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1.
West Afr J Med ; 36(3): 211-216, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31622482

RESUMO

BACKGROUND: Antiepileptic drugs are necessary for successful treatment of epilepsy. Unfortunately, epilepsy itself and some antiepileptic drugs have been documented to provoke or worsen seizure frequency by altering blood levels of some oxidants and antioxidants in persons with epilepsy. OBJECTIVE: This study investigated the effect of epilepsy and antiepileptic drugs on blood levels of some oxidants and antioxidants. METHODOLOGY: This was a cross-sectional case-control study. Blood samples were obtained from 35 antiepileptic drug-experienced persons with epilepsy; 35 antiepileptic-naive persons with epilepsy; and 35 age- and- sex matched apparently healthy controls; and analysed for malondialdehyde and antioxidants (uric acid, superoxide dismutase, glutathione peroxidase and catalase) using enzyme-linked immunosorbent assay. RESULTS: One-hundred and five (105) subjects (35 patients on antiepileptic drugs, 35 newly diagnosed, antiepileptic drug-naive and 35 healthy controls) were investigated. The median ages of antiepileptic drug-experienced, antiepileptic drug-naive and healthy participants were 30.0, 26.0 and 37.0 years respectively. Persons with epilepsy had significantly higher blood levels of malondialdehyde and uric acid and lower levels of enzymatic antioxidants than healthy controls. Also, persons with epilepsy on antiepileptic drug polytherapy had signi-ficantly higher blood levels of malondialdehyde and uric acid and lower levels of enzymatic antioxidants than antiepileptic drug-naive persons with epilepsy and persons with epilepsy on antiepileptic drug monotherapy respectively. CONCLUSION: Epilepsy and antiepileptic drug significantly altered blood levels of malondialdehyde, uric acid and enzymatic antioxidants and/or their homeostatic kinetics.


Assuntos
Anticonvulsivantes/uso terapêutico , Antioxidantes/metabolismo , Epilepsia/sangue , Epilepsia/tratamento farmacológico , Eritrócitos/metabolismo , Malondialdeído/sangue , Adulto , Antioxidantes/análise , Estudos de Casos e Controles , Estudos Transversais , Humanos
2.
Exp Parasitol ; 205: 107747, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31442454

RESUMO

Development of new chemotherapeutic agents is an essential issue in the treatment and control of a disease. This study aimed to evaluate the anti-leishmanial activity of amiodarone, an antiarrhythmic class III drug, against Leishmania major, the most prevalent etiological agent of cutaneous leishmaniasis in the old world. The proliferation of promastigotes and intracellular amastigotes in the absence or presence of amiodarone was estimated, in an in vitro study. For in vivo study, five weeks after infection of BALB/c mice with L. major, when the lesions appeared at the injection site, the mice were divided into four groups (n = 6 each); treatment was conducted for 28 consecutive days with vehicle, amiodarone at 40 mg/kg orally and glucantime at 60 mg/kg intraperitoneally. Therapy with amiodarone reduced the size of lesions compared to the untreated group after 12 days. Amiodarone decreased the parasite load and inflammatory responses, particularly the macrophages containing amastigotes, and enhanced granulation tissue formation in the dermis and subcutaneous area. The Tumor necrosis factor-α and Interleukin-6 levels were significantly lower in the cell culture supernatants of the inguinal lymph node in the amiodarone treated group compared to the vehicle and untreated groups. Amiodarone significantly increased the activity of glutathione peroxidase in comparison to the vehicle and untreated groups but did not affect the plasma levels of superoxide dismutase, malondialdehyde, adiponectin, and ferric reducing ability of plasma. Therefore, the anti- L. major activity and immunomodulatory effects of amiodarone reduced the parasitic load and enhanced wound healing in cutaneous leishmaniasis in BALB/c mice. Amiodarone reduced the lesion surface area, but it did not cure it completely.


Assuntos
Amiodarona/uso terapêutico , Antiprotozoários/uso terapêutico , Leishmania major/efeitos dos fármacos , Leishmaniose Cutânea/tratamento farmacológico , Adiponectina/sangue , Amiodarona/farmacologia , Animais , Antiarrítmicos/farmacologia , Antiarrítmicos/uso terapêutico , Antiprotozoários/farmacologia , Linhagem Celular , Feminino , Glutationa Peroxidase/metabolismo , Concentração Inibidora 50 , Interleucina-6/análise , Leishmania major/ultraestrutura , Leishmaniose Cutânea/parasitologia , Linfonodos/química , Linfonodos/imunologia , Macrófagos/parasitologia , Malondialdeído/sangue , Antimoniato de Meglumina/farmacologia , Antimoniato de Meglumina/uso terapêutico , Camundongos , Camundongos Endogâmicos BALB C , Carga Parasitária , Distribuição Aleatória , Pele/parasitologia , Pele/patologia , Pele/ultraestrutura , Superóxido Dismutase/metabolismo , Fator de Necrose Tumoral alfa/análise
3.
Artigo em Chinês | MEDLINE | ID: mdl-31446727

RESUMO

Objective:The aim of this study is to analyze the effects of glutathione on oxidative stress, leptin and adiponectin in patients with obstructive sleep apnea(OSA) complicated with metabolic syndrome. Method:One hundred and fifty-nine patients with OSA and MS were enrolled in the group A according to the exclusion criteria. One hundred and fifty-nine patients with MS group were not included in the OSA group, and 159 patients were included in the control group. Before and after treatment, the levels of serum malondialdehyde (MDA), superoxide dismutase (SOD), Leptin and ADP were respectively detected, and the clinical effects of the three groups were compared. Result:Compared with the control group, the contents of MDA and Leptin in the case A and B groups were significantly higher than that of the control group, and the contents of SOD and ADP were significantly lower than that of the control group, and the difference was statistically significant, especially in case group A. The level of SOD and ADP was significantly higher in the group after treatment than before treatment, and the level of MDA and Leptin was significantly lower than before treatment. The difference was statistically significant, especially in case group A, too. Conclusion:Patients with OSA and MS are associated with oxidative stress. Glutathione can effectively improve the body's ability to resist oxidative stress, reduce oxidative damage, reduce leptin, and increase ADP levels.


Assuntos
Adiponectina/administração & dosagem , Glutationa/farmacologia , Leptina/administração & dosagem , Síndrome Metabólica/complicações , Estresse Oxidativo , Apneia Obstrutiva do Sono/sangue , Estudos de Casos e Controles , Humanos , Malondialdeído/sangue , Apneia Obstrutiva do Sono/complicações , Superóxido Dismutase/sangue
4.
Arch Endocrinol Metab ; 63(4): 376-384, 2019 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-31365624

RESUMO

OBJECTIVE: To test the influence of oral fructose and glucose dose-response solutions in blood glucose (BG), glucagon, triglycerides, uricaemia, and malondialdehyde in postprandial states in type 1 diabetes mellitus (T1DM) patients. SUBJECTS AND METHODS: The study had a simple-blind, randomized, two-way crossover design in which T1DM patients were selected to receive fructose and glucose solutions (75g of sugars dissolved in 200 mL of mineral-water) in two separate study days, with 2-7 weeks washout period. In each day, blood samples were drawn after 8h fasting and at 180 min postprandial to obtain glucose, glucagon, triglycerides, uric acid, lactate, and malondialdehyde levels. RESULTS: Sixteen T1DM patients (seven men) were evaluated, with a mean age of 25.19 ± 8.8 years, a mean duration of disease of 14.88 ± 4.73 years, and glycated hemoglobin of 8.13 ± 1.84%. Fructose resulted in lower postprandial BG levels than glucose (4.4 ± 5.5 mmol/L; and 12.9 ± 4.1 mmol/L, respectively; p < 0.01). Uric acid levels increased after fructose (26.1 ± 49.9 µmol/L; p < 0.01) and reduced after glucose (-13.6 ± 9.5 µmol/L; p < 0.01). The malondialdehyde increased after fructose (1.4 ± 1.6 µmol/L; p < 0.01) and did not change after glucose solution (-0.2 ± 1.6 µmol/L; p = 0.40). Other variables did not change. CONCLUSIONS: Fructose and glucose had similar sweetness, flavor and aftertaste characteristics and did not change triglycerides, lactate or glucagon levels. Although fructose resulted in lower postprandial BG than glucose, it increased uric acid and malondialdehyde levels in T1DM patients. Therefore it should be used with caution. ClinicalTrials.gov registration: NCT01713023.


Assuntos
Diabetes Mellitus Tipo 1/metabolismo , Frutose/metabolismo , Glucose/metabolismo , Período Pós-Prandial/efeitos dos fármacos , Edulcorantes/metabolismo , Adolescente , Adulto , Glicemia/análise , Glicemia/efeitos dos fármacos , Estudos Cross-Over , Relação Dose-Resposta a Droga , Tolerância a Medicamentos , Feminino , Frutose/farmacologia , Glucose/farmacologia , Humanos , Masculino , Malondialdeído/sangue , Pessoa de Meia-Idade , Projetos Piloto , Método Simples-Cego , Soluções/farmacologia , Edulcorantes/farmacologia , Paladar/efeitos dos fármacos , Triglicerídeos/sangue , Ácido Úrico/sangue , Adulto Jovem
5.
J Int Soc Sports Nutr ; 16(1): 31, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31366352

RESUMO

BACKGROUND: Cashew apple juice (CAJ) was shown to improve immunological mechanisms by regulating a balance between reactive oxygen species and antioxidant concentrations. However, no study exploring the effects of the CAJ and training status on the immune system and oxidative stress induced by exercise. Therefore, we investigated the effects of CAJ supplementation primarily on leukocyte counts and secondary on oxidative stress and cortisol changes after high-intensity exercise in trained and untrained men. METHODS: Ten moderately (endurance) trained (Age = 21.5 ± 0.97 yr., VO2max = 45.6 ± 4.12 mL/kgBM/min) and ten sedentary men (Age = 20.4 ± 2.72 yr., VO2peak = 32.2 ± 7.26 mL/kgBM/min) were randomized to ingest either daily CAJ or a placebo at 3.5 mL/kgBM/day for 4 weeks, with a four-week washout period. Before and after each period, they performed 20-min, high-intensity cycling (85% VO2max), with blood samples collected immediately preceding and the following exercise. Samples were analyzed to determine leukocyte counts, malondialdehyde, 8-isoprostane, and cortisol concentrations. A repeated measures analysis of variance was used to examine the effects of supplement and training status over time with an alpha level of 0.05. RESULTS: There was no interaction between supplement and training status on those variables before and after exercise. However, CAJ raised resting neutrophil counts and exercise-induced leukocyte counts in the trained group (all p < 0.05). Besides, CAJ significantly reduced plasma malondialdehyde concentrations at rest and after exercise and reduced the post-exercise plasma 8-isoprostane concentration in both groups of subjects (p < 0.05). Moreover, CAJ reduced plasma cortisol after exercise in the untrained subjects. CONCLUSIONS: We suggest that 4-week CAJ supplementation can enhance exercise-induced leukocyte and resting neutrophil counts in trained men. The possible mechanism is a reduction in oxidative stress. However, the supplementation did not change the immune responses of untrained men, but it did reduce stress hormone concentrations. TRIAL REGISTRATION NUMBER: TCTR20181127002 Registered 26 November 2018 "retrospectively registered".


Assuntos
Suplementos Nutricionais , Exercício , Sucos de Frutas e Vegetais , Contagem de Leucócitos , Estresse Oxidativo , Anacardium , Estudos Cross-Over , Dinoprosta/análogos & derivados , Dinoprosta/sangue , Método Duplo-Cego , Humanos , Hidrocortisona/sangue , Masculino , Malondialdeído/sangue , Malus , Neutrófilos/citologia , Espécies Reativas de Oxigênio/metabolismo , Fenômenos Fisiológicos da Nutrição Esportiva , Adulto Jovem
6.
J Int Soc Sports Nutr ; 16(1): 33, 2019 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-31375128

RESUMO

BACKGROUND: Affective responses experienced during exercise are a significant determinant on exercise adherence. We have previously demonstrated that consumption of New Zealand (NZ) blackcurrants preserves cognition by attenuating the feeling of fatigue. This positive affective response correlated with the ability of blackcurrant polyphenols to support monoamine neurotransmission via inhibition of monoamine oxidase-B (MAO-B) activity. Here we explore how the consumption of a NZ blackcurrant juice (BJ) influenced affective responses and potential ergogenic action on the motivation to adhere to a low impact walking exercise. METHODS: In a parallel randomized controlled study (Trial registration #: ACTRN12617000319370p, registered 28th February 2017, http://www.anzctr.org.au/ ), 40 healthy sedentary male and female participants drank a BJ or matched placebo (PLA) (n = 20 per group), 1 h prior to a self-motivated treadmill walk, where heart rate and affective responses (exertion [ES] or feeling / mood [FS]) scores) were recorded at 3 or 5 min intervals. Blood glucose, lactate, malondialdehyde (MDA) and platelet MAO-B activity were measured pre- and post-exercise and comparisons were conducted using with Student's t-tests. Subjective data were analysed using 2-way ANOVA with appropriate post hoc tests. RESULTS: Consuming a BJ 1 h prior to exercise caused a 90% decline in platelet MAO-B activity. The exercise had no significant (p > 0.05) effect on blood lactate, glucose or plasma MDA levels. Assessment of affective responses over the first 60 mins (adjusting for participant drop-out) revealed a time-dependent ES increase in both groups, with ES reported by participants in the BJ group consistently lower than those in the PLA group (p < 0.05). FS declined in PLA and BJ groups over 60 mins, but an inverse relationship with ES was only observed within the PLA group (r2 = 0.99, p = 0.001). Whilst the average time walked by participants in the BJ group was 11 mins longer than the PLA group (p = 0.3), and 30% of the BJ group achieving > 10 km compared to only 10% for the PLA group (p = 0.28), statistical significance was not achieved. CONCLUSION: Our findings demonstrate that drinking a polyphenolic-rich NZ blackcurrant juice 1 h prior to exercise supports positive affective responses during a self-motivated exercise.


Assuntos
Afeto , Sucos de Frutas e Vegetais , Motivação , Caminhada , Adulto , Glicemia/análise , Plaquetas/enzimologia , Feminino , Frequência Cardíaca , Humanos , Ácido Láctico/sangue , Masculino , Malondialdeído/sangue , Monoaminoxidase/metabolismo , Nova Zelândia , Polifenóis/administração & dosagem , Ribes , Fenômenos Fisiológicos da Nutrição Esportiva , Fatores de Tempo
7.
Life Sci ; 232: 116608, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31254583

RESUMO

Preventing vascular damage is considered an effective strategy in patients who suffer from chronic obstructive pulmonary disease (COPD) with hypertension. Here, we investigated vascular damage in COPD-like and hypertensive rats, which demonstrated the presence of the three related factors of COPD with hypertension. These include elevated systolic blood pressure (SBP), serum malondialdehyde (MDA) and serum lactate dehydrogenase (LDH), which are positively correlated with vascular damage in patients. In addition to increases in these three related factors, COPD-like and hypertensive rats exhibited increased levels of pro-inflammatory mediators, such as tumor necrosis factor-α, interleukin-6, and matrix metallopeptidase-9 in bronchoalveolar lavage fluid, and enlargement of alveolar airspaces, recapitulating clinical findings in previous studies of patients. Moreover, the appearance of these related factors was prevented by linalyl acetate. Our results provide novel insight into the potential of LA to prevent vascular damage and elevated SBP, serum MDA and serum LDH in COPD with hypertension, and could lead to an alternative strategy for preventing vascular damage for patients who suffered from COPD with hypertension in a clinical setting.


Assuntos
Hipertensão/tratamento farmacológico , Monoterpenos/farmacologia , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Animais , Pressão Sanguínea/fisiologia , Líquido da Lavagem Broncoalveolar , Modelos Animais de Doenças , Hipertensão/patologia , Hipertensão/fisiopatologia , L-Lactato Desidrogenase/análise , L-Lactato Desidrogenase/sangue , Pulmão/patologia , Masculino , Malondialdeído/análise , Malondialdeído/sangue , Doença Pulmonar Obstrutiva Crônica/patologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Ratos , Ratos Sprague-Dawley
8.
J Agric Food Chem ; 67(28): 7832-7843, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31242723

RESUMO

Oxidative-stress-induced senescence constitutes a great risk factor for chronic diseases. Therefore, ameliorating oxidative-stress-induced senescence is expected to prevent chronic diseases. The beneficial effects of bilberry anthocyanin (BA) on healthy aging were evaluated using 12 month old, aging female SD rats in this study. The experimental results suggested that consumption of a middle-dose of BA (MBA) appreciably increased the relative liver mass by 7.34% when compared with that of the AC group. Furthermore, BA significantly increased the total antioxidant capacity, total superoxide dismutase activity, and catalase activities; decreased malondialdehyde, serum low-density lipoprotein cholesterol (LDL-C), serum total cholesterol (TC), serum triglyceride (TG), and glycated serum protein (GSP) levels; and reduced TC/high-density lipoprotein cholesterol (HDL-C) and LDL-C/HDL-C ratios. In addition, MBA decreased the activity of fecal bacterial enzymes and increased the content of fecal short-chain fatty acids. The Western blot results showed that MBA significantly upregulated the expression of OCLN, ZO-1, and autophagy-related proteins (ATP6 V0C, ATG4D, and CTSB) in aging rats. Moreover, it also showed that MBA induced the phosphorylation of AMPK and FOXO3a and inhibited the phosphorylation of mTOR, which indicated that bilberry anthocyanin induced autophagy via the AMPK-mTOR signaling pathways. This induction of autophagy further promoted oxidative stress resistance effects and intestinal epithelial barrier function of bilberry anthocyanin in aging female rats.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Envelhecimento/fisiologia , Antocianinas/administração & dosagem , Autofagia/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Vaccinium myrtillus/química , Proteínas Quinases Ativadas por AMP/genética , Envelhecimento/sangue , Envelhecimento/efeitos dos fármacos , Envelhecimento/genética , Animais , Suplementos Nutricionais/análise , Feminino , Proteína Forkhead Box O3/genética , Proteína Forkhead Box O3/metabolismo , Humanos , Lipoproteínas LDL/sangue , Malondialdeído/sangue , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/genética , Triglicerídeos/sangue
9.
BMC Complement Altern Med ; 19(1): 127, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31196042

RESUMO

BACKGROUND: Xin-Ji-Er-Kang (XJEK) is a Chinese herbal formula, which has been reported to exert effective protection against cardiovascular diseases, including hypertension and myocarditis. METHODS: Cultured human umbilical vascular endothelial cells (HUVECs) were treated with angiotensin II (Ang II) and different concentrations of aqueous layer extracts (AqE). Subsequently nitric oxide (NO) and endothelial nitric oxide synthase (eNOS) expression levels were detected. In addition, fifty Kunming mice were randomized into control, Nω-nitro-L-arginine methyl ester (L-NAME), L-NAME+AqE, L-NAME+XJEK and L-NAME+fosinopril treatment groups. Following 8 weeks of treatment, the cardiac hemodynamic index was measured, relaxation of the aorta was examined and pathological changes were observed. Colorimetric analysis and enzyme linked immunosorbent assay (ELISA) were applied to determine the relevant indicators in plasma and cardiac tissues. RESULTS: The in vitro study results demonstrated that AqE could preserve endothelial function (NO, 21.05 ± 2.03 vs. 8.64 ± 0.59; eNOS, 1.08 ± 0.17 vs.0.73 ± 0.06). In addition, the in vivo results demonstrated that compared with the control group, treatment with AqE could enhance a high hemodynamic state (left ventricular systolic pressure, 116.76 ± 9.96 vs.114.5 ± 15.16), improve endothelial function (NO, 7.98 ± 9.64 vs. 1.66 ± 3.11; eNOS, 19.78 ± 3.18 vs.19.38 ± 3.85), suppress oxidative stress (OS) (superoxide dismutase, 178.17 ± 13.78 vs. 159.38 ± 18.86; malondialdehyde, 0.77 ± 0.13 vs.1.25 ± 0.36) and reverse cardiovascular remodeling. CONCLUSION: Polysaccharide from XJEK exerts protective effects against Ang II-induced injury in HUVECs and L-NAME-induced hypertension in mice and the underlying mechanism may be attributed to improving endothelial dysfunction, OS and the inflammation status in mice.


Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Células Endoteliais/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Hipertensão/tratamento farmacológico , Remodelação Vascular/efeitos dos fármacos , Angiotensina II , Animais , Aorta/efeitos dos fármacos , Arginina/análogos & derivados , Arginina/sangue , Pressão Sanguínea/efeitos dos fármacos , Citocinas/metabolismo , Avaliação Pré-Clínica de Medicamentos , Medicamentos de Ervas Chinesas/farmacologia , Células Endoteliais/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Hipertensão/induzido quimicamente , Inflamação/tratamento farmacológico , Masculino , Malondialdeído/sangue , Camundongos , Miocárdio/metabolismo , NG-Nitroarginina Metil Éster , Óxido Nítrico/sangue , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Fitoterapia , Superóxido Dismutase/sangue
10.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 35(2): 150-154, 2019 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-31250607

RESUMO

OBJECTIVE: To investigate the effects of aerobic exercise and glutamine (Gln) on anti-oxidative stress and inflammatory factors in type 2 diabetes mellitus (T2MD) rats. METHODS: Diabetic rat model was induced by streptozotocin (STZ). Fifty 6-week old male SD rats were randomly divided into 5 groups (n=10), including quiet control group (N), diabetes control group (D), diabetic aerobic exercise group (DE), diabetic glutamine group (DG) and diabetic aerobic exercise glutamine group (DEG). After 6 weeks, the related indicators of glucose and lipid metabolism, anti-oxidative stress and inflammatory factors in diabetic rats were detected, and the possible mechanism affecting inflammatory response were explored. RESULTS: Compared with group N, the levels of serum malondialdehyde(MDA), blood glucose, total cholesterol(TC), triglyceride(TG), insulin, leptin and tumor necrosis factor-α(TNF-α) in group D were increased significantly (P<0.01). Compared with group D, serum levels of MDA, blood glucose, TC, TG, insulin, leptin and TNF-α in three intervention groups were decreased significantly, while the levels of SOD, GSH-Px and adiponectin were increased, and the combined effect was more obvious (P<0.01). CONCLUSION: Both aerobic exercise and Gln can relieve the glucose and lipid metabolism and disturbance, oxidative stress injury and inflammation in diabetic rats.


Assuntos
Diabetes Mellitus Tipo 2/terapia , Glutamina/farmacologia , Estresse Oxidativo , Condicionamento Físico Animal , Animais , Glicemia/análise , Diabetes Mellitus Experimental , Leptina/sangue , Metabolismo dos Lipídeos , Lipídeos/sangue , Masculino , Malondialdeído/sangue , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley
11.
J Dairy Sci ; 102(8): 7000-7010, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31155245

RESUMO

This study aimed to determine the effects of supplementing pregnant heifers with the organic selenium (Se) source 2-hydroxy-4-methylselenobutanoic acid (HMSeBA) during the last 8 wk of pregnancy on dam and calf Se status. A total of 42 in-calf heifers were recruited to the study and randomly allocated to 1 of 3 treatments; a negative control (Con), sodium selenite (NaSe), or HMSeBA. Animals were blocked by body weight, body condition score, and expected calving date before treatment allocation. Following enrollment, all animals underwent a 7-wk wash-out period, after which they received their respective supplements, top-dressed daily onto a basal diet for the last 8 wk of pregnancy. Heifer blood samples were taken at weekly intervals from enrollment until 2 wk before expected calving date and as soon as possible after calving for determination of whole-blood glutathione peroxidase activity (GSH-Px) and plasma Se and malondialdehyde (MDA) concentrations. Selenized AA were determined in plasma samples taken at 3 wk precalving. A colostrum sample was taken as close to parturition as possible for determination of colostrum total Se, selenized AA, and IgG concentration. Calves were blood sampled as close to birth as possible for determination of whole-blood GSH-Px activity and plasma Se and MDA concentrations. Differences in whole-blood GSH-Px activity did not become apparent until calving; GSH-Px activity was lowest in Con heifers but similar between NaSe and HMSeBA heifers. Plasma Se was lowest in unsupplemented heifers and greatest in those supplemented with HMSeBA; this was attributable to greater selenomethionine concentrations in the plasma of HMSeBA heifers. Colostrum Se was lowest in Con heifers and greatest in HMSeBA heifers. The greater Se concentration of HMSeBA heifers was attributable to a greater proportion of total Se comprising selenocysteine; the reason for this is not known. There was no effect of supplementation on colostrum IgG concentration. Plasma Se was lowest in calves born to Con heifers and greatest in those born to HMSeBA heifers. There were no effects of treatment on calf whole-blood GSH-Px activity or plasma MDA concentration. The enhanced Se status associated with HMSeBA supplementation is likely a consequence of selenomethionine supply and may confer benefits to both the dam and her calf postpartum.


Assuntos
Bovinos/fisiologia , Colostro/química , Suplementos Nutricionais/análise , Selênio/sangue , Selenometionina/farmacologia , Selenito de Sódio/farmacologia , Animais , Animais Recém-Nascidos , Indústria de Laticínios , Dieta/veterinária , Feminino , Malondialdeído/sangue , Período Pós-Parto/efeitos dos fármacos , Gravidez , Distribuição Aleatória
12.
Molecules ; 24(9)2019 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-31086027

RESUMO

Naodesheng (NDS) is a widely used traditional Chinese medicine (TCM) prescription for the treatment of ischemic stroke. A combination of 10 components is derived from NDS. They are: Notoginsenoside R1, ginsenoside Rg1, ginsenoside b1, ginsenoside Rd, hydroxysafflor yellow A, senkyunolide I, puerarin, daidzein, vitexin, and ferulic acid. This study aimed to investigate the protective effect of the ten-component combination derived from NDS (TCNDS) on ischemic stroke rats with a middle cerebral artery occlusion (MCAO) model by integrating an NMR-based metabonomics approach with biochemical assessment. Our results showed that TCNDS could improve neurobehavioral function, decrease the cerebral infarct area, and ameliorate pathological features in MCAO model rats. In addition, TCNDS was found to decrease plasma lactate dehydrogenase (LDH) and malondialdehyde (MDA) production and increase plasma superoxide dismutase (SOD) production. Furthermore, 1H-NMR metabonomic analysis indicated that TCNDS could regulate the disturbed metabolites in the plasma, urine, and brain tissue of MCAO rats, and the possible mechanisms were involved oxidative stress, energy metabolism, lipid metabolism, amino acid metabolism, and inflammation. Correlation analysis were then performed to further confirm the metabolites involved in oxidative stress. Correlation analysis showed that six plasma metabolites had high correlations with plasma LDH, MDA, and SOD. This study provides evidence that an NMR-based metabonomics approach integrated with biochemical assessment can help to better understand the underlying mechanisms as well as the holistic effect of multiple compounds from TCM.


Assuntos
Isquemia Encefálica/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Espectroscopia de Ressonância Magnética/métodos , Metabolômica/métodos , Acidente Vascular Cerebral/tratamento farmacológico , Animais , Apigenina/uso terapêutico , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Isquemia Encefálica/sangue , Isquemia Encefálica/urina , Ginsenosídeos/uso terapêutico , Infarto da Artéria Cerebral Média , Isoflavonas/uso terapêutico , L-Lactato Desidrogenase/sangue , Masculino , Malondialdeído/sangue , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral/urina , Superóxido Dismutase/sangue
13.
Zhongguo Zhong Yao Za Zhi ; 44(7): 1357-1362, 2019 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-31090292

RESUMO

In this study, solid dispersion technology was used to develop volatile oil from Acorus tatarinowii self-nanoemulsion dropping pills(VOA-SNEDDS-DP) and its protective effect on acute myocardial ischemia injury was evaluated. Taking exterior quality, weight variation and the resolving time as comprehendsive evaluation indexes, the preparation process and formulation of the dropping pills were optimized by orthogonal design, and the dissolution rate in vitro of the optimized VOA-SNEDDS-DP was investigated. The rat model of acute myocardial ischemia was induced by intraperitoneal injection of isoproterenol hydrochloride and the serum levels of superoxide dismutase(SOD), malondialdehyde(MDA), creatine kinase(CK) and pathological changes of myocardial tissue were determined to evaluate therapeutic effect of the dropping pills on acute myocardial ischemia. The results showed that the optimal formulation and preparation process of VOA-SNEDDS-DP were as follows: PEG6000-PEG8000 was 1∶1, proportion of VOA-SNEDDS and matrix was l∶2.5, the temperature of drug fluids was 75 ℃, drop rate was 35 drops/min, drop distance was 5 cm, the condensing agent temperature was 2-10 ℃. The content of ß-asarone in the dropping pills was 42.46 mg·g~(-1). The accumulated dissolution rate of the dropping pills reached 93.85% in 10 min. The results of pharmacodynamic experiments showed that VOA-SNEDDS-DP could significantly increase the SOD content(P<0.05), reduce the levels of MDA and CK(P<0.05) in serum, and effectively improve the pathological morphology of myocardial tissue. These results revealed that the preparation of VOA-SNEDDS-DP by solid dispersion technology was stable and feasible, and VOA-SNEDDS-DP had protective effect on acute myocardial ischemia injury.


Assuntos
Acorus/química , Medicamentos de Ervas Chinesas/farmacologia , Isquemia Miocárdica/tratamento farmacológico , Óleos Voláteis/farmacologia , Óleos Vegetais/farmacologia , Animais , Creatina Quinase/sangue , Malondialdeído/sangue , Ratos , Superóxido Dismutase/sangue
14.
Int J Mol Sci ; 20(10)2019 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-31100876

RESUMO

Type 2 diabetes mellitus (DM2) leads to cardiomyopathy characterized by cardiomyocyte hypertrophy, followed by mitochondrial dysfunction and interstitial fibrosis, all of which are exacerbated by angiotensin II (AT). SIRT1 and its transcriptional coactivator target PGC-1α (peroxisome proliferator-activated receptor-γ coactivator), and heme oxygenase-1 (HO-1) modulates mitochondrial biogenesis and antioxidant protection. We have previously shown the beneficial effect of caloric restriction (CR) on diabetic cardiomyopathy through intracellular signaling pathways involving the SIRT1-PGC-1α axis. In the current study, we examined the role of HO-1 in diabetic cardiomyopathy in mice subjected to CR. METHODS: Cardiomyopathy was induced in obese diabetic (db/db) mice by AT infusion. Mice were either fed ad libitum or subjected to CR. In an in vitro study, the reactive oxygen species (ROS) level was determined in cardiomyocytes exposed to different glucose levels (7.5-33 mM). We examined the effects of Sn(tin)-mesoporphyrin (SnMP), which is an inhibitor of HO activity, the HO-1 inducer cobalt protoporphyrin (CoPP), and the SIRT1 inhibitor (EX-527) on diabetic cardiomyopathy. RESULTS: Diabetic mice had low levels of HO-1 and elevated levels of the oxidative marker malondialdehyde (MDA). CR attenuated left ventricular hypertrophy (LVH), increased HO-1 levels, and decreased MDA levels. SnMP abolished the protective effects of CR and caused pronounced LVH and cardiac metabolic dysfunction represented by suppressed levels of adiponectin, SIRT1, PPARγ, PGC-1α, and increased MDA. High glucose (33 mM) increased ROS in cultured cardiomyocytes, while SnMP reduced SIRT1, PGC-1α levels, and HO activity. Similarly, SIRT1 inhibition led to a reduction in PGC-1α and HO-1 levels. CoPP increased HO-1 protein levels and activity, SIRT1, and PGC-1α levels, and decreased ROS production, suggesting a positive feedback between SIRT1 and HO-1. CONCLUSION: These results establish a link between SIRT1, PGC-1α, and HO-1 signaling that leads to the attenuation of ROS production and diabetic cardiomyopathy. CoPP mimicked the beneficial effect of CR, while SnMP increased oxidative stress, aggravating cardiac hypertrophy. The data suggest that increasing HO-1 levels constitutes a novel therapeutic approach to protect the diabetic heart. Brief Summary: CR attenuates cardiomyopathy, and increases HO-1, SIRT activity, and PGC-1α protein levels in diabetic mice. High glucose reduces adiponectin, SIRT1, PGC1-1α, and HO-1 levels in cardiomyocytes, resulting in oxidative stress. The pharmacological activation of HO-1 activity mimics the effect of CR, while SnMP increased oxidative stress and cardiac hypertrophy. These data suggest the critical role of HO-1 in protecting the diabetic heart.


Assuntos
Restrição Calórica/métodos , Cardiomiopatias Diabéticas/tratamento farmacológico , Cardiomiopatias Diabéticas/metabolismo , Heme Oxigenase-1/metabolismo , Heme Oxigenase-1/uso terapêutico , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Angiotensina II/metabolismo , Animais , Glicemia , Carbazóis/farmacologia , Cardiomegalia/metabolismo , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2/complicações , Masculino , Malondialdeído/sangue , Mesoporfirinas/uso terapêutico , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/metabolismo , Estresse Oxidativo/efeitos dos fármacos , PPAR gama/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Protoporfirinas/metabolismo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/antagonistas & inibidores , Sirtuína 1/metabolismo
15.
Biomarkers ; 24(5): 492-498, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31099265

RESUMO

Objective: The aim of the present work is to evaluate the toxicity of titanium dioxide nanoparticles (TiO2NPs) according to their doses and particle sizes. Materials and methods: The effect of five days oral administration of TiO2NPs (21 and 80 nm) with different doses (50, 250 and 500 mg/kg body weight) was assessed in mice via measurement of oxidative stress markers; glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and nitric oxide (NO), liver function indices; aspartate and alanine aminotransferases (AST and ALT), chromosomal aberrations and liver histopathological pattern. Results: The results revealed drastic alterations in all the measured parameters and showed positive correlation with the gradual dose increment. In addition, the smaller particle size of TiO2NPS (21 nm) had more adverse effect in all the selected biochemical parameters, genetic aberrations and histological investigations. Conclusions: Toxicity of TiO2NPs increases in a dose-dependent manner and vice versa with particles size. The evaluated biomarkers are good indicators for TiO2NPs toxicity. More detailed studies are required before the recommendation of TiO2NPS as food additives.


Assuntos
Biomarcadores/sangue , Nanopartículas/toxicidade , Titânio/toxicidade , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Catalase/sangue , Aberrações Cromossômicas/efeitos dos fármacos , Relação Dose-Resposta a Droga , Glutationa/sangue , Fígado/metabolismo , Fígado/patologia , Malondialdeído/sangue , Camundongos , Nanopartículas/metabolismo , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Tamanho da Partícula , Superóxido Dismutase/sangue , Titânio/metabolismo
16.
Transplant Proc ; 51(4): 1049-1053, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31101169

RESUMO

BACKGROUND: Long-term consequences of donor nephrectomy might be reduced kidney function, increased risk for cardiovascular disease, and impaired quality of life. The purpose of the current cross-sectional study was to evaluate the relationship between clinical, laboratory, and donation-specific outcomes of living kidney donors and systemic oxidative DNA damage. METHODS: We conducted a cross-sectional study and assessed retrospectively pre- and postdonation data from 60 donors who donated between 2010 and 2015. Plasma malondialdehyde levels and 8-hydroxy-2'-deoxyguanosine/deoxyguanosine ratio (8-OHdG/dG ratio) were determined as oxidative stress markers. Catalase, carbonic anhydrase, and paraoxonase (PON) activities were measured as antioxidants. RESULTS: Approximately 3 years after donation, the hypertensive donor ratio was 12%, and 11% of the donors had glomerular filtration rate <60 mL/min/1.73 m2. Mean serum urea (P = .001) and serum creatinine levels (P = .001) were increased; creatinine clearance level (126.2 ± 35.5 vs 94.6 ± 26.8, P = .001) was decreased in the postdonation period. There was a significant positive correlation between predonation serum urea and 8-0HdG/dG ratio (r = 0.338, P = .016) and predonation serum creatinine and 8-0HdG/dG ratio (r = 0.442, P = .001), while there was a significant negative correlation between serum creatinine and PON activity (r = -0.545, P < .001). CONCLUSION: Our data have demonstrated that kidney donors exhibit increased oxidative DNA damage and decreased antioxidant activity. We propose that predonation serum creatinine is positively correlated with 8-0HdG/dG ratio and negatively correlated with antioxidant PON activity. This is the first study to demonstrate that plasma oxidative DNA damage increases in healthy kidney donors.


Assuntos
Antioxidantes , Dano ao DNA , Nefrectomia/efeitos adversos , Estresse Oxidativo , Adulto , Biomarcadores/sangue , Creatinina/sangue , Estudos Transversais , Desoxiguanosina/análogos & derivados , Desoxiguanosina/sangue , Feminino , Humanos , Doadores Vivos , Masculino , Malondialdeído/sangue , Pessoa de Meia-Idade , Estudos Retrospectivos , Coleta de Tecidos e Órgãos/efeitos adversos
17.
BMC Complement Altern Med ; 19(1): 97, 2019 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-31060549

RESUMO

BACKGROUND: Endothelial dysfunction (ED) has been observed in individuals with metabolic syndrome (MetS) and contributes to the initiation and progression of atherosclerosis. The primary management of MetS involves lifestyle modifications and treatment of its individual components with drugs all of which have side effects. Thus, it would be of advantageous if natural products would be used as adjuncts or substitutes for conventional drugs. The aim of the present study was to evaluate the effect of standardized aqueous extract of fruits of Phyllanthus emblica (P. emblica) 250 mg and 500 mg twice daily on ED, oxidative stress, systemic inflammation and lipid profile in subjects with MetS. METHODS: In this randomised, double-blind, placebo-controlled clinical study endothelial function was measured by calculating reflection index (RI) using digital plethysmograph. Oxidative stress biomarkers used were nitric oxide (NO), glutathione (GSH) and malondialdehyde (MDA). Systemic inflammation was measured by determining high sensitivity C-reactive protein (hsCRP) and dyslipidemia by lipid profile. ANOVA, paired and unpaired t-test were used. P-value < 0.05 was considered statistically significant. RESULTS: Out of 65 screened subjects all 59 enrolled completed the study. P. emblica aqueous extract (PEE), 250 mg and 500 mg twice daily dosing, showed significant reduction in mean RI, measure of endothelial function, at 8 and 12 weeks (p <  0.001) compared to baseline and placebo. Significant mean % change was seen in oxidative stress biomarkers, NO (+ 41.89%, + 50.7%), GSH (+ 24.31%, + 53.22%) and MDA (- 21.02%, - 31.44%), and systemic inflammation biomarker, hsCRP (- 39.68%, - 53.77%) (p <  0.001) at 12 weeks with 250 mg and 500 mg twice daily dosage respectively. Significant mean % change was also seen at 12 weeks with TC (- 7.71%, - 11.11%), HDL-C (+ 7.33% + 22.16%, p <  0.05), LDL-C (- 11.39%, - 21.8%) and TG (- 9.81%, - 19.22%) respectively with 250 mg and 500 mg twice daily (p <  0.001). PEE 500 mg twice daily was significantly more efficacious than the 250 mg twice daily and placebo. No participant discontinued the study because of adverse events. CONCLUSIONS: P.emblica aqueous extract significantly improved endothelial function, oxidative stress, systemic inflammation and lipid profile at both dosages tested, but especially at 500 mg twice daily. Thus, this product may be used as an adjunct to conventional therapy (lifestyle modification and pharmacological intervention) in the management of metabolic syndrome. TRIAL REGISTRATION: This study was registered with Clinical Trials Registry - India (CTRI) with the registration number of CTRI/2017/09/009606 . The study was registered retrospectively on 4th September 2017.


Assuntos
Inflamação/tratamento farmacológico , Lipídeos/sangue , Síndrome Metabólica/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Phyllanthus emblica , Extratos Vegetais , Idoso , Método Duplo-Cego , Dislipidemias/metabolismo , Feminino , Frutas/química , Glutationa/sangue , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/sangue , Síndrome Metabólica/metabolismo , Pessoa de Meia-Idade , Óxido Nítrico/sangue , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico
18.
Georgian Med News ; (287): 87-90, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30958295

RESUMO

Hormonal dysfunction of the thyroid gland causes disorder of all kinds of metabolism. Both hypothyroidism, and hyperthyroidism are the causes of imbalance in prooxidant-antioxidant system with the development of pathological process named "oxidative stress". Objective - study of oxidative stress in patients who underwent thyroid gland operation depending on its functional activity and the presence of autoimmune thyroiditis. The intensity of lipid peroxidation was examined by concentration of diene conjugates and malondialdehyde, and the activity of antioxidant system by the level of catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase of red blood cells. It was established that the activity level of the lipid peroxidation system and the activity of antioxidant defense enzymes have their specific characteristics depending on the various functional states of the thyroid gland, as well as on the presence or absence of autoimmune component in the structure of thyroid pathology. All changes occurring in patients with this pathology accompanied by hormonal imbalance and autoimmune thyroiditis, lead to development of chronic nonspecific metabolic endogenous intoxication.


Assuntos
Antioxidantes/metabolismo , Glutationa Peroxidase/sangue , Hipotireoidismo , Peroxidação de Lipídeos , Malondialdeído/sangue , Estresse Oxidativo , Glândula Tireoide/cirurgia , Catalase , Glutationa Peroxidase/metabolismo , Glutationa Redutase , Humanos , Malondialdeído/metabolismo , Superóxido Dismutase/sangue , Superóxido Dismutase/metabolismo
19.
Biomed Res Int ; 2019: 2835152, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30984778

RESUMO

Traditionally, in many countries, various parts of the Adansonia digitata (A. digitata) tree have been used in the treatment of many clinical ailments including diarrhea and dysentery. The phytochemical screening has indicated that the leaf extract of A. digitata contains flavonoids, saponins, mucilage, steroids, and alkaloids. Thus, this paper aims to evaluate the hyperglycaemic and hypolipidaemic effects of methanolic extract of A. digitata leaves (200 mg/kg and 400 mg/kg) in diabetic rats. The extract was administered orally for six weeks in the streptozotocin (STZ)-induced diabetic rats. The treatment with the extract caused a significant reduction in the blood glucose, glycosylated hemoglobin, cholesterol, triglycerides, low-density lipoprotein (LDL), interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-α), and malondialdehyde (MDA) levels by 46.7%, 46.15%, 48.91%, 43%, 60%, 66%, 45.45%, and 30.4%, respectively, as compared to the diabetic group after the sixth week of treatment. The leaf extract also mitigated the decline of high-density lipoprotein (HDL) level, RBCs count, hemoglobin level, packed cell volume (PCV %), and erythropoietin concentration in diabetic rats by 31%, 33.25%, 24.72%, 51.42%, and 220.68% with respect to the diabetic group. Also, the extract maintained the level of antioxidant enzymes, catalase (CAT) and superoxide dismutase (SOD), and reduced glutathione (GSH) in the diabetic rats. It also reduced the elevation in the white blood corpuscles (WBC) count in the STZ-induced diabetic rats. Our study, therefore, indicates that methanolic extract of A. digitata leaf exerts strong antidiabetic and hypolipidaemic properties in a dose-dependent manner by improving the hematological properties and redox parameters in the experimental diabetic rats.


Assuntos
Adansonia/química , Diabetes Mellitus Experimental/tratamento farmacológico , Hiperglicemia/tratamento farmacológico , Hiperlipidemias/tratamento farmacológico , Animais , Antioxidantes/administração & dosagem , Glicemia/efeitos dos fármacos , Catalase/sangue , LDL-Colesterol/sangue , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/patologia , Modelos Animais de Doenças , Humanos , Hiperglicemia/sangue , Hiperglicemia/patologia , Hiperlipidemias/sangue , Hiperlipidemias/patologia , Hipoglicemiantes/administração & dosagem , Interleucina-6/sangue , Malondialdeído/sangue , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Ratos , Superóxido Dismutase/sangue , Fator de Necrose Tumoral alfa/sangue
20.
An Acad Bras Cienc ; 91(1): e20171042, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30994751

RESUMO

The aims of this study were to investigate whether treatments with zinc edetate (Zn) and diphenyl diselenide ((PhSe)2) enhance performance, immune responses, protein metabolism, and oxidant/antioxidant status in calf serum and muscle. Animals were divided into four groups (n=6 each): control (without supplementation), and groups supplemented on days 50 and 70 of life with (PhSe)2, Zn, and a combination of (PhSe)2 and Zn. Animals treated with (PhSe)2 gained more weight by experimental day 220 than did the control group, but there was no difference by the end of the experiment (day 300). The absolute number of leukocytes and lymphocytes increased in groups Zn and (PhSe)2+Zn on day 20 of experiment, but decreased on day 40 in groups (PhSe)2, and (PhSe)2+Zn. The number of monocytes decreased in all groups compared with control. One of the principal findings was that (PhSe)2+Zn together had beneficial effects on protein metabolism, represented by increases total protein and globulin levels, compared with the control group. The combination of (PhSe)2 and Zn led to low levels of TBARS and ROS in serum and muscle, and stimulated antioxidant enzyme activities. Thus, supplementation with (PhSe)2+Zn may be a compelling approach to augmenting the calf antioxidant system during weaning.


Assuntos
Derivados de Benzeno/administração & dosagem , Bovinos/fisiologia , Ácido Edético/administração & dosagem , Compostos Organosselênicos/administração & dosagem , Desmame , Zinco/administração & dosagem , Fatores Etários , Animais , Antioxidantes/análise , Peso Corporal , Sistema Imunitário/efeitos dos fármacos , Injeções Subcutâneas/veterinária , Metabolismo dos Lipídeos/efeitos dos fármacos , Malondialdeído/sangue , Músculos/metabolismo , Proteínas/metabolismo , Reprodutibilidade dos Testes , Selênio/administração & dosagem , Fatores de Tempo
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