Mapping Spatiotemporal Heterogeneity in Multifocal Breast Tumor Progression by Noninvasive Ultrasound Elastography-Guided Mass Spectrometry Imaging Strategy.

Spatiotemporal heterogeneity of tumors provides an escape mechanism for breast cancer cells, which can obstruct the investigation of tumor progression. While molecular profiling obtained from mass spectrometry imaging (MSI) is rich in biochemical information, it lacks the capacity for in vivo analysis. Ultrasound diagnosis has a high diagnostic accuracy but low chemical specificity. Here, we describe a noninvasive ultrasound elastography (UE)-guided MSI strategy (UEg-MSI) that integrates physical and biochemical characteristics of tumors acquired from both in vivo and in vitro imaging. Using UEg-MSI, both elasticity histopathology metabolism "fingerprints" and reciprocal crosstalk are revealed, indicating the intact, multifocal spatiotemporal heterogeneity of spontaneous tumorigenesis of the breast from early, middle, and late stages. Our results demonstrate a gradual increase in malignant degree of primary focus in cervical and thoracic mammary glands. This progression is characterized by increased stiffness according to elasticity scores, histopathological changes from hyperplasia to increased nests of neoplastic cells and necrotic areas, and regional metabolic heterogeneity and reprogramming at the spatiotemporal level. De novo fatty acid (FA) synthesis focused on independent (such as ω-9 FAs) and dependent (such as ω-6 FAs) dietary FA intake in the core cancerous nest areas in the middle and late stages of tumor or in the peripheral microareas in the early stage of the tumor. SM-Cer signaling pathway and GPs biosynthesis and degradation, as well as glycerophosphoinositol intensity, changed in multiple characteristic microareas. The UEg-MSI strategy holds the potential to expand MSI applications and enhance ultrasound-mediated cancer diagnosis. It offers new insight into early cancer discovery and the occurrence of metastasis.

A Nomogram Based on Conventional and Contrast-Enhanced Ultrasound for Pre-operative Prediction of Nipple-Areola Complex Involvement in Breast Cancer: A Prospective Study.

OBJECTIVE: Accurately predicting nipple-areola complex (NAC) involvement in breast cancer is essential for identifying eligible patients for a nipple-sparing mastectomy. This study was aimed at developing a pre-operative nomogram for NAC involvement in breast cancer using conventional ultrasound (US) and contrast-enhanced ultrasound (CEUS). METHODS: All patients with primary breast cancer confirmed by pre-operative biopsy underwent US and CEUS examinations. Post-operative pathology was used as the gold standard in assessing NAC involvement. Lasso regression was used to select the predictors most associated with NAC involvement. A nomogram was constructed to calculate the diagnostic efficacy. The data were internally verified with 500 bootstrapped replications, and a calibration curve was generated to validate the predictive capability. RESULTS: Seventy-six patients with primary breast cancer were included in this study, which included 16 patients (21.1%) with NAC involvement and 60 patients (78.9%) without NAC involvement. Among the 23 features of US and CEUS, Lasso regression selected one US feature and two CEUS features, namely, ductal echo extending from the lesion, ductal enhancement extending to the nipple and focal nipple enhancement. A nomogram was constructed, and the results revealed that the area under the curve, sensitivity, specificity and accuracy were 0.891, 81.3%, 86.7% and 85.5%, respectively. The calibration curve exhibited good consistency between the predicted probability and the actual probability. CONCLUSION: The nomogram developed based on US and CEUS had good performance in predicting NAC involvement in breast cancer before surgery, which may facilitate the selection of suitable patients for NAC preservation with greater oncological safety.

A food-web model as a tool for the ecosystem-level management of bivalves in an Atlantic coastal lagoon.

The Ria de Aveiro is an important coastal lagoon for wildlife in Portugal, where the production of bivalves reaches approximately 2700 tons annually. However, the illegal overfishing of bivalves is frequent in this lagoon, which causes critical changes in the ecosystem. In this study, using a developed food-web model (Ecopath model), the ecological carrying capacity (ECC) and maximum sustained yield (MSY) of the Manila clam, Ruditapes philippinarum were estimated, and the effects of further increases in clam biomass on other species were investigated. The results showed that 1) the current biomass and legal catch of R. philippinarum do not yet exceed the ECC (172.40 tons km-2) or the MSY (86.20 tons km-2 year-1) in Ria de Aveiro; 2) the harvested Manila clams of the MSY represent removing from the ecosystem âˆ¼ 581 tons carbon (C) and ∼83 tons nitrogen (N) annually, with substantial ecological and economic implications; and 3) a further increase in the biomass levels of this species may cause the ecotrophic efficiency of other groups to become unrealistic, potentially leading to decreases in ecosystem transfer efficiency, biodiversity and health. The results here are expected to guide the sustainable development and management of bivalve aquaculture in Ria de Aveiro and the protection of the local environment.

Imaging biomarker for quantitative analysis of endometrial injury based on optical coherence tomography/ultrasound integrated imaging mode.

Precise evaluation of endometrial injury is significant to clinical decision-making in gynecological disease and assisted reproductive technology. However, there is a lack of assessment methods for endometrium in vivo. In this research, we intend to develop quantitative imaging markers with optical coherence tomography (OCT)/ultrasound (US) integrated imaging system through intrauterine endoscopic imaging. OCT/US integrated imaging system was established as our previous research reported. The endometrial injury model was established and after treatment, OCT/US integrated imaging and uterus biopsy was performed to evaluate the endometrial thickness, number of superficial fold, and intrauterine area. According to the results, three quantitative indexes acquired from OCT/US image and HE staining have the same trend and have a strong relationship with the severity of the endometrial injury. Accordingly, we developed three imaging markers for quantitative analysis of endometrial injury in vivo, which provided a precise mode for endometrium evaluation in clinical practice.

CO2-Induced Ocean Acidification Alters the Burrowing Behavior of Manila Clam Ruditapes philippinarum by Reversing GABAA Receptor Function.

Biological burrowing behavior is an important driver shaping ecosystems that is being threatened by CO2-induced ocean acidification; however, the effects of ocean acidification on burrowing behavior and its neurological mechanism remain unclear. This study showed that elevated pCO2 significantly affected the burrowing behaviors of the Manila clam Ruditapes philippinarum, such as increased foot contraction, burrowing time, and intrabottom movement and decreased burrowing depth. Delving deeper into the mechanism, exposure to elevated pCO2 significantly decreased extracellular pH and increased [HCO3-]. Moreover, an indicator GABAA receptor, a neuroinhibitor for movement, was found to be closely associated with behavioral changes. In situ hybridization confirmed that the GABAA receptor was widely distributed in ganglia and foot muscles, and elevated pCO2 significantly increased the mRNA level and GABA concentration. However, the increase in GABAA receptor and its ligand did not suppress the foot movement, but rather sent "excitatory" signals for foot contraction. The destabilization of acid-base homeostasis was demonstrated to induce an increase in the reversal potential for GABAA receptor and an alteration in GABAA receptor function under elevated pCO2. This study revealed that elevated pCO2 affects the burrowing behavior of Manila clams by altering GABAA receptor function from inhibitory to excitatory.

Polychaete Bioturbation Alters the Taxonomic Structure, Co-occurrence Network, and Functional Groups of Bacterial Communities in the Intertidal Flat.

Polychaetes are important benthic macrofauna that lives in sediments, usually in intertidal flats with high organic content and high sulfide. It has been suggested that polychaete bioturbation could perform environmental remediation. During the process, the microbial community plays important roles. Here, we used high-throughput sequencing technology to study the bioturbation effects on the bacterial community in the polychaete (Perinereis aibuhitensis) burrows at different tidal positions in intertidal flat. The results showed that the bacterial communities were dramatically influenced by the polychaete bioturbation. The ACE, Chao, and Shannon indices of the polychaete burrows increased in summer. Dominant phyla in the polychaete burrows were Proteobacteria, Campilobacterota, Desulfobacterota, Chloroflexi, and Bacteroidota, and the dominant bacterial families were Sulfurvaceae, Flavobacteriaceae, Rhodobacteraceae, Woeseiaceae, Desulfobulbaceae, and Sulfurimonadaceae. Results of linear discriminant analysis effect size (LEfSe) showed that groups that include organic matter degraders, such as Bacteroidota, Flavobacteriaceae, Rhodobacteraceae, Woeseiaceae, and groups that include sulfur oxidizers, such as Campilobacterota, Sulfurovaceae, Rhodobacteraceae, Desulfobulbaceae, and Sulfurimonadaceae, were significantly increased due to the polychaete bioturbation. The polychaete bioturbation reduced the complexity of the bacterial co-occurrence network while increased its modularity and homogeneity. The polychaete bioturbation also changed the functional groups, which significantly enhanced in functional groups of aerobic nitrite oxidation, nitration, dark thiosulfate oxidation, dark sulfur oxidation, and dark sulfite oxidation, while nitrogen respiration and nitrate respiration decreased. These results provide insight into the impact of bacterial communities under the intertidal polychaete bioturbation.

Influences of Substrate Grain Size on the Burrowing Behavior of Juvenile Meretrix meretrix.

The substrate is the key environmental factor that affects the growth, survival, population and distribution of dwelling mollusks in mudflat settings. To clarify the effect of the substrate grain size on soft substrate preference, burrowing ability and behavior during the selection process of juvenile Meretrix meretrix, four different grain size substrates (coarse sand, medium sand, fine sand, and natural substrate) were set up for comparison. The results indicated that: (1) the burrowing ability of juvenile specimens in fine sand was the strongest; (2) the degree (from high to low) of the juvenile's preference for the four substrates was in the order of fine sand > natural substrate > medium sand > coarse sand; and (3) the selection process of the substrate by the juveniles could be divided into four stages: preparation, selection, burrowing and end stages. These stages showed the behavioral characteristics of a longer selection time and higher percentage of movement in coarse sand. Therefore, our results demonstrated that sea areas or ponds with fine sand as the main component are more suitable for stock enhancement with M. meretrix. These results provide basic data for habitat selection and suitability evaluations for the aquaculture of M. meretrix.

In vivo evaluation of endometrium through dual-modality intrauterine endoscopy.

Female infertilities are highly associated with poor endometrial receptivity. A receptive endometrium is generally characterized by the normal uterine cavity, intact endometrial surface, appropriate endometrial thickness, and echo pattern. Acquiring comprehensive structural information is the prerequisite of endometrium assessment, which is beyond the ability of any single-modality imaging method. In this paper, we introduce a custom-made intrauterine dual-modality (OCT/ultrasound) endoscopic imaging system and achieve in vivo imaging of rabbit uteri, for the first time to our knowledge. The endometrial features of the injured uteri in both ultrasonic and OCT images are consistent with their corresponding pathology. The quantified parameters, including uterine thickness and endometrial surface roughness, show the correlation with the endometrial injury degree but with poor performance for injury classification. The combination of these parameters was proved to assess the degrees of endometrial injury more accurately. Our work shows the potential of the dual-modality system to be translated into a clinical tool, providing multiple quantitative imaging information and helping evaluate the endometrial receptivity and diagnose infertility.

Microplastics influence physiological processes, growth and reproduction in the Manila clam, Ruditapes philippinarum.

Microplastics (<5 mm) are widely distributed in marine environments and pose a serious threat to bivalves. Here, the ingestion and accumulation of polystyrene microplastics (PS microplastics, diameters 5 and 10 µm) by the Manila clam, Ruditapes philippinarum, and their impacts on physiological processes, growth and reproduction were studied. The results showed that both PS microplastics were ingested by the Manila clam and accumulated in their gills, hepatopancreases and intestines. Furthermore, the accumulation of 5 and 10 µm PS microplastics significantly increased the rates of respiration and excretion while significantly decreasing feeding and absorption efficiency (AE), leading to a dramatically reduced amount of energy available for growth (SfG) and ultimately led to slower growth. The dynamic energy budget (DEB) model predicts that PS microplastic exposure for 200 days would cause lower shell/flesh growth rates and reproductive potentiality. Transcriptomic profiles support these results, as carbon and protein metabolism and oxytocin and insulin-related signaling pathways were significantly altered in clams in response to PS microplastics. This study provides evidence that microplastics strongly affect the physiological activities, energy allocation, growth and reproduction of filter-feeding bivalves.

Application of ultrasound markers measured at different time points of COH cycle in the prediction of ovarian response for individualised ovulation induction.

The purpose of this study was to evaluate the predictive value of ultrasound markers measured at different time points of the controlled ovarian hyperstimulation (COH) cycle on ovarian response and outcome indicators in the IVF-ET cycle. According to the oestrogen level and the number of retrieved oocytes, patients who planned for COH treatment were separated into low-response group, normal and high-response group. The ovarian stromal artery flow parameters on the day of pituitary down-regulation, day 1, day 7, day 10, and the day of hCG injection were collected prospectively. We also have collected the data of cumulus oophorus count on the day of hCG injection by transvaginal sonography. Compared with the low-response group, on the first day of the COH cycle PI, RI, and S/D were lower in the high-response group than they were in the low-response group (p < .05). PSV and EDV were significantly higher in the high-response group than they were in the low-response group (p < .01), and the PSV on the first day of the COH cycle have statistical significance in predicting the number of high-quality embryos. The number of cumulus oophorus on the day of hCG injection has statistical significance in predicting the number of oocytes retrieved and fertilised oocytes. We conclude that the ovarian stromal artery flow parameters on the first day of the COH cycle and cumulus oophorus count on hCG injection day can serve as efficient indicators for an early assessment of ovarian response and individualised ovulation induction.IMPACT STATEMENTWhat is already known on this subject? AMH, AFC, and the age of the patient are well-known effective parameters for the evaluation of ovarian response, but these are insufficient and full of individual differences. Some researchers have investigated the value of colour Doppler ultrasound and cumulus oophorus in assessing ovarian response, but no definitive conclusion has been reached.What do the results of this study add? The hemodynamic parameters of ovarian stromal artery on the first day of the COH cycle and the number of cumulus oophorus on the day of hCG injection detected by Transvaginal Colour Doppler Sonography (TV-CDS) could be used to predict the ovarian response.What are the implications of these findings for clinical practice and/or further research? Ovarian stromal artery flow parameters and cumulus oophorus detected by TV-CDS can potentially be offered as a complementary parameter for ovarian reserve.

Contrastive rendering with semi-supervised learning for ovary and follicle segmentation from 3D ultrasound.

Segmentation of ovary and follicles from 3D ultrasound (US) is the crucial technique of measurement tools for female infertility diagnosis. Since manual segmentation is time-consuming and operator-dependent, an accurate and fast segmentation method is highly demanded. However, it is challenging for current deep-learning based methods to segment ovary and follicles precisely due to ambiguous boundaries and insufficient annotations. In this paper, we propose a contrastive rendering (C-Rend) framework to segment ovary and follicles with detail-refined boundaries. Furthermore, we incorporate the proposed C-Rend with a semi-supervised learning (SSL) framework, leveraging unlabeled data for better performance. Highlights of this paper include: (1) A rendering task is performed to estimate boundary accurately via enriched feature representation learning. (2) Point-wise contrastive learning is proposed to enhance the similarity of intra-class points and contrastively decrease the similarity of inter-class points. (3) The C-Rend plays a complementary role for the SSL framework in uncertainty-aware learning, which could provide reliable supervision information and achieve superior segmentation performance. Through extensive validation on large in-house datasets with partial annotations, our method outperforms state-of-the-art methods in various evaluation metrics for both the ovary and follicles.

Development of an auxiliary device for ultrasound-guided aspiration of pelvic cystic masses: a simulation study.

BACKGROUND: Pelvic cystic masses are a common gynecological condition. Ultrasound-guided aspiration is a minimally invasive surgical technique for the treatment of pelvic cystic masses. However, further developments to improve its stability and safety are wanting. This study evaluated the application and safety of a self-developed auxiliary device for pelvic cystic masses' ultrasound-guided aspiration through phantom testing. METHODS: Saline and coupling agents were used at different viscosity levels to simulate simple cysts, medium viscosity cysts (such as pelvic effusions), and ovarian, endometrial cysts. An auxiliary device consisting of a three-way valve, a negative pressure aspirator, and a pressurized infusion bag was developed. Phantom testing was performed to evaluate the application of this device in ultrasound-guided aspiration of pelvic cystic masses. The indicators, including the time of aspiration, time of injection, and the incidence of complications, were compared to cases in which ultrasound-guided aspiration was performed manually with a syringe. RESULTS: The incidence of complications in the auxiliary device group was significantly lower compared to the manual operation group (P<0.05). The ovarian cystic aspiration times and operation times were significantly shorter in the auxiliary device group compared to the traditional manual puncture group (P<0.05). CONCLUSIONS: Ultrasound-guided aspiration is repeatable and minimally invasive for the treatment of pelvic cystic masses. Using the auxiliary device designed in this report resulted in shorter operation times, definite needle fixation, and fewer complications, which may allow for a more stable and safer aspiration procedure for the treatment of pelvic cystic masses.

UTMD promoted local delivery of miR-34a-mimic for ovarian cancer therapy.

MicroRNA-mediated gene therapy is emerging as a promising method for the treatment of ovarian cancer, but the development of miRNA mimic delivery vectors is still in its infancy, where the safety and efficacy of miR-34a-mimic remain unknown. Ultrasound-targeted microbubble destruction (UTMD) can be an effective and minimally invasive tool for the delivery of miR-34a-mimic in vitro and in vivo. Here, we describe a high-efficiency gene delivery strategy by using miR-34a-mimic loaded folate modified microbubbles (miR-34a-FM) with a portable ultrasonic irradiation system. Ultrasonic parameters, including acoustic intensity (AI), exposure time (ET) and duty cycle (DC), were optimized and the optimal acoustic condition (1.0 W/cm2, 20 s, and 15% DC) was used to deliver miRNA-34a into cells in vitro. MiR-34a mimic was successfully introduced into the cytoplasm and was found to inhibit proliferation and induce apoptosis of SK-OV-3 cells. Next, miR-34a-mimic was delivered to tumor tissue via UTMD, inhibiting tumor growth and prolonging the survival time of mice. In summary, UTMD-mediated miR-34a-mimic delivery has potential application in the clinical treatment of ovarian cancer.

Integrated transcriptomics and metabolomics analyses reveal benzo[a]pyrene enhances the toxicity of mercury to the Manila clam, Ruditapes philippinarum.

Mercury (Hg2+) and benzo[a]pyrene (BaP) are ubiquitous and persistent pollutants with multiple toxicities in bivalve molluscs. Here, the toxicological responses in the gills of Manila clams, Ruditapes philippinarum, to Hg2+ (10 µg L-1), BaP (3 µg L-1), and their mixture were analysed using transcriptomics and metabolomics approaches. Comparisons of the transcriptomes and metabolomes of Hg2+-and/or BaP-treated clams with control animals revealed the involvement of the detoxification metabolism, immune defence, energy-related pathways, and osmotic regulation in the stress response of R. philippinarum. Exposure to Hg2+ alone primarily enhanced the detoxification and energy metabolic pathways by significantly increasing the expression of genes associated with heat-shock proteins and oxidative phosphorylation. However, co-exposure to Hg2+ and BaP caused greater immunotoxicity and disrupted detoxification metabolism, the TCA cycle, glycolysis, and ATP generation. The expression levels of cytochrome P450 1A1 (CYP1A1), multidrug resistance-associated protein 1 (MRP1), and myosin (MYO), and the activity of electron transport system (ETS) in gills were detected, supporting the underlying toxic mechanisms of Hg2+ and BaP. We suggest that the presence of BaP enhances the toxicity of Hg2+ by 1) hampering the detoxification of Hg2+, 2) increasing the immunotoxicity of Hg2+, and 3) constraining energy availability for clams.

The effects of teflubenzuron on mortality, physiology and accumulation in Capitella sp.

The chitin synthesis inhibitor teflubenzuron (TFB) is a feed antiparasitic agents used to impede molting of the salmon lice, an ecto-parasite that severely affects the salmon industry. Low absorption of oral administered TFB may cause elevated concentrations in the feces discharged from the salmon into the benthic environment. The polychaete Capitella sp. are often dominant in such habitats and consume organic waste deposited on the sediment. In the present study, Capitella sp. were exposed to doses of TFB in salmon feed of 1, 2 and 4 g TFB kg-1 (0 g TFB kg-1 in control group) over an experimental period of 32 days. Cumulative mortality was 12%-15% in both treatment groups with 1 and 2 g TFB kg-1 and reached 27% in the group with 4 g TFB kg-1. Only the highest dose (4 g TFB kg-1) negatively affected feed intake, growth and respiration of the polychaetes while food conversion efficiency was not affected. At the end of the experiment, the concentrations of TFB in the Capitella sp. were high, in the range of 9.24-10.32 µg g-1 for the three treatment groups. It was suggested that a maximum level of absorption rate was reached, also for the lowest dose. High concentrations of TFB in the Capitella sp. might pose a risk to crustaceans that forage for polychaetes in the vicinity of fish farms. We conclude that the effects of TFB on Capitella sp. may therefore primarily be to the predators rather than the Capitella sp.

CR-Unet-Based Ultrasonic Follicle Monitoring to Reduce Diameter Variability and Generate Area Automatically as a Novel Biomarker for Follicular Maturity.

Follicle size is closely related to ovarian function and is an important biomarker in transvaginal ultrasound examinations for assessing follicular maturity during an assisted reproduction cycle. However, manual measurement is time consuming and subject to high inter- and intra- observer variability. Based on the deep learning model CR-Unet described in our previous study, the aim of our present study was to investigate further the feasibility of using this model in clinical practice by validating its performance in reducing the inter- and intra-observer variability of follicle diameter measurement. This study also investigated whether follicular area is a better biomarker than diameter in assessing follicular maturity. Data on 106 ovaries and 230 follicles collected from 80 cases of single follicular cycles and 26 cases of multiple follicular cycles constituted the validation set. Intra-observer variability was 0.973 and 0.982 for the senior sonographer and junior sonographer in single follicular cycles and 0.979 (0.971, 0.985) and 0.920 (0.892, 0.943) in multiple follicular cycles, respectively, while CR-Unet had no intra-group variation. Bland-Altman plot analysis indicated that the 95% limits of agreement between senior sonographer and CR-Unet (-2.1 to 1.1 mm, -2.02 to 0.75 mm) were smaller than those between senior sonographer and junior sonographer (-1.51 to 1.15 mm, -2.1 to 1.56 mm) in single and multiple follicular cycles. The average operating times of diameter measurement taken by the junior sonographer, senior sonographer and CR-Unet were 7.54 ± 1.8, 4.87 ± 0.84 and 1.66 ± 0.76 s, respectively (p < 0.001). Correlation analysis indicated that both manual and automated follicular area correlated better with follicular volume than diameter. The deep learning algorithm and the new biomarker of follicular area hold potential for clinical application of ultrasonic follicular monitoring.

CR-Unet: A Composite Network for Ovary and Follicle Segmentation in Ultrasound Images.

Transvaginal ultrasound (TVUS) is widely used in infertility treatment. The size and shape of the ovary and follicles must be measured manually for assessing their physiological status by sonographers. However, this process is extremely time-consuming and operator-dependent. In this study, we propose a novel composite network, namely CR-Unet, to simultaneously segment the ovary and follicles in TVUS. The CR-Unet incorporates the spatial recurrent neural network (RNN) into a plain U-Net. It can effectively learn multi-scale and long-range spatial contexts to combat the challenges of this task, such as the poor image quality, low contrast, boundary ambiguity, and complex anatomy shapes. We further adopt deep supervision strategy to make model training more effective and efficient. In addition, self-supervision is employed to iteratively refine the segmentation results. Experiments on 3204 TVUS images from 219 patients demonstrate the proposed method achieved the best segmentation performance compared to other state-of-the-art methods for both the ovary and follicles, with a Dice Similarity Coefficient (DSC) of 0.912 and 0.858, respectively.

Biomarkers responses in Manila clam, Ruditapes philippinarum after single and combined exposure to mercury and benzo[a]pyrene.

Physiological and biochemical responses in bivalves exposed to pollutants have proved a valuable tool to assess the health of organisms in aquatic ecosystems. The single and combined effects of mercury (Hg2+, 2 and 10 µg/L) and benzo[a]pyrene (BaP, 3 µg/L) on physiological and biochemical biomarkers in Manila clam, Ruditapes philippinarum were evaluated. Results showed that significant higher oxygen consumption (OR) and ammonia-N excretion rates (NR) together with significant lower ingestion rates (IR) were observed for the 10 µg/L Hg2+ or 3 µg/L BaP treatments compared to controls (P < 0.05). However, clam NR decreased significantly in response to the binary mixtures of 10 µg/L Hg2+ and 3 µg/L BaP (P < 0.05). Moreover, the levels of superoxide dismutase (SOD), catalase (CAT), glutathione-s-transferases (GSTs), glutathione (GSH), acetylcholinesterase (AChE) and malondialdehyde (MDA) in the hepatopancreas of clams were induced substantially, whereas glycogen (GLY) contents were suppressed dramatically after Hg2+ and BaP exposure. Additionally, the integrated biomarker response (IBR) values measured showed significant increases in combination treatments and they were much higher than that in the Hg2+ treatment. This study will provide further information on the defense mechanism in the Manila clam after exposure to marine pollutants and may help evaluate the quality of the aquatic environment.

Metabolic responses to elevated pCO2 in the gills of the Pacific oyster (Crassostrea gigas) using a GC-TOF-MS-based metabolomics approach.

Rising atmospheric carbon dioxide (CO2), primarily from anthropogenic emissions, are resulting in increasing absorption of CO2 by the oceans, leading to a decline in oceanic pH in a process known as ocean acidification (OA). There is a growing body of evidence demonstrating the potential effect of OA on the energetics/physiology and consequently life-history traits of commensally important marine organisms. However, despite this little is known of how fundamental metabolic pathways that underpin changes in organismal physiology are affected by OA. Consequently, a gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) based metabolic profiling approach was applied to examine the metabolic responses of Crassostrea gigas to elevated pCO2 levels, under otherwise natural field conditions. Oysters were exposed natural environmental pCO2 (~625.40 µatm) and elevated pCO2 (~1432.94 µatm) levels for 30 days. Results indicated that 36 differential metabolites were identified. Differential metabolites were mapped in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database to search for the related metabolic pathways. Pathway enrichment analysis indicates that alanine, aspartate and glutamate metabolism and glycine, serine and threonine metabolism were the most statistically enriched pathways. Further analysis suggested that elevated pCO2 disturb the TCA cycle via succinate accumulation and C. gigas most likely adjust their energy metabolic via alanine and GABA accumulation accordingly to cope with elevated pCO2. These findings provide an understanding of the molecular mechanisms involved in modulating C. gigas metabolism under elevated pCO2.