Purpose:The aim of the study was to determine the effect of a combination of medium chain triglycerides (caprylic, capric, caproic and lauric acids), linoleic acid (essential fatty acid), vitamins A and E and soy lecithin, through a morphometric study, on the wound healing kinetics of experimental cutaneous ulcers. Methods: A total of 45 male Wistar rats were used, in which a skin flap of total thickness with an area of 4 cm 2 was removed. The animals were divided randomly into 3 groups of 15 rats each, Control, Reference and Test groups, which were treated topically with 0.9% NaCl, a preparation of clostebol combined with neomycin sulfate and the test formulation, respectively. The wound areas were measured by digital planimetry at days zero, 3, 7 and 12 postoperative. Based on the wound area, we determined the degree of tissue repair and mean rate of repair at different time intervals. Results: At day 3, an expansion of the wound area was observed in the Reference group and slight contraction in the Control and Test groups. On the subsequent days, the healing process, according to the degree of repair, proceeded in a linear manner, such that, at day 12, the healed area reached 77.95% of the initial ulcerated region in the Control group, 78.40% in the Reference group and 83.49% in the Test group, showing no significant differences. The overall mean rate of repair was equally similar at 12 days of treatment: 25.79 mm 2 /dia in the Control group, 25.42 mm 2 /dia in the Reference group and 27.38 mm 2 /dia in the Test group. Conclusion: The test preparation, applied topically on the experimentally induced cutaneous ulcers in rats, did not accelerate the process of tissue repair by secondary union. Key words: Triglycerides. Linoleic Acid. Wound Healing. Rats. RESUMO Objetivo:Avaliar o efeito da associação de triglicerídeos de cadeia média (ácidos caprílico, cáprico, capróico e láurico), ácido linoléico (ácido graxo essencial), vitaminas A e E e lecitina de soja, através de estudo morfométrico, na cinética de reparação de úlceras cutâneas experimentais. Métodos: Utilizaram-se 45 ratos, machos, da linhagem Wistar, nos quais foi removido um retalho cutâneo de espessura total com 4 cm 2 de área. Os animais foram divididos aleatoriamente em 3 grupos constituídos de 15 ratos, Controle, Referência e Teste, que foram tratados por via tópica respectivamente, com solução salina 0,9%, composto de clostebol associado a sulfato de neomicina e a formulação em teste. As áreas das feridas foram mensuradas por planimetria digital nos dias zero, 3, 7 e 12 de pós-operatório. A partir da área da ferida, calcularam-se ainda o grau de reparação e a taxa média de reparação em intervalo de tempo. Resultados: No 3o dia observou-se uma expansão da área da ferida no grupo referência e uma leve contração nos grupos controle e teste. Nos dias subseqüentes o processo de reparação, medido pela variável grau de reparação, evoluiu de forma linear, de modo que, no 12 o dia, a área reparada alcançou 77,95% da região ulcerada inicial no grupo...
Essential oil (EO) of Alpinia zerumbet leaves, at non-toxic concentrations (50-300 μg/mL), did not induce genotoxicity in human leukocytes. However, at the highest concentration (500 μg/mL) tested caused a reduction in cell proliferation and viability, and an increase in DNA damage. Moreover, in vivo experiments showed that EO (400 mg/kg) did not exert mutagenicity on peripheral blood cells and bone marrow in mice. In DPPH test, EO showed scavenging effects against DPPH radicals, and other free radicals (determination of intracellular GSH and lipid peroxidation assays). Furthermore, EO was able to reduce the intracellular levels of ROS, and prevented leukocytes DNA against oxidative damage. The ability of EO to reduce H(2)O(2) toxicity was observed only when cells were treated with EO during and after exposure to H(2)O(2). With the co- and post-treatment procedures, EO decreased the frequency of apoptotic and micronucleated leukocytes as well DNA strand breaks. However, a synergistic effect was observed in cultures exposed to 500 μg/mL EO. In conclusion, EO at concentrations up to 300 μg/mL or doses up to 400mg/kg are not mutagenic in leukocytes and in mice, but do have antioxidative and protective effects against the cytotoxicity and clastogenesis induced by H(2)O(2).
The effects of a lectin (AaL) from seeds of Araucaria angustifolia were investigated in the model of rat paw edema. In vivo anti-and pro-inflammatory activities, role of sugar residues, inflammatory mediators and systemic toxicity were assessed. Intravenous injection of AaL (0.1-1 mg/kg) dose-dependently inhibited the dextran-induced increase in edema and vascular permeability, which were prevented by association of the lectin with its binding sugar N-acetyl-glucosamine (Glyc-Nac). AaL also significantly inhibited edema induced by serotonin (18%) and compound 48/80 (33%), but not edema induced by histamine. In contrast, when applied by the s.c. route, AaL evoked a paw edema that peaked 1 h later and was partially prevented by association with Glyc-Nac (59%) or by prior i.v. administration of the lectin itself (38.8%). This AaL edematogenic activity was significantly inhibited by pentoxifylline (44.4%) or dexamethasone (51%) and also by depletion of rat paw mast cells (45.6%), but not by L-N-nitro-arginine methyl ester or indomethacin, excluding involvement of nitric oxide and prostaglandins. Treatment of animals with a single anti-inflammatory dose of AaL (1 mg/kg, i.v.) for 7 days did not affect rat corporal mass, liver, kidney, spleen or stomach wet weight, blood leukocyte count, and urea, creatinine or serum transaminase activity. Systemic toxicity was apparent only at much higher doses (LD50=88.3 mg/kg) than those required for the anti-inflammatory effect. Summarizing, AaL exerts anti-and pro-edematogenic actions via interaction with its specific lectin domain. These actions may share a common pathway involving either activation or inhibition of inflammatory mediators from resident mast cells.
A lectin-like protein from the seeds of Acacia farnesiana was isolated from the albumin fraction, characterized, and sequenced by tandem mass spectrometry. The albumin fraction was extracted with 0.5 M NaCl, and the lectin-like protein of A. farnesiana (AFAL) was purified by ion-exchange chromatography (Mono-Q) followed by chromatofocusing. AFAL agglutinated rabbit erythrocytes and did not agglutinate human ABO erythrocytes either native or treated with proteolytic enzymes. In sodium dodecyl sulfate gel electrophoresis under reducing and nonreducing conditions, AFAL separated into two bands with a subunit molecular mass of 35 and 50 kDa. The homogeneity of purified protein was confirmed by chromatofocusing with a pI = 4.0 +/- 0.5. Molecular exclusion chromatography confirmed time-dependent oligomerization in AFAL, in accordance with mass spectrometry analysis, which confers an alteration in AFAL affinity for chitin. The protein sequence was obtained by a liquid chromatography quadrupole time-of-flight experiment and showed that AFAL has 68% and 63% sequence similarity with lectins of Phaseolus vulgaris and Dolichos biflorus, respectively.
A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL -1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing α-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL -1 . The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale α-amylase over a wide range (2.4 to 7,500 U.mL -1 ) in scientific investigation as well as in teaching laboratory activities.Keywords: alfa-amylase, alternative method, quantification. Alternativa para quantificação de alfa-amilase ResumoModificações foram propostas ao método sensível de difusão em ágar para a macrodeterminação de alfa-amilase. As modificações propostas diminuem os custos, com a utilização de amido como substrato e ágar como meio solidificante. Assim, foi construída uma curva padrão utilizando uma solução de alfa-amilase de Aspergillus oryzae com concentrações variando de 2,4 a 7.500 U.mL -1. Em seguida, as zonas claras de difusão radial foram mensuradas depois de 4 horas de incubação a 20 °C. Foi obtida uma relação linear entre o logaritmo da atividade enzimática e os diâmetros das zonas claras. O método foi validado utilizando-se soluções de alfa-amilase de cevada nas concentrações de 2,4; 60; 300 e 1.500 U.mL -1. O método tornou-se mais simples, rápido, com baixo custo e passível de ser utilizado para macrodeterminação de alfa-amilase em ampla faixa (2,4 a 7.500 U.mL -1 ) na investigação científica e para fins didáticos em aulas práticas.Palavras-chave: alfa-amilase, método alternativo, quantificação.
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