1. Murder by HIV? Grades 5–8 Edition by Michèle I. Shuster Department of Biology New Mexico State University, Las Cruces, NM Naowarat (Ann) Cheeptham Department of Biological Sciences Thompson Rivers University, Kamloops, B.C., Canada Laura B. Regassa Department of Biology Georgia Southern University, Statesboro, GA

2. Key Concepts and Background Basic knowledge of HIV and what happens during an HIV infection (see next slides for a brief re-cap) Genetic relationships and phylogenetic trees – # of genetic differences indicates distance – Reading nodes/common ancestors on phylogenetic trees

3. HIV Infection Infects, replicates, infects, replicates Every time – Chance for mutation – “Typo” during replication So within the body – Many RELATED strains of HIV – All descended from original (infecting) strain – All related to each other – All related to the infecting strain

4. Over time: genetically diverse population Early Later Even Later

5. The Case

6. State of Louisiana vs. Richard J. Schmidt, MD Dr. Schmidt was having an affair with a nurse in his office On-going for 10 years During this time – He gave her injections – Vitamin B-12 She broke off the relationship (July 1994) August 4, 1994 – He showed up and gave her a “B-12” injection – Against her will

7. January 1995 She tested positive for HIV Question: Was this a new or old infection? – She was a regular blood donor (HIV testing) – Last blood donation—April 1994 – She was HIV negative Accusation…

8. April 1994 No HIV Blood donation August 1994 Injection by Doc/boyfriend January 1995 HIV positive January 1995 Nurse accused boyfriend of infecting her

9. Investigation Doctor’s office Hidden blood-draw record – HIV positive patient – Blood drawn on August 4 th, 1994 – Blood not sent to lab – Staff were told to hold the tube of blood for the doctor

10. April 1994 No HIV Blood donation August 4, 1994 Injection by Doc/boyfriend January 1995 HIV positive January 1995 Nurse accused boyfriend of infecting her August 4, 1994: Blood drawn from HIV-positive patient at Doc’s office

11. Questions Is Nurse’s (victim) HIV related to the patient’s HIV? Is Nurse’s (victim) HIV related to other HIV strains in LA? Is Nurse’s (victim) HIV related to other strains of HIV in the U.S.?

12. Based on the questions What are some hypotheses? How can these be tested? What do you need in order to test these?

13. Draw Blood and Obtain HIV Sequences HIV positive patient – Possible source Nurse – Victim HIV positive people from the same city in LA HIV positive person from the U.S. (not LA)

14. If Doctor injected Nurse with Patient HIV… Which will be most closely related? A.Victim HIV and Patient HIV B.Victim HIV and LA cases HIV C.Victim HIV and USA HIV

15. Let’s Build the Tree Go online to MUSCLE: http://www.ebi.ac.uk/Tools/msa/muscle/ http://www.ebi.ac.uk/Tools/msa/muscle/ We will import HIV sequences – The pol gene – From patient HIV, from Victim HIV, from “other” HIV

16. Copy/Paste Sequences from HIV Sequence file Paste Here Then hit “Submit” Submit You will need the.txt file from the supplemental materials for the sequences to copy and paste.

17. Wait……

18. Wait… then you will get this— click on “Result Summary”

19. Then click on “Start Jalview”

20. Then you’ll get this An alignment of all the sequences Patient (possible source) Victim (the nurse) LA (random HIV from LA) USA (random HIV from the US) Click on Calculate…

21. Calculate Calculate Tree – Average Distance/ BLOSUM 62

22. Last Common Ancestor between Patient strains and Victim (Nurse) strains? Last Common Ancestor between USA strain and LA strains? Last Common Ancestor between Patient strains and LA strains?

23. So which HIV is more closely related to the Nurse’s HIV? A. Patient HIV B. LA HIV C. USA HIV So which HIV is more closely related to the Nurse’s HIV? A. Patient HIV B. LA HIV C. USA HIV

24. So what is most likely source of Nurse’s HIV?

25. So what can we conclude?

26. Jury Agreed So did the LA Court of Appeals So did US Supreme Court Sentence Upheld – 50 years of hard labor Don’t underestimate the power of the phylogenetic tree!

27. April 1994 No HIV Blood donation August 4, 1994 Injection by Doc/boyfriend January 1995 HIV positive January 1995 Nurse accused boyfriend of infecting her August 4 th, 1994: Blood drawn from HIV-positive patient at Doc’s office 1998 Doc convicted 50 yrs of hard labor 1998 Doc convicted 50 yrs of hard labor 2000 LA Court of Appeals Upheld conviction 2000 LA Court of Appeals Upheld conviction 2002 US Supreme Court Upheld conviction 2002 US Supreme Court Upheld conviction