Effect of carboxymethyl chitosan on the detoxification and biotransformation of paralytic shellfish toxins in oyster Ostrea rivularis.

Economic bivalve ingested toxic algae causes frequent human poisoning events. To explore new compounds that can accelerate the depuration of toxins in shellfish, we investigated the detoxification of the paralytic shellfish toxins (PSTs) and the biotransformation pathway of PSTs during detoxification by the application of three treatments to a toxic bloom, Alexandrium minutum (A. minutum). The detoxification effect of Platymonas subcordiformis (PS) mixed with carboxymethyl chitosan (CMC) group is significantly better than the starving group in each oyster tissues. The toxicity of viscera which occupied 78.95% of total toxicity reduced to 155 MU/100g after 13 days' depuration experiment. And adding CMC could significantly achieve rapid detoxification and effectively reduce the STX to 0.07 µmol/100 g in viscera. Meanwhile, PSTs underwent biotransformation during the depuration process, which mainly manifested as GTX1/4→GTX2/3→STX, GTX2→dcGTX2. This study explored a new strategy for toxin depuration in shellfish.

Biocompatibility of Alexandrium minutum in the growth and paralytic shellfish toxins production under five cultivation methods.

This study evaluated the performance of an easy-cultivation device for the mass culture of Alexandrium minutum (A. minutum), a dinoflagellate that produces paralytic shellfish toxins (PSTs). Five culture conditions including three different sizes of containers (250 mL conical flask, 500 mL beaker, and 20 L jar) in two different environments (out-incubator and incubator) were compared in terms of growth and PSTs production. Compared with the incubator environment, the out-incubator environment had more fluctuations in temperature and light intensity. Results showed that the cell densities of A. minutum increased in all groups, especially in the conical flask (I, out-incubator, 6.29×106 cells/mL) and the beaker (IV, incubator, 7.28×106 cells/mL). When cultured in the 20 L jar under out-incubator condition, the algae had the lowest cell density (2.82×106 cells/mL) but the highest toxicity (93.42 ± 2.55×10-6 MU/cell). The negative correlation between average growth rate and single-cell toxicity could be explained by biocompatibility, thereby indicating that the low growth rate led to high toxicity. HPLC-FLD showed that the cellular toxicity increased due to the quantitative increase in GTX1/4, which are the more toxic derivatives. The PSTs types consistently contained GTX1/4 and GTX2/3. The differences in algae growth and toxin-production could be due to changes in bacteria (out-incubator) and CO2 (incubator) with vessel size. The effects of environmental factors on algae are strain specific. The out-incubator device can be applied for large-scale cultivation of A. minutum considering the algae density and toxin-producing ability.

15N Stable Isotope Labeling PSTs in Alexandrium minutum for Application of PSTs as Biomarker.

The dinoflagellate Alexandrium minutum (A. minutum) which can produce paralyticshellfish toxins (PSTs) is often used as a model to study the migration, biotransformation,accumulation, and removal of PSTs. However, the mechanism is still unclear. To provide a new toolfor related studies, we tried to label PSTs metabolically with 15N stable isotope to obtain 15N-PSTsinstead of original 14N, which could be treated as biomarker on PSTs metabolism. We then culturedthe A. minutum AGY-H46 which produces toxins GTX1-4 in f/2 medium of different 15N/Pconcentrations. The 15N-PSTs' toxicity and toxin profile were detected. Meanwhile, the 15N labelingabundance and 15N atom number of 15N-PSTs were identified. The 14N of PSTs produced by A.minutum can be successfully replaced by 15N, and the f/2 medium of standard 15N/P concentrationwas the best choice in terms of the species' growth, PST profile, 15N labeling result and experimentcost. After many (>15) generations, the 15N abundance in PSTs extract reached 82.36%, and the 15Natom number introduced into GTX1-4 might be 4⁻6. This paper innovatively provided the initialevidence that 15N isotope application of labeling PSTs in A. minutum is feasible. The 15N-PSTs asbiomarker can be applied and provide further information on PSTs metabolism.

Inhibition of 5-HT(3) receptors-activated currents by cannabinoids in rat trigeminal ganglion neurons.

This study investigated the modulatory effect of synthetic cannabinoids WIN55,212-2 on 5-HT(3) receptor-activated currents (I(5-HT3)) in cultured rat trigeminal ganglion (TG) neurons using whole-cell patch clamp technique. The results showed that: (1) The majority of examined neurons (78.70%) were sensitive to 5-HT (3-300 µmol/L). 5-HT induced inward currents in a concentration-dependent manner and the currents were blocked by ICS 205-930 (1 µmol/L), a selective antagonist of the 5-HT(3) receptor; (2) Pre-application of WIN55,212-2 (0.01-1 µmol/L) significantly inhibited I(5-HT3) reversibly in concentration-dependent and voltage-independent manners. The concentration-response curve of 5-HT(3) receptor was shifted downward by WIN55,212-2 without any change of the threshold value. The EC(50) values of two curves were very close (17.5±4.5) µmol/L vs. (15.2±4.5) µmol/L and WIN55,212-2 decreased the maximal amplitude of I(5-HT3) by (48.65±4.15)%; (3) Neither AM281, a selective CB1 receptor antagonist, nor AM630, a selective CB2 receptor antagonist reversed the inhibition of I(5-HT3) by WIN55,212-2; (4) When WIN55,212-2 was given from 15 to 120 s before 5-HT application, inhibitory effect was gradually increased and the maximal inhibition took place at 90 s, and the inhibition remained at the same level after 90 s. We are led to concluded that-WIN55,212-2 inhibited I(5-HT3) significantly and neither CB1 receptor antagonist nor CB2 receptor antagonist could reverse the inhibition of I(5-HT3) by WIN55,212-2. Moreover, WIN55,212-2 is not an open channel blocker (OCB) of 5-HT(3) receptor. WIN55,212-2 significantly inhibited 5-HT-activated currents in a non-competitive manner. The inhibition of I(5-HT3) by WIN55,212-2 is probably new one of peripheral analgesic mechanisms of WIN55,212-2, but the mechanism by which WIN55,212-2 inhibits I(5-HT3) warrants further investigation.

Inhibition of 5-HT3 Receptors-activated Currents by Cannabinoids in Rat Trigeminal Ganglion Neurons / 华中科技大学学报（医学）（英德文版）

This study investigated the modulatory effect of synthetic cannbinoids WIN55,212-2 on 5-HT3 receptor-activated currents (I5-Hr3) in cultured rat trigeminal ganglion (TG) neurons using whole-cell patch clamp technique.The results showed that:(!) The majority of examined neurons (78.70％) were sensitive to 5-HT (3-300 μmol/L).5-HT induced inward currents in a concentrationdependent manner and the currents were blocked by ICS 205-930 (1 μmol/L),a selective antagonist of the 5-HT3 receptor; (2) Pre-application of WIN55,212-2 (0.01-1 μmol/L) significantly inhibited I5-HT3 reversibly in concentration-dependent and voltage-independent manners.The concentration-response curve of 5-HT3 receptor was shifted downward by WIN55,212-2 without any change of the threshold value.The EC50values of two curves were very close (17.5±4.5) μmol/L vs.(15.2±4.5) μmol/L and WIN55,212-2 decreased the maximal amplitude of I5-HI3 by (48.65±4.15)％; (3) Neither AM281,a selective CBI receptor antagonist,nor AM630,a selective CB2 receptor antagonist reversed the inhibition of I5-HT3by WIN55,212-2; (4) When WIN55,212-2 was given from 15 to 120 s before 5-HT application,inhibitory effect was gradually increased and the maximal inhibition took place at 90 s,and the inhibition remained at the same level after 90 s.We are led to concluded thatWIN55,212-2 inhibited I5-Hr3 significantly and neither CB1 receptor antagonist nor CB2 receptor antagonist could reverse the inhibition of I5-HT3 by WIN55,212-2.Moreover,WIN55,212-2 is not an open channel blocker (OCB) of 5-HT3 receptor.WIN55,212-2 significantly inhibited 5-HT-activated currents in a non-competitive manner.The inhibition of I5-HT3 by WIN55,212-2 is probably new one of peripheral analgesic mechanisms of WIN55,212-2,but the mechanism by which WIN55,212-2 inhibits I5-HT3 warrants further investigation.