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Hypothesis: Depletion of RANKL decreases pulmonary M cell differentiation and protects against prion infection by the inhaled route. 1. Summary of proposed research ( 400 words) 2. Aims: 1. Depletion of M cells in the FAE of Peyer’s patches by RANKL neutralization. 2. M cell-depletion blocks prion neuroinvasion from the inhaled route 3. Background: Provide figures and data, review of previous research in the area and justification for further research. 4. Experimental design (with table) and methods, expected outcomes and their importance • Animals: C57BL/6, Edinburgh, treatment with anti-RANKL, anti-IgG(not specific) as a control. • Inoculation and tissue collection: Prion exposure by an inhalation chambers containing a nebulizer device. • Disease monitoring: Behavior test ( memory and motor tests) • Histological analysis: Immunohistochemistry and immunoflorescent 5. Prevention of Bias: Blinding, Randomization etc… 6. Data Analysis 7. Future Direction 8. Justify staff, materials and consumables requested providing estimates of total cost 9. Timetable plan 10. References Please read the following papers for ideas and methods; • http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3282432/ • http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497678/ • http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1001257

Hypothesis: Depletion of RANKL decreases pulmonary M cell differentiation and protects against prion infection by the inhaled route.

1. Summary of proposed research ( 400 words)

2. Aims: 1. Depletion of M cells in the FAE of Peyer’s patches by RANKL neutralization. 2. M cell-depletion blocks prion neuroinvasion from the inhaled route

3. Background: Provide figures and data, review of previous research in the area and justification for further research.

4. Experimental design (with table) and methods, expected outcomes and their importance

• Animals: C57BL/6, Edinburgh, treatment with anti-RANKL, anti-IgG(not specific) as a control.
• Inoculation and tissue collection: Prion exposure by an inhalation chambers containing a nebulizer device.
• Disease monitoring: Behavior test ( memory and motor tests)
• Histological analysis: Immunohistochemistry and immunoflorescent

5. Prevention of Bias: Blinding, Randomization etc…

6. Data Analysis

7. Future Direction

8. Justify staff, materials and consumables requested providing estimates of total cost

9. Timetable plan

10. References

Please read the following papers for ideas and methods;
• http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3282432/
• http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497678/
• http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1001257

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