Mitochondrial respiration promotes Cdc37-dependent stability of the Cdk1 homolog Cdc28.

Cdc28, the homolog of mammalian Cdk1, is a conserved key regulatory kinase for all major cell cycle transitions in yeast. We have found that defects in mitochondrial respiration (including deletion of ATP2, an ATP synthase subunit) inhibit growth of cells carrying a degron allele of Cdc28 (cdc28td) or Cdc28 temperature-sensitive mutations (cdc28-1 and cdc28-1N) at semi-permissive temperatures. Loss of cell proliferation in the atp2Δcdc28td double mutant is associated with aggravated cell cycle arrest and mitochondrial dysfunction, including mitochondrial hyperpolarization and fragmentation. Unexpectedly, in mutants defective in mitochondrial respiration, steady-state protein levels of mutant cdc28 are strongly reduced, accounting for the aggravated growth defects. Stability of Cdc28 is promoted by the Hsp90-Cdc37 chaperone complex. Our results show that atp2Δcdc28td double-mutant cells, but not single mutants, are sensitive to chemical inhibition of the Hsp90-Cdc37 complex, and exhibit reduced levels of additional Hsp90-Cdc37 client kinases, suggesting an inhibition of this complex. In agreement, overexpression of CDC37 improved atp2Δcdc28td cell growth and Cdc28 levels. Overall, our study shows that simultaneous disturbance of mitochondrial respiration and Cdc28 activity reduces the capacity of Cdc37 to chaperone client kinases, leading to growth arrest.

Redox Homeostasis Disclosed in the Saltmarsh Plant Halimione portulacoides upon Short Waterborne Exposure to Inorganic Mercury.

The saltmarsh plant Halimione portulacoides was shortly exposed to realistic levels of inorganic mercury (iHg) with the aim of investigating the adaptative processes of the roots and leaves regarding redox homeostasis, physiology, and Hg accumulation. Plants were collected at a contaminated (CONT) and a reference (REF) site to address the interference of contamination backgrounds. The influence of major abiotic variables (i.e., temperature and light) was also examined. Total Hg levels, antioxidant enzymes, lipid peroxidation (LPO), and photosynthetic activity were analyzed after 2 and 4 h of exposure. A poor accumulation of Hg in the roots was noticed, and no translocation to the stems and leaves was found, but plants from the CONT site seemed more prone to iHg uptake (in winter). Despite this, antioxidant modulation in the roots and leaves was found, disclosing, in winter, higher thresholds for the induction of enzymatic antioxidants in CONT leaves compared to REF plants, denoting that the former are better prepared to cope with iHg redox pressure. Consistently, CONT leaves exposed to iHg had remarkably lower LPO levels. Exposure did not impair photosynthetic activity, pinpointing H. portulacoides' ability to cope with iHg toxicity under very-short-term exposure. Biochemical changes were noticed before enhancements in accumulation, reinforcing the relevance of these responses in precociously signaling iHg toxicity.

Spectroscopic Detection of Biosignatures in Natural Ice Samples as a Proxy for Icy Moons.

Some of the icy moons of the solar system with a subsurface ocean, such as Europa and Enceladus, are the targets of future space missions that search for potential extraterrestrial life forms. While the ice shells that envelop these moons have been studied by several spacecrafts, the oceans beneath them remain unreachable. To better constrain the habitability conditions of these moons, we must understand the interactions between their frozen crusts, liquid layers, and silicate mantles. To that end, astrobiologists rely on planetary field analogues, for which the polar regions of Earth have proven to be great candidates. This review shows how spectroscopy is a powerful tool in space missions to detect potential biosignatures, in particular on the aforementioned moons, and how the polar regions of the Earth are being used as planetary field analogues for these extra-terrestrial environments.

Roles of Oxidative Stress and Nrf2 Signaling in Pathogenic and Non-Pathogenic Cells: A Possible General Mechanism of Resistance to Therapy.

The coordinating role of nuclear factor erythroid-2-related factor 2 (Nrf2) in cellular function is undeniable. Evidence indicates that this transcription factor exerts massive regulatory functions in multiple signaling pathways concerning redox homeostasis and xenobiotics, macromolecules, and iron metabolism. Being the master regulator of antioxidant system, Nrf2 controls cellular fate, influencing cell proliferation, differentiation, apoptosis, resistance to therapy, and senescence processes, as well as infection disease success. Because Nrf2 is the key coordinator of cell defence mechanisms, dysregulation of its signaling has been associated with carcinogenic phenomena and infectious and age-related diseases. Deregulation of this cytoprotective system may also interfere with immune response. Oxidative burst, one of the main microbicidal mechanisms, could be impaired during the initial phagocytosis of pathogens, which could lead to the successful establishment of infection and promote susceptibility to infectious diseases. There is still a knowledge gap to fill regarding the molecular mechanisms by which Nrf2 orchestrates such complex networks involving multiple pathways. This review describes the role of Nrf2 in non-pathogenic and pathogenic cells.

Mercury methylation rates in Deception Island (Maritime Antarctica) waters and pyroclastic gravel impacted by volcanic mercury.

Deception Island is an active volcano in the Antarctica being volcanism a source of mercury. To improve the understanding of the Hg cycle in this remote ecosystem, pyroclastic gravel and water samples were collected and total (THg) and monomethylmercury (MMHg) concentrations were measured as well as the potential for Hg methylation. Gravel samples collected close to active fumaroles showed the highest THg levels (72 ng/g) while in water samples the highest concentrations of THg (1.2 ng/L) and MMHg (0.45 ng/L) where found. Methylation activity was barely observable in gravel samples. Biotic methylation rates in water were up to 13 times higher compared to those recorded previously in other polar waters. Abiotic methylation processes also play an important role, with up to 0.54 ± 0.43% of added Hg converted instantaneously to MMHg. These results suggest that Deception Island presents favourable conditions for MMHg explaining the elevated concentrations of both THg and MMHg in this ecosystem.

Evaluation of the potential of the common cockle (Cerastoderma edule L.) for the ecological risk assessment of estuarine sediments: bioaccumulation and biomarkers.

Common cockles (Cerastoderma edule, L. 1758, Bivalvia: Cardiidae) were subjected to a laboratory assay with sediments collected from distinct sites of the Sado Estuary (Portugal). Cockles were obtained from a mariculture site of the Sado Estuary and exposed through 28-day, semi-static, assays to sediments collected from three sites of the estuary. Sediments from these sites revealed different physico-chemical properties and levels of metals and organic contaminants, ranging from unimpacted (the reference site) to moderately impacted, when compared to available sediment quality guidelines. Cockles were surveyed for bioaccumulation of trace elements (Ni, Cu, Zn, As, Cd and Pb) and organic contaminants (PAHs, PCBs and DDTs). Two sets of potential biomarkers were employed to assess toxicity: whole-body metallothionein (MT) induction and digestive gland histopathology. The bioaccumulation factor and the biota-to-soil accumulation factor were estimated as ecological indices of exposure to metals and organic compounds. From the results it is inferred that C. edule responds to sediment-bound contamination and might, therefore, be suitable for biomonitoring. The species was found capable to regulate and eliminate both types of contaminants. Still, the sediment contamination levels do not account for all the variation in bioaccumulation and MT levels, which may result from the moderate metal concentrations found in sediments, the species' intrinsic resistance to pollution and from yet unexplained xenobiotic interaction effects.

Metal and nutrient dynamics in a eutrophic coastal lagoon (Obidos, Portugal): the importance of observations at different time scales.

Water and sediment quality was monitored at four sites of Obidos coastal lagoon (Portugal) in February, May, July and October 2006, covering different hydrological conditions. Concentrations of nutrients and metals increased in autumn/winter, particularly in an inner branch with symptoms of eutrophication that receives a small tributary contaminated by agro-industrial activities. Moreover, concentrations of PO(4)(3-), Si(OH)(4) and Mn (diffusive gradients of thin films (DGT)-measured) varied inversely with salinity. Additionally, that branch was monitored over 26-h in July 2006 to assess variations of water quality parameters, nutrients and metals on short timescale. During the night, O(2) in water reached a minimum of 40% saturation followed by a pronounced increase of DGT-measured metals and nutrients in water column: Fe and Mn (ten times); Cr, Co, PO(4)(3-) and Si(OH)(4) (six times). Enhancements were also registered for metal/Al ratios in suspended particulate matter: Mn, Cr and Cd (four to six times); Fe, Ni and Co (1.5 times). The metal distribution coefficients calculated along the 26-h survey showed a maximum at daylight suggesting a preferential association of metals with suspended particles. Data recorded under different hydrological conditions and over the 26-h survey allowed to address the influence of external and internal sources on water quality. The results of this study highlight the importance of day/night cycles on the availability of nutrients and metals in eutrophic environments.

Mercury mobility and effects in the salt-marsh plant Halimione portulacoides: Uptake, transport, and toxicity and tolerance mechanisms.

The plant Halimione portulacoides, an abundant species widely distributed in temperate salt-marshes, has been previously assessed as bioindicator and biomonitor of mercury contamination in these ecosystems. The present study aims to assess uptake and distribution of total mercury (THg) and methylmercury (MMHg) within H. portulacoides, potential mercury release by volatilization through leaves, and toxicity and tolerance mechanisms by investigating plant photochemical responses. Stem cuttings of H. portulacoides were collected from a salt-marsh within the Tagus estuary natural protected area, and grown under hydroponic conditions. After root development, plants were exposed to 199HgCl2 and CH3201HgCl, and sampled at specific times (0, 1, 2, 4, 24, 72, 120, 168 (7 days) and 432 h (18 days)). After exposure, roots, stems and leaves were analysed for total 199Hg (T199Hg) and MM201Hg content. Photobiology parameters, namely efficiency and photoprotection capacity, were measured in leaves. Both THg and MMHg were incorporated into the plant root system, stems and leaves, with roots showing much higher levels of both isotope enriched spikes than the other plant tissues. Presence of both mercury isotopes in the stems and leaves and high significant correlations found between roots and stems, and stems and leaves, for both THg and MMHg concentrations, indicate Hg translocation between the roots and above-ground organs. Long-term uptake in stems and leaves, leading to higher Hg content, was more influenced by temperature and radiation than short-term uptake. However, the relatively low levels of both THg and MMHg in the aerial parts of the plant, which were influenced by temperature and radiation, support the possibility of mercury release by stems and leaves, probably via stomata aperture, as a way to eliminate toxic mercury. Regarding photochemical responses, few differences between control and exposed plants were observed, indicating high tolerance of this salt marsh plant to THg and MMHg.

Evidence for preferential depths of metal retention in roots of salt marsh plants.

Depth variation (2-cm resolution) of Fe, Mn, Zn, Cr, Ni, Cu, As and Cd concentrations were determined in belowground biomass of Spartina maritima and Sarcocornia fruticosa and in sediments between roots from two marshes in Tagus (Rosário) and Guadiana (Castro Marim) estuaries in Portugal with different anthropogenic pressures. Levels of metals were also determined in aboveground plant parts. Metal concentrations in belowground material were 2-4 orders of magnitude greater than levels in aboveground plant parts providing evidence of weak upward translocation. Although both studied species showed poor extraction of Cr and Ni from sediments, S. fruticosa exhibited a large capability to remove Zn, Cu, As and Cd from contaminated sediments and stabilised them in belowground biomass. Accumulated metals showed a sub-surface concentration maximum or increase to basal roots. To evaluate whether these preferential layers of accumulation resulted from availability in sediments or controlled by plant activity, Enrichment Factors (EF=[Me](root)/[Metal](sediment)) were calculated for each sediment layer. Maximum values in Rosário plant species (Zn=9.3, Cu=18, As=20, Cd=46) exceeded those obtained in Castro Marim (Zn=1.3, Cu=4.3, As=6.1, Cd=18). Moreover, EFs varied with the depth indicating the presence of preferential layers of metal accumulation in roots of both plants, but depth zonation was not the same as in the sediments. These results suggested that levels in belowground biomass either integrated in time changes that occurred in solid sediments and pore water, or metal uptake by roots was not proportional to levels in sediments. The same sequence of metals transferred from sediment to belowground biomass for the two plants was obtained for the two marshes (Cd > As > Cu, Zn), although metals differed from mining ore to industrial/urban sources.

Mercury and methylmercury transport and fate in the water column of Tagus estuary (Portugal).

Six campaigns were performed in North Channel (CNOR), Barcas Channel (BC) and lower zones (EZ) of Tagus estuary to better understand methylmercury (MMHg) and mercury (Hg) transport and fate. Highest concentrations of particulate and dissolved MMHg were observed in CNOR in bottom waters and in the warmest months. The MMHg distribution coefficients between particulate and dissolved fractions were mainly influenced by particulate matter and dissolved organic carbon. The values were slightly higher in summer than in winter and in CNOR. Overall, results established that the tidal effect is a main driver on the transport and fate of Hg and MMHg from CNOR to outer areas, evidenced by the exportation of the Hg species from CNOR to the upstream station in high tide and to the downstream one in low tide. Therefore, CNOR may be considered a source of Hg and MMHg to the outer estuary.

Dissolved gaseous mercury formation and mercury volatilization in intertidal sediments.

Intertidal sediments of Tagus estuary regularly experiences complex redistribution due to tidal forcing, which affects the cycling of mercury (Hg) between sediments and the water column. This study quantifies total mercury (Hg) and methylmercury (MMHg) concentrations and fluxes in a flooded mudflat as well as the effects on water-level fluctuations on the air-surface exchange of mercury. A fast increase in dissolved Hg and MMHg concentrations was observed in overlying water in the first 10min of inundation and corresponded to a decrease in pore waters, suggesting a rapid export of Hg and MMHg from sediments to the water column. Estimations of daily advective transport exceeded the predicted diffusive fluxes by 5 orders of magnitude. A fast increase in dissolved gaseous mercury (DGM) concentration was also observed in the first 20-30min of inundation (maximum of 40pg L-1). Suspended particulate matter (SPM) concentrations were inversely correlated with DGM concentrations. Dissolved Hg variation suggested that biotic DGM production in pore waters is a significant factor in addition to the photochemical reduction of Hg. Mercury volatilization (ranged from 1.1 to 3.3ngm-2h-1; average of 2.1ngm-2h-1) and DGM production exhibited the same pattern with no significant time-lag suggesting a fast release of the produced DGM. These results indicate that Hg sediment/water exchanges in the physical dominated estuaries can be underestimated when the tidal effect is not considered.

Evaluation of mercury methylation and methylmercury demethylation rates in vegetated and non-vegetated saltmarsh sediments from two Portuguese estuaries.

Neurotoxic methylmercury (MMHg) is formed from inorganic divalent mercury (Hg2+). However, it is poorly understood to what extent different mercury (Hg) pools contribute to existent MMHg levels. In this study, ambient concentrations of total Hg (THg) and MMHg as well as rates of methylation and demethylation were measured simultaneously in sediments with and without salt-marsh plant vegetation, which were collected in Guadiana and Tagus estuaries, Portugal. Concurrent processes of Hg methylation and MMHg demethylation were directly monitored and compared by spiking sediments cores with stable isotope tracers of 199Hg2+ and CH3201Hg+ followed by gas chromatographic separation and isotope-specific detection using inductively coupled plasma mass spectrometry. Compared to the Guadiana estuary, where concentrations were comparatively low, THg and MMHg levels varied between vegetated and non-vegetated sediments collected at the Rosário site (ROS) of the Tagus estuary. Methylation (KM) and demethylation rates (KD) were also different between estuaries being dependent on the presence of vegetation. In addition, the type of macrophyte species influenced KM and KD values. In fact, the highest KM value was found in Sarcocornia fruticosa vegetated sediments at the Castro Marim site in Guadiana (CM, 0.160 day-1) and the lowest KM was observed in non-vegetated sediments at the Alcochete site in Tagus (ALC, 0.009 day-1). KD varied by a factor of three among sites with highest rates of demethylation observed in non-vegetated sediments in Guadiana (12 ± 1.3 day-1, corresponding to a half-life of 1.4 ± 0.2 h). This study clearly shows that the presence of vegetation in sediments favors the formation of MMHg. Moreover, this effect might be site specific and further studies are needed to confirm the findings reported here.

Seasonal variation of methylmercury in sediment cores from the Tagus Estuary (Portugal).

Seasonal and spatial variations of dissolved and particulate methylmercury were evaluated for the first time in sediment cores from the Tagus Estuary. Results showed the highest MeHg concentrations in summer months indicating that the "seasonally" methylation process occurs not only at the topmost layers of the sediments but also in the deeper layers of the sediment column. The proportion of MeHg (up to 92%) in some of our pore water samples was higher than values reported in the literature for other estuaries suggesting that the sedimentary environment in the Tagus tends to favour Hg methylation. This work points to the importance of seasonal variation of the MeHg production in sediment cores. In physically dominated estuaries this enhances seasonal MeHg production in deeper sediments that can have serious ecological impacts due to resuspension or advection processes under extreme events by the increase of MeHg transported to the water column.

The sub-cellular fate of mercury in the liver of wild mullets (Liza aurata)--Contribution to the understanding of metal-induced cellular toxicity.

Mercury is a recognized harmful pollutant in aquatic systems but still little is known about its sub-cellular partitioning in wild fish. Mercury concentrations in liver homogenate (whole organ load) and in six sub-cellular compartments were determined in wild Liza aurata from two areas - contaminated (LAR) and reference. Water and sediment contamination was also assessed. Fish from LAR displayed higher total mercury (tHg) organ load as well as in sub-cellular compartments than those from the reference area, reflecting environmental differences. However, spatial differences in percentage of tHg were only observed for mitochondria (Mit) and lysosomes plus microsomes (Lys+Mic). At LAR, Lys+Mic exhibited higher levels of tHg than the other fractions. Interestingly, tHg in Mit, granules (Gran) and heat-denaturable proteins was linearly correlated with the whole organ. Low tHg concentrations in heat stable proteins and Gran suggests that accumulated levels might be below the physiological threshold to activate those detoxification fractions.

Evidence for elevated production of methylmercury in salt marshes.

Depth variations of total mercury (Hg) and methylmercury (MeHg) concentrations were obtained in cores from nonvegetated sediments, sediments colonized by Sarcocornia fruticosa, Halimione portulacoides, and Spartina maritima and below-ground biomass in three Portuguese estuaries. Similar analyses were also performed on the above-ground plant tissues. Concentrations in below-ground biomass exceeded up to 9 (Hg) and 44 (MeHg) times the levels in sediments. Mercury and MeHg in below-ground biomass were up to 400 (Hg) and 4700 (MeHg) times higher than those found in above-ground parts, indicating a weak upward translocation. Methylmercury in colonized sediments reached 18% of the total Hg, which was 70 times above the maximum values found in nonvegetated sediments. Concentrations of MeHg in vegetated sediments were not related to plant type but were linearly proportional to the total mercury levels. The analysis of below-ground biomass at high depth resolution (2 cm) provided evidence that Hg and MeHg were elevated. The higher enrichment factors were found where the shifting of redox conditions suggested high microbial activity. Mercury and MeHg in below-ground tissues were a function of total levels in sediments and again were not plant-specific. These results suggest that the bioremediation of mercury-contaminated sediments is likely to increase the formation of methylmercury.