Hydralazine inhibits compression and acrolein-mediated injuries in ex vivo spinal cord

We have previously shown that acrolein, a lipid peroxidation byproduct, is significantly increased following spinal cord injury in vivo , and that exposure to neuronal cells results in oxidative stress, mitochondrial dysfunction, increased membrane permeability, impaired axonal conductivity, and eventually cell death. Acrolein thus may be a key player in the pathogenesis of spinal cord injury, where lipid peroxidation is known to be involved. The current study demonstrates that the acrolein scavenger hydralazine protects against not only acrolein-mediated injury, but also compression in guinea pig spinal cord ex vivo . Specifically, hydralazine (500 μmol/L to 1 mmol/L) can significantly alleviate acrolein (100–500 μmol/L)-induced superoxide production, glutathione depletion, mitochondrial dysfunction, loss of membrane integrity, and reduced compound action potential conduction. Additionally, 500 μmol/L hydralazine significantly attenuated compression-mediated membrane disruptions at 2 and 3 h following injury. This was consistent with our findings that acrolein-lys adducts were increased following compression injury ex vivo , an effect that was prevented by hydralazine treatment. These findings provide further evidence for the role of acrolein in spinal cord injury, and suggest that acrolein-scavenging drugs such as hydralazine may represent a novel therapy to effectively reduce oxidative stress in disorders such as spinal cord injury and neurodegenerative diseases, where oxidative stress is known to play a role.     

小花盾叶薯蓣甙的酶降解

从小花盾叶薯蓣 (DioscoreaparvifloraC .T .Ting)的新鲜根状茎中分离到一个新的呋甾烷型配糖体 ,命名为小花盾叶薯蓣甙 (parvifloside) (1) ,其结构通过波谱和化学方法鉴定为 :(2 5R)_2 6_O_β_glucopyranosyl_furost_5_en_3β ,2 2 ξ ,2 6_triol 3_O_β_D_glucopyranosyl (1→ 3)_β_D_glucopyranosyl (1→ 4 )_[α_L_rhamnopyranosyl (1→ 2 ) ]_β_D_glucopyra noside。化合物 1在纤维素酶粗酶和 β_葡萄糖苷酶中进行水解 ,得到降解产物 2 - 7。对 1的酶解现象进行了讨论。同时 ,对所分离的甾体皂甙的抗稻瘟霉菌活性进行了初步筛选。     

生态转换系统中土壤有机质变化的稳定碳同位素示踪研究进展

由于气候变异、人类活动等因素影响 ,全球各生态系统间短期内正经历着前所未有的区域性转变 ,由此导致的一系列生态环境问题如区域气候变迁、荒漠化、水土流失等广受关注[2 ] ,而其中土壤有机质的变化起着极为关键的作用。据估计 ,地表土壤层中有机碳的总量为大气的 2～ 4倍 ,土壤有机质的变化对大气ＣＯ2 浓度的影响甚大[15] ;同时 ,土壤有机质又是维护土壤结构 ,保持土壤肥力的重要组成部分。因此 ,生态系统的转变对土壤有机质产生的影响为各国科学家所重视。尽管稳定碳同位素是示踪有机质变化过程最有效的手段之一 ,并且Ｎｉｓ ｓｅｎｂ…     

红棕象甲室内生物学特性及形态观察

在室内对红棕象甲Rhynchophorus ferrugineus Fab.生物学特性和形态进行系统观察。结果表明在26℃、RH80%、寄主食料华盛顿棕榈饲养条件下,红棕象甲世代历期平均114.77d,最短87.00d、最长166.00d,其中卵期平均3.50d,幼虫期平均36.68d,结茧到产卵前历期平均29.96d。幼虫以8龄为主,1~2龄幼虫的体重小于0.01g,头壳宽短于1.0mm,3龄后幼虫体重、头壳宽度、体长和体宽开始增加,8龄时幼虫平均体重为3.36g,头壳宽度6.0mm。成虫雌雄性比1.78∶1,补充营养能显著延长成虫寿命,在无补充营养时,雌雄成虫寿命只有7.00d左右,而在补充营养后,雌雄成虫寿命最长可分别达130.00d和244.00d。雌虫产卵量最大376粒,最小89粒,平均216粒。     

Screening and distributing features of bacteria with hydantoinase and carbamoylase

Hydantoinase and carbamoylase are key biocatalysts for the production of optically pure amino acids from dl-5-substituted hydantoins (SSH). Out of 364 isolated strains with hydantoinase and carbamoylase at 45 degrees C, 24 strains showed efficient hydantoinase and carbamoylase activities. Among them, 17 produced l-amino acids, and 7 produced d-amino acids from both aromatic dl-5-benzylhydantoin and aliphatic dl-5-isopropylhydantoin. Most of the strains that were able to form l-amino acid belonged to genera Bacillus, Geobacillus, Brevibacillus, Aneurinibacillus, Microbacterium, and Kurthia. Phylogenetic relationships were investigated based on 16S rRNA from the hydantoinase-producing bacteria. Distinct tendencies toward certain genera were observed between most of the strains forming l-amino acids and d-amino acids from SSH. The results from this study can be utilized to develop new isolation technology of hydantoinase-producing microorganisms, and to understand metabolism and evolutionary origins of hydantoinase and carbamoylase among different bacteria.     

Structure-based shape pharmacophore modeling for the discovery of novel anesthetic compounds

Current anesthetics, especially the inhaled ones, have troublesome side effects and may be associated with durable changes in cognition. It is therefore highly desirable to develop novel chemical entities that reduce these effects while preserving or enhancing anesthetic potency. In spite of progress toward identifying protein targets involved in anesthesia, we still do not have the necessary atomic level structural information to delineate their interactions with anesthetic molecules. Recently, we have described a protein target, apoferritin, to which several anesthetics bind specifically and in a pharmacodynamically relevant manner. Further, we have reported the high resolution X-ray structure of two anesthetic/apoferritin complexes (Liu, R.; Loll, P. J.; Eckenhoff, R. G. FASEB J. 2005, 19, 567). Thus, we describe in this paper a structure-based approach to establish validated shape pharmacophore models for future application to virtual and high throughput screening of anesthetic compounds. We use the 3D structure of apoferritin as the basis for the development of several shape pharmacophore models. To validate these models, we demonstrate that (1) they can be used to effectively recover known anesthetic agents from a diverse database of compounds; (2) the shape pharmacophore scores afford a significant linear correlation with the measured binding energetics of several known anesthetic compounds to the apoferritin site; and (3) the computed scores based on the shape pharmacophore models also predict the trend of the EC₅₀ values of a set of anesthetics. Therefore, we have now obtained a set of structure-based shape pharmacophore models, using ferritin as the surrogate target, which may afford a new way to rationally discover novel anesthetic agents in the future.     

Imatinib and Nilotinib Reverse Multidrug Resistance in Cancer Cells by Inhibiting the Efflux Activity of the MRP7 (ABCC10)

Background

One of the major mechanisms that could produce resistance to antineoplastic drugs in cancer cells is the ATP binding cassette (ABC) transporters. The ABC transporters can significantly decrease the intracellular concentration of antineoplastic drugs by increasing their efflux, thereby lowering the cytotoxic activity of antineoplastic drugs. One of these transporters, the multiple resistant protein 7 (MRP7, ABCC10), has recently been shown to produce resistance to antineoplastic drugs by increasing the efflux of paclitaxel. In this study, we examined the effects of BCR-Abl tyrosine kinase inhibitors imatinib, nilotinib and dasatinib on the activity and expression of MRP7 in HEK293 cells transfected with MRP7, designated HEK-MRP7-2.

Methodology and/or Principal Findings

We report for the first time that imatinib and nilotinib reversed MRP7-mediated multidrug resistance. Our MTT assay results indicated that MRP7 expression in HEK-MRP7-2 cells was not significantly altered by incubation with 5 µM of imatinib or nilotinib for up to 72 hours. In addition, imatinib and nilotinib (1-5 µM) produced a significant concentration-dependent reversal of MRP7-mediated multidrug resistance by enhancing the sensitivity of HEK-MRP7-2 cells to paclitaxel and vincristine. Imatinib and nilotinib, at 5 µM, significantly increased the accumulation of [³H]-paclitaxel in HEK-MRP7-2 cells. The incubation of the HEK-MRP7-2 cells with imatinib or nilotinib (5 µM) also significantly inhibited the efflux of paclitaxel.

Conclusions

Imatinib and nilotinib reverse MRP7-mediated paclitaxel resistance, most likely due to their inhibition of the efflux of paclitaxel via MRP7. These findings suggest that imatinib or nilotinib, in combination with other antineoplastic drugs, may be useful in the treatment of certain resistant cancers.     

Identification of a Topological Characteristic Responsible for the Biological Robustness of Regulatory Networks

Attribution of biological robustness to the specific structural properties of a regulatory network is an important yet unsolved problem in systems biology. It is widely believed that the topological characteristics of a biological control network largely determine its dynamic behavior, yet the actual mechanism is still poorly understood. Here, we define a novel structural feature of biological networks, termed ‘regulation entropy’, to quantitatively assess the influence of network topology on the robustness of the systems. Using the cell-cycle control networks of the budding yeast (Saccharomyces cerevisiae) and the fission yeast (Schizosaccharomyces pombe) as examples, we first demonstrate the correlation of this quantity with the dynamic stability of biological control networks, and then we establish a significant association between this quantity and the structural stability of the networks. And we further substantiate the generality of this approach with a broad spectrum of biological and random networks. We conclude that the regulation entropy is an effective order parameter in evaluating the robustness of biological control networks. Our work suggests a novel connection between the topological feature and the dynamic property of biological regulatory networks.     

Mechanism of the Rpn13-induced activation of Uch37

Uch37 is a de-ubiquitinating enzyme that is activated by Rpn13 and involved in the proteasomal degradation of proteins. The full-length Uch37 was shown to exhibit low iso-peptidase activity and is thought to be auto-inhibited. Structural comparisons revealed that within a homodimer of Uch37, each of the catalytic domains was blocking the other’s ubiquitin (Ub)-binding site. This blockage likely prevented Ub from entering the active site of Uch37 and might form the basis of auto-inhibition. To understand the mode of auto-inhibition clearly and shed light on the activation mechanism of Uch37 by Rpn13, we investigated the Uch37-Rpn13 complex using a combination of mutagenesis, biochemical, NMR, and smallangle X-ray scattering (SAXS) techniques. Our results also proved that Uch37 oligomerized in solution and had very low activity against the fluorogenic substrate ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) of de-ubiquitinating enzymes. Uch37Δ^Hb,Hc,KEKE, a truncation removal of the C-terminal extension region (residues 256–329) converted oligomeric Uch37 into a monomeric form that exhibited iso-peptidase activity comparable to that of a truncation-containing the Uch37 catalytic domain only. Wealso demonstrated that Rpn13C (Rpn13 residues 270–407) could disrupt the oligomerization of Uch37 by sequestering Uch37 and forming a Uch37-Rpn13 complex. Uch37 was activated in such a complex, exhibiting 12-fold-higher activity than Uch37 alone. Time-resolved SAXS (TR-SAXS) and FRET experiments supported the proposed mode of auto-inhibition and the activation mechanism of Uch37 by Rpn13. Rpn13 activated Uch37 by forming a 1:1 stoichiometric complex in which the active site of Uch37 was accessible to Ub.     

Improved production of phenylethanoid glycosides by Cistanche deserticola cells cultured in an internal loop airlift bioreactor with sifter riser

An internal loop airlift bioreactor with sifter riser (ILABSR) was composed of a bubble column and a draught-tube rolled with 40-mesh sifter that placed 5 cm above the bottom at the center of the column. A 2 L ILABSR was used for the suspension cultivation of Cistanche deserticola cells and its performance was compared with shake flask culture and a bubble column. Under the optimum culture conditions with the air flowrate of 0.075 m³/h and the inoculation size of 4.7%, about one-fifth cells were attached to the sifter draught-tube. PeG content in these cells was 16.3%, which was 104% higher than that of suspension cells. The production of phenylethanoid glycosides reached 0.85 g/L, which was 102 and 4% higher than those cultured in a 2 L bubble column and shake flasks respectively under their optimal culture conditions.