Types and distribution of mucous cells of the abalone Haliotis diversicolor

The types and distribution of mucous cells of Haliotis diversicolorwere observed and analyzed using the alcian blue and periodic acid schiffs (AB-PAS) reaction and histological procedures. According to the color of the mucous cells, they were divided into four types: Type I, pure red; type II, pure blue; type III, purple reddish; type IV, blue purple. Most of the mucous cells in the mantle were type II, with cup- or stick-like shape. Gill axis and gill filament epithelia were rich in mucous cells and most of them were type II and III, with circle-, cup- and stick-like shapes. There were a few mucous cells in the pedal epithelia, mainly type II, whereas, the pedal gland had a great density of mucous cells, which were large or small and mainly type II and IV. There were many mucous cells in the epithelia around the mouth, most of which were medium-sized cup-shaped type II. In the esophagus, the number of mucous cells decreased gradually from the anterior to posterior, whereas in the distal intestine mucous cells, there were more than in the proximal one and they were more cup–shaped and circle–shaped, mainly type II.Keywords: Haliotis diversicolor; mucous cells, types, distributio

Differential proteomic profiles and characterizations between hyalinocytes and granulocytes in ivory shell Babylonia areolata

Abstract(#br)The haemocytes of the ivory shell, Babylonia areolata are classified by morphologic observation into the following types: hyalinocytes (H) and granulocytes (G). Haemocytes comprise diverse cell types with morphological and functional heterogene and play indispensable roles in immunological homeostasis of invertebrates. In the present study, two types of haemocytes were morphologically identified and separated as H and G by Percoll density gradient centrifugation. The differentially expressed proteins were investigated between H and G using mass spectrometry. The results showed that total quantitative proteins between H and G samples were 1644, the number of up-regulated proteins in G was 215, and the number of down-regulated proteins in G was 378. Among them, cathepsin, p38 MAPK, toll-interacting protein-like and beta-adrenergic receptor kinase 2-like were up-regulated in G; alpha-2-macroglobulin-like protein, C-type lectin, galectin-2-1, galectin-3, β-1,3-glucan-binding protein, ferritin, mega-hemocyanin, mucin-17-like, mucin-5AC-like and catalytic subunit of protein kinase A were down-regulated in G. The results showed that the most significantly enriched KEGG pathways were the pathways related to ribosome, phagosome, endocytosis, carbon metabolism, protein processing in endoplasmic reticulum and oxidative phosphorylation. For phagosome and endocytosis pathway, the number of down-regulation proteins in G was more than that of up-regulation proteins. For lysosome pathway, the number of up-regulation proteins in G was more than that of down-regulation proteins. These results suggested that two sub-population haemocytes perform the different immune functions in B. areolata

Morphologic, cytometric, quantitative transcriptomic and functional characterisation provide insights into the haemocyte immune responses of Pacific abalone (Haliotis discus hannai)

In recent years, the abalone aquaculture industry has been threatened by the bacterial pathogens. The immune responses mechanisms underlying the phagocytosis of haemocytes remain unclear in Haliotis discus hannai. It is necessary to investigate the immune mechanism in response to these bacterial pathogens challenges. In this study, the phagocytic activities of haemocytes in H. discus hannai were examined by flow cytometry combined with electron microscopy and transcriptomic analyses. The results of Vibrio parahaemolyticus, Vibrio alginolyticus and Staphylococcus aureu challenge using electron microscopy showed a process during phagosome formation in haemocytes. The phagocytic rate (PP) of S. aureus was higher than the other five foreign particles, which was about 63%. The PP of Vibrio harveyi was about 43%, the PP peak of V. alginolyticus in haemocyte was 63.7% at 1.5 h. After V. parahaemolyticus and V. alginolyticus challenge, acid phosphatase, alkaline phosphatase, total superoxide dismutase, lysozyme, total antioxidant capacity, catalase, nitric oxide synthase and glutathione peroxidase activities in haemocytes were measured at different times, differentially expressed genes (DEGs) were identified by quantitative transcriptomic analysis. The identified DEGs after V. parahaemolyticus challenge included haemagglutinin/amebocyte aggregation factor-like, supervillin-like isoform X4, calmodulin-like and kyphoscoliosis peptidase-like; the identified DEGs after V. alginolyticus challenge included interleukin-6 receptor subunit beta-like, protein turtle homolog B-like, rho GTPase-activating protein 6-like isoform X2, leukocyte surface antigen CD53-like, calponin-1-like, calmodulin-like, troponin C, troponin I-like isoform X4, troponin T-like isoform X18, tumor necrosis factor ligand superfamily member 10-like, rho-related protein racA-like and haemagglutinin/amebocyte aggregation factor-like. Some immune-related KEGG pathways were significantly up-regulated or down-regulated after challenge, including thyroid hormone synthesis, Th17 cell differentiation signalling pathway, focal adhesion, melanogenesis, leukocyte transendothelial migration, inflammatory mediator regulation of TRP channels, ras signalling pathway, rap1 signalling pathway. This study is the first step towards understanding the H. discus hannai immune system by adapting several tools to gastropods and providing a first detailed morpho-functional study of their haemocytes

Improved Sampling of Hemolymph and Screening of Anti-Coagulants of Hemocytes in the Snail <em>Babylonia areolata</em>

Our aim was to improve sampling of hemolymph from the snail Babylonia areolata in order to evaluate the physiological and immune capacities of hemocytes in aquaculture. We also identified appropriate types of hemolymph anti-coagulants for B. areolata. Hemolymph samples were collected using an improved foot plantaris puncture method. We screened five types of anti-coagulants from Penaeid shrimp (A), marine decapods (B), abalone (C), and oyster (D), as well as a home-made anti-coagulant (E), to act against Babylonia areolata hemocytes on the basis of cell death rate and hemocyte aggregation. We improved the former foot plantaris puncture method and identified the home-made anti-coagulant as the best anti-coagulant from the five which we tested

Phagocytosis and respiratory burst activity of haemocytes from the ivory snail, Babylonia areolata

Earmarked Fund for Modern Agro-industry Technology Research System [CARS-48]; PCSIRT [IRT0941]; Program of Introducing Talents of Discipline to Universities [B07034]Haemocytes from the ivory snail, Babylonia areolata phagocytized Saccharomyces cerevisiae and Vibrio parahaemolyticus after 30 min. Haemocytes phagocytized V parahaemolyticus at a greater rate than they phagocytized S. cerevisiae. The phagocytic rate (PP) of V. parahaemolyticus by granulocytes to was a little higher than that of S. cerevisiae. The phagocytic index (PI) of V parahaemolyticus by granulocytes was significantly higher than that of S. cerevisiae. The same was true of hyalinocytes. The PP of granulocytes was significantly higher than that of hyalinocytes for each pathogen. No difference in PI was observed in granulocytes and hyalinocytes. Two defense mechanisms of B. areolata were quantified using flow cytometry. Haemocyte phagocytosis was quantified using fluorescent microbeads and respiratory burst activity was measured using H2O2 increases detected by 2', 7'-dichlorofluorescein diacetate. Both phagocytosis and respiratory burst activity of the haemocytes increased over time. After 90 min the phagocytic rate no longer increased. In the case of respiratory burst, the greatest increase in fluorescence occurred between 30 and 120 min, no further increase was seen after 120 min. These results showed unequivocally that a native (unstimulated) haemocyte oxidative burst was active in B. areolata. The aim of this study was to further the knowledge of immunology in gastropods. (C) 2013 Elsevier Ltd. All rights reserved

Pulmonary Hypertension in a Patient with Non-cirrhotic Portal Hypertension and Scleroderma Sine Scleroderma: A Case Report &quot;Pulmonary Hypertension in a Patient with Non-cirrhotic Portal Hypertension and Scleroderma Sine Scleroderma: A Case Report

Abstract Porto-pulmonary hypertension is a known complication of liver cirrhosis but its association with noncirrhotic portal hypertension patients is rare. We report a case of pulmonary hypertension in a patient with noncirrhotic portal hypertension and scleroderma sine scleroderma. The two latter conditions have been shown to be independently associated with pulmonary arterial hypertension. DLCO is expected to decrease in scleroderma patients due to pulmonary vascular disease, which will result in an increased FVC/DLCO ratio. The low FVC/DLCO ratio in our patient suggests that pulmonary arterial hypertension was more likely to have been due to non-cirrhotic portal hypertension than scleroderma sine scleroderma

Morphological and biomechanical response to eutrophication and hydrodynamic stresses

Eutrophication and hydrodynamics determine the final distribution patterns of aquatic macrophytes; however, there is limited available knowledge regarding their interactive effects. Morphological and biomechanical responses to eutrophication and hydrodynamic stresses were assessed by sampling five abundant and dominant species, Potamogeton maackianus, P. pectinatus, P. lucens, Ceratophyllum demersum and Myriophyllum spicatum, in three macrophyte beds in Lake Erhai, Yunnan Province, China: one exposed to eutrophication and moderate southeast (SE) wind; one with mesotrophication, but sheltered by the lakeshore, with weak wind disturbance; and one with meso-eutrophication and strong SE wind. The results showed significant interactive effects of eutrophication and hydrodynamics on most biomechanical traits and some morphological traits, suggesting that aquatic macrophytes preferentially undergo biomechanical adjustments to resist the coexisting eutrophication and hydrodynamic stresses. In particular, hydrodynamics increased both the tensile force and tensile strain of P. maackianus under meso-eutrophication and dramatically decreased them in eutrophic areas, suggesting that eutrophication triggers mechanical failure in this species. Additionally, P. pectinatus, C. demersumand M. spicatum showed the lowest and highest values for the biomechanical variables (greater values for M. spicatum) in the most eutrophic and hydrodynamic areas, respectively, implying that increases in hydrodynamics primarily induce mechanical damage in eutrophic species. The plants generally exhibited greater tensile strain in both shallow and deep waters and the greatest tensile force at moderate depths. The stem cross-sectional area, plant height, stem length, internode length, and branch traits were all responsible for determining the biomechanical variables. This study reveals that hydrodynamic changes primarily induce mechanical damage in eutrophic species, whereas eutrophication triggers mechanical damage in sensitive species. (c) 2017 Published by Elsevier B.V

Proteomic analysis of trochophore and veliger larvae development in the small abalone Haliotis diversicolor

Abstract Background Haliotis diversicolor is commercially important species. The trochophore and veliger are distinct larval stages in gastropod development. Their development involves complex morphological and physiological changes. We studied protein changes during the embryonic development of H. diversicolor using two dimensional electrophoresis (2-DE) and label-free methods, tandem mass spectrometry (MS/ MS), and Mascot for protein identification. Results A total of 150 2-DE gel spots were identified. Protein spots showed upregulation of 15 proteins and downregulation of 28 proteins as H. diversicolor developed from trochophore to veliger larvae. Trochophore and veliger larvae were compared using a label-free quantitative proteomic approach. A total of 526 proteins were identified from both samples, and 104 proteins were differentially expressed (> 1.5 fold). Compared with trochophore larvae, veliger larvae had 55 proteins upregulated and 49 proteins downregulated. These differentially expressed proteins were involved in shell formation, energy metabolism, cellular and stress response processes, protein synthesis and folding, cell cycle, and cell fate determination. Compared with the 5 protein (fructose-bisphosphate aldolase, 14–3-3ε, profilin, actin-depolymerizing factor (ADF)/cofilin) and calreticulin) expression patterns, the mRNA expression exhibited similar patterns except gene of fructose-bisphosphate aldolase. Conclusion Our results provide insight into novel aspects of protein function in shell formation, torsion, and nervous system development, and muscle system differentiation in H. diversicolor larvae. “Quality control” proteins were identified to be involved in abalone larval development

Analysis of evolutionary relationships provides new clues to the origins of weedy rice

Abstract Weedy rice (WR) (Oryza sativa f. spontanea) is considered to be a pest in modern rice production systems because it competes for resources, has poor yield characteristics, and subsequently has a negative effect on rice grain yield. The evolutionary relationships among WR, landrace rice (LR), improved rice (IR) cultivars, and wild rice are largely unknown. In this study, we conducted a population genetic analysis based on neutral markers and gene haplotypes in 524 rice accessions and a comparative transcriptomic analysis using 15 representative samples. The results showed that WR populations have the highest level of genetic diversity (He = 0.8386) and can be divided into two groups (japonica‐type and indica‐type). The japonica‐type WR accessions from Heilongjiang province (HLJ), Jilin province (JL), Liaoning province (LN), and NX provinces clustered with the landraces grown in these same provinces. The indica‐types from Jiangsu province (JS) also clustered with the indica‐type landraces from JS province. Comparative transcriptome analysis of WR‚ IR and LR from HLJ, JL, and LN provinces showed that the WR still clustered with the LR, and that the IR lines comprise a single population. Thirty‐two differentially expressed genes were shared by the IR and LR groups as well as between the IR and WR groups. Using Gene ontology (GO) analysis, we identified 19 shared GO terms in the IR and LR groups as well as between the IR and WR groups. Our results suggest that WR populations in China have diverse origins, and comparative transcriptome analysis of different types of rice from HLJ, JL, and LN provinces suggests that IR populations have become a end point in the evolution of WR, which provides a new perspective for the study of WR origins and lays a solid foundation for rice breeding