Functional mutation of SMAC/DIABLO, encoding a mitochondrial proapoptotic protein, causes human progressive hearing loss DFNA64

SMAC/DIABLO is a mitochondrial proapoptotic protein that is released from mitochondria during apoptosis and counters the inhibitory activities of inhibitor of apoptosis proteins, IAPs. By linkage analysis and candidate screening, we identified a heterozygous SMAC/DIABLO mutation, c.377C>T (p.Ser126Leu, refers to p.Ser71Leu in the mature protein) in a six-generation Chinese kindred characterized by dominant progressive nonsyndromic hearing loss, designated as DFNA64. SMAC/DIABLO is highly expressed in human embryonic ears and is enriched in the developing mouse inner-ear hair cells, suggesting it has a role in the development and homeostasis of hair cells. We used a functional study to demonstrate that the SMAC/DIABLO(S71L) mutant, while retaining the proapoptotic function, triggers significant degradation of both wild-type and mutant SMAC/DIABLO and renders host mitochondria susceptible to calcium-induced loss of the membrane potential. Our work identifies DFNA64 as the human genetic disorder associated with SMAC/DIABLO malfunction and suggests that mutant SMAC/DIABLO(S71L) might cause mitochondrial dysfunction.     

Regulation of tolerance of Chlamydomonas reinhardtii to heavy metal toxicity by heme oxygenase-1 and carbon monoxide

Investigation of heavy metal tolerance genes in green algae is of great importance because heavy metals have become one of the major contaminants in the aquatic ecosystem. In plants, accumulation of heavy metals modifies many aspects of cellular functions. However, the mechanism by which heavy metals exert detrimental effects is poorly understood. In this study, we identified a role for HO-1 (encoding heme oxygenase-1) in regulating the response of Chlamydomonas reinhardtii, a unicellular green alga, to mercury (Hg). Transgenic algae overexpressing HO-1 showed high tolerance to Hg exposure, with a 48.2% increase in cell number over the wild type, but accumulated less Hg. Physiological analysis revealed that expression of HO-1 suppressed the Hg-induced generation of reactive oxygen species. We further identified the effect of carbon monoxide (CO), a product of HO-1-mediated heme degradation, on growth and physiological parameters. Interestingly, administration of exogenous CO at non-toxic levels also conferred the tolerance of algae to Hg exposure. The CO-mediated alleviation of Hg toxicity was closely related to the lower accumulation of Hg and free radical species. These results indicate that functional identification of HO-1 is useful for molecular breeding designed to improve plant tolerance to heavy metals and reduce heavy metal accumulation in plant cells.     

Sequential binding of αVβ3 and ICAM-1 determines fibrin-mediated melanoma capture and stable adhesion to CD11b/CD18 on neutrophils

Fibrin (Fn) deposition defines several type 1 immune responses, including delayed-type hypersensitivity and autoimmunity in which polymorphonuclear leukocytes (PMNs) are involved. Fn monomer and fibrinogen are multivalent ligands for a variety of cell receptors during cell adhesion. These cell receptors provide critical linkage among thrombosis, inflammation, and cancer metastasis under venous flow conditions. However, the mechanisms of Fn-mediated interactions among immune cells and circulating tumor cells remain elusive. By using a cone-plate viscometer shear assay and dual-color flow cytometry, we demonstrated that soluble fibrinogen and Fn had different abilities to enhance heterotypic aggregation between PMNs and Lu1205 melanoma cells in a shear flow, regulated by thrombin levels. In addition, the involvement of integrin α(v)β(3), ICAM-1, and CD11b/CD18 (Mac-1) in fibrin(ogen)-mediated melanoma-PMN aggregations was explored. Kinetic studies provided evidence that ICAM-1 mediated initial capture of melanoma cells by PMNs, whereas α(v)β(3) played a role in sustained adhesion of the two cell types at a shear rate of 62.5 s(-1). Quantitative analysis of the melanoma-PMN interactions conducted by a parallel-plate flow chamber assay further revealed that at a shear rate of 20 s(-1), α(v)β(3) had enough contact time to form bonds with Mac-1 via Fn, which could not otherwise occur at a shear rate higher than 62.5 s(-1). Our studies have captured a novel finding that leukocytes could be recruited to tumor cells via thrombin-mediated Fn formation within a tumor microenvironment, and α(v)β(3) and ICAM-1 may participate in multistep fibrin(ogen)-mediated melanoma cell adhesion within the circulation.     

Efficient synthesis and biological evaluation of epiceanothic acid and related compounds

Epiceanothic acid (1) is a naturally occurring, but very rare pentacyclic triterpene with a unique pentacyclic triterpene (PT) structure. An efficient synthesis of 1 starting from betulin (3) has been accomplished in 12-steps with a total yield of 10% in our study. Compound 1 and selected synthetic intermediates were further evaluated as anti-HIV-1 agents, inhibitors of glycogen phosphorylase (GP), and cytotoxic agents. Compound 1 exhibited moderate HIV-1 inhibition. Most importantly, compound 5, with an opened A-ring, showed significant GP inhibitory activity with an IC₅₀ of 0.21 μM, suggesting a potential for development as an anti-diabetic agent. On the other hand, compound 12, with a closed A-ring, showed potent cytotoxicity against A549 and MCF-7 human tumor cell lines, with IC₅₀ values of 0.89 and 0.33 μM, respectively. These results suggest that the A-ring of PTs is an important pharmacophore that could be modified to involve different biological activities.     

Efficacy switching SAR of mGluR5 allosteric modulators: highly potent positive and negative modulators from one chemotype

A series of metabotropic glutamate 5 receptor (mGluR5) allosteric ligands with positive, negative or no modulatory efficacy is described. The ability of this series to yield both mGluR5 PAMs and NAMs with single-digit nanomolar potency is unusual, and the underlying SAR is detailed.     

覆膜与沟垄种植模式对旱作马铃薯产量形成及水分运移的影响

研究了覆膜及不同沟垄种植模式对黄土高原西部半干旱区旱作马铃薯产量形成和水分运移的影响.结果表明:平畦覆膜(T2)、全膜双垄沟播(T3)、全膜双垄垄播(T4)、半膜膜侧种植(T5)和半膜沟垄垄播(T6)种植方式的产量分别比传统平畦不覆膜(T1)方式高50.1%、75.9%、86.8%、69.6%和60.6%;水分利用效率(WUE)分别提高47.0%、82.7%、84.0%、75.2%和54.3%,其中,T4、T3产量和WUE增加幅度最大.与传统方式相比,各覆膜及沟垄处理普遍优化了马铃薯各产量构成性状,其中T4和T3最有利于大薯率和中薯率的提高、绿薯率和烂薯率的降低,其单株结薯数和单株薯产量也较高.因此,全膜双垄垄播和全膜双垄沟播为半干旱区马铃薯适宜的抗旱节水高产种植模式.     

长期施用含硫和含氯化肥对稻田杂草生长动态的影响

利用湖南祁阳红壤稻田长期定位试验,研究了在等量氮磷钾养分条件下,长期施用含Cl-、SO2-4和Cl-+SO2-4化肥水稻生育期间杂草种类和生物量的变化.结果表明:施肥34年后,施用含Cl-化肥处理水稻生育期间杂草的种类最多、总生物量(浮生杂草和湿生杂草的生物量之和)最大,早稻期间杂草平均总干物质量分别比含SO2-4和Cl-+SO2-4化肥处理增加了51.4%和17.6%,晚稻期间分别增加了144%和242%.含SO2-4和Cl-+SO2-4化肥处理稻田中浮生杂草生物量较大,而含Cl-化肥处理田间几乎没有浮生杂草生长.杂草总干物质量和湿生杂草干物质量均与土壤Cl-含量呈显著正相关(相关系数分别为0.764**和0.948**),与土壤SO2-4-S含量呈显著负相关(相关系数分别为0.849**和0.641*).土壤碱解氮和有效磷受土壤SO2-4-S、Cl-及pH的共同作用对杂草总干物质量产生影响.通过各种施肥措施维持土壤适宜pH及碱解氮、有效磷含量,提高土壤SO2-4-S含量、降低Cl-含量,能有效抑制南方红壤稻田中湿生杂草的生长,降低杂草总生物量.     

Microsatellite analysis revealed genetic diversity and population structure among Chinese cashmere goats

Most cashmere goats are found in northern China and Mongolia. They are regarded as precious resources for their production of high quality natural fibre for the textile industry. It was the first time that the genetic diversity and population structure of nine Chinese cashmere populations has been assessed using 14 ISAG/FAO microsatellite markers. In addition, two Iranian populations and one West African goat population were genotyped for comparison. Results indicated that the genetic diversity of Chinese cashmere goats was rich, but less than those of the Iranian goat populations. All pairwise F_ST values between the Chinese cashmere goat populations reached a highly significant level (P < 0.001), suggesting that they should all be considered as separate breeds. Finally, clustering analysis divided Chinese cashmere goats into at least two clusters, with the Tibetan Hegu goats alone in one cluster. An extensive admixture was detected among the Chinese goat breeds (except the Hegu), which have important implications for breeding management.     

A、O型口蹄疫T/B细胞多抗原表位融合基因植物表达载体的构建

通过对(pMD-xsB/xsT/xsBT/xsBTT)4个克隆载体进行PCR,在(xsB/xsT/xsBT/xsBTT)目的基因前接入引导肽序列(用Y表示),克隆中间载体(pM D-xsY B/xsYT/xsYBT/xsYBTT),再利用同尾酶的酶切及连接得到4个重组植物表达载体(pBI-xsYB/xsYT/xsYBT/xsYBTT).经限制性酶切鉴定和测序分析证实表达载体构建正确,为下一步热研2号柱花草转基因植株的获得奠定了基础.