Exposure to the mixotrophic dinoflagellate Lepidodinium sp. and its cues increase toxin production of Pseudo-nitzschia multiseries.

The present study examined the defense responses of toxigenic Pseudo-nitzschia species (P. multiseries) to a mixotrophic dinoflagellate, Lepidodinium sp., and its associated cues. We evaluated their responses to different predation risks, including direct physical contact and indirect interactions facilitated by cues from Lepidodinium sp. during active feeding on heterospecific prey (Rhodonomas salina), limited feeding on conspecific prey (P. multiseries) and non-feeding (autotrophic growth in f/2 medium) states. This study is the first investigation of these trophic interactions. Our results demonstrated a significant increase in cellular domoic acid (cDA) in P. multiseries when exposed to Lepidodinium sp. and its associated cues, which was 1.38 to 2.42 times higher than the non-induced group. Notably, this increase was observed regardless of Lepidodinium sp. feeding on this toxic diatom and nutritional modes. However, the most significant increase occurred when they directly interacted. These findings suggest that P. multiseries evaluates predation risk and increases cDA production as a defensive strategy against potential grazing threats. No morphological changes were observed in P. multiseries in response to Lepidodinium sp. or its cues. P. multiseries cultured in flasks of Group L+P-P showed a decrease in growth, but Group L-P and Group L+R-P did not exhibit any decrease. These results suggest a lack of consistent trade-offs between the defense response and growth, thus an increase in cDA production may be a sustainable and efficient defense strategy for P. multiseries. Furthermore, our findings indicate that P. multiseries had no significant impact on the fitness (cell size, growth and/or grazing) of Lepidodinium sp. and R. salina, which suggests no evident toxic or allelopathic impacts on these two phytoplankton species. This study enhances our understanding of the trophic interactions between toxic diatoms and mixotrophic dinoflagellates and helps elucidate the dynamics of Harmful Algal Blooms, toxin transmission, and their impact on ecosystem health.

Two-Trait Predictor of Venous Invasion on Contrast-Enhanced CT as a Preoperative Predictor of Outcomes for Early-Stage Hepatocellular Carcinoma After Hepatectomy.

OBJECTIVES: This study aimed to assess the effectiveness of the two-trait predictor of venous invasion (TTPVI) on contrast-enhanced computed tomography (CECT) for the preoperative prediction of clinical outcomes in patients with early-stage hepatocellular carcinoma (HCC) after hepatectomy. METHODS: This retrospective study included 280 patients with surgically resected HCC who underwent preoperative CECT between 2012 and 2013. CT imaging features of HCC were assessed, and univariate and multivariate Cox regression analyses were used to evaluate the CT features associated with disease-free survival (DFS) and overall survival (OS). Subgroup analyses were used to summarized the hazard ratios (HRs) between patients in whom TTPVI was present and those in whom TTPVI was absent using a forest plot. RESULTS: Capsule appearance [HR, 0.504; 95% confidence interval (CI), 0.341-0.745; p < 0.001], TTPVI (HR, 1.842; 95% CI, 1.319-2.572; p < 0.001) and high level of alanine aminotransferase (HR, 1.620; 95% CI, 1.180-2.225, p = 0.003) were independent risk factors for DFS, and TTPVI (HR, 2.509; 95% CI, 1.518-4.147; p < 0.001), high level of alpha-fetoprotein (HR, 1.722; 95% CI, 1.067-2.788; p = 0.026), and gamma-glutamyl transpeptidase (HR, 1.787; 95% CI, 1.134-2.814; p = 0.026) were independent risk factors for OS. A forest plot revealed that the TTPVI present group had lower DFS and OS rates in most subgroups. Patients in whom TTPVI was present in stages I and II had a lower DFS and OS than those in whom TTPVI was absent. Moreover, there were significant differences in DFS (p < 0.001) and OS (p < 0.001) between patients classified as Barcelona Clinic Liver Cancer stage A in whom TTPVI was absent and in whom TTPVI was present. CONCLUSIONS: TTPVI may be used as a preoperative biomarker for predicting postoperative outcomes for patients with early-stage HCC.

TRPP2 and STIM1 form a microdomain to regulate store-operated Ca2+ entry and blood vessel tone.

BACKGROUND: Polycystin-2 (TRPP2) is a Ca2+ permeable nonselective cationic channel essential for maintaining physiological function in live cells. Stromal interaction molecule 1 (STIM1) is an important Ca2+ sensor in store-operated Ca2+ entry (SOCE). Both TRPP2 and STIM1 are expressed in endoplasmic reticular membrane and participate in Ca2+ signaling, suggesting a physical interaction and functional synergism. METHODS: We performed co-localization, co-immunoprecipitation, and fluorescence resonance energy transfer assay to identify the interactions of TRPP2 and STIM1 in transfected HEK293 cells and native vascular smooth muscle cells (VSMCs). The function of the TRPP2-STIM1 complex in thapsigargin (TG) or adenosine triphosphate (ATP)-induced SOCE was explored using specific small interfering RNA (siRNA). Further, we created TRPP2 conditional knockout (CKO) mouse to investigate the functional role of TRPP2 in agonist-induced vessel contraction. RESULTS: TRPP2 and STIM1 form a complex in transfected HEK293 cells and native VSMCs. Genetic manipulations with TRPP2 siRNA, dominant negative TRPP2 or STIM1 siRNA significantly suppressed ATP and TG-induced intracellular Ca2+ release and SOCE in HEK293 cells. Inositol triphosphate receptor inhibitor 2-aminoethyl diphenylborinate (2APB) abolished ATP-induced Ca2+ release and SOCE in HEK293 cells. In addition, TRPP2 and STIM1 knockdown significantly inhibited ATP- and TG-induced STIM1 puncta formation and SOCE in VSMCs. Importantly, knockdown of TRPP2 and STIM1 or conditional knockout TRPP2 markedly suppressed agonist-induced mouse aorta contraction. CONCLUSIONS: Our data indicate that TRPP2 and STIM1 are physically associated and form a functional complex to regulate agonist-induced intracellular Ca2+ mobilization, SOCE and blood vessel tone. Video abstract.

Increased TRPP2 expression in vascular smooth muscle cells from high-salt intake hypertensive rats: The crucial role in vascular dysfunction.

SCOPE: High-salt intake is a major risk factor in the development of hypertension. The underlying mechanism of high sodium on the cardiovascular system has received extensive attention. TRPP2 (Polycystin-2) is a Ca(2+) permeable nonselective cation channel that mediates Ca(2+) mobilization to control vascular smooth muscle cells (VSMCs) contraction. Here, we investigated TRPP2 expression change in VSMCs from high-salt intake hypertensive rats and role of TRPP2 in the development of high-salt diet-induced hypertension. METHODS AND RESULTS: After 4 ws of dietary treatment, systolic blood pressure was significantly elevated in high-salt intake rats (132 ± 3 mmHg) compared with regular diet control rats (104 ± 2 mmHg). Results from vessel tension and diameter measurements show that high-salt intake potentiated phenylephrine-induced contraction in denuded mesenteric artery and thoracic aorta. Immunoblot and immunofluorescence data indicate that TRPP2 expression in VSMCs from mesenteric artery and thoracic aorta was significantly increased in high-salt intake-induced hypertensive rats. However, agonist-induced contractions in denuded mesenteric artery and thoracic aorta were markedly decreased if TRPP2 was knocked down by specific shRNA. CONCLUSION: Our data demonstrate that high-salt intake increased TRPP2 expression in VSMCs, which in turn potentiated blood vessel response to contractors; this may participate in the development of hypertension.