Chemical profile, anti 5-lipoxygenase and cyclooxegenase inhibitory effects of ginger (Zingiber officinale) rhizome, callus and callus treated with elicitors / Perfil químico, anti-5-lipoxigenase e efeitos inibitórios da ciclooxegenase do rizoma de gengibre (Zingiber officinale), do calo e calos tratados com elicitores

The present study investigated the chemical profiles and evaluated the inhibitory effect against 5-Lipoxygenase (5-Lox) activity for extracts of ginger rhizome, callus, and callus treated with the elicitors; yeast extract (100, 300 and 500 mg/L), glycine (100, 200 and 300 mg/L) and salicylic acid (100 and 200 mg/L). Oils and chloroform: methanol (CM) extracts were prepared by maceration in petroleum ether and CM (1:1, v/v), respectively. Chemical profiles were determined by gas chromatography/mass spectrometry (GC/MS) analysis. Oil of the callus recorded higher 5-Lox inhibitory effect (IC50 58.33±4.66 µg/mL) than the oil of rhizome (IC50168.34±15.64 µg/mL) and comparable to that of the positive control; Nordihydroguaiaretic acid (IC50 61.25±1.02 µg/mL). The chemical profile of the callus oil contained large amounts of fatty acids, mainly the unsaturated fatty acid oleic acid (31.11%) and saturated fatty acid palmitic acid (28.56%). Elicitors modified the chemical profile of the callus and ameliorated the anti-5-Lox activity of CM extract of the callus. CM extracts of callus treated with 100 and 300 mg/L yeast extract and 50 mg/L salicylic acid significantly suppressed (P ≤ 0.05) the 5-Lox activity by 33.16%, 25.46% and 16%, respectively as compared to the CM extract of untreated callus. In conclusion, ginger callus could be considered as a valuable dietary supplement in the treatment of various inflammatory disorders.

O presente estudo teve como objetivo investigar os perfis químicos e avaliar o efeito inibitório da atividade da 5-Lipoxigenase (5-Lox) em extratos de rizoma, calo e calo de gengibre tratados com os eliciadores; extrato de levedura (100, 300 e 500 mg / L), glicina (100, 200 e 300 mg / L) e ácido salicílico (100 e 200 mg / L). Extratos de óleos e clorofórmio: metanol (CM) foram preparados por maceração em éter e CM (1: 1, v / v), respectivamente. Os perfis químicos foram determinados por análise de cromatografia gasosa / espectrometria de massa (GC / MS). O óleo do calo registrou maior efeito inibitório de 5-Lox (IC50 58,33 ± 4,66 µg / mL) do que o óleo de rizoma (IC50168,34 ± 15,64 µg / mL) e comparável ao do controle positivo; Ácido nordi-hidroguaiarético (IC50 61,25 ± 1,02 µg / mL). O perfil químico do óleo de calo continha grandes quantidades de ácidos graxos, principalmente o ácido graxo insaturado ácido oleico (31,11%) e ácido graxo saturado palmítico (28,56%). Os elicitores modificaram o perfil químico do calo e melhoraram a atividade anti-5-Lox do extrato de CM do calo. Extratos de CM de calos tratados com 100 e 300 mg / L de extrato de levedura e 50 mg / L de ácido salicílico suprimiram significativamente (P ≤ 0,05) a atividade de 5-Lox em 33,16%, 25,46% e 16%, respectivamente, em comparação com o extrato de CM de calo não tratado. Em conclusão, o calo de gengibre pode ser considerado um suplemento dietético valioso no tratamento de vários distúrbios inflamatórios.

Beneficial effects of methanolic extract of Indigofera linnaei Ali. on the inflammatory and nociceptive responses in rodent models

ABSTRACT Indigofera linnaei Ali. (Tamil Name: Cheppu Nerinjil) belongs to the family Fabaceae, used for the treatment of various ailments in the traditional system of medicine. In the present study, the beneficial effects of methanol extract of whole plant of I. linnaei (MEIL) were evaluated on inflammation and nociception responses in rodent models. In vitro nitric oxide (NO), lipoxygenase (LOX) and cyclooxygense (COX) inhibitory activities were also performed to understand the mode of action. MEIL at the dose of 200 & 400 mg/kg, p.o. significantly inhibited carrageenan induced rat paw volume and reduced the weight of granuloma in cotton pellet granuloma model. The results obtained were comparable with the standard drug aceclofenac. The anti-nociceptive effect of MEIL in mice was evaluated in hot plate and acetic acid induced writhing model. The plant extract significantly reduced the number of writhes and the analgesic effect was higher than that of the standard drug aspirin. However, the extract fails to increase the latency period in hot plate method suggesting that the extract produce nociception by peripheral activity. The extract produced inhibitory effect on NO, LOX and COX in concentration dependent manner. The extract exhibited pronounced and selective COX-2 inhibition. Altogether, these results suggested that the methanol extract of Indigofera linnaei could be considered as a potential anti-inflammatory and analgesic agent.

RESUMO Indigofera linnaei Ali pertence à família Leguminosae e é utilizada para o tratamento de várias doenças na medicina tradicional. No presente estudo, os efeitos benéficos do extrato metanólico da planta inteira de I. linnaei (MEIL) foram avaliados em respostas inflamatórias e nocicepção em modelos de roedores. Testes in vitro de atividade inibitória do óxido nítrico (NO), lipoxigenase (LOX) e ciclooxigenase (COX) também foram realizados para compreender o modo de ação. MEIL nas doses de 200 e 400 mg/kg, p.o. inibiu significativamente o volume da pata de rato induzido por carragenana e reduziu o peso do granuloma no modelo de pélete de algodão. Os resultados obtidos foram comparáveis ao do fármaco padrão, aceclofenaco. O efeito anti-nociceptivo de MEIL foi avaliado em camundongos no modelo de placa quente e de contorção induzida por ácido acético. O extrato da planta reduziu significativamente o número de contorções e o efeito analgésico foi maior do que o do fármaco padrão, ácido acetilsalicílico. Porém, o extrato não conseguiu aumentar o período de latência no método da placa quente, sugerindo que este produz nocicepção por atividade periférica. O extrato produziu efeito inibitório sobre o NO, LOX e COX dependente da concentração. O extrato exibiu inibição acentuada e seletiva da COX-2. No seu conjunto, estes resultados sugerem que o extrato metanólico de Indigofera linnaei poderia ser considerado como agente anti-inflamatório e analgésico potencial.

Panificación con harina de arvejas (Pisum sativum) previamente sometidas a inactivación enzimática / Inactivated pea flour (Pisum sativum) in bread making

La harina de arveja (Pisum sativum) es una fuente proteica de relativo bajo costo y escasamente utilizada en la elaboración de productos de consumo masivo. Su incorporación a la harina de trigo, en la elaboración de productos panificados, ofrece una buena alternativa para complementar un perfil de aminoácidos. En este trabajo se evalúa el efecto de la inactivación enzimática de la arveja sobre las características de los panes de molde formulados con niveles de sustitución de 5 por ciento, 10 por ciento y 15 por ciento. Se efectuaron determinaciones de proteínas y de lisina y por cálculo se obtuvieron los valores de score químico, siendo la lisina el aminoácido limitante. La evaluación sensorial se realizó con un panel de seis evaluadores entrenados y se aplicó el Análisis Descriptivo Cuantitativo. Los puntajes asignados por el panel fueron procesados estadísticamente mediante Análisis de Varianza al nivel de significación p = 0,05. La actividad residual de lipoxigenasa para el tratamiento térmico de un minuto fue de 48.6 por ciento mientras que para un minuto y medio resultó prácticamente nula (2.1 por ciento). En cuanto al efecto del tratamiento térmico de la arveja sobre el volumen específico, se aprecia que el mejor resultado se obtiene cuando el tiempo de tratamiento térmico es de un minuto. A su vez la valoración sensorial del panel asignó los mayores puntajes a los panes con nivel de sustitución de 5 por ciento de harina de arveja, para ambos tiempos de tratamiento térmico (1.0 y 1.5 min). Mayores porcentajes de harina de arveja producen un efecto negativo sobre el volumen y sobre los atributos sensoriales del pan.

Pea flour (Pisum sativum) is a relatively cheap protein source and it is scarcely utilized in making widely consumed products. It provides a good opportunity to improve the amino acidic profile. The purpose of this study was to determine the effect of the enzymatic inactivation of pea on bread characteristics, made with levels of 5, 10 and 15 percent of pea flour. Protein and lysine contents were determined and then chemical score obtained considering lysine as limiting amino acid. Sensory evaluation was carry out by six trained panelists using quantitative descriptive analysis (QDA) and analysis of variance (ANOVA) at p = 0.05. Residual lipoxygenase activity was 48.6 percent when heat treatment was made during 1 minute, and only 2.1 percent when the heat treatment was carry out during 1.5 minutes. Highest specific volumes of bread were obtained with pea flour treated during 1 minute. The sensory evaluation by panel determined that pea flour at a level of 5 percent could be successfully substituted for both heat treatments. But pea flour substitution at levels of 10 and 15 percent had adverse effects on specific volume and sensory characteristics.

ELISA na determinaçäo quantitativa e análise genética nas lipoxigenases L1 e L2 em soja / ELISA in the quantification and genetic analysis of lipoxygenases L1 and L2 from soybeans

A soybean breeding program has been undertaken at the Federal University of Viçosa aiming to reduce lipoxygenase lelvels ib the seeds of commercial varieties. A direct ELISA (Enzyme Linked Immunosorbent Assay) procedure has been developed to quantify lipoxygenases L1 and L2 in soybean seeds. Polyclonal antibodies were reised in rabbits against pure L1 and L2 and purified further by affinity chromatography. The best conditions to perform the analaysis were determined as foloow: coating antibody dilution 1:100, crude soybean protein extract 1:2 x 105 and conjugate 1:100 for both L1 and L2 isozymes. The method proved to be sensible enough to identify homozygous and heterozygous genotypes in the breeding program performing single non destructive analysis of seeds or even cotileldones right after germination. Besides, L1 was quantified in about 120 soybean Brazilian cultivars an L2 in some of them. The results showed a significant variability in the level of lipoxygenase-1 among the cultivars analysed. With respect to L2, it was found that the level of this isozyme was significantly greater in the seeds than observed for L1, in spite of its lower "in vitro" specific activity

Dupla difusäo radial na análise genética das lipoxigenases L1 e L2 em soja / Double gel immunodiffusion in the genetic analysis of lipoxygenases L1 and L2 from soybeans

In the effort to genetically manipulate soybean seeds aiming to improve flavor quality toward human consuption, lipoxygenase null alleles are being transferred to a brazilian commercial variety. Immunochemical approach has been taken to perform analysus of hundred of seeds derived from the breeding program. For that reason lipoxygenases L1 and L2 were puridied to hemogeneity by gel filtration and affinity chromatography. Policlonal antibodies were prepared in rabbits and purified further by affinity chromatogrphy. By using the purified anti-L1 and anti-L2 an Ouchterlony double immunodifusion method has been developed in our lab to detect the presence/absence of the isozymes simultaneously in soybean seeds. The method proved to be highly specific and efficient enought to perform single non destructive seed analysis