent for hydroxyl radicals before sacrifice. Atrophied and contralateral soleus, typical slow red muscles were collected and their levels of 2,3-dihydroxybenzoic acid (2,3-DHB), one of the main reaction products formed by the attack of hydroxyl radical on salicylate, were determined using high-performance liquid chromatography with an electrochemical detector. There was a significant increase in 2,3-DHB in the atrophied muscle. This result strongly suggests the enhanced generation in vivo of hydroxyl radicals in atrophied muscle.
Asunto(s)
Radical Hidroxilo/metabolismo , Músculos/metabolismo , Atrofia Muscular/metabolismo , Animales , Depuradores de Radicales Libres , Hidroxibenzoatos/metabolismo , Inmovilización , Masculino , Atrofia Muscular/inducido químicamente , Ratas , Ratas Wistar , Salicilatos , Ácido SalicílicoRESUMEN
Single ankle joints of male Wistar rats (15-week-old) were immobilized in the extended position for 7 days and remobilized for 5 days after the immobilization period. Atrophic and contralateral soleus, typical slow red muscles, were collected and their levels of thiobarbituric acid-reactive substance (TBARS) and glutathione were measured. Five-day remobilization did not increase muscle weight significantly. However, there were significant increases in TBARS and oxidized glutathione in the recovering muscle, which strongly suggested that enhanced oxidative stress occurred during the recovery from disuse muscle atrophy. Vitamin E injection accelerated the recovery from atrophy, thus showing that oxidative stress slowed it down.
Asunto(s)
Músculos/metabolismo , Atrofia Muscular/metabolismo , Animales , Glutatión/metabolismo , Inmovilización , Masculino , Músculos/patología , Atrofia Muscular/patología , Tamaño de los Órganos , Oxidación-Reducción , Ratas , Ratas Wistar , Tiobarbitúricos , Vitamina E/farmacologíaRESUMEN
To estimate the nutritional and the pathological states in thiamin-deficiency-related diseases, especially Wernicke-Korsakoff syndrome, we studied the relationship among transketolase activity, transketolase concentration, and thiamin phosphate esters in rats chronically fed alcohol. In the brain of alcohol-fed rats, the enzyme activity and concentration decreased although there was no positive correlation between the two. On the contrary, transketolase activity in the liver correlated positively with concentration, and both transketolase activity and concentration were decreased in the thiamin-deficient groups. These findings suggest that transketolase in the brain may be different from that in the liver and that the alteration of the enzyme activity in the brain may be based on the conformational change of the protein molecule caused by chronic alcohol administration.
Asunto(s)
Encéfalo/enzimología , Etanol/farmacología , Hígado/enzimología , Deficiencia de Tiamina/enzimología , Transcetolasa/efectos de los fármacos , Animales , Masculino , Ratas , Ratas Endogámicas , Tiamina Monofosfato/metabolismo , Tiamina Pirofosfato/metabolismo , Tiamina Trifosfato/metabolismo , Transcetolasa/metabolismoRESUMEN
Drifting snow samples were collected at Asuka Station (71 degrees 32'S, 24 degrees 08'E, 930 m above sea level) over a period from July to December 1991; 36 elements (including Na, Mg, K, Ca, Fe, Al, Li, V, Cr, Mn, Co, Ni, Cu, Zn, Ga, Se, Rb, Sr, Cd, Pb, Y, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, and Th) in snow were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) by direct sample introduction. Concentrations of Cl-, NO3-, and SO(4)2- in the snow were also determined by ion chromatography. In late September to early October, there was a pronounced peak concentration of most of the elements together with non-sea salt sulfate. Enrichment factor analyses suggest that Na, Mg, Ca, K, and Sr are of marine origin and Al, Fe, Mn, Rb, Cr, Ni, Ga, V, and all the rare earth elements are of crustal origins. Volcanic eruption of Mt. Pinatubo (June 1991) and Mt. Hudson (August 1991) could be the reason for the precipitation of Pb, Cd, Cu, Zn, and Se together with non-sea salt sulfates in the austral spring at Asuka Station.
Asunto(s)
Nieve , Oligoelementos/análisis , Conceptos Meteorológicos , Estaciones del AñoRESUMEN
In convulsive pigeons on a diet of polished rice, a significant decrease of thiamine and calcium in the central nervous system rather than in peripheral nervous system was observed. In subcellular fractions of telencephalon in these same pigeons, thiamine and calcium levels decreased in the myelin-membrane and synaptosomal fractions. When radioactive thiamine was injected into the pieon during a convulsion, radio-activity was most prominent in the telencephalon and in subcellular fractions, in the myelin-membrane and synaptosomal fractions of telencephalon immediately after recovery from the convulsion. When calsium was added to the polished rice, the pigeons never had such convulsions. Thiamine and calsium concentrations in myelin-membrane and synaptosomal fractions of telencephalon of calcium-rice fed pigeons were significantly higher than in convulsive pigeons. It is suggested that calsium plays a role in binding the protein and thiamine which is related to the sodium transport system of excitable membranes.
Asunto(s)
Calcio/deficiencia , Enfermedades del Sistema Nervioso/etiología , Deficiencia de Tiamina/complicaciones , Tiamina/metabolismo , Animales , Calcio/metabolismo , Calcio de la Dieta/metabolismo , Columbidae , Magnesio/metabolismo , Convulsiones/dietoterapia , Convulsiones/etiología , Telencéfalo/metabolismo , Deficiencia de Tiamina/enzimología , Transcetolasa/metabolismoRESUMEN
Xanthine oxidase activity in the rat brain was measured by means of high-performance liquid chromatography with electrochemical detection of uric acid. Cerebral ischemia was produced by a four-vessel occlusion method. In the control rat, the enzyme activity was 0.87 +/- 0.13 nmol/gm wet weight/min at 25 degrees C (mean +/- standard deviation), of which 92.4% was associated with the nicotinamide adenine dinucleotide (NAD)-dependent dehydrogenase form and only 7.6% with the oxygen-dependent superoxide-producing oxidase form. However, the ratio of the latter form increased to 43.7% after 30 minutes of global ischemia, despite the total xanthine oxidase activity remaining the same. Thus, it was revealed that uric acid can be synthesized in the rat brain and that cerebral ischemia induced the conversion of xanthine oxidase from an NAD-dependent dehydrogenase to an oxygen-dependent superoxide-producing oxidase. Although the xanthine oxidase pathway has been proposed as a source of oxygen-derived free radicals in various ischemic organs other than brain, the results of the present study suggest the involvement of the oxygen free radicals generated from this pathway in the pathogenesis of the ischemic injury of the rat brain.
Asunto(s)
Isquemia Encefálica/enzimología , Encéfalo/enzimología , Xantina Oxidasa/metabolismo , Animales , Isoenzimas/metabolismo , Masculino , NAD/farmacología , Oxígeno/farmacología , Ratas , Ratas Endogámicas , Superóxidos/biosíntesis , Xantina Deshidrogenasa/metabolismoRESUMEN
To verify whether lipid peroxidation is associated with focal cerebral ischemia, a unilateral middle cerebral artery occlusion was carried out in rats. The concentrations of various endogenous antioxidants in the ischemic center were measured, including alpha-tocopherol and ubiquinones as lipid-soluble antioxidants and ascorbate as a water-soluble antioxidant. At 30 minutes after ischemia, alpha-tocopherol decreased to 79% of baseline, reduced ubiquinone-9 to 73%, ubiquinone-10 to 66%, and reduced ascorbate to 76%. Six hours after ischemia, alpha-tocopherol decreased to 63% and reached a plateau, whereas reduced ubiquinones and reduced ascorbate declined further to 16% and 10%, respectively, 12 hours after ischemia and then reached plateau levels. These results suggest functional and durational differences between antioxidants and lipid peroxidation in this ischemic model. Although the reciprocal increase in oxidized ubiquinones during ischemia was not observed, that of oxidized ascorbate was noted. The complementary antioxidant system between cytoplasmic and membranous components, the combination alpha-tocopherol/ascorbate, was estimated from the calculated consumption ratio of these antioxidants on the basis that the loss of these reduced antioxidants is due to neutralization of free radicals. This system is suggested to play an important role in the early ischemic period. Urate also increased during ischemia. The possible involvement of the xanthine-xanthine oxidase system in initiating free radical reactions in cerebral ischemia is also discussed.
Asunto(s)
Isquemia Encefálica/metabolismo , Peróxidos Lipídicos/biosíntesis , Animales , Ácido Ascórbico/metabolismo , Encéfalo/metabolismo , Masculino , Ratas , Ratas Endogámicas , Ubiquinona/metabolismo , Ácido Úrico/metabolismo , Vitamina E/metabolismoRESUMEN
The influence of dietary protein levels on taste sensitivity was studied in adult rats. Low protein diets of 0.0, 2.5, or 5.0% purified egg protein (PEP) were fed to animals for 28 days. Two bottle choice preference tests between aqueous solutions of either 2, 9, 17, or 86 mM sodium chloride and deionized water were conducted in an ascending order on days 14, 16, 18, and 20. Urine samples were collected for zinc and creatinine analysis. Blood samples were also collected for measuring serum zinc and creatinine concentrations. Scanning electron microscopy was performed to observe rats' tongue epithelia. Protein free diet group showed significantly lower taste sensitivity and renal reabsorption rate than other protein containing diet groups, while serum zinc and creatinine concentrations, and creatinine clearance were not affected by dietary protein level. Degeneration of filiform papillae and imperforation of taste pore of fungiform papillae were observed in protein free diet group. This experiment implies at least 2.5% dietary protein is required to manifest normal taste function in the adult.
Asunto(s)
Deficiencia de Proteína/fisiopatología , Umbral Gustativo/fisiología , Factores de Edad , Animales , Creatina/sangre , Femenino , Preferencias Alimentarias/fisiología , Tasa de Filtración Glomerular/fisiología , Microscopía Electrónica de Rastreo , Degeneración Nerviosa/fisiología , Deficiencia de Proteína/patología , Ratas , Ratas Sprague-Dawley , Papilas Gustativas/patología , Papilas Gustativas/fisiopatología , Zinc/sangreRESUMEN
Standard reference ranges for all laboratory test values are mandatory. This study was designed to establish a reference range for blood vitamin B1 levels, since the normal range has not been determined in the Japanese population. We founded the Japan Committee for Vitamin Laboratory Standards, which was incorporated with the Vitamin Society of Japan and the Japanese Society of Nutrition and Food Science. We standardized whole blood vitamin B1 levels using three HPLC techniques (post-column reverse-phase HPLC, pre-column reverse-phase HPLC, and precolumn GP-HPLC). The reference range was obtained in 54 volunteers administered a 1,800 kcal diet with 2 mg of vitamin B1 (1.74 mg measured) daily to avoid marginal vitamin B1 deficiency in the population. The range for each assay was 26-47, 28-51, and 28-56 ng/ml, respectively. Our data suggest that 26-28 ng/ml is the lower limit of normal for whole blood vitamin B1, but further studies in a larger population are needed in order to obtain more definitive results.
Asunto(s)
Deficiencia de Tiamina/sangre , Tiamina/sangre , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Líquida de Alta Presión/normas , Ingestión de Energía , Humanos , Japón , Garantía de la Calidad de Atención de Salud , Valores de ReferenciaRESUMEN
We have clarified that thiamine is cleaved by residual chlorine in proportion to the rise in temperature, pH and concentration of residual chlorine. When rice was boiled in an electric rice cooker, the thiamine in the rice was cleaved by residual chlorine. It is assumed that thiamine is cleaved into hydroxymethylpyrimidine and the thiazole moiety.
Asunto(s)
Cloro/efectos adversos , Tiamina/antagonistas & inhibidores , Abastecimiento de Agua , Fenómenos Químicos , Química , Culinaria , Calor , Concentración de Iones de Hidrógeno , OryzaRESUMEN
We performed a pharmacokinetic analysis of the blood thiamin profile after oral administration of thiamin tetrahydrofurfuryl disulfide (TTFD) to healthy adults. To distinguish between thiamin derived from TTFD ingestion and that from previous dietary intake, the baseline thiamin level was subtracted from the apparent blood vitamin levels measured after administration. Following administration of 100 mg of TTFD, the peak blood thiamin level was almost 10 times the baseline level and the blood thiamin profile could be simulated by a two-compartment model to obtain reasonable pharmacokinetic parameters. When the blood thiamin profile for a 10-mg dose of TTFD was estimated using scaled-down pharmacokinetic parameters derived at the 100-mg dose level, a reasonable fit for the raw data obtained at 10-mg dose was obtained. Therefore, the parameters derived from the data at a dose of 100 mg appeared to be reliable. Since even 180 mg of TTFD is completely absorbed and the absorption ratio is independent of the dose, it can be concluded that gastrointestinal absorption of TTFD is good within the dose range.
Asunto(s)
Fursultiamina/administración & dosificación , Tiamina/farmacocinética , Administración Oral , Adulto , Fursultiamina/farmacocinética , Humanos , Modelos Biológicos , Tiamina/sangreRESUMEN
To clarify the cooking losses of minerals (sodium, potassium, phosphorus, calcium, magnesium, iron, zinc, manganese, copper), various food materials were analyzed before and after cooking, and the following results were obtained. (1) The mineral contents of cooked foods in mass cooking were on an average about 60-70 percent of those in raw or uncooked foods. (2) Cooking losses were particularly high in minerals of vegetables. (3) Among various cooking methods, loss of mineral was largest in squeezing after boil and in soaking in water after thin slice, followed by parching, frying and stewing. (4) Cooking losses of minerals in meals cooked in home brought about the similar results as those by the mass cooking procedures. (5) The measures to prevent cooking loss are (a) eating the boiled food with the soup, (b) addition of small amount of salt (about 1% NaCl) in boiling, (c) avoidance of too much boiling, (d) selection of a cooking method causing less mineral loss (stewing, frying or parching).
Asunto(s)
Culinaria , Análisis de los Alimentos , Minerales/análisis , Animales , Calcio/análisis , Cobre/análisis , Humanos , Hierro/análisis , Magnesio/análisis , Manganeso/análisis , Carne , Valor Nutritivo , Oryza , Fósforo/análisis , Potasio/análisis , Sodio/análisis , Porcinos , Verduras , Zinc/análisisRESUMEN
The role of thiamin in the catabolism of ethanol and acetaldehyde has been investigated. When thiamin and subsequently ethanol were administered orally to rabbits, the thiamin concentration in blood increased slightly during the first 3 h and then decreased gradually. After 12 h, it became lower than the value before thiamin administration. Finally, it reached the lowest value after 24 h and then increased slowly to revert to normal in 72 h. It is suggested that thiamin participates in the catabolic pathway of ethanol. An oral administration of pyrazole, an inhibitor of alcohol dehydrogenase, followed by ethanol to rabbits caused a delay in ethanol elimination from blood. When acetaldehyde was injected intravenously to rabbits, thiamin concentration and the transketolase activity in blood decreased gradually and after 12 h the thiamin level reached its lowest value, then increased slowly and normalized in 72 h. Thus, it could be postulated that the decrease in thiamin after an acute ethanol ingestion linked greatly to the acetaldehyde catabolism.
Asunto(s)
Acetaldehído/farmacología , Etanol/farmacología , Tiamina/metabolismo , Administración Oral , Animales , Interacciones Farmacológicas , Eritrocitos/enzimología , Masculino , Pirazoles/farmacología , Conejos , Tiamina/farmacología , Transcetolasa/sangreRESUMEN
A technique for the rapid and accurate estimation of nonheme iron using inductively coupled plasma-atomic emission spectrometry is described. Yttrium was used as an internal standard. An external calibration method was used. The standards were prepared in a matrix composed of 2.5N HCl in 10% (w/v) trichloroacetic acid. The supernatant and coagulum fractions of liver nonheme iron were separated by the method of Drysdale and Ramsay with minor modification. The data determined by this procedure was compared and found to be agreement with data determined by the method of Hallgren. To evaluate the iron status of rats, hemoglobin and liver nonheme iron were determined. Hemoglobin and all of the nonheme iron fractions of the rats fed an iron-deficient diet were significantly lower than those of the rats fed an iron-sufficient diet. The blood content in the liver was estimated to be 80 microL/g from the blood iron concentration, and the difference between total and nonheme iron concentration in liver.
Asunto(s)
Hierro/análisis , Hígado/química , Animales , Hemo , Indicadores y Reactivos , Hierro/sangre , Deficiencias de Hierro , Ratas , Valores de Referencia , Análisis EspectralRESUMEN
Female Wistar rats were separated into 9 groups, and 9 different synthetic diets (each diet contains different level of protein and calcium) were given to each group. After 5 weeks of these dietary regimens, all rats were sacrificed and calcium, magnesium, and phosphorus levels in plasma and various tissues were determined. In calcium-deficient groups, calcium, magnesium, and phosphorus level in bone decreased, plasma calcium level decreased and there was a tendency that magnesium levels in brain and liver and phosphorus level in brain increased. When comparison was made among the calcium-deficient groups, calcium, magnesium, and phosphorus levels in bone were higher in low protein/calcium-deficient group than higher protein/calcium-deficient groups. It is probable that protein deficiency inhibits calcium depletion and consequently the influence of calcium deficiency is less significant in the condition of deficiency in both protein and calcium.
Asunto(s)
Huesos/metabolismo , Calcio/deficiencia , Calcio/metabolismo , Magnesio/metabolismo , Fósforo/metabolismo , Deficiencia de Proteína/metabolismo , Animales , Encéfalo/metabolismo , Calcio/sangre , Femenino , Hígado/metabolismo , Músculos/metabolismo , Ratas , Ratas Wistar , Aumento de PesoRESUMEN
To clarify the mineral status in selenium (Se)-deficient rats fed a vitamin-free casein (VFC)-based or torula yeast (TY)-based diet, 24 weanling male Wistar rats were divided into 4 groups fed diets using VFC or TY as the protein source and containing Se at sufficient (0.5 microgram/g, +Se) or deficient (0.019 microgram/g for VFC-based and < 0.005 microgram/g for TY-based diets, -Se) level for 8 wk. TY supplied a larger amount of extra minerals (Na, K, Ca, Mg, Fe, Mn, Zn, and Cu) except Se than VFC. Se concentration and glutathione peroxidase activity were significantly lower in TY-fed rats than in VFC-fed rats, as well as in -Se rats compared to +Se rats. Compared to +Se rats, Fe concentration was higher in liver and muscle of -Se rats fed the VFC-based diet and in plasma, heart, liver, and tibia of -Se rats fed the TY-based diet. Compared to +Se rats, decreases of Mn concentration appeared in plasma, heart, and tibia of VFC-fed -Se rats and in brain, heart, liver and tibia of TY-fed -Se rats. There was also a little imbalance in Ca, Mg, Na, K, and Cu caused by Se deficiency. The results indicated that Se deficiency induced the mineral imbalance in rats, especially an increase in Fe and decrease in Mn, which was more severe in TY-fed rats than VFC-fed rats. However, TY cannot be used as a model for both Se and other mineral deficiency because of the extra minerals except Se found in TY. Instead, VFC can be employed, which contains fewer minerals except Se than TY and also can produce a severe degree of Se deficiency.