New synaptic bouton formation is disrupted by misregulation of microtubule stability in aPKC mutants

The Baz/Par-3-Par-6-aPKC complex is an evolutionarily conserved cassette critical for the development of polarity in epithelial cells, neuroblasts, and oocytes. aPKC is also implicated in long-term synaptic plasticity in mammals and the persistence of memory in flies, suggesting a synaptic function for this cassette. Here we show that at Drosophila glutamatergic synapses, aPKC controls the formation and structure of synapses by regulating microtubule (MT) dynamics. At the presynapse, aPKC regulates the stability of MTs by promoting the association of the MAP1Brelated protein Futsch to MTs. At the postsynapse, aPKC regulates the synaptic cytoskeleton by controlling the extent of Actin-rich and MT-rich areas. In addition, we show that Baz and Par-6 are also expressed at synapses and that their synaptic localization depends on aPKC activity. Our findings establish a novel role for this complex during synapse development and provide a cellular context for understanding the role of aPKC in synaptic plasticity and memory.     

Conservation of fragmented small populations: endemic species persistence on California's smallest channel island

We examined resilience to extreme reduction of habitat, and long-term and long-distance isolation for an endemic species using California's Santa Barbara Island as a natural model. The island is smaller than 260ha, has been isolated by 40km of ocean from the nearest other island for more than 10,000 years and was severely impacted by human activities and feral browsers during at least the past 90 years. Less than 0.2km² of native plant cover remains, yet seven apparently endemic plants and animals persist, including a moth, Argyrotaenia isolatissima (Tortricidae). Using mitochondrial DNA sequence we examined the levels of genetic divergence between the moth and its closest relatives on the other Channel Islands and the California mainland. A. isolatissima has 15bp differences, including one non-synonymous substitution, from the most closely related taxon, on San Nicolas Island. Both parsimony and likelihood-based molecular systematic analyses confirm the evolutionary independence of A. isolatissima and indicate that portions of an endemic flora and fauna may persist in small fragments, despite long-term isolation and disturbance. Habitat conservation planning may underestimate the value of very small, temporally and spatially isolated fragments of native habitat not only to maintain, but also to generate endemic biodiversity.     

采用共聚焦显微术对家兔呼吸道感受器进行观察

呼吸道感受器在机体对肺部物理和化学环境变化作出反应时起到重要作用,它们引起的反射具有调节或保护作用,或产生病理效应。基于电生理研究,可将呼吸道感受器分为四类,但它们的组织结构不详。由于对感受器形态的认识不足,阻碍了对其生理功能的理解。近来,我们采用共聚焦显微术与免疫组织化学方法（用钠钾ATP酶作为标记),首次高清晰度地展示了呼吸感受器结构。本文采用这种新方法在家兔中进行了系统观察。结果显示,各级气道通均有多种不同的感受器结构。大气道中的结构往往比小气道中的复杂,虽然它们的大小、所在部位与走向不尽相同,但都有多个终端末梢。有些感受器埋置在气道平滑肌中,其结构便于感受对气道的机械性刺激：有些覆盖在气道上皮的表面,其形态适合于感受通过气道的刺激性物质;另一些则位于气道粘膜下,在上皮与平滑肌之间。有些传入神经轴突可支配多个感受器结构。据此,传入单纤维中记录到的电活动是来自一个感觉单位。后者可以由多个感受器组成。除了感受器之外,我们还观察到气道内神经节,它们与感受神经的轴突密切相关。本文证实气道内存在不同形态结构的感受器,并探讨了其生理分类。     

4-Amino cyclohexylglycine analogues as potent dipeptidyl peptidase IV inhibitors

Substituted 4-amino cyclohexylglycine analogues were evaluated for DP-IV inhibitory properties. Bis-sulfonamide 15e was an extremely potent 2.6 nM inhibitor of the enzyme with excellent selectivity over all counterscreens. 2,4-difluorobenzenesulfonamide 15b and 1-naphthyl amide 16b, however, combined an acceptable in vitro profile with good pharmacokinetic properties in the rat, and 15b was orally efficacious at 3 mpk in an OGTT in lean mice.     

Synthesis and activity of 1-aryl-1'-imidazolyl methyl ethers as non-thiol farnesyltransferase inhibitors

A series of imidazole-containing methyl ethers (4-5) have been designed and synthesized as potent and selective farnesyltransferase inhibitors (FTIs) by transposition of the D-ring to the methyl group on the imidazole of the previously reported FTIs 3. Several compounds such as 4h and 5b demonstrate superior enzymatic activity to the current benchmark compound tipifarnib (1) with IC(50) values in the lower subnanomolar range, while maintaining excellent cellular activity comparable to tipifarnib. The compounds are characterized as being simple, easier to make, and possess no chiral center involved.     

Design, synthesis, and activity of 4-quinolone and pyridone compounds as nonthiol-containing farnesyltransferase inhibitors

As a part of our efforts to identify potent inhibitors of farnesyltransferase (FTase), modification of the structure of tipifarnib through structure-based design was undertaken by replacing the 2-quinolones with 4-quinolones and pyridones, and subsequent relocation of the D-ring to the N-methyl group on the imidazole ring. This study has yielded a novel series of potent and selective FTase inhibitors. The X-ray structure of tipifarnib (1) in complex with FTase was described.     

Synthesis of acyloxymethyl ester prodrugs of the transferable protein farnesyl transferase substrate farnesyl methylenediphosphonate

Three isoprenoid diphosphate analogues of farnesyl diphosphate (FPP) where the diphosphate has been replaced by methylene diphosphonate and the negative charges masked by frangible pivaloyloxymethyl (POM) esters were prepared. Farnesyl methylenediphosphonate is a sub-micromolar substrate for protein farnesyl transferase. The tripivaloyloxymethyl esters of isoprenoid methylenediphosphonate have significantly increased lipophilicity and may act as important farnesyl diphosphate prodrugs.     

Micropatterned agarose gels for stamping arrays of proteins and gradients of proteins

We describe a method for repetitive and rapid formation of planar microarrays and gradients of proteins using patterned agarose stamps. It demonstrates: (i) micropatterning of agarose gels with feature sizes as small as 2 microm; (ii) inking of posts (diameter 50-1000 microm) on patterned agarose stamps with one or multiple (here, eight) proteins and repetitive stamping of patterns (>100 times in the case of one protein) and arrays (20 times in the case of eight proteins) without the need for intermediate re-inking; (iii) transferring spots of proteins with good homogeneity in surface coverage to glass slides; (iv) applying this technique to surface-based immunoassays; (v) stamping that requires only sub-nanomolar amounts of protein (typically approximately 3 microg in approximately 0.6 microL of solution); (vi) stamping without the need for drying of the proteins, as opposed to stamping with stamps made of poly(dimethylsiloxane); and (vii) patterning gradients of proteins by allowing two proteins to diffuse toward each other in an agarose stamp, followed by printing the protein gradients onto a surface.