Hoss S et al. (2012) Environ Toxicol Chem "Interlaboratory comparison of a standardized toxicity test using the nematode Caenorhabditis elegans (ISO 10872)."

Pluta HJ, Bergtold M, Bluebaum-Gronau E, Menzel R, Hoss S, Mohlenkamp C, Traunspurger W, Brinke M, Ratte HT, Ahlf W, von Danwitz B, Donnevert G

[Environ Toxicol Chem, 2012]

A ring test was carried out within the standardization process of ISO 10872 to evaluate the precision of the toxicity test for the nematode Caenorhabditis elegans. Eight different laboratories tested aqueous solutions of the reference substance benzylcetyldimethylammonium chloride as well as native sediments and soils for toxic effects on the growth and reproduction of C. elegans. Validity criteria were met in all laboratories. Average median- and low-effect concentrations were determined to be 15.1 mg L(-1) (EC50) and 8.7 mg L(-1) (EC10) for growth and 7.5 mg L(-1) (EC50) and 3.8 mg L(-1) (EC10) for reproduction of C. elegans, with ECx values showing a high degree of reproducibility (CV(R) : <21% and <11% for EC10 and EC50, respectively) and repeatability (CV(r) : <20% and <7% for EC10 and EC50, respectively). The toxic effects of the sediments and soils revealed by the different laboratories were well related to each samples' degree of chemical contamination. Moreover, the effects showed an acceptable reproducibility (CV(R) : 5-33% and 0-28% for growth and reproduction, respectively) and repeatability (CV(r) : 3-13% and 0-12% for growth and reproduction, respectively). The present study confirms that the toxicity test with C. elegans according to ISO 10872 is a reliable and precise tool to assess the toxicity of aqueous media, freshwater sediments, and soils.

Zhao X et al. (2000) Phytopathology "Species-dependent effects of border cell and root tip exudates on nematode behavior."

Zhao X, Hawes MC, Schmitt M

[Phytopathology, 2000]

ABSTRACT Effects of border cell and root tip exudates on root knot nematode (Meloidogyne incognita) behavior were examined. In whole-plant assays using pea, M. incognita second-stage juveniles (J2) accumulated rapidly around the 1- to 2-mm apical region ensheathed by border cells, but not in the region of elongation. Within 15 to 30 min, J2 which had accumulated within detached clumps of border cells lost motility and entered into a quiescent state. When border cells (and associated root tip exudates) were washed from pea roots prior to challenge with nematodes, no such accumulation and quiescence was induced. Attraction of nematodes by roots was species dependent: no attraction or accumulation occurred in snap bean. Using a quantitative assay, three categories of chemotaxis responses occurred: attraction (pea and alfalfa cv. Thor), repulsion (alfalfa cv. Moapa 69), and no response (snap bean and alfalfa cv. Lahonton). In contrast, total root tip exudates from all three plant species acted as a repellent for M. incognita in the sand assay. An in vitro assay was developed to characterize the induced quiescence response. When total root tip exudate from the tested legumes (as well as corn) was incubated with J2 populations, >80% of the nematodes lost motility. A similar response occurred in Caenorhabditis elegans. Border cell exudates did not induce or contribute to the induction of quiescence. Cocultivation of pea border cells with M. incognita resulted in changes in border cell shape similar to those observed in response to exogenous plant hormones. No such changes occurred in snap bean border cells. Understanding the cell- and host-specific extracellular recognition that occurs between roots and pathogenic nematodes in the early stages before infection occurs could lead to new avenues for disease control.

Saini AK et al. (2011) BMC Res Notes "Allyl isothiocyanate induced stress response in Caenorhabditis elegans."

Saini AK, Reaney MJ, Tyler RT, Shim YY

[BMC Res Notes, 2011]

UNLABELLED: ABSTRACT: BACKGROUND: Allyl isothiocyanate (AITC) from mustard is cytotoxic; however the mechanism of its toxicity is unknown. We examined the effects of AITC on heat shock protein (HSP) 70 expression in Caenorhabditis elegans. We also examined factors affecting the production of AITC from its precursor, sinigrin, a glucosinolate, in ground Brassica juncea cv. Vulcan seed as mustard has some potential as a biopesticide. FINDINGS: An assay to determine the concentration of AITC in ground mustard seed was improved to allow the measurement of AITC release in the first minutes after exposure of ground mustard seed to water. Using this assay, we determined that temperatures above 67C decreased sinigrin conversion to AITC in hydrated ground B. juncea seed. A pH near 6.0 was found to be necessary for AITC release. RT-qPCR revealed no significant change in HSP70A mRNA expression at low concentrations of AITC (< 0.1 M). However, treatment with higher concentrations (> 1.0 M) resulted in a four- to five-fold increase in expression. A HSP70 ELISA showed that AITC toxicity in C. elegans was ameliorated by the presence of ground seed from low sinigrin B. juncea cv. Arrid. CONCLUSIONS: AITC induced toxicity in C. elegans, as measured by HSP70 expression. Conditions required for the conversion of sinigrin to AITC in ground B. juncea seed were determined. The use of C. elegans as a bioassay to test AITC or mustard biopesticide efficacy is discussed.

Hamilton RG et al. (1984) J Immunol Methods "Limitations of the radioimmunoprecipitation polyethylene glycol assay (RIPEGA) for detection of filarial antigens in serum."

Adkinson NF, Alexander E, Hamilton RG, Hussain R

[J Immunol Methods, 1984]

The performance of the radioimmunoprecipitation polyethylene glycol assay (RIPEGA) was examined for quantitation of filarial antigens (Brugia malayi and Dirofilaria immitis) in serum from infected human and animal hosts and non-infected controls. Multiple PEG concentrations were employed to determine the level of non-specific binding (NSB) in non-exposed human sera (NEHS) containing no filarial antigen. The NSB observed when 3 different 125I-labeled IgG antibodies were added to 26 NEHS varied 3-fold and was correlated significantly with total serum IgM (r = 0.80, P less than 0.005, n = 24) but not with serum IgA (r = 0.37) or IgG (r = 0.45). NSB levels were significantly reduced when a Fab'2 fragment of the 125I-labeled antibody was used, but the correlation of NSB with total serum IgM remained significant (r = 0.57, P less than 0.01). The presence of rheumatoid factor in NEHS sera also significantly increased NSB by an average of 3-fold. These effects eliminated the assay's ability to detect in sera from infected hosts filarial antigen the presence of which could be readily demonstrated by an immunoradiometric assay. The RIPEGA's precision (intra-assay coefficient of variation (CV) = 21% at 35% Bmax) and reproducibility (inter-assay CV = 29% at 35% Bmax) are less satisfactory than many alternative immunoassays. In many cases, positive sera failed to dilute out in parallel with each other or with an antigen-spiked standard reference curve. We conclude that poor performance characteristics currently limit the utility of the RIPEGA for quantitating filarial antigen in human and animal serum.

Meier F et al. (2021) Nat Commun "Deep learning the collisional cross sections of the peptide universe from a million experimental values."

Mann M, Brunner AD, Meier F, Wanka JH, Theis FJ, Kohler ND, Voytik E, Strauss MT

[Nat Commun, 2021]

The size and shape of peptide ions in the gas phase are an under-explored dimension for mass spectrometry-based proteomics. To investigate the nature and utility of the peptide collisional cross section (CCS) space, we measure more than a million data points from whole-proteome digests of five organisms with trapped ion mobility spectrometry (TIMS) and parallel accumulation-serial fragmentation (PASEF). The scale and precision (CV < 1%) of our data is sufficient to train a deep recurrent neural network that accurately predicts CCS values solely based on the peptide sequence. Cross section predictions for the synthetic ProteomeTools peptides validate the model within a 1.4% median relative error (R > 0.99). Hydrophobicity, proportion of prolines and position of histidines are main determinants of the cross sections in addition to sequence-specific interactions. CCS values can now be predicted for any peptide and organism, forming a basis for advanced proteomics workflows that make full use of the additional information.

Mukai D et al. (2008) J Nat Med "Potential anthelmintics: polyphenols from the tea plant Camellia sinensis L. are lethally toxic to Caenorhabditis elegans."

Yamasaki T, Sato M, Matsuda N, Mukai D, Yoshioka Y

[J Nat Med, 2008]

A novel gallate of tannin, (-)-epigallocatechin-(2 beta--&gt;O--&gt;7'',4 beta--&gt;8'')-epicatechin-3''-O-gallate (8), together with (-)-epicatechin-3-O-gallate (4), (-)-epigallocatechin (5), (-)-epigallocatechin-3-O-gallate (6), and (+)-gallocatechin-(4 alpha--&gt;8'')-epigallocatechin (7), were isolated from the tea plant Camellia sinensis (L.) O. Kuntze var. sinensis (cv., Yabukita). The structure of 8, including stereochemistry, was elucidated by spectroscopic methods and hydrolysis. The compounds, along with commercially available pyrogallol (1), (+)-catechin (2), and (-)-epicatechin (3), were examined for toxicity towards egg-bearing adults of Caenorhabditis elegans. The anthelmintic mebendazole (9) was used as a positive control. Neither 2 nor 3 were toxic but the other compounds were toxic in the descending order 8, 7 approximately 6, 9, 4, 5, 1. The LC(50) (96 h) values of 8 and 9 were evaluated as 49 and 334 micromol L(-1), respectively. These data show that many green tea polyphenols may be potential anthelmintics.

Varoqui H et al. (1994) FEBS Lett "Cloning and expression of the vesamicol binding protein from the marine ray Torpedo. Homology with the putative vesicular acetylcholine transporter UNC-17 from Caenorhabditis elegans."

Varoqui H, Usdin TB, Rand JB, Meunier FM, Diebler MF, Erickson JD, Bonner TI, Eiden LE

[FEBS Lett, 1994]

Complementary DNA clones corresponding to a messenger RNA encoding a 56 kDa polypeptide have been obtained from Torpedo marmorata and Torpedo ocellata electric lobe libraries, by homology screening with a probe obtained from the putative acetylcholine transporter from the nematode Caenorhabditis elegans. The Torpedo proteins display approximately 5036777764577verall identity to the C. elegans unc-17 protein and 43-134223489dentity to the two vesicle monoamine transporters (VMAT1 and VMAT2). This family of proteins is highly conserved within 12 domains which potentially span the vesicle membrane, with little similarity within the putative intraluminal glycosylated loop and at the N- and C-termini. The approximately 3.0 kb mRNA species is specifically expressed in the brain and highly enriched in the electric lobe of Torpedo. The Torpedo protein, expressed in CV-1 fibroblast cells, possesses a high-affinity binding site for vesamicol (Kd = 6 nM), a drug which blocks in vitro and in vivo acetylcholine accumulation in cholinergic vesicles.

Schinkmann K et al. (1994) Brain Res Mol Brain Res "Comparison of two Caenorhabditis genes encoding FMRFamide(Phe-Met-Arg-Phe-NH2)-like peptides."

Schinkmann K, Li C

[Brain Res Mol Brain Res, 1994]

We have identified a gene encoding multiple FMRFamide-like peptides in the necromenic nematode Caenorhabditis vulgaris. This gene, Cv-flp-1, shares strong sequence homology in the coding regions with the flp-1 gene from the related free-living soil nematode C. elegans. The predicted neuropeptide precursor proteins differ by only four conservative amino acid changes, none of which affects sequences of the predicted neuropeptides. DNA sequences in the non-coding areas are less conserved, but areas of sequence homology are found in introns and in 3' and 5' non-translated regions, suggesting some functional significance for these conserved regions. In C. vulgaris, as was found in C. elegans, two transcripts are presumably produced as a result of use of an alternative 3' splice acceptor site. Lastly, an antibody specific for the RF-moiety of FMRFamide stains a similar subset of cells in C. elegans and C. vulgaris. These results indicate that the function and regulation of the peptides are likely to be conserved in both species.

Cabruja M et al. (2021) Anal Chim Acta "In-depth triacylglycerol profiling using MS3 Q-Trap mass spectrometry."

Contrepois K, Kroetz DL, Papsdorf K, Brunet A, Snyder MP, Priotti J, Cabruja M, Domizi P

[Anal Chim Acta, 2021]

Total triacylglycerol (TAG) level is a key clinical marker of metabolic and cardiovascular diseases. However, the roles of individual TAGs have not been thoroughly explored in part due to their extreme structural complexity. We present a targeted mass spectrometry-based method combining multiple reaction monitoring (MRM) and multiple stage mass spectrometry (MS3) for the comprehensive qualitative and semiquantitative profiling of TAGs. This method referred as TriP-MS3 - triacylglycerol profiling using MS3 - screens for more than 6,700 TAG species in a fully automated fashion. TriP-MS3 demonstrated excellent reproducibility (median interday CV &#8764; 0.15) and linearity (median R2 = 0.978) and detected 285 individual TAG species in human plasma. The semiquantitative accuracy of the method was validated by comparison with a state-of-the-art reverse phase liquid chromatography (RPLC)-MS (R2 = 0.83), which is the most commonly used approach for TAGs profiling. Finally, we demonstrate the utility and the versatility of the method by characterizing the effects of a fatty acid desaturase inhibitor on TAG profiles in vitro and by profiling TAGs in Caenorhabditis elegans.

Prithika U et al. (2021) Microbes Infect "Evaluation of changes in C. elegans immune response during bacterial infection: A single nematode approach."

Balamurugan K, Prithika U

[Microbes Infect, 2021]

Routinely, studies were performed using age-synchronized group of C. elegans as host which suggested a collective response by the host system. Here, we report the modulation of immune response in a single nematode against Staphylococcus aureus and Proteus mirabilis. Initially, the survival of wild-type N2 was tested and was found that S. aureus killed single nematode at 42 h while P. mirabilis failed to provoke infection but colonized the nematode's intestine. With this milieu, the pathogenicity of the bacteria was assessed by Fourier Transform Infra-Red (FTIR) spectroscopy and Cyclic Voltammetry (CV) and was found that S. aureus in the presence of host elicited its virulence while P. mirabilis and Escherichia coli OP50 did not show any alteration. Vertical transmission of infection was also deduced by colony forming unit assay using Cyanine dyes. The MALDI-TOF/TOF analysis was also performed to identify the proteome changes in the single nematode that showcased different proteins related to various immune pathways. This study suggested the importance of understanding the infection pathology and traits of individual nematode which could help our understanding on otherwise the disordered processes during host and microbe interactions.