Chagas disease in a domestic transmission cycle in southern Texas, USA

Centers for Disease control and Prevention http://www.cdc.gov/ncidod/EID/vol9no1/02-0217.htmAfter three dogs died from acute Chagas cardiomyopathy at one location, an investigation was conducted of the home, garage, and grounds of the owner. A serologic study was conducted on stray dogs, and an ecologic niche model was developed to predict areas where the vector Triatoma gerstaeckeri might be expected

Molecular Typing of Trypanosoma cruzi Isolates, United States

Studies have characterized Trypanosoma cruzi from parasite-endemic regions. With new human cases, increasing numbers of veterinary cases, and influx of potentially infected immigrants, understanding the ecology of this organism in the United States is imperative. We used a classic typing scheme to determine the lineage of 107 isolates from various hosts

Chagas Disease in a Domestic Transmission Cycle in Southern Texas, USA

After three dogs died from acute Chagas cardiomyopathy at one location, an investigation was conducted of the home, garage, and grounds of the owner. A serologic study was conducted on stray dogs, and an ecologic niche model was developed to predict areas where the vector Trypanosoma gerstaeckeri might be expected

Identification of Potential Sites for Tryptophan Oxidation in Recombinant Antibodies Using tert-Butylhydroperoxide and Quantitative LC-MS

Amino acid oxidation is known to affect the structure, activity, and rate of degradation of proteins. Methionine oxidation is one of the several chemical degradation pathways for recombinant antibodies. In this study, we have identified for the first time a solvent accessible tryptophan residue (Trp-32) in the complementary-determining region (CDR) of a recombinant IgG1 antibody susceptible to oxidation under real-time storage and elevated temperature conditions. The degree of light chain Trp-32 oxidation was found to be higher than the oxidation level of the conserved heavy chain Met-429 and the heavy chain Met-107 of the recombinant IgG1 antibody HER2, which have already been identified as being solvent accessible and sensitive to chemical oxidation. In order to reduce the time for simultaneous identification and functional evaluation of potential methionine and tryptophan oxidation sites, a test system employing tert-butylhydroperoxide (TBHP) and quantitative LC-MS was developed. The optimized oxidizing conditions allowed us to specifically oxidize the solvent accessible methionine and tryptophan residues that displayed significant oxidation in the real-time stability and elevated temperature study. The achieved degree of tryptophan oxidation was adequate to identify the functional consequence of the tryptophan oxidation by binding studies. In summary, the here presented approach of employing TBHP as oxidizing reagent combined with quantitative LC-MS and binding studies greatly facilitates the efficient identification and functional evaluation of methionine and tryptophan oxidation sites in the CDR of recombinant antibodies

Evolutionary relationships among mismatch-repair class 2 gene (<i>MSH2</i>) and the thiol-disulfide oxido-reductase Tc52 gene (<i>Tc52</i>) from 50 and 51 <i>Trypanosoma cruzi</i> isolates, respectively.

<p>Three phylogenetic trees were created by neighbor-joining (NJ), minimum evolution (ME), and maximum parsimony (MP) methods from the alignment of each gene target and a consensus tree was interpreted. Numbers at the branches are bootstrap values >50% (500 replicates) for the same nodes of the NJ, ME, MP trees. Evolutionary distances were computed using the Kimura 2-parameter method <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056198#pone.0056198-Saitou1" target="_blank">[29]</a>. ▴ = the 17 US TcI isolates that were identical; • = the 24 or 27 US TcIIa isolates that were identical. * = reference strains: SilvioX10 cl1, Colombiana, P209 (TcI); X110/8 (TcIII); CANIII cl1, Dog Theis, Ecua6 (TcIV); CL Brener (TcVI).</p

Nucleotide sequence variations within the Tc52 gene sequence of 51 <i>T. cruzi</i> isolates from the United States compared to reference strains.

<p>Genotype of each isolate precedes the isolate name. Nucleotide positions correspond to sites from P209 (Genbank EF065175). Dots represent nucleotide site identical to reference strain (either P209 for TcI or CANIII cl1 for TcIV). Dashes represent missing nucleotides.</p>*<p>The following 17 US TcI sequences were identical: Human isolates (TC CC, CA R), domestic dogs (USA Dog Y), Virginia opossums (USA Opossum, Opossum 1970, 92101601P cl2, FH4, 93070103P cl2, FL Opo 3, FL Opo 15), armadillos (Armadillo 1973, USA Armadillo), triatomine bugs (Florida C1F8, Florida), and raccoons (GA Rac 143, 93053103R cl3, FL Rac 13).</p>†<p>The following 26 US TcIV sequences were identical: Raccoons (STC 10R cl3, FL Rac 9, 92122102R, 93071502R cl2, 93040701R cl1, 93072805R cl3, FL Rac 15, FL Rac 46, FL Rac 5, FL Rac 30, GA Rac 134, GA Rac 69, GA Rac 107, Maryland Rac, STC 35R), domestic dogs (Samantha Dog, Caesar Dog, Dog Theis, OK Dog, Smokey), ring-tailed lemurs (Clarence, Meg, Nilda), striped skunk (GA Sk 1), triatomine bug (T sang5 cl1), and armadillo (GA Arm 20).</p