With the publication of the sequence of the human genome, we are challenged to identify the functions of an estimated 70,000 human genes and the much larger number of proteins encoded by these genes. Of particular interest is the identification of gene products that play a role in human disease pathways, as these proteins include potential new targets that may lead to improved therapeutic strategies. This requires the direct measurement of gene function on a genomic scale in cell-based, functional assays. We have constructed and validated an individually arrayed, replication-defective adenoviral library harboring human cDNAs, termed PhenoSelect library. The adenoviral vector guarantees efficient transduction of diverse cell types, including primary cells. The arrayed format allows screening of this library in a variety of cellular assays in search for gene(s) that, by overexpression, induce a particular disease-related phenotype. The great majority of phenotypic assays, including morphological assays, can be screened with arrayed libraries. In contrast, pooled-library approaches often rely on phenotype-based isolation or selection of single cells by employing a flow cytometer or screening for cell survival. An arrayed placental PhenoSelect library was screened in cellular assays aimed at identifying regulators of osteogenesis, metastasis, and angiogenesis. This resulted in the identification of known regulators, as well as novel sequences that encode proteins hitherto not known to play a role in these pathways. These results establish the value of the PhenoSelect platform, in combination with cellular screens, for gene function discovery.

No abstract

A reverse line blot (RLB) assay was developed to identify different Trichinella genotypes. The RLB assay accomplishes detection and specific identification of the different Trichinella genotypes and relies on hybridization of the amplified 5S ribosomal DNA intergenic spacer regions to specific, membrane-bound oligonucleotide probes. After one single amplification, we were able to detect and genetically identify six sibling species, i.e., T. spiralis, T. britovi, T. nativa, T. murrelli, T. nelsoni, and T. pseudospiralis, and two additional Trichinella genotypes, T6 and T8. Twenty-four Trichinella strains of different genotypes were unequivocally identified evaluated using one simple PCR-based assay based on single larvae. This assay allows the specific identification of Trichinella species without the need to passage larvae in laboratory animals.

JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact support@jstor.org.

Summary:Trichinella infections in foxes and wild boars were studied to determine the prevalence of infection in wildlife in the Netherlands. Muscles of 429 forelegs of foxes (Vulpes vulpes) and diaphragms of 11 wild boars (Sus scrofa) were artificially digested. Single larvae of Trichinella were identified at species level using random amplified polymorphic DNA (RAPD-PCR). In addition, an ELISA using ES antigen was used to test serum samples of 458 wild boars. The prevalence of Trichinella In foxes ranged from 3.9 % in the the eastern part of the country, 13.1% in the central part of the country and 1.3 % in the most western part of the country. Trichinella larvae of foxes were identified as T. britovi. In most samples, identification of larvae did not show reproducible results. The serological prevalence of Trichinella infections in wild boars was 6.8 %. Wild boar populations are located in the central and In the southern part of the country. Trichinella larvae of wild boar were identified as T. spiralis. These results show that two Trichinella species are involved in the epidemiology of trichinellosis among wildlife. On the basis of previous reports, the present results suggest that the prevalence of Trichinella infection in wildlife is increasing in the last 20 years.KEY WORDS : wildlife, T. britovi, T. spiralis, prevalence, the Netherlands. I n Europe, four Trichinella species have been described in wildlife, T. spiralis, T. britovi, T. nativa and T. pseudospiralis.Of these, T. spiralis is also able to be maintained by a domestic cycle (Pozio, 1998 showed a prevalence of 1.8 % (Franchimont et al., 1993).At that time larvae could not be identified at species level.The aim of the present study was the evaluation of the