Directionally selective calcium signals in dendrites of starburst amacrine cells

The detection of image motion is fundamental to vision. In many species, unique classes of retinal ganglion cells selectively respond to visual stimuli that move in specific directions. It is not known which retinal cell first performs the neural computations that give rise to directional selectivity in the ganglion cell. A prominent candidate has been an interneuron called the 'starburst amacrine cell'. Using two-photon optical recordings of intracellular calcium concentration, here we find that individual dendritic branches of starburst cells act as independent computation modules. Dendritic calcium signals, but not somatic membrane voltage, are directionally selective for stimuli that move centrifugally from the cell soma. This demonstrates that direction selectivity is computed locally in dendritic branches at a stage before ganglion cells.     

RhoA/Rho激酶在大鼠阴茎勃起机制中的调节作用

一般认为一氧化氮（nitric oxide,NO）释放增加促进小动脉和海绵体平滑肌舒张是导致男性阴茎勃起的主要机制。研究发现内源性血管收缩因子对维持阴茎萎状态有重要作用。应用大鼠模型活体检测使用Y-27632拮抗Rho-激酶活性后对阴茎勃起生理机制的影响;应用免疫转印技术检测 阴茎海绵体内RhoA和Rho激酶蛋白的表达。结果显示在大鼠海绵体组织内有内源性RhoA和Rho激酶蛋白的表达和存在;Y-27632海绵体内注射阻滞RhoA/Rho激酶活性使ICP和CCP/MAP比值明显升高;局部小剂量应用Y-27632对MAP没有明显影响;Y-27632可增强系列电刺激引起的由NO介导的CCP/MAP比值的增加;NO合成酶和鸟苷酸环化酶抑制剂的作用不能阻滞RhoA/Rho激酶抑制剂对大鼠阴茎海绵体平滑肌的松弛作用和CCP/MAP的增加。说明RhoA/Rho激酶信号系统发挥了维持海绵体萎软状态重要作用,这是与NO介导途径不同的阴茎勃起生理调节机制。RhoA/Rho激酶抑制剂可能是ED治疗的新领域新方法。     

Re-evaluation of the phylogenetic position of the genus Dexiotrichides (Protozoa, Ciliophora, Scuticociliatida) inferred from stomatogenetic and molecular information for Dexiotrichides pangi

The divisional process and systematic position of the marine scuticociliate Dexiotrichides pangi are studied. Based on both stomatogenetic data and 18S rDNA gene sequences, the phylogeny and morphogenetic characteristics of this taxon, and of other related genera, are analyzed and discussed. Both the divisionary events and the molecular biological data indicate that this speciesgenus, together with certain other genera in the Dexiotricha-complex, occupies an intermediate position between the tetrahymenids and the “typical” scuticociliate, which suggests that the Dexiotricha-like taxa should be excluded from the “true” scuticociliates. As a further contribution, the process of stomatogenesis in D. pangi can be summarized as follows: (1) The oral primordia in the opisthe are formed only by the proliferation of basal bodies in the scutica field, which subsequently develop into three membranelles, while the new paroral membrane seems to be generated by the sub-anterior portion of somatic kinety 1 (the 1st postoral intercalary kinety). The latter character exhibits a mode similar to Tetrahymena. (2) In the proter the parental membranelles are retained and remain unchanged throughout the entire division process; only the old paroral membrane is disassembled and differentiated into the anlage, which then gives rise to the new paroral membrane and the scutica of the proter. The 18S rRNA gene sequence reported here is the first one for a ciliate in the Dexiotricha-complex.     

Steady-state kinetic studies with the polysulfonate U-9843, an HIV reverse transcriptase inhibitor

The tetramer of ethylenesulfonic acid (U-9843) is a potent inhibitor of HIV-1 RT^* and possesses excellent antiviral activity at nontoxic doses in HIV-1 infected lymphocytes grown in tissue culture. Kinetic studies of the HIV-1 RT-catalyzed RNA-directed DNA polymerase activity were carried out in order to determine if the inhibitor interacts with the template: primer or the deoxyribonucleotide triphosphate (dNTP) binding sites of the polymerase. Michaelis-Menten kinetics, which are based on the establishment of a rapid equilibrium between the enzyme and its substrates, proved inadequate for the analysis of the experimental data. The data were thus analyzed using steady-state Briggs-Haldane kinetics assuming that the template:primer binds to the enzyme first, followed by the binding of the dNTP and that the polymerase is a processive enzyme. Based on these assumptions, a velocity equation was derived which allows the calculation of all the specific forward and backward rate constants for the reactions occurring between the enzyme, its substrates and the inhibitor. The calculated rate constants are in agreement with this model and the results indicated that U-9843 acts as a noncompetitive inhibitor with respect to both the template:primer and dNTP binding sites. Hence, U-9843 exhibits the same binding affinity for the free enzyme as for the enzyme-substrate complexes and must inhibit the RT polymerase by interacting with a site distinct from the substrate binding sites. Thus, U-9843 appears to impair an event occurring after the formation of the enzyme-substrate complexes, which involves either an event leading up to the formation of the phosphoester bond, the formation of the ester bond itself or translocation of the enzyme relative to its template:primer following the formation of the ester bond.     

The use of complexes of lymphocyte antigens with antibody as immunogens for the preparation of antilymphocytic sera

Résumé Des complexes antigène-anticorps préparées par réaction du sérum antilymphocytaire (ALS) avec des lymphocytes ou avec des broyats lymphocytaires furent injectées aux lapins. Les ALS obtenu ainsi des lapins furent immunodépresseurs. Cette technique peut être utilisée pour obtenir des ALS plus specifique et moins toxique.

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Acknowlegments. We are indebted to our colleagues in the ALS Project Group of these Laboratories for much help and advice.     

The alkaloids ofAspidosperma neblinae. An application of a directly coupled gas chromatograph-mass spectrometer

Zusammenfassung Die Analyse der Basenfraktion vonAspidosperma neblinae mit Hilfe eines mit einem Massenspektrometer gekuppelten Gas-Chromatographen führte zur Identifizierung der folgenden Alkaloide: Aspidospermidin, 1, 2-Dehydroaspidospermidin, Deacetylpyrifolidin, 1, 2-Dehydrodeacetylpyrifolidin, Demethoxyaspidospermin, Aspidospermin, Demethylaspidospermin, Pyrifolidin, Aspidocarpin, Eburnamonin, und Neblinin.     

Transmission of cutaneous leishmaniasis by sand flies is enhanced by regurgitation of fPPG

Sand flies are the exclusive vectors of the protozoan parasite Leishmania, but the mechanism of transmission by fly bite has not been determined nor incorporated into experimental models of infection. In sand flies with mature Leishmania infections the anterior midgut is blocked by a gel of parasite origin, the promastigote secretory gel. Here we analyse the inocula from Leishmania mexicana-infected Lutzomyia longipalpis sand flies. Analysis revealed the size of the infectious dose, the underlying mechanism of parasite delivery by regurgitation, and the novel contribution made to infection by filamentous proteophosphoglycan (fPPG), a component of promastigote secretory gel found to accompany the parasites during transmission. Collectively these results have important implications for understanding the relationship between the parasite and its vector, the pathology of cutaneous leishmaniasis in humans and also the development of effective vaccines and drugs. These findings emphasize that to fully understand transmission of vector-borne diseases the interaction between the parasite, its vector and the mammalian host must be considered together.     

Mechanisms of gene silencing by double-stranded RNA

Double-stranded RNA (dsRNA) is an important regulator of gene expression in many eukaryotes. It triggers different types of gene silencing that are collectively referred to as RNA silencing or RNA interference. A key step in known silencing pathways is the processing of dsRNAs into short RNA duplexes of characteristic size and structure. These short dsRNAs guide RNA silencing by specific and distinct mechanisms. Many components of the RNA silencing machinery still need to be identified and characterized, but a more complete understanding of the process is imminent.