Avaliação do potencial mutagênico, genotóxico, estrogênico e modulação da expressão gênica pelo nemorosone, isolado da resina de Clusia rosea

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of nemorosone, isolated from the plant Clusia rosea, on the cell cycle and gene expression in MCF-7 BUS breast cancer cell lines

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Assessment of estrogenic, mutagenic and antimutagenic activity of nemorosone

Currently, a wide range of research involving natural products is focused on the discovery of new drugs in many different therapeutic areas. A great number of the synthetic compounds on the market were derived from natural products, especially plants. Nemorosone is the major constituent of the floral resin of Clusia rosea Jacq., Clusiaceae, and in Cuban propolis. In vitro studies have shown cytotoxic activity in this substance against various tumor cell lines, including those resistant to various cytotoxic drugs, whereas it has low cytotoxicity to non-tumoral cells. Therefore, in order to characterize the biological activity of nemorosone, a substance with potential antitumor activity, and in view of preclinical testing of the toxicity of drug candidate compounds, the main aim of this study was to determine the mutagenic and antimutagenic activity of nemorosone by the Ames test, using the strains TA97a, TA98, TA100 and TA102 of Salmonella typhimurium. Secondly, to characterize the estrogenic activity in an experimental recombinant yeast model (Recombinant Yeast Assay) mutagenic activity was observed at in any of the concentrations in any of the test strains. To evaluate the antimutagenic potential, direct and indirect mutagenic agents were used: 4 nitro-o-phenylenediamine (NPD), mitomycin C (MMC) and aflatoxin B1 (AFL). Nemorosone showed moderate antimutagenic activity (inhibition level 31%), in strain TA100 in the presence of AFL, and strong antimutagenic activity in TA102 against MMC (inhibition level 53%). Estrogenic activity was observed, with an EEq of 0.41±0.16 nM at various tested concentrations.

Evaluation of estrogenic, antiestrogenic and genotoxic activity of nemorosone, the major compound found in brown Cuban propolis

Background: Brown propolis is the major type of propolis found in Cuba; its principal component is nemorosone, the major constituent of Clusia rosea floral resins. Nemorosone has received increasing attention due to its strong in vitro anti-cancer action. The citotoxicity of nemorosone in several human cancer cell lines has been reported and correlated to the direct action it has on the estrogen receptor (ER). Breast cancer can be treated with agents that target estrogen-mediated signaling, such as antiestrogens. Phytoestrogen can mimic or modulate the actions of endogenous estrogens and the treatment of breast cancer with phytoestrogens may be a valid strategy, since they have shown anti-cancer activity.Methods: The aim of the present investigation was to assess the capacity of nemorosone to interact with ERs, by Recombinant Yeast Assay (RYA) and E-screen assays, and to determine by comet assay, if the compound causes DNA-damaging in tumoral and non-tumoral breast cells.Results: Nemorosone did not present estrogenic activity, however, it inhibited the 17-β-estradiol (E2) action when either of both methods was used, showing their antiestrogenicity. The DNA damage induced by the benzophenone in cancer and normal breast cells presented negative results.Conclusion: These findings suggest that nemorosone may have therapeutic application in the treatment of breast cancer. © 2013 Camargo et al.; licensee BioMed Central Ltd.

The mutagenic potential of Clusia alata (Clusiaceae) extract based on two short-term in vivo assays

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Bouchardia rosea, a vanishing brachiopod species of the Brazilian platform: taphonomy, historical ecology and conservation paleobiology

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Taphonomy of Bouchardia rosea (Rhynchonelliformea, Brachiopoda) shells from Ubatuba Bay, Brazil: implications for the use of taphonomic signatures in (paleo)environmental analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Germinação de sementes e emergência de plântulas de Tabebuia rosea (Bertoloni) a.p. de Candolle (Bignoniaceae), uma espécie exótica com potencial invasor

Este estudo teve por objetivo determinar os efeitos da temperatura e da luz na germinação de sementes de Tabebuia rosea, bem como avaliar a influência dos ambientes de pleno sol e sob dossel na emergência de suas plântulas. Foram testadas temperaturas constantes de 10 a 45 ºC com intervalos de 5 ºC, sob luz branca e escuro. Verificou-se que a faixa ótima de temperatura para a germinação foi de 20 a 40 ºC na luz e de 20 a 35 ºC no escuro. As sementes apresentaram maior sincronização da germinação a 25 ºC na luz e 30 ºC no escuro. Os resultados indicam que as sementes de Tabebuia rosea podem germinar em ampla faixa de temperatura, tanto em ambientes abertos, com disponibilidade de luz, quanto em locais com ausência de luz, enterradas no solo ou, mesmo, no interior das florestas. Essa germinação, bem como o recrutamento das plântulas, tanto a pleno sol quanto sob a sombra da vegetação, indica o potencial invasor da espécie.

Efeito da radiação UV -B na interação Botrytis cinerea – clonostachys rosea em morangueiro e do ácido 4 - aminobenzóico no controle do patógeno em tabaco

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Androecia in two Clusia species: development, structure and resin secretion

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of UV-B radiation on the antagonistic ability of Clonostachys rosea to Botrytis cinerea on strawberry leaf disc

Human activities are altering the concentrations of ozone in the troposphere and hence in the incidence of ultraviolet-B (UV-B) on Earth’s surface. Although representing only five percent of UV-B radiation striking the Earth's surface, this radiation has the potential to cause effects on biologically active molecules. Sensitivity to UV-B radiation is one of the limitations of biological control of plant pathogens in the field. The objectives of this work were to evaluate the effects of UV-B on several isolates of Clonostachys rosea, and the ability of an isolate of C. rosea, previously selected for its tolerant to UV-B radiation, to control Botrytis cinerea on strawberry leaves in controlled conditions (strawberry leaf discs). The germination of C. rosea conidia was inversely proportional to the irradiance. The most tolerant strain (LQC62) had relative germination of about 60% after irradiation of 4.2kJ/m2, and this strain was selected to be used in the subsequent studies. The data showed that even with exposure to UV-B radiation, C. rosea LQC62 controlled the pathogen. Conidial concentrations of strain LQC62 above 105 conidia/ml showed higher tolerance to UV-B radiation and increased ability to control more than 75% of the B. cinerea even with exposure to radiation. According to our results, in addition to showing less growth under UV-B, conidia of C. rosea had lower antagonistic ability. Further studies are needed to observe the tolerance of B. cinerea conidia to UV-B radiation and thereby prove that an environment with increased UV-B radiation may be favoring the pathogen due to a lower ability of C. rosea to control the pathogen in conditions of increased UV-B.

Avaliação da esporulação de Bionectria ochroleuca em diferentes meios de culturas

The biological control of the plants diseases, caused by fungi, is carried out using others organisms (predator, parasite or pathogen). Among the possible agents of biocontrol, a fungus has been highlighting as promising and it is known as Clonostachys rosea, asexual form of Bionectria ochroleuca. For that, it is necessary the in vitro production of spores of this fungus. In this study were tested several culture media to select those with better conidia production. The study was conducted at Plant Protection Division, in the Plant Production Department, FCA - UNESP, Botucatu campus, São Paulo state, Brazil. We used the isolated CCR64 (Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)-CNPMA). The means of crops were: BDA; Oats-Agar; Mazeina-Agar; Rice-Agar; V8-5%; V8-10%; V8-20%; TJ-5 %; TJ-10, TJ-20%. The sporulation of the fungus in different culture media was estimated at 8 days after of the incubation. The data were analyzed using method of comparing averages, using the Tukey test, at 5% probability, and the data processed, using (X + 1) 0.5 transformation. All culture media tested were able to produce conidia. It was found that the best culture media for production of conidia of Bionectria ochroleuca is the TJ-5%, followed by TJ-20%, with an sporulation average of 3,5 x 106 conidia / ml.

Quantitative comparisons and models of time-averaging in bivalve and brachiopod shell accumulations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Viés Analítico em Estudos Tafonômicos com Macroinvertebrados: Implicações (Paleo)Ambientais e (Paleo)Ecológicas

A review of recent literature shows that most taphonomic studies of Holocene and fossil macrovertebrates are not methodologically standardized. Hence, results from distinct studies are not comparable, even among researches sharing virtually identical goals, targeting the same biological group of similar age and depositional environment. The effects of the shell size in the taphonomic analysis are still poorly understood. In order to study this issue, the taphonomic signatures (articulation, valve type, fragmentation, abrasion, corrosion, edge modification, color alteration, bioerosion and encrustation) of brachiopod shells (Bouchardia rosea (Mawe)), from Ubatuba Bay in the northern coast of São Paulo State, were investigated according to the sieve sizes. In the study area, 14 collecting stations were sampled via Van Veen grab sampler, along a bathymetric gradient, ranging from 0 to 35 m of depth. Bulk samples were sieved through 8 mm, 6 mm, and 2 mm mesh sizes, yielding a total of 5.204 shells. The results indicate that, when taphonomic signatures were independently analyzed per size classes (8 mm, 6 mm, and 2 mm), the taphonomic signatures are recorded in a complex and random way. Additionally, cluster analysis showed that the similarity among the clusters vary according to the considered sieve size. Thus, the sieve size plays an important role in the distribution of taphonomic signatures in shells of distinct sizes. These results suggest that the concentration of the taphonomic analysis on one class (e.g., the largest sieve size, 8 mm) is not always the best method. Rather, the total data (all sieves included) seems more accurate in recording the whole spectrum of taphonomic processes recorded in shells of a given assemblage.

Braquiópodes (Rhynchonelliformea, Bouchardioidea) neógenos da bacia de Pelotas (RS) e seu significado paleoambiental

The occurrence of brachiopods in Cenozoic rocks of the Pelotas Basin is known since 1862. In spite of that, detailed systematic and taphonomic studies are still missing. Investigations made a half century ago, have suggested that these brachiopods could belong to Bouchardia cf. zitteli, a species found in the San Julian Formation, Late Oligocene, Argentina. Our data suggest that those brachiopods may resemble Bouchardia transplatina. In the Uruguayan portion of the Pelotas Basin B. transplatina is known in rocks of the Camacho Formation, Miocene. In addition, small recrystallized shells of brachiopods were also recovered from three Petrobras boreholes (2PJ-1-RS, 2PN-1-RS, and 2GA-1-RS) from the Pelotas Basin. Brachiopods come from the interval of 130 to 150 meters within the Miocene Henryhowella evax Zone. Despite the degree of taphonomic modiication of those brachiopod shells they indubitably belong to Bouchardia sp. This is noteworthy for various reasons: 1- Bouchardia is a brachiopod with warm water afinities. Presently, extant members of this genus are unknown in latitudes up to 34[degree]S, with the main records at 23[degree]S. 2- Although occurring in depths down to 200 meters, the living member (Bouchardia rosea) of this genus is most abundant in shallow platformal, nutrient-rich waters. 3- The occurrence of Bouchardia in the Miocene of the Pelotas Basin indicates that, at least to the interval of Henryhowella evax Zone, warm waters of the Brazilian currents prevail. This interpretation is in strong accordance with other paleoeoceanographic and paleoclimatic data offered by various groups of co-occurring microfossils, such as ostracodes and foraminifers.