Activated PAK4 regulates cell adhesion and anchorage-independent growth

The serine/threonine kinase PAK4 is an effector molecule for the Rho GTPase Cdc42. PAK4 differs from other members of the PAK family in both sequence and function. Previously we have shown that an important function of this kinase is to mediate the induction of filopodia in response to activated Cdc42. Since previous characterization of PAK4 was carried out only with the wild-type kinase, we have generated a constitutively active mutant of the kinase to determine whether it has other functions. Expression of activated PAK4 in fibroblasts led to a transient induction of filopodia, which is consistent with its role as an effector for Cdc42. In addition, use of the activated mutant revealed a number of other important functions of this kinase that were not revealed by studying the wild-type kinase. For example, activated PAK4 led to the dissolution of stress fibers and loss of focal adhesions. Consequently, cells expressing activated PAK4 had a defect in cell spreading onto fibronectin-coated surfaces. Most importantly, fibroblasts expressing activated PAK4 had a morphology that was characteristic of oncogenic transformation. These cells were anchorage independent and formed colonies in soft agar, similar to what has been observed previously in cells expressing activated Cdc42. Consistent with this, dominant-negative PAK4 mutants inhibited focus formation by oncogenic Dbl, an exchange factor for Rho family GTPases. These results provide the first demonstration that a PAK family member can transform cells and indicate that PAK4 may play an essential role in oncogenic transformation by the GTPases. We propose that the morphological changes and changes in cell adhesion induced by PAK4 may play a direct role in oncogenic transformation by Rho family GTPases and their exchange factors.     

A novel matrix derivatized from hydrophilic gigaporous polystyrene-based microspheres for high-speed immobilized-metal affinity chromatography

Agarose coated gigaporous polystyrene microspheres were evaluated as a novel matrix for immobilized-metal affinity chromatography (IMAC). With four steps, nickel ions were successfully immobilized on the microspheres. The gigaporous structure and chromatographic properties of IMAC medium were characterized. A column packed with the matrix showed low column backpressure and high column efficiency at high flow velocity. Furthermore, this matrix was used for purifying superoxide dismutase (SOD), which was expressed in Escherichia coli (E. coli) in submerged fermentation, on an Äkta purifier 100 system under different flow velocities. The purity of the SOD from this one-step purification was 79% and the recovery yield was about 89.6% under the superficial flow velocity of 3251 cm/h. In conclusion, all the results suggested that the gigaporous matrix has considerable advantages for high-speed immobilized-metal affinity chromatography.     

Comparative Effects of Hispidulin,Genistein, and Icariin with Estrogen on Bone Tissue in Ovariectomized Rats

Icariin, Genistein, and Hispidulin have been proven to have estrogen-like and antiosteoporotic activity and can be potentially used for the treatment of osteoporosis. The present study found that Icariin, Genistein, and Hispidulin treatments, emulating estrogen, significantly contributed to bone density. Comparative effects of Icariin, Genistein, and Hispidulin with estrogen on in ovariectomized rats were investigated. Our results showed that genistein was found to have superior bone protective effects against osteoporosis among genistein, Icariin, and Hispidulin.     

一个新水稻温敏感叶色突变体的遗传分析及其基因分子定位

在粳稻品种嘉花1号(Oryza sativa L. ssp. japonica ‘Jiahua No.1’)种子经⁶⁰Co γ射线辐照处理的后代中, 发现了1个低温敏感叶色突变体mr21。在较低温度(<25.0°C)条件下, 该突变体幼苗叶色呈黄色; 随着温度逐渐升高, 叶色由黄转绿,其临界温度约为27.5°C; 在低温条件下, 突变体幼苗总叶绿素含量以及叶绿素a、b的含量均较野生型嘉花1号明显下降, 表明该突变体的叶色性状具有明显的温敏感性。遗传分析表明, 该突变体叶色性状受1对隐性核基因控制, 暂将该突变基因命名为thermo-sensitive leaf-color 1(tsl-1)。以该突变体与籼稻9311(Oryza sativa L. ssp. indica ‘9311’)杂交的F₂代分离群体作为定位群体, 利用SSR分子标记将tsl-1基因初步定位在水稻(Oryza sativa)第1号染色体短臂上的MM1799与RM8132分子标记之间, 其遗传距离分别为2.4 cM和3.0 cM; 然后, 进一步利用扩大F₂代群体及新发展的分子标记将tsl-1基因定位在分子标记InDel2与InDel4之间的198 kb内。研究结果为今后对该基因的克隆和功能分析奠定了基础。     

In vivo effects of adenosine A1 receptor agonist and antagonist on neuronal and astrocytic intermediary metabolism studied with ex vivo 13C NMR spectroscopy

Adenosine is a neuromodulator, and it has been suggested that cerebral acetate metabolism induces adenosine formation. In the present study the effects that acetate has on cerebral intermediary metabolism, compared with those of glucose, were studied using the adenosine A1 receptor agonist 2-chloro-N6-cyclopentyladenosine (CCPA) and antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). Fasted rats received an intravenous injection of CCPA, DPCPX, or vehicle. Fifteen minutes later either [1,2-13C]acetate or [1-13C]glucose was given intraperitoneally; after another 30 min the rats were decapitated. Cortical extracts were analyzed with 13C NMR spectroscopy and HPLC analysis. DPCPX affected neuronal and astrocytic metabolism. De novo synthesis of GABA from neuronal and astrocytic precursors was significantly reduced. De novo syntheses of glutamate and aspartate were at control levels, but their degradation was significantly elevated. In glutamine the anaplerotic activity and the amount of label in the position representing the second turn in the tricarboxylic acid cycle were significantly increased, suggesting elevated metabolic activity in astrocytes. CCPA did not influence GABA, aspartate, or glutamine synthesis. In glutamate the contribution from the astrocytic anaplerotic pathway was significantly decreased. In the present study the findings in the [1,2-13C]acetate and [1-13C]glucose control, CCPA, and DPCPX groups were complementary, and no adenosine A1 agonist effects arising from cerebral acetate metabolism were detected.     

盐度和CO2倍增环境下碱蓬幼苗呼吸酶活性的变化

研究了生长在正常大气ＣＯ２和ＣＯ２倍增环境中的盐生植物碱蓬（Ｓｕａｅｄａｓａｌｓａ）幼苗呼吸酶活性对ＫＣｌ和ＮａＣｌ的反应．结果表明,在ＣＯ２倍增（７００μｌ·Ｌ－１）和正常大气ＣＯ２（３５０μｌ·Ｌ－１）下,３００ｍｍｏｌ·Ｌ－１ＫＣｌ和ＮａＣｌ均能抑制琥珀酸脱氢酶（ＳＤＨ）和苹果酸脱氢酶（ＭＤＨ）活性,而异柠檬酸脱氢酶（ＩＤＨ）活性为ＮａＣｌ抑制、ＫＣｌ促进;ＮａＣｌ和ＫＣｌ明显抑制细胞色素氧化酶（ＣＯ）和光呼吸中乙醇酸氧化酶（ＧＯ）、羟基丙酮酸还原酶（ＨＰＲ）活性;并指出在ＫＣｌ胁迫下,ＣＯ２使三羧酸循环（ＴＣＡＣ）的运行变慢,ＮａＣｌ胁迫下使其加快,ＴＣＡＣ运行限速步骤与ＭＤＨ无关,ＣＯ为盐对呼吸代谢影响的重要位点．另外,Ｋ＋、Ｎａ＋对蛋白表达的影响有差异,ＣＯ２可使盐胁迫下的碱蓬幼苗蛋白表达降低．     

Diversity surveys and evolutionary relationships of aoxB genes in aerobic arsenite-oxidizing bacteria

A new primer set was designed to specifically amplify ca. 1,100 bp of aoxB genes encoding the As(III) oxidase catalytic subunit from taxonomically diverse aerobic As(III)-oxidizing bacteria. Comparative analysis of AoxB protein sequences showed variable conservation levels and highlighted the conservation of essential amino acids and structural motifs. AoxB phylogeny of pure strains showed well-discriminated taxonomic groups and was similar to 16S rRNA phylogeny. Alphaproteobacteria-, Betaproteobacteria-, and Gammaproteobacteria-related sequences were retrieved from environmental surveys, demonstrating their prevalence in mesophilic As-contaminated soils. Our study underlines the usefulness of the aoxB gene as a functional marker of aerobic As(III) oxidizers.