Critical roles of reactive oxygen species in mitochondrial permeability transition in mediating evodiamine-induced human melanoma A375-S2 cell apoptosis

Previous studies have shown that evodiamine could trigger apoptosis in human malignant melanoma A375-S2 cells within 24 h. To further investigate the biochemical basis of this activity, the roles of reactive oxygen species (ROS) and mitochondrial permeability transition (MPT) were evaluated. Exposure to evodiamine led to a rapid increase in intracellular ROS followed by an onset of mitochondrial depolarization. ROS scavenger rescued the ΔΨm dissipation and cell death induced by evodiamine, whilst MPT inhibitor blocked the second-time ROS formation as well as cell death. Expressions of key proteins in Fas- and mitochondria-mediated pathways were furthermore examined. Both pathways were activated and regulated by ROS and MPT and were converged to a final common pathway involving the activation of caspase-3. These data suggested that a phenomenon termed ROS-induced ROS release (RIRR) was involved in evodiamine-treated A375-S2 cells and greatly contributed to the apoptotic process through both extrinsic and intrinsic pathways.     

Reactive oxygen species and nitric oxide regulate mitochondria-dependent apoptosis and autophagy in evodiamine-treated human cervix carcinoma HeLa cells

The redox environment of the cell is currently thought to be extremely important to control either apoptosis or autophagy. This study reported that reactive oxygen species (ROS) and nitric oxide (NO) generations were induced by evodiamine time-dependently; while they acted in synergy to trigger mitochondria-dependent apoptosis by induction of mitochondrial membrane permeabilization (MMP) through increasing the Bax/Bcl-2 or Bcl-x_L ratio. Autophagy was also stimulated by evodiamine, as demonstrated by the positive autophagosome-specific dye monodansylcadaverine (MDC) staining as well as the expressions of autophagy-related proteins, Beclin 1 and LC3. Pre-treatment with 3-MA, the specific inhibitor for autophagy, dose-dependently decreased cell viability, indicating a survival function of autophagy. Importantly, autophagy was found to be promoted or inhibited by ROS/NO in response to the severity of oxidative stress. These findings could help shed light on the complex regulation of intracellular redox status on the balance of autophagy and apoptosis in anti-cancer therapies.     

Annual survey of planktonic dinoflagellates and related cysts in the Gulf of Trieste (Northern Adriatic Sea)

Abstract

A survey of planktonic dinoflagellates and related cysts was carried out in the Gulf of Trieste throughout one year from April 1992 to March 1993. 113 taxa were recovered by the analysis of phytoplankton net samples. The most represented genera were Protoperidinium (34 species), Ceratium (24 species), Dinophysis (15 species), Gonyaulax (11 species) and Prorocentrum (8 species). A particular attention was given to potentially toxic species belonging to the genera Dinophysis, Alexandrium and Prorocentrum. The highest number of species (67 species) was recorded in July, and the lowest one (18 species) in February.

33 cyst morphotypes were recorded by the analysis of sediment samples. The most represented genera were Protoperidinium (8 morphotypes), Scrippsiella (3 morphotypes), Gonyaulax (3 morphotypes) and Alexandrium (2 morphotypes); the cysts most frequently found were those of Conyaulax polyedra and Alexandrium pseudogonyaulax.     

Breeding systems in two species of the genus Helleborus (Ranunculaceae)

ABSTRACT

The results of various types of manual and free pollination are reported in Helleborus bocconei and Helleborus foetidus, two sympatric species of the understorey of submediterranean woods that flower in January–March and are pollinated by bumble-bees of the genus Bombus. Both species were found to be partially self-compatible even if at different rates; geitonogamy seems to be more frequent in H. foetidus and spontaneous self-pollination does not usually occur for morphological reasons, and the species are not apomictic. Efficiency in seed production after free pollination was very high indicating that in spite of harsh climatic condition, there were no limitations imposed by pollen or pollinators.     

Assessing the effects of forest management on epiphytic lichens in coppiced forests using different indicators

Abstract

Epiphytic lichens are one of the taxonomic groups most sensitive to forest management. Nevertheless, they have not yet been exhaustively included in the assessment of Sustainable Forest Management. This work aimed at evaluating the effects of forest management on epiphytic lichens in coppiced forests, exploring the spatial patterns of diversity and the composition of communities. Moreover, the goal was to compare the performance of four potential indicators for monitoring the effects of forest management on epiphytic lichens: total lichen diversity, species associated with intensive management, species associated with aged coppiced woodlands and Indicator Species Ratio (ISR). In humid Mediterranean Liguria, 50 sampling units were chosen in Castanea sativa and deciduous Quercus spp. forests subjected to different forest management practices: intensively managed coppice and aged coppice/high forest. The effect of forest management was evident in terms of species composition, since it was possible to find significantly associated species for each of the two management types. At each sampling site, the four indicators were calculated using Indicator Value Analysis and compared through correspondence analysis. The ISR was shown to be a more effective indicator, being independent of floristic composition and the occurrence of rare species.     

Forest vascular plants as indicators of plant species richness: A data analysis of a flora atlas from northwestern Germany

Abstract

Our study had the objective to examine whether the number of forest vascular plants in a forest-poor region may be indicative of total plant species richness and of the number of threatened plant species. We also related forest plant species richness to geological and soil variables. The analysis was based on a regional flora atlas from the Weser-Elbe region in northwestern Germany including incidence data of species in a total of 1109 grid cells (each ca. 2.8 × 2.8 km²). All taxa were classified either as forest or non-forest species. Total species richness in the grid cells ranged from 65 to 597, with a mean value of 308. The number of forest species varied between 20 and 309 (mean 176). Grid cells with or without particular geological units differed in total and forest species richness, with those containing peatland and marshland being particularly species-poor. Indicator value analysis showed that both total and forest species richness in the grid cells were related to soil acidity and nitrogen in a hump-backed manner, with the highest number of species found at moderately low values for nitrogen and at intermediate values of pH. Forest species richness was highly positively correlated with the number of non-forest species and threatened non-forest species. Indicators for high species richness were primarily those species that are confined to closed semi-natural forests with a varied topography and relatively base- and nutrient-rich soils. Grid cells including historically ancient forest exhibited a higher species richness than grid cells lacking ancient forest, indicating the importance of a long habitat continuity for a high phytodiversity. The “habitat coincidence” of high species richness is best explained by similar responses of forest species and species of other habitats to the main environmental gradients. It is suggested that the regional patterns found for the Weser-Elbe region can be transferred also to other forest-poor regions in Central Europe.     

Germination response of four pasture species to temperature,light, and post-harvest period

Abstract

The purpose of this research was to explore the effect of temperature, light, and post-harvest period, and their interactions, on seed germination ecology of four common pasture species in the Mediterranean environment. Mature seeds of Diplotaxis erucoides, Hirschfeldia incana, Hyoseris scabra (Mediterranean distribution) and Sonchus oleraceus (cosmopolitan distribution) were subjected to seven constant temperatures (10–40°C, at intervals of 5°C) under continuous darkness, or a 12 h/12 h light/dark photoperiod at 30, 150 and 270 days after harvest (DAH). Cumulative germination and germination speed were determined. In all the tested species, except S. oleraceus, light significantly enhanced germination. S. oleraceus seeds maintained germination values over 90%, in a wide range of temperatures (10–35°C), in the dark as well as in light. Seeds of H. incana germinated well soon after seed dispersal. In D. erucoides and H. scabra, germination increased with storage period, while in S. oleraceus there was no effect of seed age. In all the species, moreover, no germination was recorded at 40°C. Temperature, light, and post- harvest requirement may be regarded as an adaptation strategy to ensure optimal conditions for seedling development and survival in Mediterranean species, while the species with a cosmopolitan distribution germinates under almost all tested conditions.     

Ecogeographic and genetic evaluation of endemic species in the Maritime Alps: the case of Moehringia lebrunii and M. sedoides (Caryophyllaceae)

Abstract

The Maritime Alps are one of the ‘hot spots’ in the Mediterranean basin. This study investigated two endemic plants, Moehringia lebrunii and Moehringia sedoides (Caryophyllaceae) in order to increase knowledge of the vegetation of this region, and to investigate possible conservation strategies. Ecogeographic surveys and molecular analyses were undertaken. Gene diversity, the Shannon index and G_ST were calculated within and among populations of the two species based on ISSR data. The populations of M. lebrunii had a density ranging between 0.04 and 0.86 individual/m² and a rather low inner genetic variability value. According to IUCN Red List Criteria, the current status of M. lebrunii is Endangered [EN B2ab(ii, iv)]. M. sedoides is an endemic of the SW Alps (not exclusive of the Maritime Alps), and is very abundant within the core of the range. Its range of occurrence is smaller than previously reported; nevertheless, the species is not under threat. This taxon showed a population density ranging between 0.03 and 0.58 individual/m². Genetic variability values revealed a high variation among the species. Only peripheral populations seemed to suffer from their segregated position. Thus, M. sedoides is to be considered Critically Endangered [CR B1ab(i, ii, iii, iv) + 2ab(i, ii, iii, iv)] for Italy according to Regional Guidelines.     

Exercise training provides cardioprotection via a reduction in reactive oxygen species in rats submitted to myocardial infarction induced by isoproterenol

Exercise training has demonstrated cardioprotection effects. However, the exact mechanism behind this effect is not is clear. The present study evaluated the effects of 12 weeks of previous treadmill training on the levels of oxidative damage, antioxidant enzyme activity and injury in the myocardium of rats submitted to infarction induced by isoproterenol (ISO). Isoproterenol treatment (80 mg/kg given over 2 days in two equal doses) caused arrhythmias and 60% mortality within 24 h of the last injection in the control group (C + ISO) group when compared with the saline control group (saline). Creatine Kinase ? MB levels were markedly increased in hearts from ISO-treated animals in the C + ISO group. Twelve weeks of treadmill training reduced superoxide production, lipid peroxidation levels and protein carbonylation in these animals, as well as increasing the activities and expressions of SOD and CAT. Previous training also reduced CK-MB levels and numbers of deaths by 40%, preventing the deleterious effects of ISO. Based on the data obtained in this study, it is suggested that 12-week treadmill training increases antioxidant enzymes, decreases oxidative damage and reduces the degree of infarction induced by ISO in the hearts of male rats.     

Arguments against the significance of the Fenton reaction contributing to signal pathways under in vivo conditions

One of the common explanations for oxidative stress in the physiological milieu is based on the Fenton reaction, i.e. the assumption that radical chain reactions are initiated by metal-catalyzed electron transfer to hydrogen peroxide yielding hydroxyl radicals. On the other hand — especially in the context of so-called “iron switches” — it is postulated that cellular signaling pathways originate from the interaction of reduced iron with hydrogen peroxide.

Using fluorescence detection and EPR for identification of radical intermediates, we determined the rate of iron complexation by physiological buffer together with the reaction rate of concomitant hydroxylations of aromatic compounds under aerobic and anaerobic conditions. With the obtained overall reaction rate of 1,700 M^-1s^-1 for the buffer-dependent reactions and the known rates for Fenton reactions, we derive estimates for the relative reaction probabilities of both processes.

As a consequence we suggest that under in vivo conditions initiation of chain reactions by hydroxyl radicals generated by the Fenton reaction is of minor importance and hence metal-dependent oxidative stress must be rather independent of the so-called “peroxide tone”. Furthermore, it is proposed that — in the low (subtoxic) concentration range — hydroxylated compounds derived from reactions of “non-free” (crypto) OH radicals are better candidates for iron-dependent sensing of redox-states and for explaining the origin of cellular signals than the generation of “free” hydroxyl radicals.