Project Proposal

My project will be an entry of a paper that relays my research into a scientific journal for
publication. The paper will target doctors who prescribe Entresto, a blood pressure medication, and
will be beneficial to patients who are currently using this medicine or are planning on using it.

January: Expand fly line DGRP 229 to collect approximately 200-250 male flies. Virgin collection
might be needed and I will be using males who have less of a physiological change after mating. I
will expand the lines by feeding them food that is equivalent to an unhealthy human diet- high with
sodium- by adding salt/oily components to the food. Options for such a diet include coconut oil or
maple syrup. Also, complete additional training for food preparation. The drug order will be
completed by the end of January.

February: Continue expanding lines and feeding a high sodium, high carb diet to the flies.
Calculate dosages of drug that correspond to human dosages which the flies will be able to
metabolize without it being toxic. Make the food with the different dosages of drug. Transfer the
expanded lines to the drug food. Complete methodology, introduction, and material section of the
paper by February 28th.

March: Continue experiment by feeding flies different dosages of Entresto. In mid march I will
perform the cold stress test on the flies. As data is collected, I will consolidate it into a data table and
begin looking for trends in data. By the end of March, I will be able to begin the data/results section
of the paper.

April: Finish consolidating data on the amount of flies that survived by April 15th. Complete
data/results section by April 20th. Analyze the data for trends and draw conclusions,Write the
conclusion/discussion section for the paper by April 25th. Submit paper to mentor for review on
April 26th. Depending on when Dr. Leips returns the paper to me with his corrections, I will
complete the final changes.

May: On May 1st, I will submit the paper to the journal.

https://hal.archives-ouvertes.fr/hal-00894183/document
Cold Stress test:
Place vials of flies in 0 degrees Celsius for 48 hours (decreasing initial temperature of 20-22 degrees
celsius in 15 minutes to the desired 0 degrees).- either in a fridge or a cold bath. After 48 hours, pull
the flies out and check the flies reactivity when they are touched with forceps. (ask about
quantitative way to measure this)

Entresto how it works:

Valsartan: decreases blood vessel tightening and the buildup of sodium and fluid
Sacubitril- blocks the activity of an enzyme that breaks down peptides. These peptides actually relax
blood vessels and decrease sodium and fluid in the body

Pilot studies were performed and a prefreezing temperature of 3.3 C for 120 h was chosen for the
assessment of mortalities. All experimental flies were aged between 2 and 3.5 days and manipulation
was carried out without anaesthesia. Cold resistance was measured using shell vials with culture
medium in a horizontal position. The number of survivors to stress was scored (at 25 C) as the
number of flies that could walk 24 h after the cold treatment.

Flies both from mass populations and isofemale lines were subjected to cold treatment of 0C for 48
h in a thermostatic chamber with saturated humidity. The initial temperature was 20-22C, and the
temperature declined to 0C in about 15 min.
Resistance to cold was measured by flies ability to react when touched with forceps

Resilience- cold/shock therapy and their ability to do something afterwards like climbing or strength
Pilot projects: gonna have to test the assays before doing it with the drug

Chill coma recovery time- individual flies placed in ice water mixture for 6 hours and they measured
how soon it took for each individual fly to stand on its legs
Can also get survivorship data
4 flies/vial
Blocking-
Set up experiment in the warm room bc of humidty and fluctuating temperature
20/30 flies per treatment for observation

Double sided tape

Flies
Timer
10 flies
Knock the flies out by putting them in the fridge
Time when they first move a leg
Put petri dish on top of it to prevent it from flying away