Three-Dimensional Image Visualization under Photon-Starved Conditions Using N Observations and Statistical Estimation.

In this paper, we propose a method for the three-dimensional (3D) image visualization of objects under photon-starved conditions using multiple observations and statistical estimation. To visualize 3D objects under these conditions, photon counting integral imaging was used, which can extract photons from 3D objects using the Poisson random process. However, this process may not reconstruct 3D images under severely photon-starved conditions due to a lack of photons. Therefore, to solve this problem, in this paper, we propose N-observation photon-counting integral imaging with statistical estimation. Since photons are extracted randomly using the Poisson distribution, increasing the samples of photons can improve the accuracy of photon extraction. In addition, by using a statistical estimation method, such as maximum likelihood estimation, 3D images can be reconstructed. To prove our proposed method, we implemented the optical experiment and calculated its performance metrics, which included the peak signal-to-noise ratio (PSNR), structural similarity (SSIM), peak-to-correlation energy (PCE), and the peak sidelobe ratio (PSR).

Image Processing Techniques for Improving Quality of 3D Profile in Digital Holographic Microscopy Using Deep Learning Algorithm.

Digital Holographic Microscopy (DHM) is a 3D imaging technology widely applied in biology, microelectronics, and medical research. However, the noise generated during the 3D imaging process can affect the accuracy of medical diagnoses. To solve this problem, we proposed several frequency domain filtering algorithms. However, the filtering algorithms we proposed have a limitation in that they can only be applied when the distance between the direct current (DC) spectrum and sidebands are sufficiently far. To address these limitations, among the proposed filtering algorithms, the HiVA algorithm and deep learning algorithm, which effectively filter by distinguishing between noise and detailed information of the object, are used to enable filtering regardless of the distance between the DC spectrum and sidebands. In this paper, a combination of deep learning technology and traditional image processing methods is proposed, aiming to reduce noise in 3D profile imaging using the Improved Denoising Diffusion Probabilistic Models (IDDPM) algorithm.

Three-Dimensional (3D) Visualization under Extremely Low Light Conditions Using Kalman Filter.

In recent years, research on three-dimensional (3D) reconstruction under low illumination environment has been reported. Photon-counting integral imaging is one of the techniques for visualizing 3D images under low light conditions. However, conventional photon-counting integral imaging has the problem that results are random because Poisson random numbers are temporally and spatially independent. Therefore, in this paper, we apply a technique called Kalman filter to photon-counting integral imaging, which corrects data groups with errors, to improve the visual quality of results. The purpose of this paper is to reduce randomness and improve the accuracy of visualization for results by incorporating the Kalman filter into 3D reconstruction images under extremely low light conditions. Since the proposed method has better structure similarity (SSIM), peak signal-to-noise ratio (PSNR) and cross-correlation values than the conventional method, it can be said that the visualization of low illuminated images can be accurate. In addition, the proposed method is expected to accelerate the development of autonomous driving technology and security camera technology.

3D photon counting integral imaging by using multi-level decomposition.

In this paper, we propose three-dimensional (3D) photon counting integral imaging by using multi-level decomposition such as discrete wavelet transform to improve the visual quality and measurement accuracy under photon-starved conditions. Conventional 3D integral imaging can visualize 3D objects and acquire their depth information. However, the amount of irradiated light on the object causes the degradation of visual quality for 3D images under photon-starved conditions. To visualize 3D objects, photon counting integral imaging has been utilized. It can detect photons from 3D scenes by using a computational photon counting model, which is modelled by the Poisson random process. However, photons occur not only from objects but also in areas where objects do not exist. Moreover, photon fluctuation may occur in the scene through shot noise. Since these noise photons are measurement errors, it may decrease the image quality and accuracy. In contrast, our proposed method uses 2D discrete wavelet transform, which can emphasize the object photons effectively. Finally, our proposed method can enhance the visual quality of 3D images and provide more accurate depth information under photon-starved conditions. To prove the feasibility of our proposed method, we implement the optical experiment and calculate various image quality metrics.

Noise Filtering Method of Digital Holographic Microscopy for Obtaining an Accurate Three-Dimensional Profile of Object Using a Windowed Sideband Array (WiSA).

In the image processing method of digital holographic microscopy (DHM), we can obtain a phase information of an object by windowing a sideband in Fourier domain and taking inverse Fourier transform. In this method, it is necessary to window a wide sideband to obtain detailed information on the object. However, since the information of the DC spectrum is widely distributed over the entire range from the center of Fourier domain, the window sideband includes not only phase information but also DC information. For this reason, research on acquiring only the phase information of an object without noise in digital holography is a challenging issue for many researchers. Therefore, in this paper, we propose the use of a windowed sideband array (WiSA) as an image processing method to obtain an accurate three-dimensional (3D) profile of an object without noise in DHM. The proposed method does not affect the neighbor pixels of the filtered pixel but removes noise while maintaining the detail of the object. Thus, a more accurate 3D profile can be obtained compared with the conventional filter. In this paper, we create an ideal comparison target i.e., microspheres for comparison, and verify the effect of the filter through additional experiments using red blood cells.

Effects of low temperature on longevity and lipid metabolism in the marine rotifer Brachionus koreanus.

Low-temperature exposure prolongs lifespans and changes lipid metabolism but the relationship between longevity and lipids is largely unknown. Here, we examine the relationship between longevity and lipid metabolism at low temperatures (20 °C and 15 °C) compared with a 25 °C control. Life parameters, fatty acid composition, and transcriptome changes were analyzed in the monogonont rotifer Brachionus koreanus. In vivo life-parameter data indicate that lifespan and fecundity exhibit opposite correlations at low temperatures. The amount of total fatty acids decreased significantly at low temperatures but areas stained with Nile red increased at 15 °C compared with the control. From RNA-seq-based transcriptional analysis, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway-enrichment analysis were conducted. GO analysis shows that telomeres were positively regulated at low temperatures. KEGG pathway-enrichment results indicate that gene expression involved in lipid metabolism was activated with increased glycerol and/or choline synthesis at low temperatures. We suggest that reduced reproductive rates are associated with a decrease of lecithin, which is involved in the conversion of glycerol to triacylglycerol in response to low temperatures by lowering the temperature of body fluid.

Exercise-induced AMPK activation is involved in delay of skeletal muscle senescence.

Accumulation of senescent cells leads to aging related phenotypes in various organs. Sarcopenia is a frequently observed aging-related disease, which is associated with the loss of muscle mass and functional disability. Physical activity represents the most critical treatment method for preventing decreased muscle size, mass and strength. However, the underlying mechanism as to how physical activity provides this beneficial effect on muscle function has not yet been fully understood. In particular, one unresolved question about aging is how the boost in catabolism induced by aerobic exercise affects skeletal muscle atrophy and other senescence phenotypes. Here we show that pre-activation of AMPK with the AMPK activator, AICAR can mitigate the diminished cellular viability of skeletal muscle cells induced by doxorubicin, which accelerates senescence through free radical production. Pre-incubation for 3 h with AICAR decreased doxorubicin-induced phosphorylation of AMPK in a differentiated skeletal muscle cell line. Accordingly, cellular viability of skeletal muscle cells was recovered in the cells pre-treated with AICAR then administered doxorubicin as compared to that of doxorubicin-only treatment. In accordance with the results of cellular experiments, we verified that 4 weeks of treadmill exercise decreased the senescence marker, p16 and p21 in 19-month-old mice compared to sedentary mice. In this study, we provide new evidence that prior activation of AMPK can reduce doxorubicin induced cell senescence phenotypes. The evidence in this paper suggest that aerobic exercise-activated catabolism in the skeletal muscle may prevent cellular senescence, partially through the cell cycle regulation.

Tributyltin Affects Retinoid X Receptor-Mediated Lipid Metabolism in the Marine Rotifer Brachionus koreanus.

To examine how tributyltin (TBT), a model obesogen, affects the lipid metabolism in the marine rotifer Brachionus koreanus, we carried out life-cycle studies and determined the in vitro and in silico interactions of retinoid X receptor (RXR) with TBT, the transcriptional levels of RXR and lipid metabolic genes, and the fatty acid content. The lethal concentration 10% (LC10) was determined to be 5.12 µg/L TBT, and negative effects on ecologically relevant end points (e.g., decreased lifespan and fecundity) were detected at 5 µg/L TBT. On the basis of these findings, subsequent experiments were conducted below 1 µg/L TBT, which did not show any negative effects on ecologically relevant end points in B. koreanus. Nile red staining analysis showed that after exposure to 1 µg/L TBT, B. koreanus stored neutral lipids and had significantly increased transcriptional levels of RXR and lipid metabolism-related genes compared to the control. However, the content of total fatty acids did not significantly change at any exposure level. In the single fatty acids profile, a significant increase in saturated fatty acids (SFAs) 14:0 and 20:0 was observed, but the contents of omega-3 and omega-6 fatty acids were significantly decreased. Also, a transactivation assay of TBT with RXR showed that TBT is an agonist of Bk-RXR with a similar fold-induction to the positive control. Taken together, these results demonstrate that TBT-modulated RXR signaling leads to increase in transcriptional levels of lipid metabolism-related genes and the synthesis of SFAs but decreases the content of polyunsaturated fatty acids (PUFAs). Our findings support a wider taxonomic scope of lipid perturbation due to xenobiotic exposure that occurs via NRs in aquatic animals.

Interrelationship of salinity shift with oxidative stress and lipid metabolism in the monogonont rotifer Brachionus koreanus.

Salinity is a critical key abiotic factor affecting biological processes such as lipid metabolism, yet the relationship between salinity and lipid metabolism has not been studied in the rotifer. To understand the effects of salinity on the monogonont rotifer B. koreanus, we examined high saline (25 and 35psu) conditions compared to the control (15psu). In vivo life cycle parameters (e.g. cumulative offspring and life span) were observed in response to 25 and 35psu compared to 15psu. In addition, to investigate whether high salinity induces oxidative stress, the level of reactive oxygen species (ROS) and glutathione S-transferase activity (GST) were measured in a salinity- (15, 25, and 35psu; 24h) and time-dependent manner (3, 6, 12, 24h; 35psu). Furthermore composition of fatty acid (FA) and lipid metabolism-related genes (e.g. elongases and desaturases) were examined in response to different salinity conditions. As a result, retardation in cumulative offspring and significant increase in life span were demonstrated in the 35psu treatment group compared to the control (15psu). Furthermore, ROS level and GST activity have both demonstrated a significant increase (P<0.05) in the 35psu treatment. In general, the quantity of FA and mRNA expression of the lipid metabolism-related genes was significantly decreased (P<0.05) in response to high saline condition with exceptions for both GST-S4 and S5 demonstrated a significant increase in their mRNA expression. This study demonstrates that high salinity induces oxidative stress, leading to a negative impact on lipid metabolism in the monogonont rotifer, B. koreanus.

Microplastic Size-Dependent Toxicity, Oxidative Stress Induction, and p-JNK and p-p38 Activation in the Monogonont Rotifer (Brachionus koreanus).

In this study, we evaluated accumulation and adverse effects of ingestion of microplastics in the monogonont rotifer (Brachionus koreanus). The dependence of microplastic toxicity on particle size was investigated by measuring several in vivo end points and studying the ingestion and egestion using 0.05-, 0.5-, and 6-µm nonfunctionalized polystyrene microbeads. To identify the defense mechanisms activated in response to microplastic exposure, the activities of several antioxidant-related enzymes and the phosphorylation status of mitogen-activated protein kinases (MAPKs) were determined. Exposure to polystyrene microbeads of all sizes led to significant size-dependent effects, including reduced growth rate, reduced fecundity, decreased lifespan and longer reproduction time. Rotifers exposed to 6-µm fluorescently labeled microbeads exhibited almost no fluorescence after 24 h, while rotifers exposed to 0.05- and 0.5-µm fluorescently labeled microbeads displayed fluorescence until 48 h, suggesting that 6-µm microbeads are more effectively egested from B. koreanus than 0.05- or 0.5-µm microbeads. This observation provides a potential explanation for our findings that microbead toxicity was size-dependent and smaller microbeads were more toxic. In vitro tests revealed that antioxidant-related enzymes and MAPK signaling pathways were significantly activated in response to microplastic exposure in a size-dependent manner.

Genomic organization and transcriptional modulation in response to endocrine disrupting chemicals of three vitellogenin genes in the self-fertilizing fish Kryptolebias marmoratus.

Vitellogenin (Vtg) is the precursor of egg yolk proteins, and its expression has been used as a reliable biomarker for estrogenic contamination in the aquatic environment. To examine the biomarker potential of the self-fertilizing killifish Kryptolebias marmoratus Vtgs (Km-Vtgs), full genomic DNAs of Km-Vtgs-Aa, Km-Vtgs-Ab, and Km-Vtgs-C were cloned, sequenced, and characterized. Three Vtg genes in K. marmoratus are tandemly placed in a 550 kb section of the same chromosome. In silico analysis of promoter regions revealed that both the Km-Vtgs-Aa and Km-Vtgs-Ab genes had an estrogen response element (ERE), but the Km-Vtgs-C gene did not. However, all three Km-Vtgs genes had several ERE-half sites in their promoter regions. Phylogenetic analysis demonstrated that the three deduced amino acid residues were highly conserved with conventional Vtgs protein, forming distinctive clades within teleost Vtgs. Liver tissue showed the highest expression of Km-Vtg transcripts in all tested tissues (brain/pituitary, eye, gonad, intestine, skin, and muscle) in response to endocrine disrupting chemical (EDC)-exposed conditions. Km-Vtg transcripts were significantly increased in response to 17ß-estradiol (E2), tamoxifen (TMX), 4-n-nonylphenol (NP), bisphenol A (BPA), and octylphenol (OP) over 24hr exposure. The Km-Vtg-A gene was highly expressed compared to the control in response to NP and OP. EDC-induced modulatory patterns of Km-Vtg gene expression were different depending on tissue, gender, and isoforms.

Genome-wide identification of nuclear receptor (NR) superfamily genes in the copepod Tigriopus japonicus.

BACKGROUND: Nuclear receptors (NRs) are a large superfamily of proteins defined by a DNA-binding domain (DBD) and a ligand-binding domain (LBD). They function as transcriptional regulators to control expression of genes involved in development, homeostasis, and metabolism. The number of NRs differs from species to species, because of gene duplications and/or lineage-specific gene losses during metazoan evolution. Many NRs in arthropods interact with the ecdysteroid hormone and are involved in ecdysone-mediated signaling in arthropods. The nuclear receptor superfamily complement has been reported in several arthropods, including crustaceans, but not in copepods. We identified the entire NR repertoire of the copepod Tigriopus japonicus, which is an important marine model species for ecotoxicology and environmental genomics. RESULTS: Using whole genome and transcriptome sequences, we identified a total of 31 nuclear receptors in the genome of T. japonicus. Nomenclature of the nuclear receptors was determined based on the sequence similarities of the DNA-binding domain (DBD) and ligand-binding domain (LBD). The 7 subfamilies of NRs separate into five major clades (subfamilies NR1, NR2, NR3, NR4, and NR5/6). Although the repertoire of NR members in, T. japonicus was similar to that reported for other arthropods, there was an expansion of the NR1 subfamily in Tigriopus japonicus. The twelve unique nuclear receptors identified in T. japonicus are members of NR1L. This expansion may be a unique lineage-specific feature of crustaceans. Interestingly, E78 and HR83, which are present in other arthropods, were absent from the genomes of T. japonicus and two congeneric copepod species (T. japonicus and Tigriopus californicus), suggesting copepod lineage-specific gene loss. CONCLUSIONS: We identified all NR receptors present in the copepod, T. japonicus. Knowledge of the copepod nuclear receptor repertoire will contribute to a better understanding of copepod- and crustacean-specific NR evolution.

Effects of bisphenol A on reproduction, oxidative stress, and lipid regulation in the marine rotifer Brachionus plicatilis.

This study reports the effects of bisphenol A (BPA) on the rotifer Brachionus plicatilis, focusing on growth performance, reproductive output, oxidative stress responses, and lipid metabolism genes. High BPA levels disrupted peak daily offspring production and led to oxidative stress and increased superoxide dismutase and catalase activity. The research identified distinctive monoacylglycerol O-acyltransferase (MGAT) and diacylglycerol O-acyltransferase (DGAT) genes in B. plicatilis, B. rotundiformis, and B. koreanus, enhancing understanding of lipid metabolism in these species. BPA exposure significantly altered MGAT and DGAT expression, and feeding status affected these regulatory patterns. When food was unavailable, BPA reduced DGAT2 and MGAT2a expression. However, under feeding conditions, DGAT2 and MGAT1 levels increased, indicating that nutritional status and BPA exposure interact to affect gene expression.

Multigenerational resilience of the marine rotifer Brachionus plicatilis to high temperature after additive exposure to high salinity and nanoplastics.

To study multigenerational resilience to high temperature (HT) conditions, we exposed Brachionus plicatilis marine rotifers to HT, high salinity (HS), and nanoplastics (NPs), and measured reproductive and life-cycle endpoints. After exposure to HT, rotifer lifespans were reduced, but daily production of offspring increased. However, both combined HT/HS and HT/HS/NP exposure led to additional decreases in longevity and reproductive ability; the antioxidant defense mechanisms of the rotifers were also notably upregulated as measured by reactive oxygen species levels. Fatty-acid profiles were reduced in all conditions. In multigenerational experiments, the negative effects of HT dissipated rapidly; however, the effects of HT/HS and HT/HS/NPs required four generations to disappear completely. The findings indicated that B. plicatilis were able to recover from these environmental stressors. This study demonstrated the resilience of aquatic organisms in response to changing environmental conditions and provides insights into the complex interactions of different abiotic stressors.

Oxidative stress-mediated deleterious effects of hypoxia in the brackish water flea Diaphanosoma celebensis.

In this study, we investigated the acute toxicity, in vivo effects, oxidative stress, and gene expression changes caused by hypoxia on the brackish water flea Diaphanosoma celebensis. The no-observed-effect concentration (NOEC) of 48 h of hypoxia exposure was found to be 2 mg/L O2. Chronic exposure to NOEC caused a significant decline in lifespan but had no effect on total fecundity. The induction of reactive oxygen species increased in a time-dependent manner over 48 h, whereas the content of antioxidant enzymes (superoxide dismutase and catalase) decreased. The transcription and translation levels were modulated by hypoxia exposure. In particular, a significant increase in hemoglobin level was followed by up-regulation of hypoxia-inducible factor 1α gene expression and activation of the mitogen-activated protein kinase pathway. In conclusion, our findings provide a better understanding of the molecular mechanism of the adverse effects of hypoxia in brackish water zooplankton.

Toxicity and speciation of inorganic arsenics and their adverse effects on in vivo endpoints and oxidative stress in the marine medaka Oryzias melastigma.

Here, we investigate the effects of acute and chronic exposure to arsenate (AsV) and arsenite (AsIII) in the marine medaka Oryzias melastigma. In vivo effects, biotransformation, and oxidative stress were studied in marine medaka exposed to the two inorganic arsenics for 4 or 28 days. An investigation of embryonic development revealed no effect on in vivo parameters, but the hatching rate increased in the group exposed to AsIII. Exposure to AsIII also caused the greatest accumulation of arsenic in medaka. For acute exposure, the ratio of AsV to AsIII was higher than that of chronic exposure, indicating that bioaccumulation of inorganic arsenic can induce oxidative stress. The largest increase in oxidative stress was observed following acute exposure to AsIII, but no significant degree of oxidative stress was induced by chronic exposure. During acute exposure to AsV, the increase in the enzymatic activity of glutathione-S-transferase (GST) was twice as high compared with exposure to AsIII, suggesting that GST plays an important role in the initial detoxification process. In addition, an RNA-seq-based ingenuity pathway analysis revealed that acute exposure to AsIII may be related to cell-cycle progression. A network analysis using differentially expressed genes also revealed a potential link between the generation of inflammatory cytokines and oxidative stress due to arsenic exposure.

A Novel Image Processing Method for Obtaining an Accurate Three-Dimensional Profile of Red Blood Cells in Digital Holographic Microscopy.

Recently, research on disease diagnosis using red blood cells (RBCs) has been active due to the advantage that it is possible to diagnose many diseases with a drop of blood in a short time. Representatively, there are disease diagnosis technologies that utilize deep learning techniques and digital holographic microscope (DHM) techniques. However, three-dimensional (3D) profile obtained by DHM has a problem of random noise caused by the overlapping DC spectrum and sideband in the Fourier domain, which has the probability of misjudging diseases in deep learning technology. To reduce random noise and obtain a more accurate 3D profile, in this paper, we propose a novel image processing method which randomly selects the center of the high-frequency sideband (RaCoHS) in the Fourier domain. This proposed algorithm has the advantage of filtering while using only recorded hologram information to maintain high-frequency information. We compared and analyzed the conventional filtering method and the general image processing method to verify the effectiveness of the proposed method. In addition, the proposed image processing algorithm can be applied to all digital holography technologies including DHM, and in particular, it is expected to have a great effect on the accuracy of disease diagnosis technologies using DHM.

Brain glycogen supercompensation following exhaustive exercise.

Brain glycogen localized in astrocytes, a critical energy source for neurons, decreases during prolonged exhaustive exercise with hypoglycaemia. However, it is uncertain whether exhaustive exercise induces glycogen supercompensation in the brain as in skeletal muscle. To explore this question, we exercised adult male rats to exhaustion at moderate intensity (20 m min(-1)) by treadmill, and quantified glycogen levels in several brain loci and skeletal muscles using a high-power (10 kW) microwave irradiation method as a gold standard. Skeletal muscle glycogen was depleted by 82-90% with exhaustive exercise, and supercompensated by 43-46% at 24 h after exercise. Brain glycogen levels decreased by 50-64% with exhaustive exercise, and supercompensated by 29-63% (whole brain 46%, cortex 60%, hippocampus 33%, hypothalamus 29%, cerebellum 63% and brainstem 49%) at 6 h after exercise. The brain glycogen supercompensation rates after exercise positively correlated with their decrease rates during exercise. We also observed that cortical and hippocampal glycogen supercompensation were sustained until 24 h after exercise (long-lasting supercompensation), and their basal glycogen levels increased with 4 weeks of exercise training (60 min day(-1) at 20 m min(-1)). These results support the hypothesis that, like the effect in skeletal muscles, glycogen supercompensation also occurs in the brain following exhaustive exercise, and the extent of supercompensation is dependent on that of glycogen decrease during exercise across brain regions. However, supercompensation in the brain preceded that of skeletal muscles. Further, the long-lasting supercompensation of the cortex and hippocampus is probably a prerequisite for their training adaptation (increased basal levels), probably to meet the increased energy demands of the brain in exercising animals.

Muscle fiber type-dependence effect of exercise on genomic networks in aged mice models.

Skeletal muscles are made up of various muscle fiber type including slow and fast-twitch fibers. Because each muscle fiber has its own physiological characteristics, the effects of aging and exercise vary depending on the type of muscle fiber. We used bioinformatics screening techniques such as differentially expressed gene analysis, gene ontology analysis and gene set enrichment analysis, to try to understand the genetic differences between muscle fiber types. The experiment and gene expression profiling in this study used the soleus (SOL, slow-twitch muscle) and gastrocnemius (GAS, fast-twitch muscle). According to our findings, fatty acid metabolism is significantly up-regulated in SOL compared to GAS, whereas the glucose metabolism pathway is significantly down-regulated in SOL compared to GAS. Furthermore, apoptosis and myogenesis patterns differ between SOL and GAS. SOL did not show differences in apoptosis due to the aging effect, but apoptosis in GAS was significantly up-regulated with age. Apoptosis in GAS of old groups is significantly reduced after 4 weeks of aerobic exercise, but no such finding was found in SOL. In terms of myogenesis, exercise intervention up-regulated this process in GAS of old groups but not in SOL. Taken together, muscle fiber type significantly interacts with aging and exercise. Despite the importance of the interaction between these factors, large-scale gene expression data has rarely been studied. We hope to contribute to a better understanding of the relationship between muscle fiber type, aging and exercise at the molecular level.

Effects of temperature and combinational exposures on lipid metabolism in aquatic invertebrates.

Studies of changes in fatty acids in response to environmental temperature changes have been conducted in many species, particularly mammals. However, few studies have considered aquatic invertebrates, even though they are particularly vulnerable to changes in environmental temperature. In this review, we summarize the process by which animals synthesize common fatty acids and point out differences between the fatty acid profiles of vertebrates and those of aquatic invertebrates. Unlike vertebrates, some aquatic invertebrates can directly synthesize polyunsaturated fatty acids (PUFAs), which can be used to respond to temperature changes. Various studies have shown that aquatic invertebrates increase the degree of saturation in their fatty acids through an increase in saturated fatty acid production or a decrease in PUFAs as the temperature increases. In addition, we summarize recent studies that have examined the complex effects of temperature and combinational stressors to determine whether the degree of saturation in aquatic invertebrates is influenced by other factors. The combined effects of carbon dioxide partial pressure, food quality, starvation, salinity, and chemical exposures have been confirmed, and fatty acid profile changes in response to high temperature were greater than those from combinational stressors.