Effect of Suspended Particulate Matter on the Accumulation of Dissolved Diarrhetic Shellfish Toxins by Mussels (Mytilus galloprovincialis) under Laboratory Conditions.

In recent years, detection of trace amounts of dissolved lipophilic phycotoxins in coastal waters has been possible using solid phase adsorption toxin tracking (SPATT) samplers. To explore the contribution of dissolved diarrhetic shellfish toxins (DST) to the accumulation of toxins by cultivated bivalves, mussels (Mytilus galloprovincialis) were exposed to different concentrations of purified okadaic acid (OA) and dinophysistoxin-1 (DTX1) in filtered (0.45 µm) seawater for 96 h. Accumulation and esterification of DST by mussels under different experimental conditions, including with and without the addition of the food microalga Isochrysis galbana, and with the addition of different size-fractions of suspended particulate matter (SPM) (<75 µm, 75⁻150 µm, 150⁻250 µm) were compared. Results showed that mussels accumulated similar amounts of OA and DTX1 from seawater with or without food microalgae present, and slightly lower amounts when SPM particles were added. Mussels preferentially accumulated OA over DTX1 in all treatments. The efficiency of the mussel’s accumulation of OA and DTX1 from seawater spiked with low concentrations of toxins was higher than that in seawater with high toxin levels. A large proportion of OA (86⁻94%) and DTX1 (65⁻82%) was esterified to DTX3 by mussels in all treatments. The proportion of I. galbana cells cleared by mussels was markedly inhibited by dissolved OA and DTX1 (OA 9.2 µg L−1, DTX1 13.2 µg L−1) in seawater. Distribution of total OA and DTX1 accumulated in the mussel tissues ranked in all treatments as follows: digestive gland > gills > mantle > residual tissues. However, the percentage of total DST in the digestive gland of mussels in filtered seawater (67%) was higher than with the addition of SPM particles (75⁻150 µm) (51%), whereas the gills showed the opposite trend in filtered seawater with (27%) and without (14.4%) SPM particles. Results presented here will improve our understanding of the mechanisms of DST accumulation by bivalves in marine aquaculture environments.

Ubiquity of the neurotoxin ß-N-methylamino-L-alanine and its isomers confirmed by two different mass spectrometric methods in diverse marine mollusks.

In recent years, the neurotoxin ß-N-methylamino-L-alanine (BMAA) has been reported in some marine mollusk species. To further discover BMAA in marine animals, a total of 59 samples belonging to 3 phyla, 22 families, and 43 species, were collected from Dalian, Rongcheng, and Zhoushan cities, China, in April 2017. All samples were quantified by a hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) analysis of underivatized extract, and ten samples were also analyzed by a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis using a precolumn AQC (6-aminoquinolyl-N-hydroxysuccinimidyl carbamate)-derivatization method. Results indicated that 48 mollusk samples contained BMAA with concentrations above the limit of detection (0.31 µg g-1 wet weight), and the isomers of BMAA, ß-amino-N-methylalanine (BAMA) and 2,4-diaminobutyric acid (DAB) were universally present in most samples. However, N-(2-aminoethyl) glycine (AEG) was not found in any sample. Comparison of both analytical methods showed that BMAA and BAMA were not completely separated by the HILIC column although they still could be identified by specific transitions. In contrast the C18 column provided good separation for the AQC-derivatives of BMAA and all of its isomers. Development of analytical methods and stable isotope tracing of BMAA should be carried out in the future.

Four mononuclear platinum(II) complexes: synthesis, DNA/BSA binding, DNA cleavage and cytotoxicity.

Four new platinum(II) complexes: PtII L1·H2O (C1, H2 L1 = C20H16N2O2), PtII L2Cl2 (C2, L2 = C22H16N2O2), PtII L3Cl2·H2O (C3, L3 = C20H16N2), PtII L4Cl2·0.4H2O (C4, L4 = C18H14N4) have been synthesized and characterized by using various physico-chemical techniques. The binding interaction of the four platinum(II) complexes C1-C4 with calf thymus (CT)-DNA has been investigated by UV-Vis and fluorescence emission spectrometry. The apparent binding constant (K app) values follow the order: C3 > C1 > C2 > C4. In addition, fluorescence spectrometry of bovine serum albumin (BSA) with the four platinum(II) complexes C1-C4 showed that the quenching mechanism might be a static quenching procedure. For C1-C4, the number of binding sites was about one for BSA and the binding constants follow the order: C3 (7.08 × 105M-1) > C1 (2.82 × 105M-1) > C2 (0.85 × 105M-1) > C4 (0.15 × 105M-1). With the single condition change such as absence of an external agent, the DNA cleavage abilities of C3 exhibit remarkable changes. In addition, the cytotoxicity of C3 in vitro on tumor cells lines (MCF-7, HepG2 and HT29) were examined by MTT and showed better antitumor effects on the tested cells.

A highly selective and sensitive turn-on fluorescent probe for the detection of Al3+ and its bioimaging.

A novel fluorescent sensor, 1-((2-hydroxynaphthalen-1-yl)methylene)urea (ocn) has been designed and applied as a highly selective and sensitive fluorescent probe for recognition of Al3+ in Tris-HCl (pH = 7.20) solution. The probe ocn exhibits an excellent selectivity to Al3+ over other examined metal ions, anions and amino acids with a prominent fluorescence 'turn-on' at 438 nm. ocn binds to Al3+ with a 2:1 binding stoichiometry and the detection limit was 0.3 µM. Furthermore, its capability of biological application was evaluated and the results showed that the sensor could be used to detect Al3+ in living cells.

New Typical Vector of Neurotoxin ß-N-Methylamino-l-Alanine (BMAA) in the Marine Benthic Ecosystem.

The neurotoxin ß-N-methylamino-l-alanine (BMAA) has been identified as an environmental factor triggering neurodegenerative diseases such as Amyotrophic Lateral Sclerosis (ALS) and Alzheimer's disease (AD). We investigated the possible vectors of BMAA and its isomers 2,4-diaminobutyric acid (DAB) and N-2(aminoethyl)glycine (AEG) in marine mollusks collected from the Chinese coast. Sixty-eight samples of marine mollusks were collected along the Chinese coast in 2016, and were analyzed by an HILIC-MS/MS (hydrophilic interaction liquid chromatography with tandem quadrupole mass spectrometer) method without derivatization. BMAA was detected in a total of five samples from three species: Neverita didyma, Solen strictus, and Mytilus coruscus. The top three concentrations of free-form BMAA (0.99~3.97 µg·g-1 wet weight) were detected in N. didyma. DAB was universally detected in most of the mollusk samples (53/68) with no species-specific or regional differences (0.051~2.65 µg·g-1 wet weight). No AEG was detected in any mollusk samples tested here. The results indicate that the gastropod N. didyma might be an important vector of the neurotoxin BMAA in the Chinese marine ecosystem. The neurotoxin DAB was universally present in marine bivalve and gastropod mollusks. Since N. didyma is consumed by humans, we suggest that the origin and risk of BMAA and DAB toxins in the marine ecosystem should be further investigated in the future.

A humanized anti-VEGF rabbit monoclonal antibody inhibits angiogenesis and blocks tumor growth in xenograft models.

Rabbit antibodies have been widely used in research and diagnostics due to their high antigen specificity and affinity. Though these properties are also highly desirable for therapeutic applications, rabbit antibodies have remained untapped for human disease therapy. To evaluate the therapeutic potential of rabbit monoclonal antibodies (RabMAbs), we generated a panel of neutralizing RabMAbs against human vascular endothelial growth factor-A (VEGF). These neutralizing RabMAbs are specific to VEGF and do not cross-react to other members of the VEGF protein family. Guided by sequence and lineage analysis of a panel of neutralizing RabMAbs, we humanized the lead candidate by substituting non-critical residues with human residues within both the frameworks and the CDR regions. We showed that the humanized RabMAb retained its parental biological properties and showed potent inhibition of the growth of H460 lung carcinoma and A673 rhabdomyosarcoma xenografts in mice. These studies provide proof of principle for the feasibility of developing humanized RabMAbs as therapeutics.