Inhibition of Eimeria tenella CDK‐Related Kinase 2: From Target Identification to Lead Compounds

Apicomplexan parasites encompass several human‐ and animal‐pathogenic protozoans such as Plasmodium falciparum, Toxoplasma gondii, and Eimeria tenella. E. tenella causes coccidiosis, a disease that afflicts chickens, leading to tremendous economic losses to the global poultry industry. The considerable increase in drug resistance makes it necessary to develop new therapeutic strategies against this parasite. Cyclin‐dependent kinases (CDKs) are key molecules in cell‐cycle regulation and are therefore prominent target proteins in parasitic diseases. Bioinformatics analysis revealed four potential CDK‐like proteins, of which one—E. tenella CDK‐related kinase 2 (EtCRK2)—has already been characterized by gene cloning and expression. 1 By using the CDK‐specific inhibitor flavopiridol in EtCRK2 enzyme assays and schizont maturation assays (SMA), we could chemically validate CDK‐like proteins as potential drug targets. An X‐ray crystal structure of human CDK2 (HsCDK2) served as a template to build protein models of EtCRK2 by comparative homology modeling. Structural differences in the ATP binding site between EtCRK2 and HsCDK2, as well as chicken CDK3, were addressed for the optimization of selective ATP‐competitive inhibitors. Virtual screening and “wet‐bench” high‐throughput screening campaigns on large compound libraries resulted in an initial set of hit compounds. These compounds were further analyzed and characterized, leading to a set of four promising lead compounds for development as EtCRK2 inhibitors.     

Reported food intake and distribution of body fat: a repeated cross-sectional study

Background  

Body mass, as well as distribution of body fat, are predictors of both diabetes and cardiovascular disease. In Northern Sweden, despite a marked increase in average body mass, prevalence of diabetes was stagnant and myocardial infarctions decreased. A more favourable distribution of body fat is a possible contributing factor.     

Aziridide-based inhibitors of cathepsin L: synthesis, inhibition activity, and docking studies

A comprehensive screening of N-acylated aziridine (aziridide) based cysteine protease inhibitors containing either Boc-Leu-Caa (Caa=cyclic amino acid), Boc-Gly-Caa, or Boc-Phe-Ala attached to the aziridine nitrogen atom revealed Boc-(S)-Leu-(S)-Azy-(S,S)-Azi(OBn)(2) (18 a) as a highly potent cathepsin L (CL) inhibitor (K(i)=13 nM) (Azy=aziridine-2-carboxylate, Azi=aziridine-2,3-dicarboxylate). Docking studies, which also accounted for the unusual bonding situations (the flexibility and hybridization of the aziridides) predict that the inhibitor adopts a Y shape and spans across the entire active site cleft, binding into both the nonprimed and primed sites of CL.     

Metal partitioning in a sulfidic canal sediment: metal solubility as a function of pH combined with EDTA extraction in anoxic conditions

The chemical forms of low concentrations of metals (Zn, Pb, Cu, Cd, Co and Mn) and main components (Fe and Ca) were determined under the original reducing conditions of a sulfide-rich sediment from the Gent-Terneuzen Canal (Belgium). Therefore, dissolution experiments as a function of pH were made in a salt solution mimicking the canal water. The centrifugates remaining after the dissolution vs. pH experiments were subsequently further extracted with 0.1 M EDTA/0.5 M NaAc (pH 4.65) to determine eventually readsorbed and/or reprecipitated metal ions. The experimental dissolution vs. pH edge of calcium, iron, manganese and cobalt had a lower slope than theoretically expected on the basis of the solubility of, respectively, calcium carbonate, iron sulfide/iron carbonate, manganese sulfide/manganese carbonate and cobalt sulfide and was explained by the combination of (a) the solubilities of the various minerals and (b) metal readsorption onto clay minerals and organic matter. Higher metal recoveries were measured in the 0.1 M EDTA/0.5 M NaAc mixture and proved that in addition coprecipitate formation with iron sulfide/iron carbonate minerals may occur. The solubility of zinc, lead, cadmium and copper was very low in the mimicking salt solution even at very low pH values (up to pH 1) in agreement with the theoretical solubility of their discrete metal sulfides. However, by using an additional 0.1 M EDTA/0.5 M NaAc extraction on the centrifugates remaining after the dissolution vs. pH experiments, it was qualitatively shown that zinc and lead were partly associated with iron sulfide/iron carbonate phases in the real sediment (in addition to their presence in discrete metal sulfides). Cadmium was present solely in discrete cadmium sulfide phases. It was not possible to verify whether copper was present in discrete copper sulfide phases and/or in mixed coprecipitates with iron sulfide/iron carbonate minerals.     

City make-overs: The place marketing of melbourne during the kennett years, 1992-99

Over the past two decades, the emergence of a new global economic order has triggered an intensification of competition between cities for footloose investment capital and tourism. Some have proven to be more successful than others at this form of civic boosterism through the adoption of place promotion and place marketing. Unfortunately, most research on this contemporary urban phenomenon has focused on Northern Europe and the United States, not Australia. To redress this imbalance, this paper examines why intercity competition has intensified, outlining the main features of place promotion and marketing and then examining how the Kennett Coalition government place-marketed Melbourne and Victoria within the Asia-Pacific Rim region. It attempts to assess whether the place marketing campaign succeeded and highlights a number of limitations with this mode of urban analysis.     

Book reviews

TOWARDS COSMOPOLIS: Planning for Multicultural Cities. Leonie Sandercock. John Wiley & Sons, Chichester, UK, 1998. ISBN 0471971987 (paperback). 258 pp.; bibliography. RRP $41.15.

THE MYTH OF THE POWERLESS STATE: Governing the Economy in a Global Era. Linda Weiss. Polity/Blackwell, Cambridge, 1998. ISBN 0745615821 (paperback). 260 pp.; index and bibliography. RRP $37.95.

SOCIAL CHANGE, SUBURBAN LIVES: An Australian Newtown 1960s to 1990s. Lois Bryson and Ian Winter. Allen & Unwin, Sydney, 1998. ISBN 1864486996 (paperback). 238 pp.; index and bibliography. RRP $29.95.

CONTESTING THE AUSTRALIAN WAY: States, Markets and Civil Society. Paul Smyth and Bettina Cass (eds). Cambridge University Press, Cambridge, 1998. ISBN 0521633060 (hardback). 280 pp.; bibliography and index. RRP $90.00.

THE ECOLOGICAL CITY AND THE CITY EFFECT: Essays on the Urban Planning Requirements for the Sustainable City. Franco Archibugi. Ashgate, Aldershot, UK, 1997. ISBN 1859726534 (hardback). 243 pp.; bibliography and index. RRP $96.50.

CITIES FOR TOMORROW: Integrating Land Use, Transport and the Environment. Hans L. Westerman. Austroads, Sydney, 1998. ISBN 0855885033 (paperback). Resource Document 202 pp.; Better Practice Guide. 394 pp.; index. RRP $180.00.

EAST WEST PERSPECTIVES ON 21ST CENTURY URBAN DEVELOPMENT: Sustainable Eastern and Western Cities in the New Millennium. John Brotchie, Peter Newton, Peter Hall and John Dickey (eds). Ashgate, Aldershot, UK, 1999. ISBN 1840143177 (hardback). 432 pp.; bibliography and index. RRP $122.00.

MADE IN LANCASHIRE: A History of Regional Industrialisation. Geoffrey Timmins. Manchester University Press, Manchester, 1998. ISBN 0719045398. 334 pp.; bibliography and index. RRP $145.00.     

Characterization of a Schistosoma mansoni cDNA encoding a B-like cyclophilin and its expression in Escherichia coli

A cDNA encoding a Schistosoma mansoni cyclophilin (SmCyP) has been cloned by polymerase chain reaction amplification using degenerate oligonucleotides based on known conserved cyclophilin (CyP) sequences and by screening an expression cDNA library. The cDNA sequence encodes a 21.5-kDa protein, which shares 59% sequence identity with human CyP B. The SmCyP protein was expressed in Escherichia coli with a hexahistidine affinity tag at its amino terminus and antibodies to the purified (His6)-SmCyP fusion protein were raised in a rabbit. Fractionation of parasite material followed by immunoblot analysis revealed that schistosome CyP is a soluble protein. The N-terminus of the predicted protein contains a hydrophobic region, suggestive of a signal sequence. Accordingly, a recombinant SmCyP protein, lacking the first 23 amino acids was found to share the same gel electrophoretic mobility as the parasite-derived CyP protein, suggesting cleavage of a leader sequence. Hybridization of genomic DNA to a full-length cDNA probe indicates that the SmCyP gene is present as a single copy. Immunohistological experiments in conjunction with confocal scanning laser microscopy and immune electron microscopy show that SmCyP is present in abundance in the adult worm as well as in the schistosomula. The function of CyP in the schistosome is presently unclear, but since its ligand, cyclosporin A, has antischistosomal activity, its function is expected to be a vital one.     

Localizing and identifying living bacteria in an abiotic environment by a combination of Raman and fluorescence microscopy

A fast, easy, and reliable identification of microorganisms is indispensable in many fields such as medicine, food production, or the pharmaceutical industry. However, in native samples, biotic particles often appear together with abiotic particles. Therefore, it is a prerequisite that biotic particles can be differentiated from abiotic particles appearing in the identification setup. In addition, for many applications, not all microorganisms are of interest but only the living ones. Therefore, in this contribution, different bacteria species were stained with a live/dead staining kit (SYTO 9 and propidium iodide) prior to Raman spectroscopic identification. Since only living and dead microorganisms are getting stained by SYTO 9 or PI, biotic particles can easily be spotted and localized in-between abiotic particles. By using a Raman laser excitation wavelength outside the absorption band of the dye, fluorescence-free Raman spectra were obtained. The living cells were identified by means of Raman spectroscopy in combination with a support vector machine. Furthermore, the localization of bacterial cells in a mix of abiotic particles is demonstrated.     

Cross-language plagiarism detection

Cross-language plagiarism detection deals with the automatic identification and extraction of plagiarism in a multilingual setting. In this setting, a suspicious document is given, and the task is to retrieve all sections from the document that originate from a large, multilingual document collection. Our contributions in this field are as follows: (1) a comprehensive retrieval process for cross-language plagiarism detection is introduced, highlighting the differences to monolingual plagiarism detection, (2) state-of-the-art solutions for two important subtasks are reviewed, (3) retrieval models for the assessment of cross-language similarity are surveyed, and, (4) the three models CL-CNG, CL-ESA and CL-ASA are compared. Our evaluation is of realistic scale: it relies on 120,000 test documents which are selected from the corpora JRC-Acquis and Wikipedia, so that for each test document highly similar documents are available in all of the six languages English, German, Spanish, French, Dutch, and Polish. The models are employed in a series of ranking tasks, and more than 100 million similarities are computed with each model. The results of our evaluation indicate that CL-CNG, despite its simple approach, is the best choice to rank and compare texts across languages if they are syntactically related. CL-ESA almost matches the performance of CL-CNG, but on arbitrary pairs of languages. CL-ASA works best on “exact” translations but does not generalize well.