Rosmarinic and chlorogenic acid, isolated from ferns, suppress stem cell damage induced by hydrogen peroxide.

OBJECTIVES: Evaluating the effects of rosmarinic (RA) and cryptochlorogenic (CGA) acids isolated from Blechnum binervatum extract on stem cell viability, toxicity and the protective effect on oxidative cell damage. METHODS: MTT and LDH methods were employed, using stem cells from teeth. RA and CGA were evaluated at 100, 250 and 500 µM. The negative effect of hydrogen peroxide (H2O2) (200-2200 µM) and the capacity of RA and CGA (10-100 µM) as protective agents were also evaluated. DAPI followed by fluorescent microscopy was employed to photograph the treated and untreated cells. KEY FINDINGS: At all tested concentrations, RA and CGA demonstrated the ability to maintain cell viability, and with no cytotoxic effects on the treated stem cells. RA also induced an increase of the cell viability and a reduction in cytotoxicity. H2O2 (1400 µM) induced >50% of cytotoxicity, and both compounds were capable of suppressing H2O2 damage, even at the lowest concentration. At 100 µM, in H2O2 presence, total cell viability was observed through microscope imaging. CONCLUSIONS: These findings contribute to the continued research into natural substances with the potential for protecting cells against oxidative injury, with the consideration that RA and CGA are useful in the regeneration of damaged stem cells.

The Wnt signaling pathway regulates Nalm-16 b-cell precursor acute lymphoblastic leukemic cell line survival and etoposide resistance.

B-cell precursor acute lymphoblastic leukemia (BCP-ALL) is the most common malignancy in children. The Wnt signaling pathway has been found to be extensively involved in cancer onset and progression but its role in BCP-ALL remains controversial. We evaluate the role of the Wnt pathway in maintenance of BCP-ALL cells and resistance to chemotherapy. Gene expression profile revealed that BCP-ALL cells are potentially sensitive to modulation of Wnt pathway. Nalm-16 and Nalm-6 cell lines displayed low levels of canonical activation, as reflected by the virtually complete absence of total beta-catenin in Nalm-6 and the beta-catenin cell membrane distribution in Nalm-16 cell line. Canonical activation with Wnt3a induced nuclear beta-catenin translocation and led to BCP-ALL cell death. Lithium chloride (LiCl) also induced a cytotoxic effect on leukemic cells. In contrast, both Wnt5a and Dkk-1 increased Nalm-16 cell survival. Also, Wnt3a enhanced the in vitro sensitivity of Nalm-16 to etoposide (VP-16) while treatment with canonical antagonists protected leukemic cells from chemotherapy-induced cell death. Overall, our results suggest that canonical activation of the Wnt pathway may exerts a tumor suppressive effect, thus its inhibition may support BCP-ALL cell survival.

Fate of copper in intensive shrimp farms: bioaccumulation and deposition in pond sediments.

We present the distribution of Cu in water, sediments and biomass from intensive shrimp (Litopenaeus vannamei Boone, 1931)) farming in northeastern Brazil. The results show no difference in dissolved Cu concentrations between waters entering and leaving the ponds. However, there was a large export of particulate Cu to adjacent environments, showed by extremely high particulate (112 microg.L-1) Cu concentrations in draining waters. Copper concentrations in the bottom sediments of the pond varied with depth from 10 to 20 microg.g-1, being 5 to 7 times higher than the local background. Pond management procedures result in a peculiar vertical distribution of Cu with peak values occurring at sub-surface depths in pond bottom sediments. Bioavailable Cu reaches about 20% of the total Cu content in sediments. Notwithstanding the relatively high Cu bioavailability, concentrations in shrimp muscle (23.2 to 63.4 microg.g-1 d.w.) were similar to reported values for reared and natural L. vannamei populations, and much lower than acceptable maximum limits for human consumption. Concentrations in the exoskeleton were always higher than in muscle. Increasing Cu mass, and to a lesser extent Cu concentrations in muscle tissues, occurred simultaneously to a decrease in Cu mass and concentrations in the exoskeleton and vice versa, suggesting a dynamical exchange of Cu between the two compartments. Although Cu fate in intensive shrimp culture does not result in increasing Cu exposure to consumers, the activity is an important source of this metal to adjacent ecosystems, in particular in the pristine environments where most aquaculture activities in NE Brazil takes place.

Fate of copper in intensive shrimp farms: bioaccumulation and deposition in pond sediments / Dinâmica do cobre na carcinicultura intensiva: bioacumulação e deposição em sedimentos de tanques de criação

We present the distribution of Cu in water, sediments and biomass from intensive shrimp (Litopenaeus vannamei Boone, 1931)) farming in northeastern Brazil. The results show no difference in dissolved Cu concentrations between waters entering and leaving the ponds. However, there was a large export of particulate Cu to adjacent environments, showed by extremely high particulate (112 µg.L-1) Cu concentrations in draining waters. Copper concentrations in the bottom sediments of the pond varied with depth from 10 to 20 µg.g-1, being 5 to 7 times higher than the local background. Pond management procedures result in a peculiar vertical distribution of Cu with peak values occurring at sub-surface depths in pond bottom sediments. Bioavailable Cu reaches about 20 percent of the total Cu content in sediments. Notwithstanding the relatively high Cu bioavailability, concentrations in shrimp muscle (23.2 to 63.4 µg.g-1 d.w.) were similar to reported values for reared and natural L. vannamei populations, and much lower than acceptable maximum limits for human consumption. Concentrations in the exoskeleton were always higher than in muscle. Increasing Cu mass, and to a lesser extent Cu concentrations in muscle tissues, occurred simultaneously to a decrease in Cu mass and concentrations in the exoskeleton and vice versa, suggesting a dynamical exchange of Cu between the two compartments. Although Cu fate in intensive shrimp culture does not result in increasing Cu exposure to consumers, the activity is an important source of this metal to adjacent ecosystems, in particular in the pristine environments where most aquaculture activities in NE Brazil takes place.

Este trabalho apresenta a distribuição de Cu em água, sedimentos e biomassa em uma fazenda de cultivo intensivo do camarão Litopenaeus vannamei Boone, 1931, no Nordeste do Brasil. Os resultados mostraram não haver diferenças entre as concentrações de Cu nas águas afluentes e efluentes da fazenda. Entretanto, foi verificada uma grande exportação de Cu para áreas adjacentes associada ao material em suspensão. Nas águas efluentes, a concentração de Cu particulado atingiu até 112 µg.L-1. Em sedimentos, as concentrações de Cu variaram com a profundidade da coluna sedimentar de 10 a 20 µg.g-1, sendo estes valores de 5 a 7 vezes mais altos que as concentrações naturais dos solos e sedimentos locais. O manejo dos tanques de criação resulta em perfis de distribuição de Cu em sedimentos com maiores concentrações em subsuperfície. A biodisponibilidade do Cu nestes sedimentos é elevada e atinge cerca de 20 por cento da concentração total. Apesar da biodisponibilidade elevada, as concentrações de Cu em músculo do L. vannamei (23.2 a 63.4 µg.g-1 peso seco) são similares aos valores reportados para populações naturais da espécie e em diversos de cultivo em outros países, e estão bem abaixo dos limites máximos permitidos para consumo humano. As concentrações de Cu no exoesqueleto foram maiores que no músculo. Um aumento da massa de Cu em músculo ocorreu simultaneamente à diminuição do Cu no exoesqueleto e vice-versa sugerindo uma troca dinâmica do Cu entre os dois compartimentos. Embora a presença do Cu no cultivo intensivo do L. vannamei não represente exposição significativa para os consumidores, a atividade é fonte importante de Cu para os sistemas aquáticos adjacentes.

Characterization of two classes of benzodiazepine binding sites in Schistosoma mansoni.

As we have recently shown that GABA should be considered a putative neurotransmitter in Schistosoma mansoni, the present work aimed to search for GABAA receptors in adult worms using [3H]-flunitrazepam to label the allosteric benzodiazepine binding site which is classically present on GABAA receptor complexes. We detected a large population (Bmax=8.25+/-1.1 pmol x mg protein(-1)) of high affinity (Kd=33.6+/-1.5 nM) binding sites for flunitrazepam. These sites harboured a singular pharmacological modulation that does not fit well with a mammalian central benzodiazepine receptor, mainly due to a very high affinity for Ro5-4864 and a very low affinity for clonazepam. We also detected a second population of benzodiazepine binding sites labelled with high affinity (IC50=85 nM) by [3H]-PK11195, a selective ligand of the mammalian peripheral benzodiazepine receptor. In conclusion, this work describes the pharmacological properties of a large population of central-like benzodiazepine receptors supporting their study as putative new targets for the development of anti-parasitic agents. We also describe, for the first time, the presence of peripheral benzodiazepine receptors in this parasite.