Winter spatial distribution of fish larvae assemblages relative to the hydrography of the waters surrounding Taiwan

The aim of this study was to investigate the species composition and distribution of fish larvae in relation to hydrographic conditions in the waters surrounding Taiwan Island (TI) in February 2003. In total, 242 kinds of fish larvae belonging to 127 genera and 75 families were recognized. Among these, 109 taxa were identified to the family or genus level, others to the species level. The 12 predominant types, which constituted 71% of the total fish larvae, were Engraulis japonica, Scomber sp., Diaphus spp., Benthosema pterotum, Carangoides ferdau, Embolichthys mitsukurii, Maurolicus sp., unidentified Myctophidae, Gonostoma gracile, Trichiurus lepturus, unidentified Gobiidae, and Myctophum asperum. The distribution of fish larvae showed a clear association with water masses around TI, with higher abundances and lower species richness northwest of TI where the China Coastal Current prevails, and lower abundances and higher species diversity east of TI where the Kuroshio Current dominates. Cluster analysis distinguished three station groups and four species groups, and the distribution patterns of fish larvae also corresponded to hydrographic conditions. The total abundances of fish larvae and eight of the 12 predominant taxa showed significant and positive correlations with zooplankton abundance, which suggests that food source might be a key factor determining the abundance and distribution of fish larvae during the winter.     

A novel and highly efficient production system for recombinant adeno-associated virus vector

Recombinant adeno-associated virus(rAAV) has proven to be a promising gene delivery vector for human gene therapy. However, its application has been limited by difficulty in obtaining enough quantities of high-titer vector stocks. In this paper, a novel and highly efficient production system for rAAV is described. A recombinant herpes simplex virus type 1(rHSV-1) designated HSV1-rc/△UL2, which expressed adeno-associated virus type2(AAV-2) Rep and Cap proteins, was constructed previously. The data confirmed that its functions were to support rAAV replication and packaging, and the generated rAAV was infectious. Meanwhile, an rAAV proviral cell line designated BHK/SG2, which carried the green fluorescent protein(GFP) gene expression cassette, was established by transfecting BHK-21 cells with rAAV vector plasmid pSNAV-2-GFP. Infecting BHK/SG2 with HSV1-rc/△UL2 at an MOI of 0.1 resulted in the optimal yields of rAAV, reaching 250 transducing unit(TU) or 4.28×104 particles per cell. Therefore, compared with the conventional transfection method, the yield of rAAV using this "one proviral cell line, one helper virus" strategy was increased by two orders of magnitude. Large-scale production of rAAV can be easily achieved using this strategy and might meet the demands for clinical trials of rAAV-mediated gene therapy.     

<Emphasis Type="Italic">Phellinus</Emphasis> extracts inhibit migration and matrix metalloproteinase secretion in porcine coronary artery endothelial cells

Phellinus linteus is a fungus which is found primarily in tropical regions of the Americas, Africa, and Asia.P. linteus has been used in traditional medical practice for the treatment of arthritis, liver damage and cancer. Angiogenesis is a process that involves migration, proliferation and cell differentiation, as well as the formation of new capillary structures. The anti-angiogenic activities evidenced by natural compounds may actually be a critical effect for the inhibition of angiogenesis-dependent disease by these agents via the blockage of vascular development. This study assessed the effects of water extracts fromP. linteus (Phellinus extracts) on primary cultured porcine coronary artery endothelial cells (PCAECs).Phellinus extracts induced no changes in DNA synthesis or cell numbers, but inhibited the migration of PCAECs.Phellinus extracts also induced a reduction in the secretion of matrix metalloproteinase-2 (MMP-2) and MMP-9. Our results show that, in endothelial cells,Phellinus extracts may inhibit angiogenesis by reducing levels of MMP-2 and MMP-9 secretion.     

辣木资源利用中的悬浮细胞培养体系研究

辣木富含多种营养成分,在食品和药物开发方面有巨大的潜在开发价值。本文提供了一种可行的辣木细胞悬浮培养技术。由辣木的根诱导形成愈伤组织和叶诱导形成愈伤组织的合适细胞悬浮培养条件分别为MS培养基(MS)+1.0mg/L 2,4-二氯苯氧乙酸(2,4-D)+1.0mg/L激动素(KT)和MS+0.5mg/L 2,4-D+0.5mg/L KT,摇床转速均为50~100r/min,将愈伤组织添加到液体悬浮培养基中20d左右可得到大量悬浮细胞。本研究为辣木细胞水平的培养和研究提供了一条途径,为辣木潜在价值的开发利用提供新的思路。     

Recovery of transgenic plants by pollen-mediated transformation in <Emphasis Type="Italic">Brassica juncea</Emphasis>

The aroA-M1 encoding the mutant of 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPS) was introduced into the Brassica juncea genome by sonication-assisted, pollen-mediated transformation. The plasmid DNA and collected pollen grains were mixed in 0.3 mol/L sucrose solution and treated with mild ultrasonication. The treated pollen was then pollinated onto the oilseed stigmas after the stamens were removed artificially. Putative transgenic plants were obtained by screening germinating seeds on a medium containing glyphosate. Southern blot analysis of glyphosate-resistant plants indicated that the aroA-M1 gene had been integrated into the oilseed genome. Western blot analysis further confirmed that the EPSPS coded by aroA-M1 gene was expressed in transgenic plants. The transgenic plants exhibited increased resistance to glyphosate compared to untransformed plants. Some of those transgenic plants had considerably high resistance to glyphosate. The genetic analysis of T₁ progeny further confirmed that the inheritance of the introduced genes followed the Mendelian rules. The results indicated that foreign genes can be transferred by pollen-mediated transformation combined with mild ultrasonication.     

Molecular Cloning of Novel Cellulase Genes <Emphasis Type="Italic">cel</Emphasis>9A and <Emphasis Type="Italic">cel</Emphasis>12A from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> GXN151 and Synergism of Their Encoded Polypeptides

A thermophilic bacterial strain GXN151 which could degrade Avicel efficiently was isolated and identified as Bacillus licheniformis. A genomic library of GXN151 was constructed and two novel endoglucanase genes designated cel9A and cel12A were isolated by screening the library on carboxylmethyl cellulase indicator plates. The analysis of amino acid sequences deduced from the genes indicated that Cel9A consisted of a catalytic domain belonging to glycosyl hydrolase family 9, a linker domain, and a carbohydrate binding module family 3 from N-terminal to C-terminal; Cel12A had only one catalytic domain belonging to glycosyl hydrolase family 12. The combinations of Cel9A and Cel12A produced by the recombinant E. coli exhibited synergistic action against substrates of carboxylmethyl cellulose as well as Avicel.     

Sediment quality criteria in use around the world

 There have been numerous sediment quality guidelines (SQGs) developed during the past 20 years to assist regulators in dealing with contaminated sediments. Unfortunately, most of these have been developed in North America. Traditionally, sediment contamination was determined by assessing the bulk chemical concentrations of individual compounds and often comparing them with background or reference values. Since the 1980s, SQGs have attempted to incorporate biological effects in their derivation approach. These approaches can be categorized as empirical, frequency-based approaches to establish the relationship between sediment contamination and toxic response, and theoretically based approaches that attempt to account for differences in bioavailability through equilibrium partitioning (EqP) (i.e., using organic carbon or acid volatile sulfides). Some of these guidelines have been adopted by various regulatory agencies in several countries and are being used as cleanup goals in remediation activities and to identify priority polluted sites. The original SQGs, which compared bulk chemical concentrations to a reference or to background, provided little insight into the ecosystem impact of sediment contaminants. Therefore, SQGs for individual chemicals were developed that relied on field sediment chemistry paired with field or laboratory-based biological effects data. Although some SQGs have been found to be relatively good predictors of significant site contamination, they also have several limitations. False positive and false negative predictions are frequently in the 20% to 30% range for many chemicals and higher for others. The guidelines are chemical specific and do not establish causality where chemical mixtures occur. Equilibrium-based guidelines do not consider sediment ingestion as an exposure route. The guidelines do not consider spatial and temporal variability, and they may not apply in dynamic or larger-grained sediments. Finally, sediment chemistry and bioavailability are easily altered by sampling and subsequent manipulation processes, and therefore, measured SQGs may not reflect in situ conditions. All the assessment tools provide useful information, but some (such as SQGs, laboratory toxicity and bioaccumulation, and benthic indices) are prone to misinterpretation without the availability of specific in situ exposure and effects data. SQGs should be used only in a “screening” manner or in a “weight-of-evidence” approach. Aquatic ecosystems (including sediments) must be assessed in a “holistic” manner in which multiple components are assessed (e.g., habitat, hydrodynamics, resident biota, toxicity, and physicochemistry, including SQGs) by using integrated approaches. Received: December 26, 2000 / Accepted: December 28, 2001     

Eutrophication assessment and bioremediation strategy in a marine fish cage culture area in Nansha Bay,China

On the basis of field data measured during four cruises from January to November 2007, variations in the characteristics of dissolved inorganic nitrogen and phosphate were analyzed in Nansha marine fish cage culture area, Ningbo City, China. Dissolved inorganic nitrogen (DIN) was selected as the parameter to balance seaweed absorption and fish DIN production. The contents of DIN and phosphate varied with different seasons, and eutrophication index (E) value ranged from 2.41 to 15.99, indicating serious eutrophication conditions; the annual average value of N/P of 32.95 indicates a nitrogen surplus in this system. The eutrophication condition in Nansha Bay was mainly caused by the fish cage culture activities. Based on their biological characteristics, Laminaria and Gracilaria were selected as the bioremediation species in winter and spring and summer and autumn, respectively. The optimal co-cultivation proportion of fish cage to Laminaria and Gracilaria in this bay was 1 cage, 450 m² and one cage, 690 m², respectively.     

Tetraspanin CD151 Stimulates Adhesion-dependent Activation of Ras, Rac, and Cdc42 by Facilitating Molecular Association between β1 Integrins and Small GTPases

Tetraspanin CD151 associates with laminin-binding α(3)β(1)/α(6)β(1) integrins in epithelial cells and regulates adhesion-dependent signaling events. We found here that CD151 plays a role in recruiting Ras, Rac1, and Cdc42, but not Rho, to the cell membrane region, leading to the formation of α(3)β(1)/α(6)β(1) integrin-CD151-GTPases complexes. Furthermore, cell adhesion to laminin enhanced CD151 association with β(1) integrin and, thereby, increased complex formation between the β(1) family of integrins and small GTPases, Ras, Rac1, and Cdc42. Adhesion receptor complex-associated small GTPases were activated by CD151-β(1) integrin complex-stimulating adhesion events, such as α(3)β(1)/α(6)β(1) integrin-activating cell-to-laminin adhesion and homophilic CD151 interaction-generating cell-to-cell adhesion. Additionally, FAK and Src appeared to participate in this adhesion-dependent activation of small GTPases. However, engagement of laminin-binding integrins in CD151-deficient cells or CD151-specific siRNA-transfected cells did not activate these GTPases to the level of cells expressing CD151. Small GTPases activated by engagement of CD151-β(1) integrin complexes contributed to CD151-induced cell motility and MMP-9 expression in human melanoma cells. Importantly, among the four tetraspanin proteins that associate with β(1) integrin, only CD151 exhibited the ability to facilitate complex formation between the β(1) family of integrins and small GTPases and stimulate β(1) integrin-dependent activation of small GTPases. These results suggest that CD151 links α(3)β(1)/α(6)β(1) integrins to Ras, Rac1, and Cdc42 by promoting the formation of multimolecular complexes in the membrane, which leads to the up-regulation of adhesion-dependent small GTPase activation.     

Study on Adsorption of Metal Ions from Water by Using Immobilized Glycine

采用环氧氯丙烷法将惰性载体SephadexG-25活化,使其与甘氨酸偶联从而得到固定羧基的离子交换吸附剂.对该吸附剂吸附金属离子性能的研究表明,在pH 9.0时,吸附剂对Ca2+等金属离子有很强的吸附,且对Fe2+、Fe3+、Mn2+的吸附力比对Ca2+和Mg2+的吸附力要强.16g(湿重)吸附剂对金属离子的饱和吸附量分别为:Ca2+ 16.99 mg,Mg2+ 6.86 mg,Fe2+ 10.06 mg,Fe3+ 4.93 mg,Mn2+ 11.51 mg.同时,该吸附剂具有稳定性好、能重复使用且制备成本低等特点,在污水处理、金属离子回收等方面有很好的应用前景.