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[
International C. elegans Meeting,
2001]
Although it is commonly stated that C. elegans is a soil nematode, hardly anyone has isolated it directly from soil extract, as opposed to from decomposing organic matter. Presumably, the dauer is the stage most likely to be encountered in soil, but it is nearly impossible to morphologically distinguish the dauer of C. elegans from that of other rhabditid species, especially when preserved in fixative and mounted in permanent slides. As a result, the ecology of C. elegans remains unknown: there are no primary data on its natural population dynamics, prevalence, dispersal or associations with phoretic vectors. PCR-based identification has now become quite easy, but is expensive to apply routinely in a survey situation, and limited in quantitative resolution. In order to study the species ecology of C. elegans and relatives, we have therefore set out to develop a protocol that allows us to combine morphological and molecular data from individual nematodes, and especially from cryptic stages such as the dauer. Our basic equipment consists of a microscope equipped with Differential Interference Contrast optics, with a thermal stage and with a video camera. The camera is connected to a personal computer equipped for Video Capture and Editing (VCE) with a generic capture card and non-linear editing software. Individual nematodes are immobilised by cooling on the microscope stage, and the morphology of different body regions is captured as videofiles on hard disk, while manually focusing through the specimen at highest magnification. The nematodes are then taken off the microscope, lysed in extraction buffer, used for PCR of ribosomal loci, and sequenced. Next, the obtained sequences are matched with known sequences, to determine the identity of each individual. Finally, the obtained identifications are used to compare the multifocal VCE files and search for morphological characters allowing consistent distinction between dauers from different species. We present the first ecological data obtained in this manner, from samples collected in central and southern California. The equipment used is similar to a stripped-down, low cost 3-D version of a 4-D microscope, consisting mostly of generic components that are widely available from various suppliers and easily configured with minimal expertise. The system is highly versatile, and can e.g. also be used for non-destructively capturing the detailed morphology of mutants, recording ephemeral staining patterns, exchanging data across the internet, teaching, and assembling a VCE reference archive. Each series of videofiles from one nematode basically represents a "virtual microscope slide", bypassing the need for permanent slides. The size and contents of the captured files can be optimized with VCE, and the resulting compressed files combine the ease of storage and distribution of still images, with the depth of information and representation of multifocal videofiles.
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[
J Immunoassay,
1987]
Paired serum, urine, and finger-prick whole blood dried on filter paper were analyzed by immunoradiometric assay (IRMA) for filarial antigen using Brugia malayi-specific rabbit antibody. Nine sera and 6 urines from the 10 paired serum-urine samples obtained from individuals with microfilaremia contained IRMA detectable filarial antigen. In contrast, all serum and urine specimens from patients with chronic infections, endemic and non-endemic controls were negative. Whole blood eluted from filter paper spots contained IRMA detectable material; their degree of positivity agreed well with IRMA binding levels obtained with paired urines. Reduced recovery of antigen dried on filter paper was observed at antigen levels less than 10 ng/ml equivalents, presumably due to irreversible absorption onto the filter paper. Urine and finger-prick filter paper blood specimens can be used in the diagnosis of microfilaremic infections that have been associated with circulating antigen in the blood.
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[
Worm Breeder's Gazette,
1994]
mab-3 YAC rescue David Zarkower, Mario de Bono, and Jonathan Hodgkin MRC Laboratory of Molecular Biology, Cambridge, England
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[
Vet Parasitol,
2008]
Strongyloides sp. (Nematoda) are very wide spread small intestinal parasites of vertebrates that can form a facultative free-living generation. Most authors considered all Strongyloides of farm ruminants to belong to the same species, namely Strongyloides papillosus (Wedl, 1856). Here we show that, at least in southern Germany, the predominant Strongyloides found in cattle and the Strongyloides found in sheep belong to separate, genetically isolated populations. While we did find mixed infections in cattle, one form clearly dominated. This variety, in turn, was never found in sheep, indicating that the two forms have different host preferences. We also present molecular tools for distinguishing the two varieties, and an analysis of their phylogenetic relationship with the human parasite Strongyloides stercoralis and the major laboratory model species Strongyloides ratti. Based on our findings we propose that Strongyloides from sheep and the predominant Strongyloides from cattle should be considered separate species as it had already been proposed by [Brumpt, E., 1921. Recherches sur le determinisme des sexes et de l''evolution des Anguillules parasites (Strongyloides). Comptes rendu hebdomadaires des seances et memoires de la Societe de Biologie et de ses filiales 85, 149-152], but was largely ignored by later authors. For nomenclature, we follow [Brumpt, E., 1921. Recherches sur le determinisme des sexes et de l''evolution des Anguillules parasites (Strongyloides). Comptes rendu hebdomadaires des seances et memoires de la Societe de Biologie et de ses filiales 85, 149-152] and use the name S. papillosus for the Strongyloides of sheep and the name Strongyloides vituli for the predominant Strongyloides of cattle.
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[
Worm Breeder's Gazette,
1994]
Mutagenesis of C. elegans using N-ethyl-N-nitrosourea Elizabeth De Stasio, Dinesh Stanislaus and Catherine Lephoto. Department of Biology, Lawrence University, Appleton, Wl 54911
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[
Parasitol Res,
1987]
The time-course of the detection of circulating and urinary filarial antigens was followed with a 2S-IRMA assay, using a mouse monoclonal antibody raised against Brugia malayi larvae, in Mastomys natalensis experimentally infected with Brugia malayi, Brugia pahangi, or Litomosoides carinii. In the prepatent phase of the infections, filarial antigen was detected 4-7 weeks before microfilariae appeared in the peripheral blood. Moreover, the sensitivity of the test was greater with urine than with serum. During the patent phase of infection, the level of circulating antigens detected varied considerably. However, there was a positive correlation (P less than 0.05) between antigenemia and microfilaremia. In L. carinii infection, filarial antigen could be easily detected in spite of the disappearance of microfilariae in peripheral blood, 49 weeks post infection. If these results are extrapolated to man, the 2S-IRMA should be useful for epidemiological surveys in endemic areas where transmission has been eliminated.
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[
Nature,
2002]
Behavioral ecologists have shown that many animals form social groups in conditions. Neurobiological evidence for this behaviour has now been discovered in the nematode worm, Caenorhabditis elegans. On pages 899 and 925 of this issue, de Bono et al. and Coates and de Bono present striking results on the genetic, molecular and neural mechanisms underlying nematode social feeding. These discoveries provide tantalizing insights into the effects of stress in social groupings.
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Stegmann APA, Bonati MT, Panis B, Smith-Hicks C, Lemke JR, Pepler A, Wilson C, Iascone M, McWalter K, Brasington C, Allen W, Di Donato N, Platzer K, Ramos L, Edwards SL, Jamra R, Gamble CN, Mandel H, Stobe P, Mahida S, Marquardt T, Demmer LA, Miller KG, Falik-Zaccai T, Pinz H, Hellenbroich Y, Sticht H, Kok F, Cho MT, Stumpel CTRM, Shinde DN, Angione KM
[
Am J Hum Genet,
2018]
Using exome sequencing, we have identified de novo variants in MAPK8IP3 in 13 unrelated individuals presenting with an overlapping phenotype of mild to severe intellectual disability. The de novo variants comprise six missense variants, three of which are recurrent, and three truncating variants. Brain anomalies such as perisylvian polymicrogyria, cerebral or cerebellar atrophy, and hypoplasia of the corpus callosum were consistent among individuals harboring recurrent de novo missense variants. MAPK8IP3 has been shown to be involved in the retrograde axonal-transport machinery, but many of its specific functions are yet to be elucidated. Using the CRISPR-Cas9 system to target six conserved amino acid positions in Caenorhabditis elegans, we found that two of the six investigated human alterations led to a significantly elevated density of axonal lysosomes, and five variants were associated with adverse locomotion. Reverse-engineering normalized the observed adverse effects back to wild-type levels. Combining genetic, phenotypic, and functional findings, as well as the significant enrichment of de novo variants in MAPK8IP3 within our total cohort of 27,232 individuals who underwent exome sequencing, we implicate de novo variants in MAPK8IP3 as a cause of a neurodevelopmental disorder with intellectual disability and variable brain anomalies.
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[
J Neurosci,
2003]
Thermotactic behavior in Caenorhabditis elegans is sensitive to both a worm's ambient temperature (T-amb) and its memory of the temperature of its cultivation (T-cult). The AFD neuron is part of a neural circuit that underlies thermotactic behavior. By monitoring the fluorescence of pH-sensitive green fluorescent protein localized to synaptic vesicles, we measured the rate of the synaptic release of AFD in worms cultivated at temperatures between 15 and 25degreesC, and subjected to fixed, ambient temperatures in the same range. We found that the rate of AFD synaptic release is high if either T-amb > T-cult or T-amb > T-cult, but AFD synaptic release is low if T-amb congruent to T-cult. This suggests that AFD encodes a direct comparison between T-amb and T-cult.
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[
J Am Soc Mass Spectrom,
2015]
De novo sequencing software has been widely used in proteomics to sequence new peptides from tandem mass spectrometry data. This study presents a new software tool, Novor, to greatly improve both the speed and accuracy of today's peptide de novo sequencing analyses. To improve the accuracy, Novor's scoring functions are based on two large decision trees built from a peptide spectral library with more than 300,000 spectra with machine learning. Important knowledge about peptide fragmentation is extracted automatically from the library and incorporated into the scoring functions. The decision tree model also enables efficient score calculation and contributes to the speed improvement. To further improve the speed, a two-stage algorithmic approach, namely dynamic programming and refinement, is used. The software program was also carefully optimized. On the testing datasets, Novor sequenced 7%-37% more correct residues than the state-of-the-art de novo sequencing tool, PEAKS, while being an order of magnitude faster. Novor can de novo sequence more than 300 MS/MS spectra per second on a laptop computer. The speed surpasses the acquisition speed of today's mass spectrometer and, therefore, opens a new possibility to de novo sequence in real time while the spectrometer is acquiring the spectral data. Graphical Abstract .