Assessing ecological health of mangrove ecosystems along South China Coast by the pressure-state-response (PSR) model.

The pressure-state-response (PSR) model was applied to establish a mangrove ecosystem health evaluation system combined with analytical hierarchy process (AHP) in this paper. The mangrove wetlands are divided into five ecological levels: excellent health, good health, health, sub-health and morbidity, which is based on the comprehensive health index (CHI) value. Twelve representative sites were selected for sampling to assess the ecological health condition of mangroves. As a result, the ecological health level of Gaoqiao mangrove area is excellent health; the ecological health level of Taiping mangrove area is good health; the ecological health level of Huguang and Qi'ao mangrove area is health; the ecological health level of Techeng and He'an mangrove area is sub-health; the ecological health level of Huidong mangrove area is morbidity. These results will give some advises for ecological protection and biological resource sustainable development of mangrove ecosystem in China.

Bacterial community variations in the South China Sea driven by different chemical conditions.

In this study, Illumina MiSeq sequencing of the 16 S rRNA gene was used to describe the bacterial communities in the South China Sea (SCS) during the southwest monsoon period. We targeted different regions in the SCS and showed that bacterial community was driven by the effects of the river, upwelling, and mesoscale eddy through changing the environmental factors (salinity, temperature, and nutrients). Distinct bacterial communities were observed among different chemical conditions, especially between the estuary and the open sea. The abundance of Burkholderiales, Frankiales, Flavobacteriales, and Rhodobacterales dominated the estuary and its adjacent waters. Bacteria in cyclonic eddy were dominated by Methylophilales and Pseudomonadales, whereas Prochlorococcus, SAR11 clade, and Oceanospirillales had relatively high abundance in the anticyclonic eddy. Overall, the abundance of specific phylotypes significantly varied among samples with different chemical conditions. Chemical conditions probably act as a driver that shapes and controls the diversity of bacteria in the SCS. This study suggests that the interaction between microbial and environmental conditions needs to be further considered to fully understand the diversity and function of marine microbes.

Mixture of Pb, Zn and Cu on root permeability and radial oxygen loss in the mangrove Bruguiera gymnorrhiza.

A short term pot trail was employed to evaluate the exposure of mixed heavy metals (Cu, Pb and Zn) on growth, radial oxygen loss (ROL) and root anatomy in Bruguiera gymnorrhiza. The possible function of BgC4H, a cytochrome P450 gene, on root lignification was also discussed. The exposures of mixed Cu, Pb and Zn directly reduce O2 leakage at root surface. The reduced ROL inhibited by heavy metals was mainly ascribed by the changes in root anatomical features, such as decreased root porosity together with increased lignification within the exodermis. BgC4H was found to be up-regulated after 0.5-day metal exposure, and remained higher transcript levels within 3-day metal exposure when compared to control roots. Besides, the inhibited photosynthesis may also result in less oxygen can be transported to the underground roots. In summary, the mangrove B. gymnorrhiza appeared to react to external mixed metal contaminants by developing a lignified and impermeable exodermis, and such a root barrier induced by mixed Cu, Pb and Zn appeared to be an adaptive response to block metal ions enters into the roots.

Dynamics of radial oxygen loss in mangroves subjected to waterlogging.

Tidal flooding can directly result in oxygen (O2) shortage, however the functions of root aeration in flooding tolerance and O2 dynamics within mangroves are still poorly understood. Thus, in this study, the correlations among waterlogging tolerance, root porosity and O2 movement within the plants were investigated using two mangrove species (Aegiceras corniculatum and Bruguiera gymnorrhiza) and a semi-mangrove Heritiera littoralis. Based on the present data, the species A. corniculatum and B. gymnorrhiza, which possessed higher root porosity, exhibited higher waterlogging tolerance, while H. littoralis is intolerant. Increased root porosity, leaf stoma, and total ROL were observed in the roots of A. corniculatum and B. gymnorrhiza growing in stagnant solution when compared to respective aerated controls. As for ROL spatial pattern along roots, external anaerobic condition could promote ROL from apical root regions but reduce ROL from basal roots, leading to a 'tighter barrier'. In summary, the present study indicated that the plants (e.g., A. corniculatum and B. gymnorrhiza) prioritized to ensure O2 diffusion towards root tips under waterlogging by increasing aerenchyma formation and reducing O2 leakage at basal root regions.

Phytoplankton community, structure and succession delineated by partial least square regression in Daya Bay, South China Sea.

Daya Bay is facing the influence of human activities and nature changes, which result in phytoplankton adjusting to the changing environment. The data about environmental changes and phytoplankton were obtained from four seasonal cruises in 2013 in the bay. It is helpful to explore seasonal succession of phytoplankton driven by the determining environmental factors in this bay. Temperature is a significant indicator of season change. The limiting factor of phytoplankton growth totally changed from P (PO4-P) limiting during the southwest monsoon to Si (SiO3-Si) limiting during northeast monsoon. The order of diatoms and dinoflagellates was the dominant phytoplankton groups in Daya Bay. The dominant species included chain-forming diatoms (Skeletonema, Pseudo-nitzschia, Thalassionema, Chaetoceros and Rhizosolenia) were found all the year round and filamentous cyanobacteria (Trichodesmium) in spring and autumn. Partial least square regression (PLS) found that salinity, temperature and nutrients were important driving force for phytoplankton seasonal succession.

Distribution patterns and source identification for heavy metals in Mirs Bay of Hong Kong in China.

Sediment quality caused by heavy metals was investigated in the Mirs Bay and Tolo Harbor, Hong Kong, China. Samples were collected in January and July, 2010. One-way analysis of variance showed that sediment quality variables (Fe, Zn, Mn, Pb, V, Cu, Cr, Ba, Ni and As) were significantly different (p < 0.05) among the sampling areas, whereas the average concentration of V, Eh and Ba exhibited the significant seasonal variations (p < 0.05) between January and July. The spatial pattern of heavy metals (Pb, Zn and Cu) can probably be attributed to anthropogenic and tidal flushing influence in the harbor. Both geo-accumulation index (Igeo) and enrichment factor (EF) were used to identify the metal pollution level and its related source. Pb, Zn, and Cu are considered as "polluted metal" in Tolo Harbor. Cluster analysis (CA) identified three distinct clusters with the Tolo Habor and Shatou Jiao, the inner bay and the south part of the bay. Principal component analysis (PCA) identified the spatial patterns and their affected parameters in the studying area. Results showed metals distribution in Mirs Bay and its adjacent area is principally affected by human activities such as marineculture, dumping, located mostly in Tolo Harbor and Shatou Jiao, where was closely related with anthropogenic influence. While the monitoring stations including MS13-MS16 and MS8 locating in the south part of the studying area might be corresponded to natural influence.

Assessment for water quality by artificial neural network in Daya Bay, South China Sea.

In this study, artificial neural network such as a self-organizing map (SOM) was used to assess for the effects caused by climate change and human activities on the water quality in Daya Bay, South China Sea. SOM has identified the anthropogenic effects and seasonal characters of water quality. SOM grouped the four seasons as four groups (winter, spring, summer and autumn). The Southeast Asian monsoons, northeasterly from October to the next April and southwesterly from May to September have also an important influence on the water quality in Daya Bay. Spatial pattern is mainly related to anthropogenic activities and hydrodynamics conditions. In spatial characteristics, the water quality in Daya Bay was divided into two groups by chemometrics. The monitoring stations (S3, S8, S10 and S11) were in these area (Dapeng Ao, Aotou Harbor) and northeast parts of Daya Bay, which are areas of human activity. The thermal pollution has been observed near water body in Daya Bay Nuclear Power Plant (S5). The rest of the monitoring sites were in the south, central and eastern parts of Daya Bay, which are areas that experience water exchanges from South China Sea. The results of this study may provide information on the spatial and temporal patterns in Daya Bay. Further research will be carry out more research concerning functional changes in the bay ecology with respect to changes in climatic factor, human activities and bay morphology in Daya Bay.

Spatial and vertical distribution of bacterial community in the northern South China Sea.

Microbial communities are highly diverse in coastal oceans and response rapidly with changing environments. Learning about this will help us understand the ecology of microbial populations in marine ecosystems. This study aimed to assess the spatial and vertical distributions of the bacterial community in the northern South China Sea. Multi-dimensional scaling analyses revealed structural differences of the bacterial community among sampling sites and vertical depth. Result also indicated that bacterial community in most sites had higher diversity in 0-75 m depths than those in 100-200 m depths. Bacterial community of samples was positively correlation with salinity and depth, whereas was negatively correlation with temperature. Proteobacteria and Cyanobacteria were the dominant groups, which accounted for the majority of sequences. The α-Proteobacteria was highly diverse, and sequences belonged to Rhodobacterales bacteria were dominant in all characterized sequences. The current data indicate that the Rhodobacterales bacteria, especially Roseobacter clade are the diverse group in the tropical waters.

Pb uptake and tolerance in the two selected mangroves with different root lignification and suberization.

Metal pollution has been widely reported in mangrove wetlands; however, the mechanisms involved in metal detoxification by mangroves are still poorly understood. This study aimed to investigate the possible function of root lignification/suberization on Pb uptake and tolerance in mangroves. Two mangroves, Acanthus ilicifolius and Rhizophora stylosa with different root lignification/suberization were selected as plant materials; the former exhibits a thin exodermis and low lignification/suberization, while the latter possesses a thick exodermis and high lignification/suberization. A pot trial with addition of Pb was conducted to investigate the differences in Pb uptake and tolerance between the two mangroves. The experiment of rhizobox was designed to explore Pb dynamics and availabilities in the rhizosphere soils, besides, the ability of Pb uptake by the excised roots and X-ray analysis for Pb distribution within roots were also detected. The results revealed that R. stylosa exhibited relatively higher Pb tolerance together with less Pb accumulations when compared to A. ilicifolius. For both species, lower proportion of exchangeable and Carbonate Pb and higher higher Fe-Mn oxides Pb were observed in the rhizosphere zone when compared to the respective non-rhizosphere zone. The results from metal uptake by the excised roots and X-ray analysis clearly showed that the thick lignified/suberized exodermis of R. stylosa could more efficiently delay Pb entering into the roots, leading to less Pb accumulation. In summary, the present study proposes a barrier property of the lignified/suberized exodermis in dealing with the stresses of Pb.

Distribution and sources of the polycyclic aromatic hydrocarbons in the sediments of the Pearl River estuary, China.

The Pearl River delta, one of the most prosperous economically region in China, has experienced significant contaminant inputs. However, the dynamics of pollutants in the Pearl River estuary and the adjacent coastal areas are still unclear at present. In the paper, distribution and sources of polycyclic aromatic hydrocarbons (PAHs) were investigated in the surface sediments of the Pearl River estuary. The total PAHs concentrations ranged from 126.08 to 3828.58 ng/g with a mean value of 563.52 ng/g, whereas the highest PAHs were observed in Guangzhou channel. Among the U.S. Environmental Protection Agency's 16 priority PAHs, PAHs with 3-4 rings exhibited relative higher levels. A positive relationship was found between PAHs and total organic carbon. The source analysis further showed that the major sources of PAHs in the Pearl River estuary were originated from the pyrolytic inputs, reflecting a mixed energy structure such as wood, coal and petroleum combustion. In summary, although PAHs in Lingding Bay and the adjacent coastal areas of the Pearl River estuary exhibited a relatively low pollution level, the relatively high pollution level of PAHs in Guangzhou channel will be attended.

Variation of phytoplankton community structure from the Pearl River estuary to South China Sea.

The Pearl River is located in the northern part of South China Sea. The environment of the Pearl River estuary (PRE) is significantly impacted by nutrients from anthropogenic activities. Along the anthropogenic pollution gradient from the PRE to South China Sea, the phylogenetic diversity and biomass of phytoplankton was examined in relation to physic-chemical variables. The richness of rbcL gene was higher in the open sea than the estuary, while the concentration of chlorophyll a (Chl a) was higher in the estuary than in the open sea. The cluster analysis of the sequences data resulted in seven phytoplankton community types and the dominant species of phytoplankton changed from Cryptophytes and Diatoms to Prymnesiophytes and Diatoms along the gradient. The community structure of phytoplankton was shaped by nutrients and salinity. The phytoplankton biomass was significantly positively affected by phosphorus, nitrite and ammonium (P < 0.01) but negatively by salinity (P < 0.05); the phytoplankton diversity was highly positively affected by salinity (P < 0.05) but negatively by silicate and nitrate (P < 0.01; P < 0.05, respectively). Anthropogenic activities played a critical role in the phytoplankton distribution and biomass of the study area. Further research is necessary to reveal the influence mechanism of environmental factors on the phytoplankton.

The diversity of coral associated bacteria and the environmental factors affect their community variation.

Coral associated bacterial community potentially has functions relating to coral health, nutrition and disease. Culture-free, 16S rRNA based techniques were used to compare the bacterial community of coral tissue, mucus and seawater around coral, and to investigate the relationship between the coral-associated bacterial communities and environmental variables. The diversity of coral associated bacterial communities was very high, and their composition different from seawater. Coral tissue and mucus had a coral associated bacterial community with higher abundances of Gammaproteobacteria. However, bacterial community in seawater had a higher abundance of Cyanobacteria. Different populations were also found in mucus and tissue from the same coral fragment, and the abundant bacterial species associated with coral tissue was very different from those found in coral mucus. The microbial diversity and OTUs of coral tissue were much higher than those of coral mucus. Bacterial communities of corals from more human activities site have higher diversity and evenness; and the structure of bacterial communities were significantly different from the corals collected from other sites. The composition of bacterial communities associated with same coral species varied with season's changes, geographic differences, and coastal pollution. Unique bacterial groups found in the coral samples from more human activities location were significant positively correlated to chemical oxygen demand. These coral specific bacteria lead to coral disease or adjust to form new function structure for the adaption of different surrounding needs further research.

Hierarchical structure and crystal orientation in poly(ethylene oxide)/clay nanocomposite films.

Water-cast nanocomposite films formed by poly(ethylene oxide) (PEO) and Laponite clay were found to display three characteristic levels of structure with large-scale orientation. The first level with the length scale of ca. 30-50 nm was the clay lamellar bundles, which tended to stack perpendicularly to the film surface. The second level with the characteristic length of 1.8 nm was associated with the alternating stacking of the silicate layers and the PEO chains sandwiched between them. The preferred orientations of these two levels of structure were independent of clay content, solvent removal rate for the film preparation, and the crystallization temperature of the PEO chains situating outside the clay bundles. The third level of structure was characterized by the preferred orientation of the PEO crystalline stems with respect to the surface of the silicate layers. Perpendicular orientation always dominated in the nanocomposite films prepared by slow solvent removal irrespective of crystallization temperature. In the films prepared by fast solvent removal, however, parallel crystal orientation set in as the clay concentration exceeded ca. 33 wt %. The preferred crystal orientation was ascribed to the confinement effect imposed by the clay bundles to the crystallization of the PEO chains situating in the interbundle region. In the films cast by slow solvent removal, the weaker confinement associated with the larger interbundle distance led to perpendicular crystal orientation. When the interbundle distance was reduced to ca. 30 nm in the films prepared by rapid solvent evaporation, the strong confinement directed the crystals to form parallel orientation.

Bacterial polycyclic aromatic hydrocarbon ring-hydroxylating dioxygenases in the sediments from the Pearl River estuary, China.

Bacterial community compositions were characterized using denaturing gradient gel electrophoresis analysis of bacterial 16S rRNA gene in the sediments of the Pearl River estuary. Sequencing analyses of the excised bands indicated that Gram-negative bacteria, especially Gammaproteobacteria, were dominant in the Pearl River estuary. The diversity of polycyclic aromatic hydrocarbon ring-hydroxylating dioxygenase (PAH-RHD) gene in this estuary was then assessed by clone library analysis. The phylogenetic analyses showed that all PAH-RHD gene sequences of Gram-negative bacteria (PAH-RHD[GN]) were closely related to the nagAc gene described for Ralstonia sp. U2 or nahAc gene for Pseudomonas sp. 9816-4, while the PAH-RHD gene sequences of Gram-positive bacteria (PAH-RHD[GP]) at sampling site A1 showed high sequence similarity to the nidA gene from Mycobacterium species. Meanwhile, molecular diversity of the two functional genes was higher at the upstream of this region, while lower at the downstream. Redundancy analysis indicated that environmental factors, such as NH4--N, ∑PAHs, pH, SiO3--Si, and water depth, affected the distribution of the PAH-RHD[GN] gene in the Pearl River estuary.

Shifts in species dominance related to spatial assemblages and variation in environmental parameters in a tropical mangrove estuary.

A shift in species dominance in the mangrove microalgae community in Cross River Estuary System (CRES), Nigeria, was carried out during the wet season to highlight the microalgae structure, identify potential indicators, and evaluate the water quality variation. Plankton samples were collected at sixteen selected sampling sites along the river. Diatoms showed the highest number of species (240 species), contributing more than 70% of the total microalgae abundance. Cluster analysis of the microalgae community delimits the CRES into three sections; upper (UCRS), middle (MCRS), and lower (LCRS) communities. The spatial shift in the microalgae community showed that the green algae (Eudorina elegans and Hylotheca dissiliens) dominated the UCRS communities, while diatoms dominated the MCRS (Polymyxus coronalis and Actinocyclus normanii) and LCRS (A. normanii and A. octonarius). The principal component analysis showed that the UCRS was influenced by turbidity and nitrate, while the MCRS and LCRS were characterized by phosphate, electrical conductivity, and salinity/total dissolved solids. The similarity percentage (SIMPER) analysis showed that Surirella tenera and Hylotheca dissiliens contributed >3.5% inter site dissimilarity between the UCRS and LCRS microalgae community. The redundancy analysis revealed that some microalgae species such as Odontella mobiliensis, Coscinodiscopsis jonesiana, A. normanii, and A. octonarius responded positively to salinity change and ammonia concentration in the estuary, while S. tenera, P. coronalis, Bacillaria paxilifer, Navicula transitans, Cyclotella meneghiniana, Humidophila contenta, Melosira granulata, Aulacoseira herzogii, Eudorina elegans, H. dissiliens, Mougeotia sp., and Dinobryon sertularia responded positively to turbidity, nitrate, and phosphate concentration. This study illustrates that CRES inhabit rich biodiversity of microalgae which some taxa could be used as indicators of the environmental changes in marine waters.

Polymeric carbohydrates utilization separates microbiomes into niches: insights into the diversity of microbial carbohydrate-active enzymes in the inner shelf of the Pearl River Estuary, China.

Polymeric carbohydrates are abundant and their recycling by microbes is a key process of the ocean carbon cycle. A deeper analysis of carbohydrate-active enzymes (CAZymes) can offer a window into the mechanisms of microbial communities to degrade carbohydrates in the ocean. In this study, metagenomic genes encoding microbial CAZymes and sugar transporter systems were predicted to assess the microbial glycan niches and functional potentials of glycan utilization in the inner shelf of the Pearl River Estuary (PRE). The CAZymes gene compositions were significantly different between in free-living (0.2-3 µm, FL) and particle-associated (>3 µm, PA) bacteria of the water column and between water and surface sediments, reflecting glycan niche separation on size fraction and selective degradation in depth. Proteobacteria and Bacteroidota had the highest abundance and glycan niche width of CAZymes genes, respectively. At the genus level, Alteromonas (Gammaproteobacteria) exhibited the greatest abundance and glycan niche width of CAZymes genes and were marked by a high abundance of periplasmic transporter protein TonB and members of the major facilitator superfamily (MFS). The increasing contribution of genes encoding CAZymes and transporters for Alteromonas in bottom water contrasted to surface water and their metabolism are tightly related with particulate carbohydrates (pectin, alginate, starch, lignin-cellulose, chitin, and peptidoglycan) rather than on the utilization of ambient-water DOC. Candidatus Pelagibacter (Alphaproteobacteria) had a narrow glycan niche and was primarily preferred for nitrogen-containing carbohydrates, while their abundant sugar ABC (ATP binding cassette) transporter supported the scavenging mode for carbohydrate assimilation. Planctomycetota, Verrucomicrobiota, and Bacteroidota had similar potential glycan niches in the consumption of the main component of transparent exopolymer particles (sulfated fucose and rhamnose containing polysaccharide and sulfated-N-glycan), developing considerable niche overlap among these taxa. The most abundant CAZymes and transporter genes as well as the widest glycan niche in the abundant bacterial taxa implied their potential key roles on the organic carbon utilization, and the high degree of glycan niches separation and polysaccharide composition importantly influenced bacterial communities in the coastal waters of PRE. These findings expand the current understanding of the organic carbon biotransformation, underlying the size-fractionated glycan niche separation near the estuarine system.

Spatial variation of bacterial community structure of the Northern South China Sea in relation to water chemistry.

Spatial distribution, diversity and composition of bacterial communities of the northern South China Sea (SCS) surface water and the relationship with the in situ environmental chemistry were investigated. Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) was used to investigate the bacterial community structure. The DGGE gel showed that each sample harbored a distinct bacterial community structure and spatial variations of bacterial community composition among all samples were obviously. A total of 17 intensive bands were excised and the sequence analysis of these DGGE bands revealed that Proteobacteria were the dominant bacterial group of surface water in the north part of SCS. Results of the taxonomic analysis showed that the communities consisted of Proteobacteria (α-subdivision, ß-subdivision, γ-subdivision), Actinobacteria, Cyanobacteria, Bacteroidetes and Firmicutes. Unweighted pair group method with arithmetic averages clustering of the sampling stations indicated that all stations were classified mainly based on geographical proximity. Canonical correspondence analysis (CCA) was employed to further investigate the relationships between DGGE band pattern and the environmental variables and the first two CCA ordination axes suggested that the structure of the bacterial community was significantly correlated with the variables of nitrate (F = 1.24, P < 0.05).

Identification of anthropogenic effects and seasonality on water quality in Daya Bay, South China Sea.

Thirteen water quality parameters from 12 monitoring stations were collected to study the effects caused by climate and anthropogenic activities on water quality in 2003 in Daya Bay, South China Sea. The data matrix has been built according to the results, which were analyzed by fuzzy c-mean cluster (FCM) and cluster analysis (CA). This analysis has identified anthropogenic effects and seasonal characters of water quality. The dry and wet seasonality have been demonstrated with FCM and CA. The precipitation and the Southeast Asian monsoons, northeasterly from October to the next April and southwesterly from May to September have also an important influence on water quality in Daya Bay. In the spatial pattern, two groups have been classified with FCM and CA. Cluster I consisted of the sites S3, S8, S10 and S11 in the west and north coastal parts of Daya Bay. Cluster I is mainly related to anthropogenic activities such as fish-farming. Cluster II consisted of the rest of the stations in the center, east and south parts of Daya Bay. Cluster II is mainly related to seawater input from South China Sea. Thus, fuzzy c-mean cluster and cluster analysis can be an important tool for the successful characterization of regional-scale water quality.

HMG-CoA reductase inhibitors activate the unfolded protein response and induce cytoprotective GRP78 expression.

AIMS: Since apoptosis of macrophages induced by stress to the endoplasmic reticulum (ER) contributes to advanced atherosclerotic lesions, we sought to understand the effects of statins on the unfolded protein response (UPR). METHODS AND RESULTS: We used pharmacological, biochemical, and siRNA (small interfering ribonucleic acid) approaches to determine the signalling cascades of statin-induced 78 kDa glucose-regulated protein (GRP78) gene transcription and its role in cytoprotection. Exposure of RAW264.7 macrophages to statins increased the expression of GRP78, activating transcription factor 6, X box protein-1, and phosphorylated eukaryotic translation initiation factor 2alpha, while it had no effect on CCAAT/enhancer binding protein-homologous protein. GRP78 induction was abolished by co-treatment with mevalonate and 1,2-bis(o-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid, indicating the involvement of both 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase-dependent and -independent mechanisms. Studies on promoter activity measurements indicated that phosphoinositide turnover, cellular homologue of v-src (c-Src), protein kinase C (PKC), extracellular signal-regulated kinase (ERK), and p38 are involved in upregulating GRP78 gene transcription. We also observed that elevation of intracellular Ca(2+) and interruption of small G proteins are two bifurcated but cooperative signalling pathways for c-Src activation, leading to downstream activation of phospholipase C, PKC, ERK, and p38. Functionally we demonstrated that fluvastatin could protect macrophages from hypoxia-induced cell death through GRP78 induction. CONCLUSION: We demonstrate a novel action of statins of inducing a cytoprotective UPR, providing new insights into the clinical potential of statins for ameliorating ER stress-related diseases.

Free fatty acids act as endogenous ionophores, resulting in Na+ and Ca2+ influx and myocyte apoptosis.

AIMS: Disturbances in lipid metabolism have been suggested to play an important role in myocardial damage. Marked accumulation of free fatty acids (FFAs), including arachidonic acid (AA), palmitic acid, oleic acid, and linoleic acid, occurs during post-ischaemia and reperfusion (post-I/R). Possible cellular mechanisms of AA/FFAs-induced myocyte apoptosis were investigated. METHODS AND RESULTS: In neonatal rat ventricular myocytes, AA/FFAs activate a novel non-selective cation conductance (NSCC), resulting in both intracellular Ca(2+) and Na(+) overload. AA caused sustained cytosolic [Na(+)](cyt) and [Ca(2+)](cyt) overload, resulting in mitochondrial [Na(+)](m) and [Ca(2+)](m) overload, which induced caspase-3-mediated apoptosis. Similar apoptotic effects were seen using Na(+) ionophore cocktail/Ca(2+)-free medium, which induced [Na(+)](cyt) and [Na(+)](m), but not [Ca(2+)](cyt) and [Ca(2+)](m) overload. Electron microscopy showed that inhibition of [Na(+)](m) overload prevented disruption of the mitochondrial membrane, showing that [Na(+)](m) overload is an important upstream signal in AA- and FFA-induced myocyte apoptosis. CONCLUSION: AA and FFAs, which accumulate in the myocardium during post-I/R, may therefore act as naturally occurring endogenous ionophores and contribute to the myocyte death seen during post-I/R.