Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model

Cha's disease is the clinical condition that ensues chronic infections by the ate prot and tno mausi. The disease attacks the heart of chronicled infected people several decades after acquisition of the infections. A across kingdom model system shows that chicken eggs are permissive to the Taz macro cruise infections.

During the first week of embryo growth, the experiments show that infection free chickens retain the parasite KA into their genomes die of chagas heart disease, excellent housing, food and water supply secures animal welfare. The cruise triple mass, good swimming Forms are obtained from tissue culture. Trans illuminate for tio and eggs to show their bubble Control.

Eggs do not receive para inoculation. Mock control eggs are inoculated with 10 microliter of culture medium. The hole is seals with essive tape 100 viral FL mania.

Brazilians is pro master. Goats are inoculating the air chamber of fertilized chicken egg 100 viral and trap macro triple master goats in 10 microliter of calcium media are inoculating the air chamber or virtualized chicken egg. The eggs are incubated at 37, centigrade at 65%in humidity for one to one days with gentle rotation each 30 minutes, the cheeks hatch equally from renoso macro inoculate eggs and from mark controls.

The cheeks grow to adult life. Blood samples are drawn from wing veins. Blood is transferred to 15 Ml conical tubes.

Mononuclear cells are separated by centrifugation upon FICO gradient wash off white cells palate and discard separation. Medium genomic DNA is extracted from blood mononuclear cells Set 1.5 ml conical tubes in res according with PCR design. PCR is made to amplify chroy nuclear G-N-A-N-G-N-A and pls GNA kg NA.Using a specific primer sets set reaction Tubes in the thermocycler after run.

Take tubes back from Thermocycler. Analyze the PCR products. The PCR products are separated by Agarro gel electrophoresis.

The GNA bands in the gel are transferred to a positively chart nylon membrane. Hybridization is made with a specific radioactive labeled probes to separate roles play independently by the parasite persistence and by the are immunity. In the pathogenesis of chagas disease, we use a chicken model refractory to the TNO cruise infection except during the early embryonic growth prior to the development of the immune system, the chicks that had from TNO cruise inoculated eggs are infection free, but the parasite mitochondrial A KG A is retained in the body.

Only chickens hatchet from tzo macro inoculated eggs show kg a bands the any DNA is not present in the chicken genome. Chicks hatchet from ex inoculated with pro master goats of Leia Brazilian do not retain the mitochondrial in circle sequences in their genomes. Lane one blank lane two control GNA lanes three and four mock GS lanes five to 14 Gs from Czech Hatcher from Las mania Brazilians is infected eggs.

Lane 15 las mania Brazil amplicon format band 750 base pairs obtained with LB three and LB five specific KDNA primer sets. L-P-C-R-A checkerboard scan depicts the gallus guns primers that were deployed after three cycles of amplification with increasing temperature walking nest. PCR is made with different KDNA PRI set.

Louis Stringency cycle creates sites for prime and kneeling and high stringency cycle. Augment the specificity of the amplification products. The PCR products from less amplification that hybridized with KA probe were cloned and sequenced commercially.

The Sequence shows the kg NA in circle blue intermediate micro homology recombination junction yellow and the gallo ga GNA Green. The Mapping of the kg NA integrating the chicken genome is AED by a target primer type PCR R TP therapy PCR technique. The Cajun a mutated adult chickens developed chagas heart disease.

The more controls grow to a healthy life. The KDNA Mutated chickens show increased cardiac silhouette on x-ray films. The necropsy show controlled chickens have small heart size.

The Cajun, a mutated chicken show, hydro pericardium, hydro tox, and a large heart. The cardiomegaly is similar to that in Chagas heart disease. The Moog controlled chicken heart shows normal histology.

The kg a mutated chicken shows a tunnel made on myocardium by the cytotoxin lymphocytes that lies the heart fibers. The sequences Of aian circles that integrate in code regions at several chromosomes are disclosed by modify nested type PCR that combines kg NA primers with host GNA specific primer sets decades. GNA mutated chickens die of heart failure and show cardiomegaly.

The microscopy pathology show myocarditis the chagas like myocarditis, translating heart. In kg NA mutated chicken is a genetically driven disease. This is the first time that Chagas disease clinical and pathological manifestations are reproduced in a clean cross kingdom model system.

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