Evaluation of Different Types of Stimuli in an Event-Related Potential-Based Brain-Computer Interface Speller under Rapid Serial Visual Presentation.

Rapid serial visual presentation (RSVP) is currently a suitable gaze-independent paradigm for controlling visual brain-computer interfaces (BCIs) based on event-related potentials (ERPs), especially for users with limited eye movement control. However, unlike gaze-dependent paradigms, gaze-independent ones have received less attention concerning the specific choice of visual stimuli that are used. In gaze-dependent BCIs, images of faces-particularly those tinted red-have been shown to be effective stimuli. This study aims to evaluate whether the colour of faces used as visual stimuli influences ERP-BCI performance under RSVP. Fifteen participants tested four conditions that varied only in the visual stimulus used: grey letters (GL), red famous faces with letters (RFF), green famous faces with letters (GFF), and blue famous faces with letters (BFF). The results indicated significant accuracy differences only between the GL and GFF conditions, unlike prior gaze-dependent studies. Additionally, GL achieved higher comfort ratings compared with other face-related conditions. This study highlights that the choice of stimulus type impacts both performance and user comfort, suggesting implications for future ERP-BCI designs for users requiring gaze-independent systems.

Evaluation of Single-Trial Classification to Control a Visual ERP-BCI under a Situation Awareness Scenario.

An event-related potential (ERP)-based brain-computer interface (BCI) can be used to monitor a user's cognitive state during a surveillance task in a situational awareness context. The present study explores the use of an ERP-BCI for detecting new planes in an air traffic controller (ATC). Two experiments were conducted to evaluate the impact of different visual factors on target detection. Experiment 1 validated the type of stimulus used and the effect of not knowing its appearance location in an ERP-BCI scenario. Experiment 2 evaluated the effect of the size of the target stimulus appearance area and the stimulus salience in an ATC scenario. The main results demonstrate that the size of the plane appearance area had a negative impact on the detection performance and on the amplitude of the P300 component. Future studies should address this issue to improve the performance of an ATC in stimulus detection using an ERP-BCI.

Comparison of Two Paradigms Based on Stimulation with Images in a Spelling Brain-Computer Interface.

A P300-based speller can be used to control a home automation system via brain activity. Evaluation of the visual stimuli used in a P300-based speller is a common topic in the field of brain-computer interfaces (BCIs). The aim of the present work is to compare, using the usability approach, two types of stimuli that have provided high performance in previous studies. Twelve participants controlled a BCI under two conditions, which varied in terms of the type of stimulus employed: a red famous face surrounded by a white rectangle (RFW) and a range of neutral pictures (NPs). The usability approach included variables related to effectiveness (accuracy and information transfer rate), efficiency (stress and fatigue), and satisfaction (pleasantness and System Usability Scale and Affect Grid questionnaires). The results indicated that there were no significant differences in effectiveness, but the system that used NPs was reported as significantly more pleasant. Hence, since satisfaction variables should also be considered in systems that potential users are likely to employ regularly, the use of different NPs may be a more suitable option than the use of a single RFW for the development of a home automation system based on a visual P300-based speller.

Emissions of Carbonaceous Particulate Matter and Ultrafine Particles from Vehicles-A Scientific Review in a Cross-Cutting Context of Air Pollution and Climate Change.

Airborne particulate matter (PM) is a pollutant of concern not only because of its adverse effects on human health but also on visibility and the radiative budget of the atmosphere. PM can be considered as a sum of solid/liquid species covering a wide range of particle sizes with diverse chemical composition. Organic aerosols may be emitted (primary organic aerosols, POA), or formed in the atmosphere following reaction of volatile organic compounds (secondary organic aerosols, SOA), but some of these compounds may partition between the gas and aerosol phases depending upon ambient conditions. This review focuses on carbonaceous PM and gaseous precursors emitted by road traffic, including ultrafine particles (UFP) and polycyclic aromatic hydrocarbons (PAHs) that are clearly linked to the evolution and formation of carbonaceous species. Clearly, the solid fraction of PM has been reduced during the last two decades, with the implementation of after-treatment systems abating approximately 99% of primary solid particle mass concentrations. However, the role of brown carbon and its radiative effect on climate and the generation of ultrafine particles by nucleation of organic vapour during the dilution of the exhaust remain unclear phenomena and will need further investigation. The increasing role of gasoline vehicles on carbonaceous particle emissions and formation is also highlighted, particularly through the chemical and thermodynamic evolution of organic gases and their propensity to produce particles. The remaining carbon-containing particles from brakes, tyres and road wear will still be a problem even in a future of full electrification of the vehicle fleet. Some key conclusions and recommendations are also proposed to support the decision makers in view of the next regulations on vehicle emissions worldwide.

Two cetacean species reveal different long-term trends for toxic trace elements in European Atlantic French waters.

Cetaceans have been naturally exposed to toxic trace elements (TEs) on an evolutionary time scale. Hence, they have developed mechanisms to control and/or mitigate their toxic effects. These long-lived species located at high trophic positions and bioaccumulating toxic elements are assumed to be good biomonitoring organisms. However, anthropogenic emissions have strongly increased environmental levels of toxic TEs in the last decades, questioning the efficiency of the detoxication mechanisms in cetaceans. In this context, temporal trends of mercury (Hg), cadmium (Cd) and lead (Pb) concentrations were studied through the analysis of 264 individuals from two cetacean species the common dolphin (Delphinus delphis) and the harbour porpoise (Phocoena phocoena) and belonging to two different Management Units (MUs) for the latter. These individuals stranded along the French Atlantic coasts from 2000s to 2017. All the trends presented were age- and sex-corrected and stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) were measured as proxies of their feeding ecology. Results showed that Pb concentrations clearly decreased over time in both species and MUs. This decrease agrees with the lead petrol regulation after 2000s, supporting the use of these species as valuable bioindicators of changes for TE levels in the marine environment. A significant long-term increase of total Hg concentrations was only observed in common dolphins. Cadmium concentrations also revealed different trends over the period in both species. The different Hg and Cd trends observed in the two species, probably reflected a contrasted contamination of habitat and prey species than a global increase of the contamination in the environment. These results highlight the necessity and gain of using different species to monitor changes in marine environments, each of them informing on the contamination of its own ecological niche. Lastly, the Se:Hg molar ratios of species suggested a low risk for Hg toxicity over time.

Effect of Distracting Background Speech in an Auditory Brain-Computer Interface.

Studies so far have analyzed the effect of distractor stimuli in different types of brain-computer interface (BCI). However, the effect of a background speech has not been studied using an auditory event-related potential (ERP-BCI), a convenient option when the visual path cannot be adopted by users. Thus, the aim of the present work is to examine the impact of a background speech on selection performance and user workload in auditory BCI systems. Eleven participants tested three conditions: (i) auditory BCI control condition, (ii) auditory BCI with a background speech to ignore (non-attentional condition), and (iii) auditory BCI while the user has to pay attention to the background speech (attentional condition). The results demonstrated that, despite no significant differences in performance, shared attention to auditory BCI and background speech required a higher cognitive workload. In addition, the P300 target stimuli in the non-attentional condition were significantly higher than those in the attentional condition for several channels. The non-attentional condition was the only condition that showed significant differences in the amplitude of the P300 between target and non-target stimuli. The present study indicates that background speech, especially when it is attended to, is an important interference that should be avoided while using an auditory BCI.

Performance Analysis With Different Types of Visual Stimuli in a BCI-Based Speller Under an RSVP Paradigm.

Brain-Computer Interface (BCI) systems enable an alternative communication channel for severely-motor disabled patients to interact with their environment using no muscular movements. In recent years, the importance of research into non-gaze dependent brain-computer interface paradigms has been increasing, in contrast to the most frequently studied BCI-based speller paradigm (i.e., row-column presentation, RCP). Several visual modifications that have already been validated under the RCP paradigm for communication purposes have not been validated under the most extended non-gaze dependent rapid serial visual presentation (RSVP) paradigm. Thus, in the present study, three different sets of stimuli were assessed under RSVP, with the following communication features: white letters (WL), famous faces (FF), neutral pictures (NP). Eleven healthy subjects participated in this experiment, in which the subjects had to go through a calibration phase, an online phase and, finally, a subjective questionnaire completion phase. The results showed that the FF and NP stimuli promoted better performance in the calibration and online phases, being slightly better in the FF paradigm. Regarding the subjective questionnaires, again both FF and NP were preferred by the participants in contrast to the WL stimuli, but this time the NP stimuli scored slightly higher. These findings suggest that the use of FF and NP for RSVP-based spellers could be beneficial to increase information transfer rate in comparison to the most frequently used letter-based stimuli and could represent a promising communication system for individuals with altered ocular-motor function.

Highly sensitive index of cardiac autonomic control based on time-varying respiration derived from ECG.

Frequency-domain indices of heart rate variability (HRV) have been used as markers of sympathovagal balance. However, they have been shown to be degraded by interindividual or task-dependent variability, and especially variations in breathing frequency. The study introduces a method to analyze respiration-(vagally) mediated HRV, to better assess subtle variations in sympathovagal balance using ECG recordings. The method enhances HRV analysis by focusing the quantification of respiratory sinus arrhythmia (RSA) gain on the respiratory frequency. To this end, instantaneous respiratory frequency was obtained with ECG-derived respiration (EDR) and was used for variable frequency complex demodulation (VFCDM) of R-R intervals to extract RSA. The ability to detect cognitive stress in 27 subjects (athletes and nonathletes) was taken as a quality criterion to compare our method to other HRV analyses: Root mean square of successive differences, Fourier transform, wavelet transform, and scaling exponent. Three computer-based tasks from MATB-II were used to induce cognitive stress. Sympathovagal index (HFnu) computed with our method better discriminates cognitive tasks from baseline, as indicated by P values and receiver operating characteristic curves. Here, transient decreases in respiratory frequency have shown to bias classical HRV indices, while only EDR-VFCDM consistently exhibits the expected decrease in the HFnu index with cognitive stress in both groups and all cognitive tasks. We conclude that EDR-VFCDM is robust against atypical respiratory profiles, which seems relevant to assess variations in mental demand. Given the variety of individual respiratory profiles reported especially in highly trained athletes and patients with chronic respiratory conditions, EDR-VFCDM could better perform in a wide range of applications.

Bacterial periphytic communities related to mercury methylation within aquatic plant roots from a temperate freshwater lake (South-Western France).

Macrophyte floating roots are considered as hotspots for methylmercury (MeHg) production in aquatic ecosystems through microbial activity. Nevertheless, very little is known about periphyton bacterial communities and mercury (Hg) methylators in such ecological niches. The ability to methylate inorganic Hg is broadly distributed among prokaryotes; however, sulfate-reducers have been reported to be the most important MeHg producers in macrophyte floating roots. In the present work, the periphyton bacterial communities colonizing Ludwigia sp. floating roots were investigated through molecular methods. Among the 244 clones investigated, anaerobic microorganisms associated with the sulfur biogeochemical cycle were identified. Notably, members of the sulfur-oxidizing prokaryotes and the anoxygenic, purple non-sulfur bacteria (Rhodobacteraceae, Comamonadaceae, Rhodocyclaceae, Hyphomicrobiaceae) and the sulfate reducers (Desulfobacteraceae, Syntrophobacteraceae, and Desulfobulbaceae) were detected. In addition, 15 sulfate-reducing strains related to the Desulfovibrionaceae family were isolated and their Hg-methylation capacity was tested using a biosensor. The overall results confirmed that Hg methylation is a strain-specific process since the four strains identified as new Hg-methylators were closely related to non-methylating isolates. This study highlights the potential involvement of periphytic bacteria in Hg methylation when favorable environmental conditions are present in such ecological micro-niches.

Mercury bioaccumulation along food webs in temperate aquatic ecosystems colonized by aquatic macrophytes in south western France.

Mercury (Hg) is considered as an important pollutant for aquatic systems as its organic form, methylmercury (MeHg), is easily bioaccumulated and bioamplified along food webs. In various ecosystems, aquatic periphyton associated with macrophyte was identified as an important place for Hg storage and methylation by microorganisms. Our study concerns temperate aquatic ecosystems (South Western France) colonized by invasive macrophytes and characterized by high mercury methylation potentials. This work establishes original data concerning Hg bioaccumulation in organisms (plants, crustaceans, molluscs and fish) from five contrasting ecosystems. For low trophic level species, total Hg (THg) concentrations were low (from 27±2ngTHgg(-1)dw in asiatic clam Corbicula fluminea to 418±114ngTHgg(-1)dw in crayfish Procambarus clarkii). THg concentrations in some carnivorous fish (high trophic level) were close to or exceeded the International Marketing Level (IML) with values ranging from 1049±220ngTHgg(-1)dw in pike perch muscle (Sander lucioperca) to 3910±1307ngTHgg(-1)dw in eel muscle (Anguilla Anguilla). Trophic levels for the individuals were also evaluated through stable isotope analysis, and linked to Hg concentrations of organisms. A significant Hg biomagnification (r(2)= 0.9) was observed in the Aureilhan lake, despite the absence of top predator fish. For this site, Ludwigia sp. periphyton, as an entry point of Hg into food webs, is a serious hypothesis which remains to be confirmed. This study provides a first investigation of Hg transfer in the ecosystems of south western France and allows the assessment of the risk associated with the presence of Hg in aquatic food webs.

Insulin effect on lipogenesis and fat distribution in three genotypes of ducks during overfeeding.

In waterfowl, the response to overfeeding differs from one genotype to the other. Pekin ducks generally store lipids in the peripheral tissues while Muscovy and mule ducks promote hepatic lipid storage. A possible reason for these various susceptibilities to hepatic steatosis could be a difference in insulin sensitivity. We suggest a resistance to insulin in Pekin ducks. In the present work we investigate the action of insulin on glucose and lipid metabolisms for the three overfed genotypes. Regardless of the kind of genotype, all ducks appear to be sensitive to insulin: their glycemia is lower when the animals are treated with insulin. Insulin-treated Muscovy and Pekin ducks present a lower increase in total body weight (-16.5% for Muscovy; -8.3% for Pekin); and a significantly lower liver weight than the controls (-9.6% and -18.3%). The percentage of total lipids in the liver is higher in the controls than in the insulin-treated Pekin and mule ducks (respectively -40.4% and -34.7%), which means a decreased hepatic lipogenesis. Pekin ducks present a higher pectoral muscle weight when the individuals are insulin-treated (+9.7%). Lipoprotein lipase (LPL) activity appears to be significantly higher in insulin-treated Pekin and Muscovy ducks (1.39 and 3.38 times greater than controls). Insulin-treated mule ducks present a decrease of muscle and abdominal lipid storage compared to controls (-11.6% and -13.8%). In this experiment, exogenous insulin has induced an increase of lipid oxidation and has led to a less favorable use and storage of dietary glucose. The hypothesis of insulin-resistance of Pekin ducks is not verified.

Incidence of invasive macrophytes on methylmercury budget in temperate lakes: central role of bacterial periphytic communities.

Several studies demonstrated high mercury (Hg) methylation and demethylation in the periphyton associated with floating roots in tropical ecosystems. The importance of aquatic plants on methylmercury production in three temperate ecosystems from south-western France was evaluated through Hg species concentrations, and Hg methylation/demethylation activities by using stable isotopic tracers ((199)Hg(II), Me(201)Hg). Hg accumulation and high methylation and demethylation yields were detected in plant roots and periphyton, whereas results for sediment and water were low to insignificant. The presence of sulfate reducing prokaryotes was detected in all compartments (T-RFLP based on dsrAB amplified through nested PCR) and their main role in Hg methylation could be demonstrated. In turn, sulfate reduction inhibition did not affect demethylation activities. The estimation of net MeHg budgets in these ecosystems suggested that aquatic rhizosphere is the principal location for methylmercury production and may represent an important source for the contamination of the aquatic food chain.

Spatial and temporal patterns of neutral and adaptive genetic variation in the endangered African wild dog (Lycaon pictus).

Deciphering patterns of genetic variation within a species is essential for understanding population structure, local adaptation and differences in diversity between populations. Whilst neutrally evolving genetic markers can be used to elucidate demographic processes and genetic structure, they are not subject to selection and therefore are not informative about patterns of adaptive variation. As such, assessments of pertinent adaptive loci, such as the immunity genes of the major histocompatibility complex (MHC), are increasingly being incorporated into genetic studies. In this study, we combined neutral (microsatellite, mtDNA) and adaptive (MHC class II DLA-DRB1 locus) markers to elucidate the factors influencing patterns of genetic variation in the African wild dog (Lycaon pictus); an endangered canid that has suffered extensive declines in distribution and abundance. Our genetic analyses found all extant wild dog populations to be relatively small (N(e) < 30). Furthermore, through coalescent modelling, we detected a genetic signature of a recent and substantial demographic decline, which correlates with human expansion, but contrasts with findings in some other African mammals. We found strong structuring of wild dog populations, indicating the negative influence of extensive habitat fragmentation and loss of gene flow between habitat patches. Across populations, we found that the spatial and temporal structure of microsatellite diversity and MHC diversity were correlated and strongly influenced by demographic stability and population size, indicating the effects of genetic drift in these small populations. Despite this correlation, we detected signatures of selection at the MHC, implying that selection has not been completely overwhelmed by genetic drift.

Effects of dietary cadmium contamination on bird Anas platyrhynchos--comparison with species Cairina moschata.

This study aimed to assess the effect of two dietary cadmium (Cd) levels (C1: 1 mgkg(-1); C10: 10 mgkg(-1)) on bird Anas platyrhynchos exposed for 10, 20 and 40 days (5 animals per experimental condition). Ducks were able to accumulate high amounts of Cd, especially in kidneys (after 40 days: C1 8.1 ± 1 mgkg(-1), C10 37.7 ± 4.3 mgkg(-1)). After 40 days, the lowest Cd level triggered oxidative stress and stimulated mitochondrial metabolism. At the same time, highest amounts of Cd (C10 group) only triggered repression of genes encoding for catalase and acetyl-CoA carboxylase, with repression factors of 1/50 and 1/5, respectively. High dose exposures were then associated with the repression of genes encoding for antioxidant, whereas low dose exposure triggered their induction. In contrast, the onset of MT gene expression appeared quickly for the C10 group even if a time delay was observed between gene expression and protein accumulation. Through the comparison of A. platyrhynchos and Cairina moschata, the response to Cd toxicity appeared species-dependent. Discrepancies between species could be explained by differential utilization of MT. This pathway of detoxification seemed sufficient to counter Cd toxicity.

Trace element concentrations (mercury, cadmium, copper, zinc, lead, aluminium, nickel, arsenic, and selenium) in some aquatic birds of the southwest Atlantic coast of France.

Trace elements (mercury [Hg], cadmium [Cd], copper [Cu], zinc [Zn], lead [Pb], aluminium [Al], nickel [Ni], arsenic [As], and selenium [Se]) were investigated using inductively coupled plasma-mass spectrometry in liver, kidney, muscle, and feather of aquatic birds wintering or inhabiting the wetlands situated on the Southwest Atlantic coast of France. A majority of greylag geese, red knots, and grey plovers were collected from among hunter-shot animals. The relation between residue concentrations, age (juvenile vs. adult), and sex was investigated. Trace elements were lower than threshold levels of toxicity, except for Pb. Greylag geese sampled could be considered Pb-poisoned. These consequential levels of contamination could be the result of the ingestion of Pb-shot from ammunition used in hunting areas they crossed during migration. Cd accumulation increased with age, whereas Pb levels in feathers were lower in adult birds in connection with moulting. As was influenced by sex. Female birds displayed higher concentrations in liver and feathers than did male birds.

Effect of dietary cadmium on lipid metabolism and storage of aquatic bird Cairina moschata.

In environment, birds often fast in connection with breeding, migration or drastic climatic conditions and need to mobilize lipid reserves during these periods. The impairment of lipid metabolism by cadmium (Cd; 1 mg kg(-1) added in diet) was investigated on palmiped Cairina moschata. Expression levels of genes involved in lipid metabolism, mitochondrial metabolism and detoxification were investigated in liver and muscle of ducks. Lipid content in muscle and liver were analysed and plasma triglycerides were quantified. After 20 days, ducks exposed to Cd displayed a lower body weight and lower lipid content in liver than controls. In muscle, the increase of lipid content was only significant for control ducks but not for exposed ducks. Exposed ducks appeared unable to sufficiently transport and store lipids into peripheral tissues. Cd impairs lipid metabolism by several ways. First, Cd triggered the down-regulation of fatty acids synthesis in liver even if the NADPH production and the mitochondrial metabolism are enhanced, suggesting a stronger energy needs. Secondly, the associated decrease of plasma triglycerides and lipoprotein lipase activity with Cd are consistent with impairment of lipids storage in peripheral tissues.

Accuracy of exhaust emission factor measurements on chassis dynamometer.

To improve the accuracy, reliability, and representativeness of emission factors, 10 European laboratories worked together to study the influence of 20 parameters on the measurement of light-vehicle emission factors on chassis dynamometer of 4 main categories: driving patterns, vehicle-related parameters, vehicle sampling, and laboratory-related parameters. The results are based on (1) literature synthesis, (2) approximately 2700 specific tests with 183 vehicles, and (3) the reprocessing of more than 900 tests. These tests concern the regulated atmospheric pollutants and pre-Euro to Euro 4 vehicles. Of the 20 parameters analyzed, 7 seemed to have no effect, 7 were qualitatively influential, and 6 were highly influential (gearshift strategy, vehicle mileage, ambient temperature, humidity, dilution ratio, and driving cycle). The first four of the six were able to have correction factors developed for them. The results allow for the design of recommendations or guidelines for the emission factor measurement method.

Impact of cadmium on aquatic bird Cairina moschata.

The impact on palmiped Cairina moschata of two levels of dietary cadmium (Cd) contamination (C1: 1 mg kg(-1) and C10: 10 mg kg(-1)) was investigated on liver gene expression by real-time PCR. Genes involved in mitochondrial metabolism, in antioxidant defences, detoxification and in DNA damage repair were studied. Metallothionein (MT) protein levels and Cd bioaccumulation were also investigated in liver, kidneys and muscle. Male ducks were subjected to three periods of exposure: 10, 20 and 40 days. Cd was mainly bioaccumulated in kidneys first and in liver. The concentrations in liver and kidneys appeared to reach a stable level at 20 days of contamination even if the concentrations in muscle still increased. Cd triggered the enhancement of mitochondrial metabolism, the establishment of antioxidant defences (superoxide dismutase Mn and Cu/Zn; catalase) and of DNA repair from 20 days of contamination. Discrepancies were observed in liver between MT protein levels and MT gene up-regulation. MT gene expression appeared to be a late hour biomarker.

Concentrations of metals (zinc, copper, cadmium, and mercury) in three domestic ducks in France: Pekin, Muscovy, and Mule ducks.

The role of different factors such as biological material (tissues, organs) and trophic condition (overfeeding or not) in the metal accumulation was studied in three genotypes of ducks (Pekin, Muscovy, and Mule) under breeding conditions. Results showed that overfeeding decreased the concentration in Cd, Cu, and Zn through the dilution process. In contrast, mercury concentration increased with this method. A relation between lipidic metabolism of genotypes and the distribution of this metal in biological material was found. Domestic ducks were little contaminated, but a low chronic contamination in Cd was observed, probably coming from the food. Due to the low levels of contamination observed in these breeding ducks, they can be considered as a good control for further contamination studies and comparison with accumulation levels recorded in the field. The impact of feeding condition on accumulation showed the importance of taking into account the life cycle of birds before studying their contamination and the impact of pollutants.

Differential channelling of liver lipids in relation to susceptibility to hepatic steatosis in two species of ducks.

In the human, hepatic steatosis can be associated with an imbalance between synthesis, secretion and storage of hepatic lipids, and exhibits a genetic susceptibility. The effect of overfeeding on hepatic lipid channelling was investigated in two genotypes of ducks that differ in their susceptibility to fatty liver, i.e. the common duck, Anas platyrhynchos, and the Muscovy duck, Cairina moschata. Before overfeeeding, the Muscovy duck exhibited a lower subcutaneous adiposity and a higher muscular development, whereas hepatic composition was similar in both genotypes (>5% lipids and triglycerides accounting for 6-10% lipids). In the plasma lipoprotein profile, HDL predominated (5.5-7.8 g/l) over VLDL (0.09-0.25 g/l) and LDL (0.65-1.06 g/l). All lipid and lipoprotein concentrations were lower in the Muscovy duck. In response to overfeeding, the Muscovy duck exhibited a higher degree of hepatic steatosis (62 vs. 50% lipids), and a lower increase in adiposity and in the concentration of plasma triglycerides (6-fold vs. 10-fold) and VLDL (23-fold vs. 34-fold). Thus, certain genotypes may be more responsive to the dietary induction of fatty liver because of a less efficient channelling of hepatic lipids towards secretion into plasma and adipose storage, and the duck may represent a suitable model in which to study the development of hepatic steatosis and its pathogenesis.