Recruitment of Nox4 to a Plasma Membrane Scaffold Is Required for Localized Reactive Oxygen Species Generation and Sustained Src Activation in Response to Insulin-like Growth Factor-I

Nox4-derived ROS is increased in response to hyperglycemia and is required for IGF-I-stimulated Src activation. This study was undertaken to determine the mechanism by which Nox4 mediates sustained Src activation. IGF-I stimulated sustained Src activation, which occurred primarily on the SHPS-1 scaffold protein. In vitro oxidation experiments indicated that Nox4-derived ROS was able to oxidize Src when they are in close proximity, and Src oxidation leads to its activation. Therefore we hypothesized that Nox4 recruitment to the plasma membrane scaffold SHPS-1 allowed localized ROS generation to mediate sustained Src oxidation and activation. To determine the mechanism of Nox4 recruitment, we analyzed the role of Grb2, a component of the SHPS-1 signaling complex. We determined that Nox4 Tyr-491 was phosphorylated after IGF-I stimulation and was responsible for Nox4 binding to the SH2 domain of Grb2. Overexpression of a Nox4 mutant, Y491F, prevented Nox4/Grb2 association. Importantly, it also prevented Nox4 recruitment to SHPS-1. The role of Grb2 was confirmed using a Pyk2 Y881F mutant, which blocked Grb2 recruitment to SHPS-1. Cells expressing this mutant had impaired Nox4 recruitment to SHPS-1. IGF-I-stimulated downstream signaling and biological actions were also significantly impaired in Nox4 Y491F-overexpressing cells. Disruption of Nox4 recruitment to SHPS-1 in aorta from diabetic mice inhibited IGF-I-stimulated Src oxidation and activation as well as cell proliferation. These findings provide insight into the mechanism by which localized Nox4-derived ROS regulates the sustained activity of a tyrosine kinase that is critical for mediating signal transduction and biological actions.     

Dissipative particle dynamics simulation of a gold nanoparticle system

Dissipative particle dynamics (DPD) was carried out to study systems containing gold atoms, organic ether (oligohydroquinonyl ether terminated with a thiol group) and organic solvents. The components in the simulated system are very different in size and chemical nature. Our simulation showed that the reproduction of the macroscopic experimental phase separation, properly dividing the polymeric molecule into beads, selecting the size of gold bead, and choosing the appropriate interaction parameters between beads are crucial. In addition, the solvent effect was the dominant factor for the formation of spherical aggregates of Au atoms and organic ether molecules. We report the interaction strengths between the solvent and gold clusters. Our work has demonstrated that DPD methods can be applied to the study of complex meso-scale systems.     

Evaluation of housekeeping genes as references for quantitative real-time PCR analysis of European eel,Anguilla anguilla

Eels are important aquaculture species for which an increasing number of reference genes are being identified and applied. In this study, five housekeeping genes [RPL7 (ribosomal protein L7), 18 S (18 S ribosomal RNA), EF1A (elongation factor 1α), ACTB (β-actin) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase)] were chosen to evaluate their reliability as reference genes for quantitative real-time PCR (qPCR) for the study of Anguilla anguilla. The expression of the selected genes in different eel tissues was determined using qPCR at different growth stages or upon challenge by Anguillid herpesvirus (AngHV), and the expression levels of these genes were then compared and evaluated using the geNorm and NormFinder algorithms. Then, RefFinder was used to comprehensively rank the examined housekeeping genes. Interestingly, the expression of the evaluated housekeeping genes exhibited tissue-dependent and treatment-dependent variations. In different growth periods A. anguilla tissues, the most stable genes were the following: ACTB in mucus; 18 S in skin and kidney; RPL7 in muscle, gill, intestine and brain; EF1A in heart and liver; and GAPDH in spleen. In contrast, in AngHV-challenged A. anguilla tissues, the most stable genes were the following: 18 S in mucus; RPL7 in skin, gill, heart, spleen, kidney and intestine; EF1A in muscle and liver; and ACTB in brain. Further comparison analysis indicated that the expression of RPL7 and EF1A was stable in multiple A. anguilla tissues in different growth periods and in eels challenged by AngHV. Nonetheless, the expression level of GAPDH in eel tissues was lower, and it was unstable in several tissues. These results indicated that the selection of reference genes for qPCR analysis in A. anguilla should be made in accordance with experimental parameters, and both RPL7 and EF1A could be used as reference genes for qPCR study of A. anguilla at different growth stages or upon challenge by AngHV. The reference genes identified in this study could improve the accuracy of qPCR data and facilitate further studies aimed at understanding the biology of eels.     

Terpenoids and Their Possible Role in Defense Against a Fungal Pathogen Alternaria tenuissima in Chrysanthemum morifolium Cultivars

Journal of Plant Growth Regulation - Terpenoids are a class of structurally diverse natural products involved in various plant biological processes. Their primary function is believed to provide...     

有机物料对喀斯特地区石灰土有效N、Fe、Zn含量的影响

为研究不同有机物料对喀斯特石灰土元素有效性的影响,采用40 d的室内培养实验,比较了单独添加不同比例（1%、3%、5%）的生物质炭、鸡粪肥、羊粪肥对喀斯特石灰土有效N、Fe、Zn含量的影响。结果表明：添加生物质炭提高了喀斯特石灰土的pH值,而添加鸡粪肥和羊粪肥降低了喀斯特石灰土的pH值;添加3种有机物料均增加了喀斯特石灰土的有机质含量,大小关系为：生物质炭 > 鸡粪肥 > 羊粪肥,但添加鸡粪肥和羊粪肥土壤有机质的化学活性和微生物活性更高。受pH、有机质活性、碳氮比等因素的影响,添加鸡粪肥和羊粪肥能增加土壤有效N含量,但两种有机肥对土壤有效N的提高效果相差不大,而添加生物质炭反而降低了土壤有效N的含量;3种有机物料均能提高土壤的有效Fe和有效Zn含量,其中鸡粪肥效果最佳,其次为羊粪肥和生物质炭。当3种有机物料的添加比例为5%时,生物质炭处理土壤的有效N、Fe、Zn含量分别是对照处理的0.92、1.13、1.21倍;鸡粪肥处理土壤的有效N、Fe、Zn含量分别是对照处理的1.22、1.63和3.39倍;羊粪肥处理土壤的有效N、Fe、Zn含量分别是对照处理的1.27、1.34和2.59倍。所以,相对于生物质炭,有机粪肥对喀斯特地区的石灰土有更好的改良效果。     

陕西食源性沙门氏菌耐药及相关基因

【目的】研究食源性沙门氏菌对常用抗生素的药敏性及相关耐药基因,更好的了解耐药性的产生和传播途径,确保食品安全。【方法】使用the Clinical and Laboratory Standards Institute推荐的琼脂稀释法测定沙门氏菌的药敏性,PCR和基因序列测定方法确定耐药沙门氏菌中整合子及其携带的耐药基因、与头孢菌素抗性相关的基因、沙门氏菌基因岛及与氟喹诺酮类抗生素耐药相关的基因突变。【结果】359株沙门氏菌中,67%的菌株对磺胺甲恶唑产生抗性,对甲氧苄啶/磺胺甲恶唑、四环素、卡那霉素、萘啶酮酸、氨苄西林、阿莫西林/克拉维酸、链霉素、氯霉素和庆大霉素、环丙沙星、头孢曲松、头孢西丁和头孢哌酮的耐药率分别为58%、56%、37%、35%、33%、32%、29%、26%、21%、16%、9%和8%。284株耐药菌中,79%的菌株可抗至少1种抗生素,25.9%可抗10种以上抗生素,2.5%可抗14种抗生素。耐药的Ⅰ类整合子以1.4kb最为常见,携带的耐药基因有aadA1、aadA2、aadA5、tetR、blaPSE-1、blaDHA-1、blaVEB-1、dhfrⅠ、dhfrⅤ、dhfrⅦ和dhfr17等。62株耐头孢曲松和/或头孢哌酮的沙门氏菌中,blaTEM和blaCMY-2基因的检出率分别为51.6%和56.5%。13.6%的沙门氏菌中检出了沙门氏菌基因岛。35株耐氟喹诺酮类抗生素的沙门氏菌的gyrA、parC和parE基因中共检出68个点突变,gyrA基因中常见突变为Ser83Phe、Ser83Tyr、Asp87Gly和Asp87Asn,parC基因中为Ser80Arg。parE基因中检出了Lys441Ile、Lys428Gln、Asp494Asn、Lys428Gln和Gly442Ser突变,这些点突变均为首次在食源性沙门氏菌中检出。【结论】陕西食源性沙门氏菌耐药状况严重,整合子、沙门氏菌基因岛和β-内酰胺酶编码基因的存在及解旋酶和拓扑异构酶基因突变是导致沙门氏菌耐药的重要机制。     

Home range and habitat use of male Reeves’s pheasant (Syrmaticus reevesii) during winter in Dongzhai National Nature Reserve, Henan Province, China

Home range and habitat use of male Reeves’s pheasant (Syrmaticus reevesii) were studied during winter of 2001∼2002 and 2002∼2003 in the Dongzhai National Nature Reserve, Henan Province. Results from five individuals of Reeves’s pheasant with over 30 relocations, indicated that the average size of home range was 10.03 ± 1.17 hm² by Minimum Convex Polygon method, 8.60 ± 0.35 hm² by 90% Harmonic Mean Transformation method, and 9.50 ± 1.90 hm² by 95% Fixed Kernel method. It was observed that the winter range is smaller than that in the breeding season. The mean core area of the home range was found to be 1.88 ± 0.37 hm². Although the habitat composition of the core area varied greatly for individuals, a large part of the habitats used were composed of confier and broadleaf mixed forests, masson pine forests, fir forests, and shrubs. Habitat use within the study area was non-random, while habitats within home ranges were randomly used. Habitat use was dictated by tree diameter at breast height, shrub height and coverage at 2.0 m. The proximity between forests and shrubs were also found to be important in providing refuge for the birds during winter. Recommendations for conservation management include protecting the existing habitats in Dongzhai National Nature Reserve, increasing suitable habitat for Reeves’s Pheasant through artificial plantations (e.g. firs), and restoring some parts of the large shrub area into forests. __________ Translated from Biodiversity Science, 2005, 13 (5) [译自: 生物多样性, 2005,13(5)]     

Imaging and measuring the rituximab-induced changes of mechanical properties in B-lymphoma cells using atomic force microscopy

The topography and mechanical properties of single B-lymphoma cells have been investigated by atomic force microscopy (AFM). With the assistance of microfabricated patterned pillars, the surface topography and ultrastructure of single living B-lymphoma cell were visualized by AFM. The apoptosis of B-lymphoma cells induced by rituximab alone was observed by acridine orange/ethidium bromide (AO/EB) double fluorescent staining. The rituximab-induced changes of mechanical properties in B-lymphoma cells were measured dynamically and the results showed that B-lymphoma cells became dramatically softer after incubation with rituximab. These results can improve our understanding of rituximab’effect and will facilitate the further investigation of the underlying mechanisms.     

High frequency of mitochondrial DNA D-loop mutations in Ewing’s sarcoma

Somatic mutations and polymorphisms in the noncoding displacement (D)-loop of mitochondrial DNA (mtDNA) are present in a variety of human cancers. To investigate whether Ewing’s sarcoma (EWS) harbors genetic alterations within the D-loop region and their potential association with EWS carcinogenesis, we analyzed and compared the complete mtDNA D-loop sequences from 17 pairs of tumor tissues and corresponding peripheral blood samples using the direct DNA sequencing method. Our results revealed that 12 of the 17 EWS tumor specimens (70.6%) carried 19 somatic mutations in the D-loop of mtDNA, including 11 single-base substitutions, 3 insertions and 5 deletions. Among the tested 17 patients, we screened a total of 40 germline polymorphisms including one novel sequence variant in the D-loop fragment. Most of these identified mutations and germline variations were clustered within two hypervariable segments (HVS1 and HVS2) as well as the homopolymeric C stretch between nucleotide position 303 and 309. In addition, there was no significant correlation between mtDNA D-loop mutations and various clinicopathological factors of EWS. In conclusion, our study reports for the first time that mtDNA D-loop mutations occur at a high frequency in EWS. These data provide evidence of mtDNA alterations’ possible involvement in the initiation and/or progression of this rare malignancy.