RESUMEN
We investigated the toxic effects on Prototheca zopfii of indole-3-acetic acid (IAA) and 2,4-pentanedione (PD) combined with horseradish peroxidase (HRP) alongside the oxidation products of 3-methyl-2-oxindole (MOI) and indole-3-carbinol (I3C) from the IAA/HRP system and methylglyoxal (MGO) from the PD/HRP system. The microorganism was incubated in the absence (control) or presence of IAA, PD, IAA/HRP, PD/HRP, MOI, I3C and MGO and determined: (1) cytotoxicity by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay; (2) growth inhibitory concentration by resazurin assay and (3) antioxidant enzymes activities of: catalase (CAT), glutathione reductase (GR) and superoxide dismutase (SOD). P. zopfii was more susceptible to IAA at 40 mM than PD at the same concentration, which seems to indicate that IAA was more effective at initiating cell death. These data corroborate results from the resazurin assay. Concentrations of 40 mM of IAA, IAA/HRP and PD/HRP, 20 mM of PD/HRP, 10 mM of MOI, 2 mM of I3C and 8 mM of MGO inhibited the growth of P. zopfii. With sub-inhibitory concentrations of IAA and IAA/HRP at 30 mM, MOI at 8 mM and I3C at 1 mM, the activities of CAT and GR increased, whereas no statistical difference was observed for CAT activity with IAA/HRP. Thus, PD at 30 mM and MGO at 6 mM increased the activities of CAT and GR, whereas PD/HRP system at 15 mM decreased CAT activity and PD/HRP and MGO showed no statistical difference for SOD activity. In conclusion, IAA/HRP or PD/HRP systems and their oxidation products exert cytotoxic effects on P. zopffi; however, I3C and MGO appear to exert greater microbicidal effect on P. zopfii.
Asunto(s)
Catalasa/metabolismo , Glutatión Reductasa/metabolismo , Oxidantes/farmacología , Estrés Oxidativo/efectos de los fármacos , Prototheca/metabolismo , Superóxido Dismutasa/metabolismo , Antioxidantes/metabolismo , Peroxidasa de Rábano Silvestre/farmacología , Ácidos Indolacéticos/farmacología , Indoles/farmacología , Estrés Oxidativo/fisiología , Oxindoles , Pentanonas/farmacología , Prototheca/enzimología , Piruvaldehído/farmacologíaRESUMEN
The ethanolic extract of Agaricus blazei and ethyl acetate and hydroalcoholic fractions were evaluated for their antioxidant activity.
Asunto(s)
Agaricus , Antioxidantes/farmacología , Fitoterapia , Extractos Vegetales/farmacología , Antioxidantes/administración & dosificación , Antioxidantes/uso terapéutico , Benzotiazoles/química , Compuestos de Bifenilo , Cuerpos Fructíferos de los Hongos , Humanos , Picratos/química , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéutico , Ácidos Sulfónicos/químicaRESUMEN
BACKGROUND: We present new findings on liver steatosis detected in a group of 20 morbidly obese patients who were reassessed shortly after bariatric surgery (BS) by assaying hepatic markers in their serum. METHODS: We assayed aspartate aminotransferase (AST), alanine aminotransferase, lactate dehydrogenase, alkaline phosphatase, gamma-glutamyl transferase (gamma-GT), cholinesterase, cholesterol, total protein, and albumin, and measured the weight and the body mass index (BMI) of patients, before and one and three months after surgery. RESULTS: There were significant reductions in BMI following surgery and also falls in transaminases and gamma-GT activities three months after BS. No changes occurred in other parameters between periods, except that cholesterol was above reference values before BS and fell to normal levels three months after BS. CONCLUSIONS: We suggest that before undergoing surgery, the patients suffered from slight steatosis, while after BS the reduction in AST and gamma-GT indicated that this condition was corrected within three months. Moreover, these enzymes may be useful markers for excess fat in the liver.
Asunto(s)
Cirugía Bariátrica , Hígado Graso/sangre , Obesidad Mórbida/patología , Alanina Transaminasa/sangre , Fosfatasa Alcalina/sangre , Aspartato Aminotransferasas/sangre , Bilirrubina/sangre , Biomarcadores/sangre , Colinesterasas/sangre , Femenino , Estudios de Seguimiento , Humanos , L-Lactato Deshidrogenasa/sangre , Masculino , Análisis Multivariante , Obesidad Mórbida/complicaciones , Obesidad Mórbida/cirugía , Periodo Posoperatorio , Estudios Retrospectivos , Resultado del Tratamiento , gamma-Glutamiltransferasa/sangreRESUMEN
Maytenus ilicifolia is an important plant with potential on cancer treatment and has been largely used in Brazil and other countries. We have evaluated the crude ethanolic extract of M. ilicifolia as a potential antioxidant source using an assay based on the bleaching of the radical monocation 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(*+)) and by HOCl scavenger capacity. Trolox and uric acid were used as positive controls. The results indicated M. ilicifolia root bark as a great source of antioxidants based on its potential as scavenger of radicals.
Asunto(s)
Antioxidantes/química , Depuradores de Radicales Libres/química , Maytenus/química , Benzotiazoles , Cromanos/farmacología , Etanol/química , Concentración 50 Inhibidora , Corteza de la Planta/química , Extractos Vegetales/química , Ácidos Sulfónicos/metabolismo , Ácido Úrico/farmacologíaRESUMEN
Hypochlorous acid (HOCl) released by activated leukocytes has been implicated in the tissue damage that characterizes chronic inflammatory diseases. In this investigation, 14 indole derivatives, including metabolites such as melatonin, tryptophan and indole-3-acetic acid, were screened for their ability to inhibit the generation of this endogenous oxidant by stimulated leukocytes. The release of HOCl was measured by the production of taurine-chloramine when the leukocytes (2 x 10(6) cells/mL) were incubated at 37 degrees C in 10 mM phosphate-buffered saline, pH 7.4, for 30 min with 5 mM taurine and stimulated with 100 nM phorbol-12-myristate acetate. Irrespective of the group substituted in the indole ring, all the compounds tested including indole, 2-methylindole, 3-methylindole, 2,3-dimethylindole, 2,5-dimethylindole, 2-phenylindole, 5-methoxyindole, 6-methoxyindole, 5-methoxy-2-methylindole, melatonin, tryptophan, indole-3-acetic acid, 5-methoxy-2-methyl-3-indole-acetic acid, and indomethacin (10 microM) inhibited the chlorinating activity of myeloperoxidase (MPO) in the 23-72% range. The compounds 3-methylindole and indole-3-acetic acid were chosen as representative of indole derivatives in a dose-response study using purified MPO. The IC50 obtained were 0.10 +/- 0.03 and 5.0 +/- 1.0 microM (N = 13), respectively. These compounds did not affect the peroxidation activity of MPO or the production of superoxide anion by stimulated leukocytes. By following the spectral change of MPO during the enzyme turnover, the inhibition of HOCl production can be explained on the basis of the accumulation of the redox form compound-II (MPO-II), which is an inactive chlorinating species. These results show that indole derivatives are effective and selective inhibitors of MPO-chlorinating activity.
Asunto(s)
Ácido Hipocloroso/metabolismo , Indoles/farmacología , Leucocitos/efectos de los fármacos , Peroxidasa/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Humanos , Concentración 50 Inhibidora , Leucocitos/fisiología , Oxidación-ReducciónRESUMEN
Hypochlorous acid (HOCl) released by activated leukocytes has been implicated in the tissue damage that characterizes chronic inflammatory diseases. In this investigation, 14 indole derivatives, including metabolites such as melatonin, tryptophan and indole-3-acetic acid, were screened for their ability to inhibit the generation of this endogenous oxidant by stimulated leukocytes. The release of HOCl was measured by the production of taurine-chloramine when the leukocytes (2 x 10(6) cells/mL) were incubated at 37°C in 10 mM phosphate-buffered saline, pH 7.4, for 30 min with 5 mM taurine and stimulated with 100 nM phorbol-12-myristate acetate. Irrespective of the group substituted in the indole ring, all the compounds tested including indole, 2-methylindole, 3-methylindole, 2,3-dimethylindole, 2,5-dimethylindole, 2-phenylindole, 5-methoxyindole, 6-methoxyindole, 5-methoxy-2-methylindole, melatonin, tryptophan, indole-3-acetic acid, 5-methoxy-2-methyl-3-indole-acetic acid, and indomethacin (10 æM) inhibited the chlorinating activity of myeloperoxidase (MPO) in the 23-72 percent range. The compounds 3-methylindole and indole-3-acetic acid were chosen as representative of indole derivatives in a dose-response study using purified MPO. The IC50 obtained were 0.10 ± 0.03 and 5.0 ± 1.0 æM (N = 13), respectively. These compounds did not affect the peroxidation activity of MPO or the production of superoxide anion by stimulated leukocytes. By following the spectral change of MPO during the enzyme turnover, the inhibition of HOCl production can be explained on the basis of the accumulation of the redox form compound-II (MPO-II), which is an inactive chlorinating species. These results show that indole derivatives are effective and selective inhibitors of MPO-chlorinating activity.
Asunto(s)
Humanos , Ácido Hipocloroso/metabolismo , Indoles/farmacología , Leucocitos/efectos de los fármacos , Peroxidasa/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Leucocitos/fisiología , Oxidación-ReducciónRESUMEN
The release of reactive oxygen specie (ROS) by activated neutrophil is involved in both the antimicrobial and deleterious effects in chronic inflammation. The objective of the present investigation was to determine the effect of therapeutic plasma concentrations of non-steroidal anti-inflammatory drugs (NSAIDs) on the production of ROS by stimulated rat neutrophils. Diclofenac (3.6 microM), indomethacin (12 microM), naproxen (160 microM), piroxicam (13 microM), and tenoxicam (30 microM) were incubated at 37 masculineC in PBS (10 mM), pH 7.4, for 30 min with rat neutrophils (1 x 10(6) cells/ml) stimulated by phorbol-12-myristate-13-acetate (100 nM). The ROS production was measured by luminol and lucigenin-dependent chemiluminescence. Except for naproxen, NSAIDs reduced ROS production: 58 +/- 2% diclofenac, 90 +/- 2% indomethacin, 33 +/- 3% piroxicam, and 45 +/- 6% tenoxicam (N = 6). For the lucigenin assay, naproxen, piroxicam and tenoxicam were ineffective. For indomethacin the inhibition was 52 +/- 5% and diclofenac showed amplification in the light emission of 181 +/- 60% (N = 6). Using the myeloperoxidase (MPO)/H2O2/luminol system, the effects of NSAIDs on MPO activity were also screened. We found that NSAIDs inhibited both the peroxidation and chlorinating activity of MPO as follows: diclofenac (36 +/- 10, 45 +/- 3%), indomethacin (97 +/- 2, 100 +/- 1%), naproxen (56 +/- 8, 76 +/- 3%), piroxicam (77 +/- 5, 99 +/- 1%), and tenoxicam (90 +/- 2, 100 +/- 1%), respectively (N = 3). These results show that therapeutic levels of NSAIDs are able to suppress the oxygen-dependent antimicrobial or oxidative functions of neutrophils by inhibiting the generation of hypochlorous acid.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Neutrófilos/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Animales , Antiinflamatorios no Esteroideos/sangre , Antiinflamatorios no Esteroideos/toxicidad , Mediciones Luminiscentes , Activación Neutrófila , Neutrófilos/metabolismo , Peroxidasa/efectos de los fármacos , RatasRESUMEN
The release of reactive oxygen specie (ROS) by activated neutrophil is involved in both the antimicrobial and deleterious effects in chronic inflammation. The objective of the present investigation was to determine the effect of therapeutic plasma concentrations of non-steroidal anti-inflammatory drugs (NSAIDs) on the production of ROS by stimulated rat neutrophils. Diclofenac (3.6 µM), indomethacin (12 µM), naproxen (160 µM), piroxicam (13 µM), and tenoxicam (30 µM) were incubated at 37°C in PBS (10 mM), pH 7.4, for 30 min with rat neutrophils (1 x 10(6) cells/ml) stimulated by phorbol-12-myristate-13-acetate (100 nM). The ROS production was measured by luminol and lucigenin-dependent chemiluminescence. Except for naproxen, NSAIDs reduced ROS production: 58 ± 2 percent diclofenac, 90 ± 2 percent indomethacin, 33 ± 3 percent piroxicam, and 45 ± 6 percent tenoxicam (N = 6). For the lucigenin assay, naproxen, piroxicam and tenoxicam were ineffective. For indomethacin the inhibition was 52 ± 5 percent and diclofenac showed amplification in the light emission of 181 ± 60 percent (N = 6). Using the myeloperoxidase (MPO)/H2O2/luminol system, the effects of NSAIDs on MPO activity were also screened. We found that NSAIDs inhibited both the peroxidation and chlorinating activity of MPO as follows: diclofenac (36 ± 10, 45 ± 3 percent), indomethacin (97 ± 2, 100 ± 1 percent), naproxen (56 ± 8, 76 ± 3 percent), piroxicam (77 ± 5, 99 ± 1 percent), and tenoxicam (90 ± 2, 100 ± 1 percent), respectively (N = 3). These results show that therapeutic levels of NSAIDs are able to suppress the oxygen-dependent antimicrobial or oxidative functions of neutrophils by inhibiting the generation of hypochlorous acid.
Asunto(s)
Animales , Ratas , Antiinflamatorios no Esteroideos/farmacología , Neutrófilos/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Antiinflamatorios no Esteroideos/sangre , Antiinflamatorios no Esteroideos/toxicidad , Mediciones Luminiscentes , Activación Neutrófila , Neutrófilos/metabolismo , Peroxidasa/efectos de los fármacosRESUMEN
The fruit of Indian Eugenia jambolana have been shown to have therapeutic properties, but because the therapeutic potential of a plant is related to the geographic region in which the plant was grown and to the part of the plant used, we investigated Brazilian Eugenia jambolana fruit using the same preparation and experimental methods as have been used in India. The well-established metabolic cage model was used to evaluate the physiological and metabolic parameters associated with streptozotocin-induced diabetes in rats (n=10) which had been administered, by gavage, 50 mg per day of lyophilised Eugenia jambolana fruit-pulp extract for 41 days. We found that, compared to untreated controls, rats treated with the lyophilised fruit-pulp showed no observable difference in body weight, food or water intake, urine volume, glycaemia, urinary urea and glucose, hepatic glycogen, or on serum levels of total cholesterol, HDL cholesterol or triglycerides. No change was observed in the masses of epididymal or retroperitoneal adipose tissue or of soleus or extensor digitorum longus muscles. This lack of any apparent effect on the diabetes may be attributable to the regional ecosystem where the fruit was collected and/or to the severity of the induced diabetes.
Asunto(s)
Diabetes Mellitus Experimental/tratamiento farmacológico , Frutas , Hipoglucemiantes/farmacología , Fitoterapia , Syzygium , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/patología , Animales , Glucemia/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Diabetes Mellitus Experimental/inducido químicamente , Epidídimo/efectos de los fármacos , Epidídimo/patología , Glucosuria/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Masculino , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/patología , Tamaño de los Órganos/efectos de los fármacos , Preparaciones de Plantas/farmacología , Preparaciones de Plantas/uso terapéutico , Ratas , Estreptozocina , Factores de TiempoRESUMEN
Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive disorder due to an inborn error of cholesterol metabolism, characterized by congenital malformations, dysmorphism of multiple organs, mental retardation and delayed neuropsychomotor development resulting from cholesterol biosynthesis deficiency. A defect in 3ß-hydroxysteroid-delta7-reductase (delta7-sterol-reductase), responsible for the conversion of 7-dehydrocholesterol (7-DHC) to cholesterol, causes an increase in 7-DHC and frequently reduces plasma cholesterol levels. The clinical diagnosis of SLOS cannot always be conclusive because of the remarkable variability of clinical expression of the disorder. Thus, confirmation by the measurement of plasma 7-DHC levels is needed. In the present study, we used a simple, fast, and selective method based on ultraviolet spectrophotometry to measure 7-DHC in order to diagnose SLOS. 7-DHC was extracted serially from 200 æl plasma with ethanol and n-hexane and the absorbance at 234 and 282 nm was determined. The method was applied to negative control plasma samples from 23 normal individuals and from 6 cases of suspected SLOS. The method was adequate and reliable and 2 SLOS cases were diagnosed
Asunto(s)
Preescolar , Humanos , Masculino , Lactante , Niño , Colesterol , Deshidrocolesteroles , Síndrome de Smith-Lemli-Opitz/diagnóstico , Biomarcadores , Síndrome de Smith-Lemli-Opitz/sangre , Espectrofotometría UltravioletaRESUMEN
Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive disorder due to an inborn error of cholesterol metabolism, characterized by congenital malformations, dysmorphism of multiple organs, mental retardation and delayed neuropsychomotor development resulting from cholesterol biosynthesis deficiency. A defect in 3 -hydroxysteroid-delta7-reductase (delta7-sterol-reductase), responsible for the conversion of 7-dehydrocholesterol (7-DHC) to cholesterol, causes an increase in 7-DHC and frequently reduces plasma cholesterol levels. The clinical diagnosis of SLOS cannot always be conclusive because of the remarkable variability of clinical expression of the disorder. Thus, confirmation by the measurement of plasma 7-DHC levels is needed. In the present study, we used a simple, fast, and selective method based on ultraviolet spectrophotometry to measure 7-DHC in order to diagnose SLOS. 7-DHC was extracted serially from 200 l plasma with ethanol and n-hexane and the absorbance at 234 and 282 nm was determined. The method was applied to negative control plasma samples from 23 normal individuals and from 6 cases of suspected SLOS. The method was adequate and reliable and 2 SLOS cases were diagnosed.
Asunto(s)
Colesterol/sangre , Deshidrocolesteroles/sangre , Síndrome de Smith-Lemli-Opitz/diagnóstico , Biomarcadores/sangre , Niño , Preescolar , Humanos , Lactante , Masculino , Síndrome de Smith-Lemli-Opitz/sangre , Espectrofotometría UltravioletaRESUMEN
Losartan, an AT1 angiotensin II (ANG II) receptor non-peptide antagonist, induces an increase in mean arterial pressure (MAP) when injected intracerebroventricularly (icv) into rats. The present study investigated possible effector mechanisms of the increase in MAP induced by icv losartan in unanesthetized rats. Male Holtzman rats (280-300 g, N = 6/group) with a cannula implanted into the anterior ventral third ventricle received an icv injection of losartan (90 micro g/2 micro l) that induced a typical peak pressor response within 5 min. In one group of animals, this response to icv losartan was completely reduced from 18 +/- 1 to 4 +/- 2 mmHg by intravenous (iv) injection of losartan (2.5-10 mg/kg), and in another group, it was partially reduced from 18 +/- 3 to 11 +/- 2 mmHg by iv prazosin (0.1-1.0 mg/kg), an alpha1-adrenergic antagonist (P<0.05). Captopril (10 mg/kg), a converting enzyme inhibitor, injected iv in a third group inhibited the pressor response to icv losartan from 24 +/- 3 to 7 +/- 2 mmHg (P<0.05). Propranolol (10 mg/kg), a beta-adrenoceptor antagonist, injected iv in a fourth group did not alter the pressor response to icv losartan. Plasma renin activity and serum angiotensin-converting enzyme activity were not altered by icv losartan in other animals. The results suggest that the pressor effect of icv losartan depends on angiotensinergic and alpha1-adrenoceptor activation, but not on increased circulating ANG II.
Asunto(s)
Agonistas alfa-Adrenérgicos/farmacología , Antagonistas de Receptores de Angiotensina , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Antihipertensivos/farmacología , Hipertensión/sangre , Losartán/farmacología , Peptidil-Dipeptidasa A/sangre , Renina/sangre , Animales , Antihipertensivos/administración & dosificación , Antihipertensivos/antagonistas & inhibidores , Presión Sanguínea/efectos de los fármacos , Hipertensión/inducido químicamente , Inyecciones Intraventriculares , Losartán/administración & dosificación , Losartán/antagonistas & inhibidores , Masculino , Radioinmunoensayo , Ratas , Ratas Sprague-DawleyRESUMEN
Losartan, an AT1 angiotensin II (ANG II) receptor non-peptide antagonist, induces an increase in mean arterial pressure (MAP) when injected intracerebroventricularly (icv) into rats. The present study investigated possible effector mechanisms of the increase in MAP induced by icv losartan in unanesthetized rats. Male Holtzman rats (280-300 g, N = 6/group) with a cannula implanted into the anterior ventral third ventricle received an icv injection of losartan (90 æg/2 æl) that induced a typical peak pressor response within 5 min. In one group of animals, this response to icv losartan was completely reduced from 18 ± 1 to 4 ± 2 mmHg by intravenous (iv) injection of losartan (2.5-10 mg/kg), and in another group, it was partially reduced from 18 ± 3 to 11 ± 2 mmHg by iv prazosin (0.1-1.0 mg/kg), an alpha1-adrenergic antagonist (P<0.05). Captopril (10 mg/kg), a converting enzyme inhibitor, injected iv in a third group inhibited the pressor response to icv losartan from 24 ± 3 to 7 ± 2 mmHg (P<0.05). Propranolol (10 mg/kg), a ß-adrenoceptor antagonist, injected iv in a fourth group did not alter the pressor response to icv losartan. Plasma renin activity and serum angiotensin-converting enzyme activity were not altered by icv losartan in other animals. The results suggest that the pressor effect of icv losartan depends on angiotensinergic and alpha1-adrenoceptor activation, but not on increased circulating ANG II
Asunto(s)
Animales , Masculino , Ratas , Agonistas alfa-Adrenérgicos , Inhibidores de la Enzima Convertidora de Angiotensina , Hipertensión , Losartán , Peptidil-Dipeptidasa A , Receptores de Angiotensina , Renina , Captopril , Inyecciones Intraventriculares , Losartán , Prazosina , Propranolol , Radioinmunoensayo , Ratas Sprague-DawleyRESUMEN
The effects of using Bauhinia forficata leaf decoction (150 g leaf/l water; 35.2+/-7.8 ml/100 g body weight mean daily dose) as a drinking-water substitute for about 1 month on streptozotocin-diabetes (STZ-diabetes) in male Wistar rats were investigated. The physico-metabolic parameters measured were: body weight, food and liquid intake, urinary volume, hepatic glycogen, serum triglycerides and cholesterol, plasma glucose, urinary glucose and urea, and the weight of epididymal and retroperitoneal adipose tissue and soleus and extensor digitorum longus muscles. The STZ-diabetic rats treated with decoction showed a significant reduction in serum and urinary glucose and urinary urea as compared to the STZ-diabetic control, no difference being seen between decoction-treated and -untreated non-diabetic rats. The other physico-metabolic factors showed no changes in treated STZ-diabetic rats. The improvement in carbohydrate metabolism seen in the rats treated with Bauhinia forficata decoction does not appear to be linked to the inhibition of glycogenolysis or the stimulation of glycogenesis nor does it appear to act in a way similar to insulin or the sulfonylureas, although it may act by the inhibition of neoglycogenesis in a manner similar to that of the biguanides.
Asunto(s)
Bauhinia , Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Animales , Diabetes Mellitus Experimental/sangre , Hipoglucemiantes/aislamiento & purificación , Hipoglucemiantes/farmacología , Masculino , Fitoterapia/métodos , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Hojas de la Planta , Ratas , Ratas WistarRESUMEN
In a previous study, we showed that purified commercial esterase activity can be detected in a chemiluminescent assay based on the hydrolysis of 2-methyl-1-propenylbenzoate (MPB) to 2-methyl-1-propenol, which is subsequently oxidized by the horseradish peroxidase (HRP)-H(2)O(2) system. The purpose of this study was to verify the applicability of this assay to human serum. The existence of an esterase activity capable of hydrolysing MPB is indicated by the fact that the MPB-serum-HRP-H(2)O(2) system consumes oxygen and emits light. Both signals were abolished by prior serum heat inactivation and were preserved when serum was stored at < or =4 degrees C. Addition of aliesterase inhibitors, such as fluoride ion and trichlorfon or the cholinesterase inhibitor eserine, totally prevents light emission. The butyrylcholinesterase-specific substrate benzoylcholine causes a delay in both O(2) uptake and light emission, while the specific acetylcholinesterase substrate, acetyl-beta-methylcholine, had practically no effect. Purified butyrylcholinesterase, but not acetylcholinesterase, triggered light emission. The finding that butyrylcholinesterase is responsible for the hydrolysis of MPB in serum should serve as the basis for the development of a specific chemiluminescent assay for this enzyme.
Asunto(s)
Benzoatos/química , Butanoles/química , Butirilcolinesterasa/sangre , Inhibidores de la Colinesterasa/química , Animales , Benzoatos/metabolismo , Benzoilcolina/química , Benzoilcolina/metabolismo , Butanoles/metabolismo , Bovinos , Inhibidores de la Colinesterasa/metabolismo , Eritrocitos/enzimología , Fluoruros/química , Peroxidasa de Rábano Silvestre/química , Peroxidasa de Rábano Silvestre/metabolismo , Humanos , Hidrólisis , Mediciones Luminiscentes , Cloruro de Metacolina/química , Cloruro de Metacolina/metabolismo , Fisostigmina/química , Fisostigmina/metabolismo , Triclorfón/químicaRESUMEN
Streptozotocin-diabetic rats were treated for 17 days with a decoction of Eugenia jambolana (Myrtaceae) leaves (15%, w/v) as a substitute for water. Body weight, food and fluid intake, urine volume, glycemia, urinary glucose and urea were evaluated every 5 days. The animals were sacrificed by decapitation and blood samples collected for the determination of glycemia, serum cholesterol, HDL-cholesterol, triglycerides and angiotensin-converting enzyme. The weight of adipose and muscle tissues was also determined. There were no statistically significant differences between treated and untreated rats for any of the biochemical or physiological parameters. We conclude that, at least in this experimental model, Eugenia jambolana leaf decoction has no antidiabetic activity.
Asunto(s)
Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Plantas Medicinales/química , Análisis de Varianza , Animales , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Peso Corporal , Diabetes Mellitus Experimental/sangre , Evaluación Preclínica de Medicamentos , Masculino , Extractos Vegetales/uso terapéutico , Hojas de la Planta/química , Ratas , Ratas Wistar , EstreptozocinaRESUMEN
Streptozotocin-diabetic rats were treated for 17 days with a decoction of Eugenia jambolana (Myrtaceae) leaves (15 percent, w/v) as a substitute for water. Body weight, food and fluid intake, urine volume, glycemia, urinary glucose and urea were evaluated every 5 days. The animals were sacrificed by decapitation and blood samples collected for the determination of glycemia, serum cholesterol, HDL-cholesterol, triglycerides and angiotensin-converting enzyme. The weight of adipose and muscle tissues was also determined. There were no statistically significant differences between treated and untreated rats for any of the biochemical or physiological parameters. We conclude that, at least in this experimental model, Eugenia jambolana leaf decoction has no antidiabetic activity
Asunto(s)
Animales , Masculino , Ratas , Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Extractos Vegetales/uso terapéutico , Plantas Medicinales/química , Análisis de Varianza , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Diabetes Mellitus Experimental/sangre , Hipoglucemiantes/farmacología , Extractos Vegetales/farmacología , Hojas de la Planta/química , Ratas Wistar , EstreptozocinaRESUMEN
In this work we investigate the possible toxicity of vanadyl sulfate (VOSO4), a compound capable of reducing hyperglycemia, on the following serum enzymes of diabetic young rats: alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LD) and creatine kinase (CK), as well as its effects on serum lipids. We find that at a concentration of 1 mg/mL VOSO4 has no toxic effect on the liver and muscles of diabetics young rats. These findings suggest that VOSO4 may be an alternative to insulin in the near future, due to its low cost, low toxicity and ready availability.
Asunto(s)
Diabetes Mellitus Experimental/sangre , Lípidos/sangre , Compuestos de Vanadio/farmacología , Administración Oral , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Creatina Quinasa/sangre , Diabetes Mellitus Experimental/enzimología , L-Lactato Deshidrogenasa/sangre , Masculino , Ratas , Ratas Wistar , Estreptozocina , Compuestos de Vanadio/administración & dosificaciónRESUMEN
Esterase from monocytes promotes the hydrolysis of 2-methyl-1-propenylbenzoate (MPB) yielding 2-methyl-1-propenol, which is oxidized by horseradish peroxidase/H2O2 producing triplet acetone. The chemiluminescence of this reaction can be enhanced by the addition of 9,10-dibromoanthracene-2-sulphonate. The non-specific esterase present in monocytes is responsible for MPB hydrolysis, since (a) the chemiluminescence of the reaction was inhibited by fluoride, and (b) cells that do not contain a significant amount of non-specific esterases, e.g. lymphocytes and neutrophils, did not trigger light emission. The analytical application of this reaction is considered.
Asunto(s)
Esterasas/sangre , Granulocitos/enzimología , Linfocitos/enzimología , Monocitos/enzimología , Benzoatos , Colorantes Fluorescentes , Peroxidasa de Rábano Silvestre , Humanos , Peróxido de Hidrógeno , Indicadores y Reactivos , Cinética , Mediciones LuminiscentesRESUMEN
A simple and sensitive chemiluminescence assay for the demonstration of the activity of intracellular myeloperoxidase (MPO) is described, which is useful for the distinction between myeloid and lymphoid commitment in blasts from acute leukemia patients. When the cut-off point was settled at 13 mV of chemiluminescence all cases of acute myeloid leukemia (AML) were distinguished from those of acute lymphoid leukemia. In addition, this technique was able to demonstrate MPO activity in AML poorly differentiated (FAB-M0) which usually does not stain for MPO in classical cytochemistry preparations and could be negative also by immunocytochemistry with anti-MPO monoclonal antibody. Therefore the method here described presented a higher sensitivity than the immunocytochemistry procedure with anti-MPO.