A nested high-resolution unstructured grid 3-D ocean-sea ice-ice shelf setup for numerical investigations of the Petermann ice shelf and fjord.

Three-dimensional numerical simulation of circulation in fjords hosting marine-terminating ice shelves is challenging because of the complexity of processes involved in such environments. This often requires a comprehensive model setup. The following elements are needed: bathymetry (usually unknown beneath the glacier tongue), ice shelf draft (impacting water column thickness), oceanographic state (including tidal elevation, salinity, temperature and velocity of the water masses), sea ice and atmospheric forcing. Moreover, a high spatial resolution is needed, at least locally, which may be augmented with a coarser and computationally cheaper (nested) model that provides sufficiently realistic conditions at the boundaries. Here, we describe procedures to systematically create such a setup that uses the Finite Volume Community Ocean Model (FVCOM) for the Petermann Fjord, Northwest Greenland. The first simulations are validated against temperature and salinity observations from the Petermann Fjord in September 2019. We provide•Complete bathymetry, ice-draft and water column thickness datasets of the Petermann Fjord, with an improved representation of the topography underneath the glacier tongue.•Boundary conditions for ocean, atmosphere and sea ice derived from a suite of high-resolution regional models that can be used to initialize and run the regional ocean model with realistic geophysical settings.

An ɑ2-adrenergic receptor is involved in larval metamorphosis in the mussel, Mytilus coruscus.

Metamorphosis is crucial in the life-cycle transition between the larval and juvenile stages of marine invertebrates. Although a number of agonists and antagonists of the adrenergic receptor (AR) are known to regulate larval metamorphosis in Mytilus coruscus (Mc), the molecular basis of the modulation of larval metamorphosis by the AR gene in this species remains elusive. Herein, the role of the AR gene in M. coruscus larval metamorphosis using the RNA interference technique was examined. The Mcα2AR transcript was observed to be present during the entire process of larval development and its level in the post-larvae was significantly increased compared to that in the pediveligers. Mcα2AR-knockdown resulted in a substantial reduction in the abundance of the Mcα2AR transcript and significantly inhibited the metamorphosis of M. coruscus larvae. These findings provide new insights into the molecular basis of modulation of larval metamorphosis in M. coruscus by the AR gene.

Ultrasensitive and Reversible Nanoplatform of Urinary Exosomes for Prostate Cancer Diagnosis.

Prostate cancer cell-derived exosomes in urine have been extensively studied recently and regarded as novel biomarkers for cancer diagnosis and prognosis, which presents wide prospects in clinical applications. Sensitive detection and specific capture methods are essential for exosomes analysis. Herein, a dual functional platform composed of superparamagnetic conjunctions and molecular beacons (SMC-MB) is reported. The SMC-MB platform is designed based on aptamer immunoaffinity with ultrasensitive detection efficiency and reversible isolation capacity, which, respectively, profit from nonenzymatic amplification methods and magnetic separation along with restriction cleavage. It is noteworthy that exosomes quantification was exactly amplified and transformed into single strand DNA detection. Correlated measurements evidence that the limit of detection of SMC-MB is as low as ∼100 particles/µL in urine, and a linear relationship meets between the logarithmic concentration of exosomes and fluorescence intensity of the molecular beacon. Furthermore, employing prostate specific membrane antigen (PSMA) aptamer, the platform adapted to detect and capture PMSA-positive exosomes from urine samples provides excellent diagnostic efficiency for prostate cancer (PCa). The expression of typical biomarkers of PCa, i.e., PSA and PCA3 mRNA, is significantly higher in PSMA-positive exosomes. Altogether, the platform and strategy described in this paper are promising in urinary exosomes analysis and prostate cancer detection.

Elucidation of the Molecular-Mechanisms and In Vivo Evaluation of the Anti-inflammatory Effect of Alginate-Derived Seleno-polymannuronate.

Alginate-derived polymannuronate (PM) is a type of polysaccharide found in edible brown seaweeds. Seleno-polymannuronate (Se-PM) was prepared from PM via synthesis using sulfation- and selenation-replacement reactions. The anti-inflammatory activity of Se-PM and its corresponding molecular mechanisms were investigated. In lipopolysaccharide (LPS)-activated murine macrophage RAW264.7 cells, Se-PM significantly attenuated the production of nitric oxide (NO), prostaglandin E2 (PGE2), and reactive oxygen species (ROS); the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2); and the secretion of proinflammatory cytokines. Moreover, Se-PM remarkably suppressed the LPS-induced activation of the nuclear-factor (NF)-κB and mitogen-activated-protein-kinase (MAPK) signaling pathways in RAW264.7 cells. Furthermore, Se-PM also decreased the production of proinflammatory mediators in LPS-triggered primary murine macrophages. Additionally, Se-PM inhibited the inflammatory response in the air-pouch inflammation model. These results might contribute to the overall understanding of the potential health benefits of Se-PM for food and drug applications.

Cloning and characterization of PHGPx and its synergistic role with p53 in mediating stress in Penaeus monodon.

Phospholipid-hydroperoxide glutathione peroxidase (PHGPx), a ubiquitous antioxidant enzyme in the glutathione peroxidase (GPx) family, plays multiple roles in different organisms. Here, a novel PHGPx (PmPHGPx) was identified from Penaeus monodon. The full-length PmPHGPx cDNA was 1885 bp in length with a 489-bp open reading frame (ORF) containing a selenocysteine codon, TGA177-179, and a selenocysteine insertion sequence in the 3'-UTR. The typical signature motifs of the GPx family were also detected in the PmPHGPx amino acid sequence. The PmPHGPx expression pattern showed tissue-specific variations, with the highest expression level in the heart and the lowest expression level in the muscle. To examine the relationship between Pmp53 and PmPHGPx, Pmp53 was successfully silenced with a dsRNA-p53 injection, and an obvious down-regulation in PmPHGPx expression was apparent. To clarify the functional roles of Pmp53 and PmPHGPx, their expression patterns were also assessed after pH-induced stress, salinity stress and heavy metal (Cu, Zn, and Cd) challenges. Similar trends in the expression profiles for PmPHGPx and Pmp53 were detected in both the gills and hepatopancreas in response to all stressors. Therefore, we conclude from the results that PmPHGPx acts synergistically and subsequently works cooperatively with Pmp53 toward mediating cell stress. This study improves our understanding of PmPHGPx and its synergistic role with Pmp53 in counteracting stressors in P. monodon.

Alginate enhances Toll-like receptor 4-mediated phagocytosis by murine RAW264.7 macrophages.

Alginate is a naturally acidic polysaccharide consisting alternately of ß-d-mannuronic acid and α-l-guluronic acid with 1, 4-glycosidic linkages and is derived from brown seaweeds. Herein, the effect of alginate on the promotion of macrophage phagocytosis and the corresponding molecular mechanisms were investigated in murine RAW264.7 cells. Alginate could enhance the intracellular phagocytosis of gold nanoparticles (AuNPs), fluorescent microspheres and immunoglobulin G (IgG)-opsonized Staphylococcus aureus (S. aureus). Moreover, alginate increased Toll-like receptor 4 (TLR4) expression and activated the Akt/nuclear factor-κB (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signalling pathways. Alginate-promoted phagocytosis was suppressed by the addition of inhibitors of TLR4, NF-κB and p38 MAPK and by TLR4 gene knockdown, indicating the involvement of these key components. This work is the first to propose that alginate promotes phagocytosis via upregulating TLR4 expression and stimulating the Akt/NF-κB and p38 MAPK signalling pathways, which may contribute to the capacity of alginate to activate macrophages.

Gene characteristics, immune and stress responses of PmPrx1 in black tiger shrimp (Penaeus monodon): Insights from exposure to pathogenic bacteria and toxic environmental stressors.

Peroxiredoxins (Prxs) are ubiquitous, multifunctional and evolutionarily conserved enzymes that can protect cells from oxidative damage caused by ROS and play a vital role in immune responses. Here, a full-length Prx1 cDNA sequence (PmPrx1) was isolated from Penaeus monodon. The PmPrx1 cDNA was 951 base pairs (bp), encoding 198 amino acid polypeptides. The results of qRT-PCR showed that the PmPrx1 mRNA was ubiquitously expressed in all tissues tested and had a comparatively high expression level in immune-associated tissues (gill, hepatopancreas). To explore the immune and anti-stress roles of PmPrx1, the gills and hepatopancreas were chosen as target tissues in Penaeus monodon and were challenged with bacteria (Vibrio harveyi and Streptococcus agalactiae) and toxic environmental stresses. To further clarify the immune function of PmPrx1 after bacterial challenge, the recombinant PmPrx1 protein was acquired using a prokaryotic expression method. The antioxidant activity of the recombinant PmPrx1 was assessed by the catalyzing hydrogen peroxide assay method, and the results showed obvious antioxidant activity in a dose-dependent and temperature-dependent manner. The antimicrobial activity of purified PmPrx1 protein was evaluated and further studied in vitro relying on a bacterial growth inhibition test which was conducted in both liquid and solid cultures. Furthermore, E. coli transferred with pRSET-PmPrx1 was dramatically protected in response to metal toxicity and H2O2 oxidative stress. In summary, this study provides useful information about the role of the Prx1 gene in defense against a variety of toxic factors in shrimps that help to further clarify the functional mechanism of Prx.

Silver Nanoparticles Impact Biofilm Communities and Mussel Settlement.

Silver nanoparticles (AgNPs) demonstrating good antimicrobial activity are widely used in many fields. However, the impact of AgNPs on the community structures of marine biofilms that drive biogeochemical cycling processes and the recruitment of marine invertebrate larvae remains unknown. Here, we employed MiSeq sequencing technology to evaluate the bacterial communities of 28-day-old marine biofilms formed on glass, polydimethylsiloxane (PDMS), and PDMS filled with AgNPs and subsequently tested the influence of these marine biofilms on plantigrade settlement by the mussel Mytilus coruscus. AgNP-filled PDMS significantly reduced the dry weight and bacterial density of biofilms compared with the glass and PDMS controls. AgNP incorporation impacted bacterial communities by reducing the relative abundance of Flavobacteriaceae (phylum: Bacteroidetes) and increasing the relative abundance of Vibrionaceae (phylum: Proteobacteria) in 28-day-old biofilms compared to PDMS. The settlement rate of M. coruscus on 28-day-old biofilms developed on AgNPs was lower by >30% compared to settlement on control biofilms. Thus, the incorporation of AgNPs influences biofilm bacterial communities in the marine environment and subsequently inhibits mussel settlement.

Molecular cloning, characterization and expression analysis of Toll-like receptor 5M gene in Japanese sea perch (Lateolabrax japonicas) after bacterial infection.

Toll-like receptor 5M belongs to Toll-like receptors (TLRs) family, which plays a crucial role in innate immunity due to its important role in the recognition of bacteria invasion and in the activation of immune related pathways downstream. In the present study, we firstly cloned the full-length cDNAs of TLR 5M (LjTLR 5M) from Japanese sea perch (Lateolabrax japonicas). The full-length cDNAs of LjTLR 5M include an open reading frame (ORF) of 2676 bp encoding a polypeptide of 891 amino acid residues. The deduced amino acid sequence analysis showed that LiTLR 5M contains LRRs (extracellular leucine rich repeats), transmembrane and TIR (Toll/interleukin-1 receptor) domain. Transcriptional expression analysis indicated that LiTLR 5M mRNAs were ubiquitously expressed in wide array of tissues and the peak level was observed in the head-kidney. The expression patterns of LjTLR 5M after Vibro harveyi and Streptococus agalactiae infection were detected by qRT-PCR, and the results showed that LjTLR 5M was significant up-regulated in spleen, liver and head-kidney. Additionally, the expression patterns of LjTLR 5M in infected spleen and head-kidney were further validated by in situ hybridization (ISH). In summary, these findings indicate that LjTLR 5M is significant induced after different bacterial infection and is involved in immune response. Furthermore, this study will provide foundational information for other TLRs research of L. japonicas against different bacterial pathogens invasion.

The effect of carbon nanotubes and titanium dioxide incorporated in PDMS on biofilm community composition and subsequent mussel plantigrade settlement.

This study investigated the effect of carbon nanotubes (CNTs) and titanium dioxide (TiO2) incorporated in PDMS on biofilm formation and plantigrade settlement of Mytilus coruscus. TiO2 increased bacterial density, and CNTs also increased bacterial density but reduced diatom density in biofilms after 28 days. Further analysis was conducted between bacterial communities on glass, PDMS, CNTs (0.5 wt%) and TiO2 (7.5 wt%). ANOSIM analysis revealed significant differences (R > 0.9) between seven, 14, 21 and 28 day-old bacterial communities. MiSeq sequencing showed that CNTs and TiO2 impacted the composition of 28 day-old bacterial communities by increasing the abundance of Proteobacteria and decreasing the abundance of Bacteroidetes. The maximum decreased settlement rate in 28 day-old biofilms on CNTs and TiO2 was > 50% in comparison to those on glass and PDMS. Thus, CNTs and TiO2 incorporated in PDMS altered the biomass and community composition of biofilms, and subsequently decreased mussel settlement.

Draft Genome Sequence of Pseudoalteromonas sp. Strain ECSMB14103, Isolated from the East China Sea.

Pseudoalteromonas sp. strain ECSMB14103 was isolated from marine biofilms formed on the East China Sea. The draft genome sequence comprises 4.11 Mp with a G+C content of 39.7%. The information from the draft genome will contribute to an understanding of bacteria-animal interaction.

[Anti-diatom compounds from marine bacterium Pseudomonas putida].

OBJECTIVE: In order to provide more natural antifouling compounds, marine bacterium Pseudomonas putida isolated from the sponge Haliclona sp. was explored to test its anti-diatom compounds. METHODS: The strain was identified by colonial morphology, scanning electron microscope (SEM) and 16S rDNA sequence analysis. The separation procedure was guided by bioactive (Anti-diatom) and chemical (TLC, DAD-HPLCand 1H NMR) analysis, and their structures were elucidated by spectrographic techniques. The anti-diatom activity of all purified compounds was assayed. RESULTS: Strain 272 isolated from the sponge Haliclona sp. was identified as Pseudomonas putida. Six diketopiperazine compounds were isolated from the culture of this strain and their structures were determined as cyclo(Leu-Pro) (1), cyclo (Leu-Ala) (2), cyclo(Phe-Ala) (3), cyclo(Val-Tyr) (4), cyclo(Ala-Tyr) (5), cyclo(Ala-Trp) (6); Compounds 3 and 6 displayed significant anti-diatom activity with the inhibitory rate of 50% and 85% at the concentration of 50 microg/mL, respectively. CONCLUSION: The anti-diatom compounds isolated from marine bacterium Pseudomonas putida were cyclo (Phe-Ala) and cyclo (Ala-Trp).

Larval settlement and metamorphosis of the mussel Mytilus coruscus in response to monospecific bacterial biofilms.

The effects of bacterial biofilms (BFs) on larval settlement and metamorphosis of the mussel, Mytilus coruscus, were investigated in the laboratory. Of nine different isolates, Shewanella sp.1 BF induced the highest percentage of larval settlement and metamorphosis, whereas seven other isolates had a moderate inducing activity and one isolate, Pseudoalteromonas sp. 4, had a no inducing activity. The inducing activity of individual bacterial isolates was not correlated either with their phylogenetic relationship or with the surfaces from which they were isolated. Among the eight bacterial species that demonstrated inducing activity, bacterial density was significantly correlated with the inducing activity for each strain, with the exception of Vibrio sp. 1. The Shewanella sp. 1 BF cue that was responsible for inducing larval settlement and metamorphosis was further investigated. Treatment of the BFs with formalin, antibiotics, ultraviolet irradiation, heat, and ethanol resulted in a significant decrease in their inducing activities and cell survival. BF-conditioned water (CW) did not induce larval metamorphosis, but it triggered larval settlement behavior. A synergistic effect of CW with formalin-fixed Shewanella sp. 1 BF significantly promoted larval metamorphosis. Thus, a cocktail of chemical cues derived from bacteria may be necessary to stimulate larval settlement and metamorphosis in this species.

Larval settlement and metamorphosis of the mussel Mytilus coruscus in response to natural biofilms.

Settlement and metamorphosis of pediveliger larvae of Mytilus coruscus in response to natural biofilms was investigated in the laboratory. Pediveliger larvae settled and metamorphosed in response to biofilms and post-larval settlement and metamorphosis increased with biofilm age. The activity of the biofilm was positively correlated with biofilm age, dry weight, bacterial density and diatom density, but had no apparent relationship with chlorophyll a concentration. The change in bacterial community composition corresponding to biofilm age may explain differences in the age-dependent inducing activities of biofilms, which in turn may play an important role in larval settlement in this species.

Larval metamorphosis of the mussel Mytilus galloprovincialis Lamarck, 1819 in response to neurotransmitter blockers and tetraethylammonium.

The metamorphic response of pediveliger larvae of Mytilus galloprovincialis to the neurotransmitter blockers chlorpromazine, amitriptyline, rauwolscine, idazoxan, atenolol and butoxamine, and to tetraethylammonium chloride (TEA) was investigated through a series of bioassays. Chlorpromazine, amitriptyline and idazoxin inhibited larval metamorphosis induced by 10⁻4 M epinephrine. The concentration that inhibited metamorphosis by 50% (IC50) for chlorpromazine and amitriptyline was 1.6 x 10⁻6 M and 6.6 x 10⁻5 M, respectively. Idazoxan was less effective with an IC50 of 4.4 x 10¹³ M. Moreover, these three inhibitors showed no toxicity at any of the concentrations tested. The larval metamorphic response to K+ was not inhibited by 10⁻³ M tetraethylammonium chloride after 96 h. Thus, the neurotransmitter blockers chlorpromazine and amitriptyline are inhibitors of larval metamorphosis, and will be useful tools for antifouling studies.

Copper affects biofilm inductiveness to larval settlement of the serpulid polychaete Hydroides elegans (Haswell).

Copper (Cu) contamination is a potential threat to the marine environment due to the use of Cu-based antifouling paints. Cu stress on larval settlement of the polychaete Hydroides elegans was investigated, and this was linked to Cu stress on biofilms and on the biofilm development process. The inductiveness of young biofilms was more easily altered by Cu stress than that of old biofilms, indicating the relative vulnerability of young biofilms. This might result from changes in bacterial survival, the bacterial community composition and the chemical profiles of young biofilms. Cu also affected biofilm development and the chemical high performance liquid chromatograph fingerprint profile. The results indicate that Cu affected larval settlement mainly through its effect on the process of biofilm development in the marine environment, and the chemical profile was crucial to biofilm inductiveness. It is strongly recommended that the effects of environmentally toxic substances on biofilms are evaluated in ecotoxicity bioassays using larval settlement of invertebrates as the end point.

Induction of metamorphosis of pediveliger larvae of the mussel Mytilus galloprovincialis Lamarck, 1819 using neuroactive compounds, KCl, NH4Cl and organic solvents.

Pediveliger larvae of Mytilus galloprovincialis were subjected to a series of bioassays to investigate the induction of metamorphosis using neuroactive compounds, K(+), NH(4)(+) and organic solvents. Growth and survival of post-larvae obtained using ethanol and methanol were also observed. Epinephrine, phenylephrine, clonidine and metanephrine induced larval metamorphosis at 10(-6) to 10(-4) M in both 24-h and continuous exposure assays. In 24-h exposure assays, alpha-methyldopa at 5 x 10(-5) M and methoxyphenamine at 5 x 10(-5)-10(-4) M induced 55-94% metamorphosis. Similarly, excess K(+) at 3 x 10(-2) M induced 39% metamorphosis and NH(4)(+) at 1-5 x 10(-2) M induced 63-78% metamorphosis. The EC50s of seven organic solvents ranged from 0.04 to 0.82 M. Post-larvae that metamorphosed using ethanol and methanol survived as juveniles and grew at the same rate as those from microbial biofilm. Thus, the above compounds can be useful inducers of metamorphosis for antifouling studies using larvae and juveniles of M. galloprovincialis.

Survival, growth, settlement and metamorphosis of refrigerated larvae of the mussel Mytilus galloprovincialis Lamarck and their use in settlement and antifouling bioassays.

Straight-hinge veliger and pediveliger larvae of the mussel Mytilus galloprovincialis were refrigerated for varying periods for use in bioassays. Straight-hinge veliger larvae grew to the umbo-veliger stage after 2 months in the refrigerator, but no pediveligers were observed during the 3-month refrigeration period. The average survival rate of larvae in the refrigerator was 79% after 1 month, but gradually decreased with the refrigeration period, and was as low as 22% after 3 months. All refrigerated larvae grew to the pediveliger stage in the incubator at 17 degrees C at the same rate as that of the control larvae that were not refrigerated. Settlement and metamorphosis of pediveligers from both refrigerated and control groups were facilitated by microbial film and epinephrine and inhibited by phentolamine. Thus, refrigeration can be used as an effective method of storing larvae of M. galloprovincialis for use in assays to assess candidate settlement inducers and antifouling substances.