A Comparative Study on Effects of Three Butyric Acid-Producing Additives on the Growth Performance, Non-specific Immunity, and Intestinal Microbiota of the Sea Cucumber Apostichopus japonicus.

The providers of butyric acid, Clostridium butyricum (CB), sodium butyrate (SB), and tributyrin (TB), have been extensively studied as aquafeed additives in recent years. However, no comparative study has been reported on the probiotic effects of CB, SB, and TB as feed additives on sea cucumber (Apostichopus japonicus). A 63-day feeding trial was performed to assess the effects of dietary live cells of C. butyricum (CB group, the basal diet supplemented with 1% CB), sodium butyrate (SB group, the basal diet supplemented with 1% SB), and tributyrin (TB group, the basal diet supplemented with 1% TB) on the growth, non-specific immunity, and intestinal microbiota of A. japonicus with a basal diet group as the control. Results indicated that all three additives considerably increased A. japonicus growth, with dietary CB having the optimal growth-promoting effect. Of the seven non-specific enzyme parameters measured in coelomocytes of A. japonicus (i.e., the activities of phagocytosis, respiratory burst, superoxide dismutase, alkaline phosphatase, acid phosphatase, catalase, and lysozyme), dietary CB, SB, and TB considerably increased the activities of six, five, and six of them, respectively. The immune genes (Aj-p105, Aj-p50, Aj-rel, and Aj-lys) expression in the mid-intestine tissues of A. japonicus was significantly increased by all three additives. The CB group had the highest expression of all four genes. Additionally, the relative expression of Aj-p105, Aj-p50, and Aj-lys genes was significantly up-regulated in the three additive groups after stimulation with inactivated Vibrio splendidus. Dietary CB enhanced the intestinal microbial diversity and richness in A. japonicus while dietary TB decreased them. Meanwhile, dietary CB, SB, and TB significantly enhanced the abundance of Firmicutes, unclassified_f_Rhodobacteraceae, and Proteobacteria, respectively, while dietary CB and SB reduced the abundance of Vibrio. Dietary CB and SB improved the stability of microbial ecosystem in the intestine of A. japonicus. In contrast, dietary TB appeared to have a negative effect on the stability of intestinal microbial ecosystem. All three additives improved the intestinal microbial functions associated with energy production and immunity regulation pathways, which may contribute directly to growth promotion and non-specific immunity enhancement in A. japonicus. Collectively, in terms of enhancing growth and non-specific immunity, as well as improving intestinal microbiota, dietary live cells of C. butyricum exhibited the most effective effects in A. japonicus.

Modification of osimertinib to discover new potent EGFRC797S-TK inhibitors.

The EGFRC797S mutation is a dominant mechanism of acquired resistance after the treatment of non-small cell lung cancer (NSCLC) with osimertinib in clinic. To date, there is no inhibitor approved to overcome the resistance caused by osimertinib. In this study, a series of compounds with phenylamino-pyrimidine scaffold deriving from osimertinib were designed, synthesized and evaluated as fourth-generation EGFRC797S-TK inhibitors. Consequently, compound Os30 exhibited potent inhibitory activities against both EGFRDel19/T790M/C797S TK and EGFRL858R/T790M/C797S TK with IC50 values of 18 nM and 113 nM, respectively. Moreover, Os30 can powerfully inhibit the proliferation of KC-0116 (BaF3-EGFRDel19/T790M/C797S) and KC-0122 (BaF3-EGFRL858R/T790M/C797S) cells. In addition, Os30 can suppress EGFR phosphorylation in a concentration-dependent manner in KC-0116 cells, arrest KC-0116 cells at G1 phase and induce the apoptosis of KC-0116 cells. More importantly, Os30 showed potent antitumor efficacy in the KC-0116 cells xenograft nude mice tumor model with the tumor growth inhibitory rate of 77.6% at a dosage of 40 mg/kg. These findings demonstrate that modification of osimertinib can discover new potent EGFRC797S-TK inhibitors, and compound Os30 is a potent fourth-generation EGFR inhibitor to treat NSCLC with EGFmRC797S mutation.

The effect of total alkalinity on growth performance and calcification in juvenile Pacific abalone Haliotis discus hannai.

A 45-day trial was conducted to study the effect of seawater total alkalinity (TA) level up- and downregulation on the growth performance and calcification of Haliotis discus hannai Ino, while seawater pH was maintained at pHNBS = 8.1. Although seawater was not acidified, the results showed that TA downregulation caused a significant reduction (P < 0.05) in the somatic tissue growth of juvenile abalone, while TA upregulation significantly increased growth performance (P < 0.05). Similar to the impacts of pH reduction, TA downregulation also induces a decline in CO2 buffering capacity, which may be the reason why somatic tissue growth was reduced, as lowered CO2 buffering capacity was reported to shift the acid-base balancing of abalone. Parts of the periostracum layer weremissing and exposed the inner shell layers of the individuals from the TA-downregulated group. Scanning electron microscopy (SEM) results showed calcium carbonate densely deposited onto the inner shell in the control and TA-upregulated groups, while sparsely deposited calcium carbonate was observed in the TA-downregulated group. The C: N ratio in the shell of individuals from the TA-downregulated group was significantly lower than that of the other two groups, indicating that less inorganic carbon was added to the shell. As a result, abalone grew lighter and thinner shells in TA-downregulated seawater. Although seawater was not acidified, TA downregulation also caused a reduction in the calcium carbonate saturation state (Ω), which induced the erosion of the surface shell and the interruption of calcium carbonate generation. In conclusion, although seawater pH remained at ambient levels, the lowered CO2 buffering capacity and Ω induced by seawater TA downregulation also showed a detrimental effect on the growth and calcification of Pacific abalone. The impact of ocean acidification on the growth of abalone should not be assessed using only seawater pH and/or pCO2 but rather taking into account all of carbonate chemistry, particularly the CO2 buffering capacity. Abalone cultivation is suggested to be carried out in seawater with a higher level of CO2 buffering capacity and Ω, which can be achieved through integrated culture with seaweed or increasing the seawater TA level.

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.

The Application of the Generalized Additive Model to Represent Macrobenthos near Xiaoqing Estuary, Laizhou Bay.

Macrobenthos is widely used as an indicator of ecological health in marine monitoring and assessment. The present study aimed to characterize the interrelationships between the distribution of the macrobenthos community and environmental factors near Xiaoqing Estuary, Laizhou Bay. Responses of species richness to environmental factors were studied using the generalized additive model (GAM) and the Margalef diversity index (dM) as indicators of species diversity instead of individual indicator species. Six factors were selected in the optimal model by stepwise regression: sediment factors (organic matter, phosphate, nitrate nitrogen, and ammonium nitrogen) and water factors (salinity, and ammonium nitrogen). The response curves generated by the GAM showed a unimodal relationship among taxa diversity, salinity in water, and sediment organic matter. dM was positively correlated with ammonium nitrogen in water and was negatively correlated with phosphate in the sediment. The model optimized by forward stepwise optimization explained 92.6% of the Margalef diversity index with a small residual (2.67). The model showed good performance, with the measured dM strongly correlated with the predicted dM (Pearson R2 = 0.845, p < 0.05). The current study examined the combined influence of multiple eco-factors on macrobenthos, and the Margalef diversity index of macrobenthos was predicted by the GAM model in a salinity-stressed estuary.

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.

Design, synthesis and evaluation of new pyrimidine derivatives as EGFRC797S tyrosine kinase inhibitors.

The clinical use of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in the treatment of non-small cell lung cancer was limited by the drug resistance caused by EGFRC797S mutation. Therefore, in order to overcome the drug resistance, we designed and synthesized a series of 2-aminopyrimidine derivatives as EGFRC797S-TKIs. Among these compounds, compounds A5 and A13 showed significant anti-proliferative activity against the KC-0116 (EGFRdel19/T790M/C797S) cell line with high selectivity. A5 inhibited EGFR phosphorylation and induced apoptosis of KC-0116 cell, arrested KC-0116 cell at G2/M phase. Molecular docking results showed that A5 and brigatinib bind to EGFR in a similar pattern. In addition to forming two important hydrogen bonds with Met793 residue, A5 also formed a hydrogen bond with Lys745 residues, which may play an important role for the potent inhibitory activity against EGFRdel19/T790M/C797S. Based on these results, A5 turned out to be effective reversible EGFRC797S-TKIs which can be further developed.

Haemolymph pH of two important mollusc species is susceptible to seawater buffering capacity instead of pH or pCO2.

The acid-base status and balance of molluscs are considered to be susceptible to environmental changes, especially in the context of ocean acidification (OA). Here, we studied the effects of manipulated seawater carbonate chemistry on the acid-base status of scallop Chlamys farreri and abalone Haliotis discus hannai. The haemolymph pH of the tested individuals showed a fast response to acidified seawater incubation, and the pH level was restored to a normal value within 1 h of recovery in control seawater. However, no significant correlation (P > 0.05) was found between haemolymph pH and seawater pCO2 or pH, while the squared Pearson correlation coefficient (R2) ranged from close to zero to 0.41. In addition, although the pCO2 level of total alkalinity (TA)-lowered seawater was lower than half of that in the control, molluscs eliminated less CO2 (less than 80%) to TA lowered waters than to the control waters. These findings seem to disagree with the crucial role of seawater pCO2 in influencing the acid-base balance of molluscs. CO2 elimination occurs in the microenvironment, and CO2 first diffuses to limited amounts of seawater that tightly surround the gills, causing dissolved inorganic carbon (DIC) accumulation in the ventilation sites, which leads to a sharp increase in the pCO2 of the surrounding seawater. Moreover, in this microenvironment, the pCO2 level increases much faster and more greatly if the environmental seawater is acidified or contains a lower level of TA. Therefore, mollusc acid-base status is influenced by rapidly varying pCO2 levels at the ventilation site, which is largely independent of that of the rest of the incubating seawater. In summary, CO2 elimination by molluscs relies heavily on the carbonate chemistry of environmental seawater, and seawater buffering capacity should be taken into consideration instead of considering only pCO2 or pH in studying the acid-base balance of marine molluscs.

Identification Invasion-Related Long Non-Coding RNAs in Lung Adenocarcinoma and Analysis of Competitive Endogenous RNA Regulatory Networks.

Background: Cell invasion plays a vital role in cancer development and progression. Aberrant expression of long non-coding RNAs (lncRNAs) is also critical in carcinogenesis. However, the prognostic value of invasion-related lncRNAs in lung adenocarcinoma (LUAD) remains unknown. Methods: Differentially expressed mRNAs (DEmRNAs), lncRNAs (DElncRNAs), and microRNAs (DEmiRNAs) were between LUAD and control samples. Pearson correlation analyses were performed to screen for invasion-related DElncRNAs (DEIRLs). Univariate and multivariate Cox regression algorithms were applied to identify key genes and construct the risk score model, which was evaluated using receiver operating characteristic (ROC) curves. Gene set enrichment analysis (GSEA) was used to explore the underlying pathways of the risk model. Moreover, an invasion-related competitive endogenous RNA (ceRNA) regulatory network was constructed. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to detect the expression of prognostic lncRNAs in the LUAD and control samples. Results: A total of 45 DElncRNAs were identified as DEIRLs. RP3-525N10.2, LINC00857, EP300-AS1, PDZRN3-AS1, and RP5-1102E8.3 were potential prognostic lncRNAs, the expression of which was verified by RT-qPCR in LUAD samples. Both the risk score model and nomogram used the prognostic lncRNAs. ROC curves showed the risk score model had moderate accuracy and the nomogram had high accuracy in predicting patient prognosis. GSEA results indicated that the risk score model was associated with many biological processes and pathways relevant to cell proliferation. A ceRNA regulatory network was constructed in which PDZRN3-miR-96-5p-CPEB1, EP300-AS1-miR-93-5p-CORO2B, and RP3-525N10.2-miR-130a-5p-GHR may be key invasion-related regulatory pathways in LUAD. Conclusion: Our study identified five novel invasion-related prognostic lncRNAs (RP3-525N10.2, LINC00857, EP300-AS1, PDZRN3-AS1, and RP5-1102E8.3) and established an accurate model for predicting the prognosis of patients with LUAD. These findings enrich our understanding of the relationships between cell invasion, lncRNAs, and LUAD and may provide novel treatment directions.

Seawater carbonate parameters function differently in affecting embryonic development and calcification in Pacific abalone (Haliotis discus hannai).

pH or pCO2 are usually taken to study the impact of ocean acidification on molluscs. Here we studied the different impact of seawater carbonate parameters on embryonic development and calcification of the Pacific abalone (Haliotis discus hannai). Early embryonic development was susceptible to elevated pCO2 level. Larvae hatching duration was positively and hatching rate was negatively correlated with the pCO2 level, respectively. Calcium carbonate (CaCO3) deposition of larval shell was found to be susceptible to calcium carbonate saturation state (Ω) rather than pCO2 or pH. Most larvae incubated in seawater with Ωarag = 1.5 succeeded in shell formation, even when seawater pCO2 level was higher than 3700 µatm and pHT was close to 7.4. Nevertheless, larvae failed to generate CaCO3 in seawater with Ωarag ≤ 0.52 and control level of pCO2, while seawater DIC level was lowered (≤ 852 µmol/kg). Surprisingly, some larvae completed CaCO3 deposition in seawater with Ωarag = 0.6 and slightly elevated DIC (2266 µmol/kg), while seawater pCO2 level was higher than 2700 µatm and pHT was lower than 7.3. This indicates that abalone may be capable of regulating carbonate chemistry to support shell formation, however, the capability was limited as surging pCO2 level lowered growth rate and jeopardized the integrity of larval shells. Larvae generated thicker shell in seawater with Ωarag = 5.6, while adult abalone could not deposit CaCO3 in seawater with Ωarag = 0.29 and DIC = 321 µmol/kg. This indicates that abalone may lack the ability to directly remove or add inorganic carbon at the calcifying sites. In conclusion, different seawater carbonate parameters play different roles in affecting early embryonic development and shell formation of the Pacific abalone, which may exhibit limited capacity to regulate carbonate chemistry.

A unified model for large-scale thickness measurement of lubricating film based on ultrasonic lag phase slope.

A unified model based on the ultrasonic lag phase slope is developed for measuring the lubricating film thickness at a large scale. The ultrasonic lag phase of adjacent waves instead of the phase of overlapped waves is calculated as a function of the ultrasonic frequency and film thickness. The slope of the ultrasonic lag phase is determined correspondingly, which is linearly proportional to the lubricating film thickness. Both the finite element analysis and tests on the lubricating film thickness are performed to verify the proposed method. The results show that despite the fluctuations of the lag phase, the lag phase slope can be used for measuring the lubricating film thickness at a large scale from 0.1 µm to 170 µm.

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.

Discovery of Potent PROTACs Targeting EGFR Mutants through the Optimization of Covalent EGFR Ligands.

Drug resistance caused by epidermal growth factor receptor (EGFR) mutation has largely limited the clinical use of EGFR tyrosine kinase inhibitors (EGFR-TKIs) for the treatment of non-small-cell lung cancer (NSCLC). Herein, to overcome the intractable problem of drug resistance, proteolysis targeting chimeras (PROTACs) targeting EGFR mutants were developed by optimizing covalent EGFR ligands. Covalent or reversible covalent pyrimidine- or purine-containing PROTACs were designed, synthesized, and evaluated. As a consequence, covalent PROTAC CP17, with a novel purine-containing EGFR ligand, was discovered as a highly potent degrader against EGFRL858R/T790M and EGFRdel19, reaching the lowest DC50 values among all reported EGFR-targeting PROTACs. Furthermore, CP17 exhibited excellent cellular activity against the H1975 and HCC827 cell lines with high selectivity. Mechanism investigation indicated that the lysosome was involved in the degradation process. Importantly, the covalent binding strategy was proven to be an effective approach for the design of PROTACs targeting EGFRL858R/T790M, which laid the practical foundation for further development of potent EGFR-targeting PROTACs.

The anti-neoplastic activities of aloperine in HeLa cervical cancer cells are associated with inhibition of the IL-6-JAK1-STAT3 feedback loop.

Cervical cancer (CC) is recognized as the most common neoplasm in the female reproductive system worldwide. The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients. Aloperine (ALO) is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation, allergies and infection. However, its therapeutic efficacy and underlying mechanism in CC are still unclear. In the current study, MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC. Then, the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry, respectively, while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay. Moreover, nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo. The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis. This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest. Simultaneously, ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio. Additionally, the migration and invasion of HeLa cells were attenuated by ALO treatment, which was considered to result from inhibition of epithelial-to-mesenchymal transition. For molecular mechanisms, the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment. This study indicated that ALO markedly suppresses the proliferation, migration and invasion and enhances the apoptosis of HeLa cells. In addition, these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.

A minimum energy optimization approach for simulations of the droplet wetting modes using the cellular Potts model.

Wetting modes of a droplet on a periodical grooved surface were simulated by using the Cellular Potts Model (CPM). An optimization approach based on the Synthesis Minimum Energy (SME), which is defined as the lowest energy of the simulation system, was proposed for determining the droplet wetting modes. The influence of the fluctuation parameter (T) was discussed. The results showed that the SME optimization approach increased the accuracy of the wetting mode simulation. For the values of T used in the SME, an increase in the range of T and a decrease in the step size of T will not only cause an increase in the accuracy of the SME but also will cause an increase in the total consumption of calculation time and a decrease in the ability of accuracy improvement. A high value of the fluctuation parameter T generated the Cassie mode transition for the droplet. With an increase in the pillar height, the droplet wetting mode transited from Wenzel mode to Cassie mode, while it transited from Cassie mode to Wenzel mode with an increase in the interpillar distance.

Effect of large-scale kelp and bivalve farming on seawater carbonate system variations in the semi-enclosed Sanggou Bay.

Although cultured algae and shellfish can be the dominant species in some localized coastal waters, research on the effect of large-scale mariculture on the carbonate system variations in these local waters is still lacking. We conducted five cruises from May to September and studied spatiotemporal variations in the seawater carbonate system in the semi-closed Sanggou Bay, which is famous for its large-scale mariculture. Our results showed that both kelp and bivalve farming induced significant spatiotemporal variations in the carbonate system within the bay. When cultured kelp reached its highest biomass in May, the maximum ΔDIC, ΔpCO2 and ΔpHT between the seawater from the kelp farming area and the non-farming outer bay area was -156 µmol kg-1, -102 µatm and 0.15 pH units, respectively. However, no significant effect of kelp farming on seawater total alkalinity (TA) was observed. Kelp farming also caused the carbonate system variations of seawater from the bivalve farming area. Assuming no kelp was farmed in May, the average pH and pCO2 would reduce by 0.12 pH units and increase by 179 µatm, respectively, in the bivalve farming area. Bivalve farming significantly reduced seawater TA, indicating that fast deposition of calcium carbonate occurred in the bivalve farming area. Although bivalve respiration released CO2 into seawater and elevated seawater pCO2 level and reduced seawater pHT, surprisingly, seawater dissolved inorganic carbon (DIC) reduced significantly in the bivalve farming area. These results indicated that bivalves fixed a larger amount of inorganic carbon by calcification than that released into seawater by respiration. Overall, large-scale kelp and bivalve farming are important biological drivers of variations in the carbonate system within the semi-enclosed Sanggou Bay. Altered carbonate systems by kelp farming may favour calcification of farmed bivalves and provide an essential refuge for these species during the future ocean acidification.

Polychlorinated biphenyls and organochlorine pesticides in surface sediments from the sand flats of Shuangtaizi Estuary, China: levels, distribution, and possible sources.

Levels of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in sediments from the sandy flat system of Shuangtaizi Estuary, the highest-latitude estuary in China, were investigated to identify their possible sources and potential ecological risk. The concentrations of 28 PCBs and 18 OCPs ranged from 1.83 to 36.68 ng g(-1) dw (mean 10.53 ng g(-1) dw) and from 0.02 to 14.57 ng g(-1) dw (mean 5.65 ng g(-1) dw), respectively. Generally, these organic pollutants showed an obvious spatial distribution, and relatively high levels were found at the high-tidal zone near river mouths. Compositional analyses indicated that tetra-PCBs were dominant for PCBs, whereas heptachlor was identified to be prevalent for OCPs in surficial sediment in the sand flats of Shuangtaizi Estuary. Overall, Shuangtaizi Estuary had moderate PCB and OCP levels in the sand flat sediments and posed a low ecological hazard to aquatic biota. Our results indicated that the sediment PCBs came from nonpoint deposition, such as atmospheric contribution and river input, for light chlorinated congeners and point source deposition, such as the industrial sources along river flow, for highly chlorinated congeners, whereas OCPs originate mainly from old residuals and new usage of pesticides in agriculture and aquaculture.

Spatial distribution and sources of heavy metals and petroleum hydrocarbon in the sand flats of Shuangtaizi Estuary, Bohai Sea of China.

The concentrations of heavy metals and petroleum hydrocarbons (PHCs) in surface sediments were investigated in the sand flats of Shuangtaizi Estuary, Bohai Sea of China in May, 2013. Ecological risk assessment indicated that most heavy metals cause low ecological risk to the estuarine environment, with the exception of Cd and Hg (considerable and moderate risk, respectively). Principal component analysis in combination with correlation analysis among heavy metals, PHCs and geological factors (e.g., granularity) was used to identify possible sources of pollutants in Shuangtaizi Estuary. Results showed that the main pollution sources of the area come from anthropogenic factors, such as sewage discharge and oil exploitation.

Identification and expression analysis on bactericidal permeability-increasing protein (BPI)/lipopolysaccharide-binding protein (LBP) of ark shell, Scapharca broughtonii.

Bactericidal permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP) are the numbers of the lipid transfer protein/lipopolysaccharide-binding protein family and play crucial roles in the innate immune response to Gram-negative bacteria. A novel Sb-BPI/LBP1 from ark shell Scapharca broughtonii was isolated by expressed sequence tag (EST) and RACE techniques. The Sb-BPI/LBP1 cDNA encoded a polypeptide of 484 amino acids with a putative signal peptide of 21 amino acid residues and a mature protein of 463 amino acids. The deduced amino acid sequence of Sb-BPI/LBP1 contained an N-terminal BPI/LBP/CETP domain BPI1 with three functional regions that display LPS-binding activity, and a C-terminal BPI/LBP/CETP domain BPI2. In structure and sequence, Sb-BPI/LBP1 showed highly similar to those of the BPI/LBPs from invertebrate and non-mammalian vertebrate, the LBPs and BPIs from mammal. By quantitative real-time RT-PCR, Sb-BPI/LBP1 transcripts could be detected in all normal tested tissues, including hepatopancreas, adductor muscle, mantle margin, heart, gonad, gill and hemocytes, and was universally up-regulatable at 24 h post LPS challenge. The mRNA expression of Sb-BPI/LBP1 in hemocytes was the most sensitive to LPS challenge, significantly up-regulated at 12 h post LPS challenge and peaked at 24 h (16.76-fold, P < 0.05). These results suggested that Sb-BPI/LBP1 was a constitutive and inducible acute-phase protein contributing to the host immune defense against Gram-negative bacterial infection in ark shell S. broughtonii.

The impact of the herbicide atrazine on growth and photosynthesis of seagrass, Zostera marina (L.), seedlings.

The impact of the widely used herbicide atrazine on seedling growth and photosynthesis of eelgrass was determined. The long-term impact of the herbicide atrazine (1, 10 and 100 µg/L) on growth of eelgrass Zostera marina (L.) seedlings, maintained in outdoor aquaria, was monitored over 4 weeks. Exposure to 10 µg/L atrazine resulted in significantly lower plant fresh weight and total chlorophyll concentration and up to 86.67% mortality at the 100 µg/L concentration. Short-term photosynthetic stress on eelgrass seedlings was determined and compared with adult eelgrass using chlorophyll fluorescence. The effective quantum yield in eelgrass seedlings was significantly depressed at all atrazine concentrations (2, 4, 8, 16, 32 and 64 µg/L) even within 2 h and remained at a lower level than for adult plants for each concentration. These results indicate that atrazine presents a potential threat to seagrass seedling functioning and that the impact is much higher than for adult plants.