Wang XY et al. (2007) Huan Jing Ke Xue "[Evaluation of toxification in acid and alkalescent industry liquid wastes from paper mill using C. elegans]"

Wang DY, Yu HX, Wang XY

[Huan Jing Ke Xue, 2007]

Model organism C. elegans was introduced into the toxicity identification evaluation (TIE) procedure to determine the toxification of acid and alkalescent industry liquid wastes from paper mill via analyzing the life span, days corresponding to 50% lethal, reproductive rate, reproductive capacity and locomotion behaviors in worms. Our data indicate that the toxic influence order of different samples on life span was: 2nd > 4th > 1st > 3rd, the toxic influence order of different samples on generation time was: 2nd > 1st > 3rd approximately 4th, the toxic influence order of different samples on brood size and head thrashes was similar: 1st approximately 2nd > 3rd approximately 4th, the toxic influence order of different samples on body bends was: 1st approximately 2nd > 3rd > 4th, and the 2nd sample always had more significantly toxic effects on different phenotypes. Thus, the toxicity of acid industry liquid waste from the paper mill relatively had the most severe multiple effects, whereas the toxicity of alkalescent industry liquid and grain-like wastes only exhibited limited effects.

Mills M et al. (1993) International C. elegans Meeting "Tc3, Tc4 and Tc5: Footprints in DNA and RNA."

Butze K, Parham C, Beinhorn J, Mills M, Collins JJ, Lyons D

[International C. elegans Meeting, 1993]

Mills DEK et al. (1993) International C. elegans Meeting "Deoxyuridine sensitization of near-ultraviolet radiation killing in C. elegans."

Hartman PS, Mills DEK

[International C. elegans Meeting, 1993]

HA Tissenbaum et al. (1999) International C. elegans Meeting "Using model organisms to identify genes that affect life span"

L Guarente, HA Tissenbaum

[International C. elegans Meeting, 1999]

Previous work in C. elegans has identified a number of mutants that affect life span. We are trying to identify new components of the aging mechanism by isolating mutations in genes that are conserved in C. elegans and have previously been shown be important in the aging process of other eukaryotic organisms. 1) Werner Syndrome : People with Werner Syndrome (WRN) show many signs of premature aging (reviewed in Yu et al. 1997). The WRN protein is a member of the RecQ family of helicases. In yeast, mutations in the WRN homolog, SGS1 , cause yeast to age prematurely (Sinclair et al. 1997). Four members of the RecQ family have been identified in the C. elegans database. We are currently isolating knockout mutations of all four of the family members. As well, we are generating GFP fusions and doing RNAi to help elucidate the different functions of these four genes. These studies will help to determine if any of the four genes is the true WRN homolog in C. elegans and whether they function to control aging. 2) SIR2 homologs: Studies from yeast ( S. cerevisiae ) have identified the SIR proteins (SIR2, 3, 4) as important components for yeast aging (reviewed in Sinclair et al. 1998). SIR2, however, is unique among the SIR proteins in several respects including that it is highly conserved from yeast to man. There are two SIR2 homologs identified in the C. elegans database. To understand the role of C. elegans SIR2 in aging, we are generating knockout mutations and GFP fusions as well as performing RNAi. We are also using the candidate gene approach to identify mutants in the SIR2 homologs. References: Sinclair D.A., Mills K., and Guarente L. (1997). Science 277 :1313-6 Sinclair D.A., Mills K., and Guarente L. (1998). Annu Rev Microbiol 52 :533-60 Yu C.E., Oshima J., Wijsman E.M., et al. (1997) Am J Hum Genet 60 :330-41

John Peloquin et al. (2007) International Worm Meeting "Worms and Filthy Lucre: Applying C. elegans to natural product development in industry."

Stuart Reeves, John Peloquin

[International Worm Meeting, 2007]

Diamond V Mills is a medium sized Agricultural Biotechnology company with more than 63 years of success in producing dietary supplements and ingredients. Recently the company has moved into human nutrition through its formation of Embria Health Sciences. Thus, the product repertoire has increased and is increasing. Our products arise naturally as the result of subtle alterations of fermentative processes, which are still not completely understood. Furthermore, their effects on animal biology are pleotropic. This presents some serious economic challenges in that far more product prototypes can be produced than economically tested in vertebrate models to demonstrate efficacy. Because many biochemical and signal transduction pathways are conserved between C. elegans and other animals, we have begun to develop C. elegans-based medium through-put screens to identify new product prototypes. We present preliminary data on anti-aging effects of an Embria product as an example of our future goals. (this work was initiated with technical assistance from Mark A. Wilson and Cathy Wolkow of the NIA).

D Baillie et al. (2005) International Worm Meeting "Insight into human genes of unknown function using C. elegans promoter::GFP expression constructs"

D Baillie, Rebecca Newbury, R Johnsen, D Anastis, J Lin, D Tu, A Mah, S Mckay, L Fang, C Mills

[International Worm Meeting, 2005]

The C. elegans Gene Expression Project is continuing to examine promoter based expression profiles for C. elegans genes with human orthologues. Transgenic strains containing extrachromosomal promoter::GFP constructs have been made for 3,000 of the orthologues. From this analysis we are generating spatial and temporal tissue expression profiles for specific genes during C. elegans development. This work provides insights into the function of un-annotated orthologous human genes. To facilitate high throughput identification of genomic promoter-containing regions and the design of suitable PCR primers we use AcePrimer (http://elegans.bcgsc.bc.ca/promoter_primers/). Once primers are designed, PCR stitching is used to knit the promoter-containing region to a promoter-less GFP expression cassette. This construct is then co-injected, with dpy-5(+), into Dpy-5(e907) hermaphrodites (a gift of C. Thacker and A. Rose). GFP expressing wild-type progeny are analysed at each stage of development. Strains that express GFP before the 1 1/2-fold stage, are tagged for the 4-D recording pipeline. For expression in larval and adult stages, strains expressing in a subset of neurons, or exclusively in one tissue, are sent to the confocal microscope for high-resolution imaging. Where detailed embryonic analysis is required, we integrate the extrachromosomal array, followed by outcrossing. The stabilization, and outcrossing of the strains, reduces mosaicism, and has allowed us to make 80 high quality 4-D embryonic recordings. These recordings are all of sufficient quality for detailed lineaging of GFP expression within the embryo. We have analysed 1440 of the C. elegans human orthologues in detail. 317 of these analysed strains are stable and will be sent to the CGC. All the images and expression pattern data generated by this project are available at our website (http://elegans.bcgsc.bc.ca/home/) and through WormBase.

Geier FM et al. (2011) Anal Chem "Cross-platform comparison of Caenorhabditis elegans tissue extraction strategies for comprehensive metabolome coverage."

Geier FM, Bundy JG, Leroi AM, Want E

[Anal Chem, 2011]

The nematode Caenorhabditis elegans is widely used as a model organism in many areas of the life sciences. Metabolite profiling (metabolomics/metabonomics) is a powerful means of assigning phenotypes to experimentally perturbed C. elegans samples (e.g., mutants, RNAi, or chemical treatments). Tissue extraction is a key step, and high-quality and reproducible extractions are essential to the success of metabolomics studies. We have performed an extensive comparison of different tissue extraction techniques with C. elegans, comparing two different solvent systems (chloroform/methanol and aqueous methanol) and six different tissue disruption techniques (including manual grinding in a cooled mortar, homogenization, and various grinding media in both reciprocating and orbital tissue mills). All twelve combinations were then compared by GC/MS, (1)H NMR spectroscopy, and UPLC-MS, and the results were evaluated by both overall multivariate clustering approaches as well as distributions over individual metabolites/metabolite features of coefficient of variation and yield. The choice of solvent had more influence than the disruption method used, although the homogenizer results were clearly outliers. Overall, we concluded that bead-beating with 80% methanol solution was a good trade-off, although it is important to note that the definition of the apparent "best" method depended on which analytical platform was used to evaluate the results.

Peloquin, John Joseph (2009) International Worm Meeting "Bacteria not normally pathogenic to humans evaluated in a C. elegans pathogen challenge model."

Peloquin, John Joseph

[International Worm Meeting, 2009]

Diamond V Mills produces food and feed supplements produced by our proprietary fermentation processes. The universe of products and potential new product prototypes that could be derived from our processes and available raw materials is vast. It is thus impractical to screen our many product prototypes in vertebrates to distinguish the most promising for further evaluation from prototypes of less potential. We have developed a number of initial in-vivo and in-vitro technologies for this purpose. In one of these techniques, we look for C elegans survivorship changes in the presence of our prototypes. If we were to challenge the worms with pathogen stress that we expect to significantly decrease worm survivorship, we could terminate our screening assays shortly after the deaths of the pathogen challenged controls and thus decrease the time needed to get a useful answer. We are always concerned about safety of the pathogens we use to humans and animals, so we are evaluating for use in this assay species and strains of bacteria known to infect and kill invertebrates but that are not expected to be pathogens to humans and other mammals under our experimental conditions.

Zhao Y et al. (2007) BMC Genomics "Poly-G/poly-C tracts in the genomes of Caenorhabditis."

Zhao Y, O'Neil NJ, Rose AM

[BMC Genomics, 2007]

ABSTRACT: BACKGROUND: In the genome of Caenorhabditis elegans, homopolymeric poly-G/poly-C tracts (G/C tracts) exist at high frequency and are maintained by the activity of the DOG-1 protein. The frequency and distribution of G/C tracts in the genomes of C. elegans and the related nematode, C. briggsae were analyzed to investigate possible biological roles for G/C tracts. RESULTS: In C. elegans, G/C tracts are distributed along every chromosome in a non-random pattern. Most G/C tracts are within introns or are close to genes. Analysis of SAGE data showed that G/C tracts correlate with the levels of regional gene expression in C. elegans. G/C tracts are over-represented and dispersed across all chromosomes in another Caenorhabditis species, C. briggase. However, the positions and distribution of G/C tracts in C. briggsae differ from those in C. elegans. Furthermore, the C. briggsae dog-1 ortholog CBG19723 can rescue the mutator phenotype of C. elegans dog-1 mutants. CONCLUSIONS: The abundance and genomic distribution of G/C tracts in C. elegans, the effect of G/C tracts on regional transcription levels, and the lack of positional conservation of G/C tracts in C. briggsae suggest a role for G/C tracts in chromatin structure but not in the transcriptional regulation of specific genes.

Driscoll MA et al. (2000) East Coast Worm Meeting "Contributions of WRN-related Helicases to C. elegans Aging"

Driscoll MA, Herndon LA, Nycz KM

[East Coast Worm Meeting, 2000]

Four genes in the C. elegans genome exhibit sequence similarity to the RecQ family of DNA helicases which include the human Werner (WRN), Bloom (BLM) and Rothmund-Thomson (RTS) syndrome genes. Mutation of human WRN and RTS can confer features of accelerated aging. Both WRN and RTS patients have a high rate of genome instability, which is also a key feature of patients with Bloom syndrome. Similarly, mutation of the yeast RecQ homolog, SGS1, results in a high rate of genomic instability and a lifespan 40% shorter than wild type1, 2. To determine if the RecQ family of helicases share similar functions in C. elegans, we generated deletions within the C. elegans gene most closely related to the Werners gene (F18C5.2) and in another homolog (E03A3.2) and have tested these strains for lifespan and genomic instability. Preliminary results indicate that animals lacking F18C5.2 or E03A3.2 are viable and fertile. They have normal lifespans and do not have a high rate of genomic instability. Since C. elegans has four related helicases, there may be redundancy in the activity of these helicases. To test this hypothesis, we are now analyzing animals that carry mutations in multiple DNA helicase family members. It has recently been determined that him-6 mutant animals, which have a high rate of chromosome nondisjunction, contain mutations in the C. elegans DNA helicase gene T04A11.6, which is most closely related to the human BLM gene3. We are also testing mutants for UV radiation or X-ray hypersensitivity, since it is possible that these helicases play a role in DNA repair. We are testing the expression pattern of these genes and are screening for a deletion in the fourth remaining C. elegans DNA helicase family member, K02F3.1. By establishing a nematode model for accelerated aging disorders, we hope to extend understanding of the action of helicase family members in senescence and learn more about mechanisms of aging overall. Watt, P.M., I.D. Hickson, R.H. Borts and E.J. Louis. 1996. Genetics 144: 935. Sinclair, D.A., K. Mills and L. Guarente. 1997. Science 277: 1313. Wicky, C., A. Rose and F. Mueller. 1998. European Worm Meeting Abstract.