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1.
Mol Biochem Parasitol ; 238: 111291, 2020 07.
Article in English | MEDLINE | ID: mdl-32479776

ABSTRACT

In free-living and parasitic nematodes, the methylation of phosphoethanolamine to phosphocholine provides a key metabolite to sustain phospholipid biosynthesis for growth and development. Because the phosphoethanolamine methyltransferases (PMT) of nematodes are essential for normal growth and development, these enzymes are potential targets of inhibitor design. The pine wilt nematode (Bursaphelenchus xylophilus) causes extensive damage to trees used for lumber and paper in Asia. As a first step toward testing BxPMT1 as a potential nematicide target, we determined the 2.05 Å resolution x-ray crystal structure of the enzyme as a dead-end complex with phosphoethanolamine and S-adenosylhomocysteine. The three-dimensional structure of BxPMT1 served as a template for site-directed mutagenesis to probe the contribution of active site residues to catalysis and phosphoethanolamine binding using steady-state kinetic analysis. Biochemical analysis of the mutants identifies key residues on the ß1d-α6 loop (W123F, M126I, and Y127F) and ß1e-α7 loop (S155A, S160A, H170A, T178V, and Y180F) that form the phosphobase binding site and suggest that Tyr127 facilitates the methylation reaction in BxPMT1.


Subject(s)
Ethanolamines/chemistry , Helminth Proteins/chemistry , Methyltransferases/chemistry , Nematoda/enzymology , Pinus/parasitology , Plant Diseases/parasitology , Amino Acid Sequence , Animals , Binding Sites , Cloning, Molecular , Crystallography, X-Ray , Escherichia coli/genetics , Escherichia coli/metabolism , Ethanolamines/metabolism , Gene Expression , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Helminth Proteins/genetics , Helminth Proteins/metabolism , Kinetics , Methyltransferases/genetics , Methyltransferases/metabolism , Models, Molecular , Nematoda/genetics , Protein Binding , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Sequence Alignment , Sequence Homology, Amino Acid , Substrate Specificity , Thermodynamics
2.
Phys Rev Lett ; 101(11): 112003, 2008 Sep 12.
Article in English | MEDLINE | ID: mdl-18851276

ABSTRACT

Inclusive K_{S};{0}K_{S};{0} production in ep collisions at the DESY ep collider HERA was studied with the ZEUS detector using an integrated luminosity of 0.5 fb;{-1}. Enhancements in the mass spectrum were observed and are attributed to the production of f_{2}(1270)/a_{2};{0}(1320), f_{2};{'}(1525) and f_{0}(1710). Masses and widths were obtained using a fit which takes into account theoretical predictions based on SU(3) symmetry arguments, and are consistent with the Particle Data Group values. The f_{0}(1710) state, which has a mass consistent with a glueball candidate, was observed with a statistical significance of 5 standard deviations. However, if this state is the same as that seen in gammagamma-->K_{S};{0}K_{S};{0}, it is unlikely to be a pure glueball state.

3.
Schweiz Arch Tierheilkd ; 140(2): 60-4, 1998.
Article in German | MEDLINE | ID: mdl-9492576

ABSTRACT

30 broiler flocks from two majors Swiss broiler producing companies were examined for lesions relevant to meat inspection between November 1995 and April 1996. 71.8% of condemned carcasses and viscera were assigned to ascites syndrome, bacterial infections and runting. Other diseases and lesions due to injury or processing were rare. Escherichia coli was responsible for the majority of infections. The average condemnation rate (1.0%) corresponded to findings in literature.


Subject(s)
Abattoirs/statistics & numerical data , Chickens , Food Inspection , Meat/standards , Poultry Diseases/epidemiology , Animals , Ascites/epidemiology , Ascites/pathology , Ascites/veterinary , Bacterial Infections/epidemiology , Bacterial Infections/pathology , Bacterial Infections/veterinary , Escherichia coli Infections/epidemiology , Escherichia coli Infections/pathology , Escherichia coli Infections/veterinary , Growth Disorders/epidemiology , Growth Disorders/pathology , Growth Disorders/veterinary , Poultry Diseases/pathology , Switzerland/epidemiology
4.
Vet Parasitol ; 72(2): 201-7, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9404846

ABSTRACT

Domestic pigs in Switzerland have been considered Trichinella-free for decades, despite the occurrence of Trichinella in the wildlife cycle. In order to reevaluate the present epidemiological situation, tissue samples from 11226 domestic pigs, 356 wild boars and 452 foxes were examined using the standard artificial digestion method. A simultaneous serological study, extended to include 25239 sera from sows provided by a Swiss pig serum bank, was also undertaken. The results of both studies support the conclusion that Trichinella spp. do not occur within the domestic pig population in Switzerland. Among the fox population, Trichinella was detected in four (0.9%) of the animals tested using the digestion method, and Trichinella britovi was identified as the infecting species by RAPD fingerprint analyses.


Subject(s)
Foxes/parasitology , Swine Diseases/epidemiology , Swine/parasitology , Trichinella/isolation & purification , Trichinellosis/veterinary , Animals , Animals, Domestic , Animals, Wild , DNA Fingerprinting , Enzyme-Linked Immunosorbent Assay , Random Amplified Polymorphic DNA Technique , Switzerland/epidemiology , Trichinellosis/epidemiology
5.
Schweiz Arch Tierheilkd ; 136(9): 298-308, 1994.
Article in German | MEDLINE | ID: mdl-7973563

ABSTRACT

For many decades trichinellosis has not been reported among Swiss domestic pigs. Considering the fact that Trichinella occurs in a sylvatic cycle in Switzerland, a study was designed to reevaluate the present epidemiologic situation by investigating 10,904 fattening pigs, 218 pigs with free access to pasturage or being kept on an alp, 104 domestic boars, 106 horses, 44 wild boars and 538 foxes using a direct and an indirect diagnostic technique (digestion method and serology with ELISA and an excretory/secretory antigen, respectively). The digestion method was performed according to EC-guidelines. Furthermore, 25,239 sera originating from a Swiss sow-serum bank were tested retrospectively for anti-Trichinella antibodies. Trichinella was not detectable in all domestic pigs using the digestion method. Serologically, 3 fattening pigs (0.027%) and 9 sows (0.036%) demonstrated weak antibody reactivities against the Trichinella E/S-antigen. Based upon statistical calculations for the negative-positive threshold, these antibody-reactions were considered to be within the normal range of variability of the test. Although statistically restricted, the results of the present study indicate the absence of Trichinella within the Swiss pig population. Based upon the rational applicability of the ELISA and its diagnostic sensitivity and specificity, this test appears as the most suitable method to perform large-scale screenings among slaughter pigs. Pigs with free access to pasturage and boars were all parasitologically and serologically negative for Trichinella. The digestion method showed that horses and wild boars were all parasitologically negative, whereas 1.3% of the foxes were positive for Trichinella larvae.


Subject(s)
Foxes/parasitology , Horse Diseases/epidemiology , Swine Diseases/epidemiology , Trichinellosis/veterinary , Abattoirs , Animals , Animals, Wild , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Horse Diseases/diagnosis , Horses , Male , Swine , Swine Diseases/diagnosis , Switzerland/epidemiology , Trichinellosis/diagnosis , Trichinellosis/epidemiology
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