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.

Efeito tempo-dose-resposta do montelucaste no processo de reparo alveolar em camundongos 129Sv/Ev / Time-dose-response effect of Montelukast in the process of alveolar repair in 129Sv / Ev mice

Metabólitos do ácido araquidônico são conhecidos por exercerem importante papel nos processos inflamatórios e no metabolismo do tecido ósseo. No entanto, as ações pontuais, especialmente dos leucotrienos derivados da 5-lipoxigenase (5-LO) sobre o processo de reparo ósseo intramembranoso são pouco exploradas. O presente estudo tem como objetivo analisar os efeitos tempo-dose-resposta da droga montelucaste (MTK), potente antagonista dos receptores de cisteinil leucotrienos tipo 1 (CisLT1Rs), no curso do reparo alveolar pós-exodontia em camundongos 129Sv/Ev, bem como nos níveis plasmáticos de marcadores ósseos bioquímicos. Para tanto, foram utilizados 70 camundongos machos jovens divididos em quatro grupos, de acordo com o tratamento: C - Grupo Controle (não tratados); CV - Grupo Controle Veículo, 20 µL de solução fisiológica (SF) 0,9%; MTK2 ­2 mg/kg de MTK e MTK4 ­ 4 mg/kg de MTK. Os animais dos grupos CV, MTK2 e MTK4 foram tratados diariamente por via oral, iniciando 24 horas antes do procedimento cirúrgico, continuando até o final dos períodos experimentais de 7, 14 e 21 dias pós-operatórios. Ao final dos períodos determinados, os animais foram submetidos à eutanásia para coleta de sangue para análise bioquímica dos níveis de cálcio, fosfato, fosfatase ácida resistente ao tartarato (TRAP) total e fosfatase alcalina (FAL), coleta da maxila direita contendo os alvéolos dentários para serem analisados por meio de microtomografia computadorizada (microCT), e análise histopatológica. Os resultados obtidos foram submetidos à testes estatísticos considerando-se nível de confiança de 5%. Observou-se aumento do BV/TV para os animais tratados com MTK em relação aos grupos C e CV, tanto aos 14 dias quanto aos 21 dias, sendo maior no grupo MTK4 aos 14 dias em relação ao grupo MTK2. Do mesmo modo, os animais tratados com MTK em ambas doses apresentaram aumento significativo de Tb.Th em comparação aos grupos C e CV aos 21 dias. Chamou a atenção valores de BV/TV e Tb.Th significativamente reduzidos no grupo CV em comparação ao C, indicando um efeito negativo da manipulação do animal. Na análise histopatológica observou-se reparo ósseo precoce nos animais MTK2 e MTK4 em todos os períodos avaliados, em comparação aos do grupo C, bem como atraso no processo de reparo no grupo CV aos 21 dias. Quanto aos marcadores plasmáticos, observou-se aumento do cálcio no grupo MTK4 em relação ao grupo C aos 7 dias, e aos 21 dias também em relação ao grupo MTK2. Já o fosfato mostrouse significantemente elevado nos períodos de 7 e 21 dias no grupo MTK2 em relação aos demais grupos. FAL e TRAP total não apresentaram níveis plasmáticos significativamente diferentes comparando-se os grupos e períodos. Considerando os resultados obtidos, concluiu-se que o MTK exerceu efeito tempo-dose-dependente, acelerando o processo de reparo ósseo intramembranoso alveolar e interferindo nos níveis plasmáticos de cálcio e fosfato no presente modelo animal(AU)

Arachidonic acid metabolites are known to play an important role in inflammatory processes and in bone metabolism. However, the role of these products on alveolar bone repair post tooth extraction remains to be explored, especially leukotrienes, derived from 5-lipoxygenase (5-LO). The present study aims to analyze the time-doseresponse effects of the drug montelukast (MTK), a potent type 1 leukotriene cystenyl antagonist (CisLT1Rs), in the alveolar repair process after extraction in male 129Sv/Ev mice. For this purpose, 70 young male mice were used, divided into four groups: C - Control Group (no treatment); VC - Vehicle Control Group, treated with 20 µL of 0.9% SF; MTK2 - treated with 2mg / Kg of MTK and MTK4 - treated with 4mg / Kg of MTK. The animals of the CV, MTK2 and MTK4 groups were treated daily orally (V.O.), starting 24 hours before the surgical procedure, continuing until the end of the experimental periods of 7, 14 and 21 days postoperatively. At the end of the experimental periods, the animals were euthanized for blood collection for serum markers as calcium, phosphate, tartrate-resistant acid phosphate (TRAP) and alkaline phosphatasis (FAL), and to removal of the right maxilla containing the dental socket to be analyzed under computed microtomography (microCT) and histopathology. The results obtained were subjected to statistical tests considering a confidence level of 5%. Results revealed an increase in BV/TV for MTK vs. C and CV groups, in both 14 and 21 days time points. Of note, this increase was higher in MTK4 than in the MTK2 at 14 days. Considering Tb.Th, both MTK2 e MTK4 groups presented positive effects in the BV/TV and Tb.Th increase when compared to controls groups (C and CV) at 21 days. A decrease in BV/TV and Tb.Th was observed in CV compared to C, as a negative effect of animal manipulation. As observed in H&E sections, both MTK2 and MTK4 experimental groups presented an early bone repair in comparison with C group from 7 to 21 days. CV group presented a slight delayed bone healing compared to C. Levels of calcium was increased in MTK4 in comparison to C and MTK2 at 7 and 21 days. Phosphate was significantly elevated at 7 and 21 days in MTK2 in comparison to the other groups. Despite of beneficial effects on observed on morphological levels on sites of healing (microCT and HE), no significant changes were found for bone markers of remodeling in blood plasma (FAL and TRAP). Taken together, these results indicate that MTK induced early bone healing post tooth extraction in 129Sv/Ev mice. Thus, the inhibition of CysLT is suggested to exert a positive influence on intramembranous bone repair post tooth extraction(AU)

Inflammation in Dry Eye Syndrome: Identification and Targeting of Oxylipin-Mediated Mechanisms.

Dry eye syndrome (DES) is characterized by decreased tear production and stability, leading to desiccating stress, inflammation and corneal damage. DES treatment may involve targeting the contributing inflammatory pathways mediated by polyunsaturated fatty acids and their derivatives, oxylipins. Here, using an animal model of general anesthesia-induced DES, we addressed these pathways by characterizing inflammatory changes in tear lipidome, in correlation with pathophysiological and biochemical signs of the disease. The decline in tear production was associated with the infiltration of inflammatory cells in the corneal stroma, which manifested one to three days after anesthesia, accompanied by changes in tear antioxidants and cytokines, resulting in persistent damage to the corneal epithelium. The inflammatory response manifested in the tear fluid as a short-term increase in linoleic and alpha-linolenic acid-derived oxylipins, followed by elevation in arachidonic acid and its derivatives, leukotriene B4 (5-lipoxigenase product), 12-hydroxyeicosatetraenoic acid (12-lipoxigeanse product) and prostaglandins, D2, E2 and F2α (cyclooxygenase products) that was observed for up to 7 days. Given these data, DES was treated by a novel ophthalmic formulation containing a dimethyl sulfoxide-based solution of zileuton, an inhibitor of 5-lipoxigenase and arachidonic acid release. The therapy markedly improved the corneal state in DES by attenuating cytokine- and oxylipin-mediated inflammatory responses, without affecting tear production rates. Interestingly, the high efficacy of the proposed therapy resulted from the synergetic action of its components, namely, the general healing activity of dimethyl sulfoxide, suppressing prostaglandins and the more specific effect of zileuton, downregulating leukotriene B4 (inhibition of T-cell recruitment), as well as upregulating docosahexaenoic acid (activation of resolution pathways).

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.

Convergent synthesis of new N -substituted 2-{[5-(1H -indol-3-ylmethyl)-1, 3, 4-oxadiazol-2-yl]sulfanyl}acetamides as suitable therapeutic agents

abstract A series of N-substituted 2-{[5-(1H-indol-3-ylmethyl)-1,3,4-oxadiazol-2-yl]sulfanyl}acetamides (8a-w) was synthesized in three steps. The first step involved the sequential conversion of 2-(1H-indol-3-yl)acetic acid (1) to ester (2) followed by hydrazide (3) formation and finally cyclization in the presence of CS2 and alcoholic KOH yielded 5-(1H-indole-3-yl-methyl)-1,3,4-oxadiazole-2-thiol (4). In the second step, aryl/aralkyl amines (5a-w) were reacted with 2-bromoacetyl bromide (6) in basic medium to yield 2-bromo-N-substituted acetamides (7a-w). In the third step, these electrophiles (7a-w) were reacted with 4 to afford the target compounds (8a-w). Structural elucidation of all the synthesized derivatives was done by 1H-NMR, IR and EI-MS spectral techniques. Moreover, they were screened for antibacterial and hemolytic activity. Enzyme inhibition activity was well supported by molecular docking results, for example, compound 8q exhibited better inhibitory potential against α-glucosidase, while 8g and 8b exhibited comparatively better inhibition against butyrylcholinesterase and lipoxygenase, respectively. Similarly, compounds 8b and 8c showed very good antibacterial activity against Salmonella typhi, which was very close to that of ciprofloxacin, a standard antibiotic used in this study. 8c and 8l also showed very good antibacterial activity against Staphylococcus aureus as well. Almost all compounds showed very slight hemolytic activity, where 8p exhibited the least. Therefore, the molecules synthesized may have utility as suitable therapeutic agents.

resumo Uma série de acetamidas 2-{[5-(1H-indol-3-ilmetil)-1,3,4-oxadiazol-2-il]sulfanila} N-substituídas (8a-w) foi sintetizada em três fases. A primeira etapa envolveu a conversão sequencial de ácido 2-(1H-indol-3-il)acético (1) a éster (2), seguido por hidrazida (3) e, finalmente, a e ciclização na presença de CS2 e KOH alcoólico produziu 5-(1H-indol-3-il- metil)-1,3,4-oxadiazole-2-tiol (4). Na segunda etapa, aminas arílicas/aralquílicas(5a-w) reagiram com brometo de 2-bromoacetila (6​​), em meio básico, para se obter acetamidas 2-bromo-N-substituídas (7a-w). Na terceira etapa, estes eletrófilos (7a- w) reagiram com 4, para se obter os compostos alvo (8a-w). A elucidação estrutural de todos os derivados sintetizados foi realizada por 1H-NMR, IR e técnicas de espectrometria de EI-MS. Além disso, eles foram submetidos a triagem de atividade antibacteriana e hemolítica. Análise da inibição enzimática foi bem apoiada pelos resultados de docking molecular. Por exemplo, o composto 8q exibiu melhor potencial inibitório contra α-glicosidase, e os compostos 8g e 8b exibiram, comparativamente, melhor inibição contra butirilcolinesterase (BChE) elipoxigenase (LOX), respectivamente. Do mesmo modo os compostos 8b e 8c mostraram excelente potencial antibacteriano contra SalmonellaTyphi, semelhante ao do ciprofloxacino, antibiótico padrão usado neste estudo. Os compostos 8c e 8l também mostraram excelente potencial antibacteriano contra Staphylococcus aureus . Quase todos os compostos mostraram pequena atividade hemolítica, sendo que o composto 8p apresentou menor atividade. Assim, as moléculas sintetizadas podem ter a sua utilidade como agentes terapêuticos adequados.

Effects of elevated [CO2 ] on maize defence against mycotoxigenic Fusarium verticillioides.

Maize is by quantity the most important C4 cereal crop; however, future climate changes are expected to increase maize susceptibility to mycotoxigenic fungal pathogens and reduce productivity. While rising atmospheric [CO2 ] is a driving force behind the warmer temperatures and drought, which aggravate fungal disease and mycotoxin accumulation, our understanding of how elevated [CO2 ] will effect maize defences against such pathogens is limited. Here we report that elevated [CO2 ] increases maize susceptibility to Fusarium verticillioides proliferation, while mycotoxin levels are unaltered. Fumonisin production is not proportional to the increase in F. verticillioides biomass, and the amount of fumonisin produced per unit pathogen is reduced at elevated [CO2 ]. Following F. verticillioides stalk inoculation, the accumulation of sugars, free fatty acids, lipoxygenase (LOX) transcripts, phytohormones and downstream phytoalexins is dampened in maize grown at elevated [CO2 ]. The attenuation of maize 13-LOXs and jasmonic acid production correlates with reduced terpenoid phytoalexins and increased susceptibility. Furthermore, the attenuated induction of 9-LOXs, which have been suggested to stimulate mycotoxin biosynthesis, is consistent with reduced fumonisin per unit fungal biomass at elevated [CO2 ]. Our findings suggest that elevated [CO2 ] will compromise maize LOX-dependent signalling, which will influence the interactions between maize and mycotoxigenic fungi.

DDX6 post-transcriptionally down-regulates miR-143/145 expression through host gene NCR143/145 in cancer cells.

In various human malignancies, widespread dysregulation of microRNA (miRNA) expression is reported to occur and affects various cell growth programs. Recent studies suggest that the expression levels of miRNAs that act as tumor suppressors are frequently reduced in cancers because of chromosome deletions, epigenetical changes, aberrant transcription, and disturbances in miRNA processing. MiR-143 and -145 are well-recognized miRNAs that are highly expressed in several tissues, but down-regulated in most types of cancers. However, the mechanism of this down-regulation has not been investigated in detail. Here, we show that DEAD-box RNA helicase 6, DDX6 (p54/RCK), post-transcriptionally down-regulated miR-143/145 expression by prompting the degradation of its host gene product, NCR143/145 RNA. In human gastric cancer cell line MKN45, DDX6 protein was abundantly expressed and accumulated in processing bodies (P-bodies). DDX6 preferentially increased the instability of non-coding RNA, NCR143/145, which encompasses the miR-143/145 cluster, and down-regulated the expression of mature miR-143/145. In human monocytic cell line THP-1, lipopolysaccharide treatment promoted the assembly of P-bodies and down-regulated the expression of NCR143/145 and its miR-143/145 rapidly. In these cells, cycloheximide treatment led to a loss of P-bodies and to an increase in NCR143/145 RNA stability, thus resulting in up-regulation of miR-143/145 expression. These data demonstrate that DDX6 contributed to the control of NCR143/145 RNA stability in P-bodies and post-transcriptionally regulated miR-143/145 expression in cancer cells.

A irradiação gama no tempo de cocção e na absorção de água em grãos de soja com e sem lipoxigenase / The gamma irradiation on the cooking time and absorption of water in soybean grains with and without lipoxygenase

O processamento térmico adequado contribui para aumentar a digestibilidade e eliminar fatores antinutricionais presentes em grãos. Porém, esse processo pode ser prejudicial, causando a perda de nutrientes. A irradiação de grãos pode facilitar sua cocção, reduzindo o tempo de amolecimento. O objetivo do presente trabalho foi avaliar possíveis alterações causadas pela irradiação no tempo de cocção e embebição de grãos de soja convencionais e sem lipoxigenase. As duas cultivares analisadas ('BRS 232' - com lipoxigenase e 'BRS 257' - sem lipoxigenase) diferiram entre si na quantidade de água absorvida, no tempo de embebição e no tempo de cocção dos grãos. A irradiação dos grãos, nas doses de 4kGy e 8kGy, diminuiu a absorção de água e o tempo de cocção.

The appropriate heat processing helps to improve the digestibility and eliminate antinutritional factors present in grain. However, this process can be harmful, causing nutrient losses. The irradiation on grain can facilitate their cooking, and reduces the time for its softening. The objective of this study was to evaluate possible changes caused by irradiation in the time of hydration and cooking in conventional soybean and soybeans without lipoxygenase. The two cultivars ('BRS 232' - with lipoxygenase and 'BRS 257' - without lipoxygenase) differ in the amount of water absorbed, in time of hydration and time of cooking. Irradiation caused lower water absorption, but less time for cooking.