Lipoxin A4 selectively programs the profile of M2 tumor-associated macrophages which favour control of tumor progression.

In tumor microenvironments, the macrophage population is heterogeneous, but some macrophages can acquire tumor-promoting characteristics. These tumor-associated macrophages (TAM) exhibit an M2-like profile, with deficient production of NO and ROS, characteristics of pro-inflammatory M1 cytotoxic macrophages. Lipoxins (LX) and 15-epi-lipoxins are lipid mediators which can induce certain features of M2 macrophages in mononuclear cells, but their effects on TAM remain to be elucidated. This study tested the hypothesis that ATL-1, a synthetic analogue of 15-epi-lipoxin A4 , could modulate TAM activity profile. We show that human macrophages (MΦ) differentiated into TAM-like cells after incubation with conditioned medium from MV3, a human melanoma lineage cell. Contrasting with the effects observed in other M2 subsets and M1 profile macrophages, ATL-1 selectively decreased M2 surface markers in TAM, suggesting unique behavior of this particular M2 subset. Importantly, these results were replicated by the natural lipoxins LXA4 and the aspirin induced 15-epi-LXA4 (ATL). In parallel, ATL-1 stimulated TAM to produce NO by increasing the iNOS/arginase ratio and activated NADPH oxidase, triggering ROS production. These alterations in TAM profile induced by ATL-1 led to loss of the anti-apoptotic effects of TAM on melanoma cells and increased their cytotoxic properties. Finally, ATL-1 was found to inhibit tumor progression in a murine model in vivo, which was accompanied by alterations in TAM profile and diminished angiogenesis. Together, the results show an unexpected effect of lipoxin, which induces in TAM a change from an M2- to an M1-like profile, thereby triggering tumor cell apoptosis and down-modulating the tumor progression.

Potencial fungitóxico de extratos vegetais sobre Curvularia eragrostidis (P. Henn. ) Meyer in vitro / Fungitoxic potential of plant extracts on in vitro Curvularia eragrostidis (P. Henn. ) Meyer

RESUMO: O objetivo deste trabalho foi avaliar a atividade fungitóxica dos extratos vegetais de alho, citronela, gengibre e nim no controle in vitro de Curvularia eragrostidis. Os extratos foram testados nas concentrações de 5, 15, 25, 35, e 45%, através da deposição de disco de colônia fúngica em placas de Petri contendo meio BDA, acrescido dos extratos e da testemunha. As placas de Petri foram incubadas a 25 ± 2ºC. Avaliou-se o diâmetro das colônias, a cada 24 horas, em dois sentidos opostos, utilizando régua milimetrada. Determinou-se a contagem de esporos do fungo em hemacitômetro após a incubação. Para a germinação de esporos adicionou-se ao meio BDA 0,1 mL de suspensão de 1,3 x 105 conídios/mL- 1 do fungo, acrescido de 0,1 mL de solução dos extratos espalhadas sobre o meio BDA. As placas de Petri foram divididas em quatro quadrantes e incubadas no regime de luz "claro contínuo" e "escuro contínuo". A avaliação foi realizada após 48 horas de incubação através da percentagem de germinação dos conídios nos tratamentos e na testemunha. De acordo com os resultados, concentrações de 5% dos extratos de gengibre e de nim foram eficientes na percentagem de inibição do crescimento micelial e esporulação de C. eragrostidis. A utilização de todos os extratos a partir da concentração de 25% apresentaram os maiores efeitos fitotóxicos nas análises in vitro, reduzindo o crescimento micelial, a esporulação, e germinação do fungo.

ABSTRACT: The objective of this study was to evaluate the fungitoxic activity of plant extracts from garlic, citronella, ginger and neem on the in vitro control of Curvularia eragrostidis. We used the following treatments: extracts of garlic, citronella, ginger and neem (5, 15, 25, 35, 45%) through the deposition of a fungal colony disk obtained from Petri dishes containing PDA medium supplemented with treatments. For control, only water was added. The Petri dishes were incubated at 25 ± 2ºC for seven days. The colony diameter was evaluated in two opposite directions every 24 hours using a millimeter ruler. At the end of the incubation period, the number of spores was counted using a hemocytometer. Spore germination was evaluated by adding to the PDA medium 0.1 mL collected from a suspension of 1.3 x 105 conidia/mL-1 plus 0.1 mL of a solution of each extract spread on the PDA medium. The Petri dishes were divided into four and incubated in either continuous light or continuous dark. The evaluation was performed 48 hours after incubation by determining the germination rate of conidia in comparison to the control. Concentration of 5% of ginger and neen extracts were efficient on the percentage of inhibition of mycelial growth and sporulation of C. eragrostidis. All extracts from the concentration of 25% showed the highest phytotoxic effects in in vitro assays, reducing mycelial growth, sporulation and germination of the fungus.

Determination of 2,4-dichlorophenoxyacetic acid and its major transformation product in soil samples by liquid chromatographic analysis.

The 2,4-dichlorophenoxyacetic acid (2,4-D) is one of the most applied herbicides around the world to control broad leave herbs in many crops. In this study, a method was developed for simultaneous extraction and determination of 2,4-D and its major transformation product, i.e., the 2,4-dichlorophenol (2,4-DCP), in soil samples. The herbicide and its degradation product were extracted twice from soil samples, after acidification, by dichloromethane on ultrasound system for 1 h. Both extracts were combined and filtrated in qualitative filter paper and Celite((R)). The total extract was concentrated in rotatory evaporator, dried under N(2) and finally dissolved in 1 ml of methanol. High Performance Liquid Chromatography with UV detection at 230 nm was used for analysis. Recoveries were obtained from soil samples fortified at 0.1, 1.0, 2.0, 3.0 and 4.0 mgkg(-1) levels and the results varied from 85 to 111% (for 2,4-D) and from 95 to 98% (for 2,4-DCP). For both compounds, the limits of quantification were 0.1 mgkg(-1), which were the loss level at which the accuracy and the precision were studied. Nevertheless, the limits of detection, calculated by considering the blank standard deviation and the minimum concentration level, were 0.03 and 0.02 mgkg(-1), for 2,4-D and 2,4-DCP, respectively. This proposed method was applied to soil samples of eucalyptus crops, which was previously treated with the herbicide. Despite that, neither 2,4-D nor its degradation product were detected 30 days after the herbicide application.

Determination of pesticide residues in coconut water by liquid-liquid extraction and gas chromatography with electron-capture plus thermionic specific detection and solid-phase extraction and high-performance liquid chromatography with ultraviolet detection.

Two simple methods were developed to determine 11 pesticides in coconut water, a natural isotonic drink rich in salts, sugars and vitamins consumed by the people and athletes. The first procedure involves solid-phase extraction using Sep-Pak Vac C18 disposable cartridges with methanol for elution. Isocratic analysis was carried out by means of high-performance liquid chromatography with ultraviolet detection at 254 nm to analyse captan, chlorothalonil, carbendazim, lufenuron and diafenthiuron. The other procedure is based on liquid-liquid extraction with hexane-dichloromethane (1:1, v/v), followed by gas chromatographic analysis with effluent splitting to electron-capture detection for determination of endosulfan, captan, tetradifon and trichlorfon and thermionic specific detection for determination of malathion, parathion-methyl and monocrotophos. The methods were validated with fortified samples at different concentration levels (0.01-12.0 mg/kg). Average recoveries ranged from 75 to 104% with relative standard deviations between 1.4 and 11.5%. Each recovery analysis was repeated at least five times. Limits of detection ranged from 0.002 to 2.0 mg/kg. The analytical procedures were applied to 15 samples and no detectable amounts of the pesticides were found in any samples under the conditions described.