BioBus Current Topics

1
Welcome to our professional development workshop
2
What is CURE?

• Connecticut United for Research Excellence, Inc.
• Represent biotechnology and pharmaceutical
  sectors before state legislature and policy
  makers
• Build a critical mass of biotech and
  pharmaceutical companies
• Foster relationships between academic and
  industry research that lead to technology
  transfer
• Be the go to source for information about
  bioscience in Connecticut

3
In Connecticut

• Achillion Pharmaceuticals, Inc.
• Alexion Pharmaceuticals, Inc.
• Bayer Corporation
• Boehringer Ingelheim Pharmeceuticals
• Bristol-Myers Squibb
• CGI Pharmaceuticals
• CuraGen Corporation
• HistoRx, Inc.
• Institute for Pharmaceutical Discovery
• Invitrogen Corporation
• Marinus Pharmaceuticals, Inc.
• Molecular Staging
• Neurogen Corporation
• Pfizer
• Purdue Pharma L.P.
• Rib-X Pharmaceuticals, Inc.

4
Bioscience Careers

• Connecticut Business and Industry Association
• http//www.cbia.com/ed/STC/career_explorations/car
  eer_explor_info/health_bio.htm

Biotechnology and pharmaceutical companies are
among the fastest-growing industries in
Connecticut, yet they are having difficulty
finding qualified workers with the right
technological and scientific skills to fill
high-demand, high-wage jobs. - CBIA
5
Sponsors of BioBus Educational Programs

• Platinum Sponsors CURE and Connecticut
  Innovations
• Gold Sponsors Pfizer Inc and The Pfizer
  Foundation
• Silver Sponsors Boehringer Ingelheim
• Bronze Sponsors Achillion Pharmaceuticals,
  State of CT
• Contributors Bayer Corporation
• Supporters Wesleyan University, Quinnipiac
• University,
  University of New Haven,
• Yale
  University, SMART Technologies,
• Bio-Rad,
  Invitrogen, Edvotek

6
CUREs Educational Goals

• Generate student interest in and excitement for
  bioscience to encourage career exploration
• Provide educators with laboratory experience,
  bioscience information and innovative teaching
  techniques
• Generate public understanding, enthusiasm and
  support for bioscience

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CT BioBus Educational Programs

• BioBus
• BioConnection
• Teacher Professional Development
• Public Outreach/Events

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As of 2008

• The BioBus has trained over 800 teachers
• Reached over 43,000 students
• Conducted over 1,300 experiment sessions

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Today We Will
Reinforce and increase your knowledge of
bioscience Help you to incorporate bioscience
in your curriculum Provide hands-on bioscience
opportunities for you and your students
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LOGISTICAL INFORMATION FOR YOUR BIOBUS VISIT
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Safety Protocol

• ALL STUDENTS MUST
• Wear long pants and closed-toe shoes
• Tie back long hair
• Refrain from eating, drinking or gum chewing
• Leave behind, jackets, hats, bags, purses or
  papers (everything except BioBus notebook and
  pen/pencil)
• Eat a meal and drink beforehand (breakfast is a
  must)
• (Non-latex gloves and safety glasses
• Are provided)

12
On board Teacher Responsibilities

• The BioBus is an extension of YOUR classroom
• Understand school emergency procedures and
  students medical needs
• Discipline
• Pair up students - standing chart
• Help instruct and demonstrate techniques
• Be present on bus the ENTIRE session (have a
  plan/helper if a student gets sick)
• Make staff aware of any conditions/health issues
  you feel we should know about (before visit day)
• We are wheelchair accessible

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Things to Know

• Delivery and Parking Site
• The Bus is usually delivered the day/night before
  your visit (occasionally the morning of)
• Staff cannot move the bus once its parked!
• The bus cannot be parked in fire zone
• If wheelchair lift is used, the BioBus entrance
  must be 8 feet from curb
• Very level ground is needed for parking
• 2 spaces for staff vehicles near BioBus is
  appreciated, for loading supply bins on to bus

14
Things to Know

• Fill out class list form for every session 24
  students MAX per session
• Break between sessions 30-60 minutes
• 1.5 or 2 hour sessions are booked, but we may
  finish early (depends on class size and skill
  level)
• Make sure the pre-lab activities were done
• Short tours can be arranged if requested four
  weeks beforehand
• Check in with us on BioBus 30 minutes before
  first session
• Staff arrives 1 hour before first session, will
  not check in with school

15
Press Releases

• Samples available on our website
• Press packets available on bus
• Photo releases up to you

16
Visiting Scientist Program

• From a sponsor company
• Scientific expert
• Information on career opportunities
• Demonstrate the applicability of the science
• Resource for teachers and school
• Parent scientists are welcome

Dr. Linda D. Strausbaugh, is a Professor in the
Dept. of Molecular Cell Biology at the
University of Connecticut
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After Your BioBus Visit

• Sign up again!
• Online request form
• Training is good for 2 years
• Consider BioConnection Program
• Thank You notes
• BioBus Sponsor
• Send to CURE offices

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Today BioBus Experiments I

• The Gene Screen (New!)
• recommended for grades 6-10
• A Twist of Lyme
• recommended for grades 7-12
• Hunting the Immune Villain
• recommended for grades 9-12

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The Gene Screen NEW! (grades 6-10)

• How do DNA mutations occur? Can our cells repair
  damaged DNA? What are some of the consequences of
  DNA damage that is not repaired? In this
  experiment, BioBus scientists will investigate
  the effects of ultraviolet light and various
  sunscreens on a UV-sensitive strain of
  Saccharomyces cerevisiae, or yeast, to learn
  about the processes of DNA mutation and repair.
  Specifically, students will design and carry out
  an experiment to test the effectiveness of
  various sunscreen samples on a strain of
  UV-sensitive yeast. This experiment will also
  highlight the effects of ultraviolet radiation on
  skin cells and the link between UV exposure and
  skin cancer.
•  

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DNA Structure
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DNA The Code of Life
www.accessexcellence.org
22
The Double Helix Gamewww.nobelprize.org
23
Gene

• Definition
• Segment of DNA that can be used to make proteins

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A DNA Analogy

• Genome Book
• Chromosome Chapter
• Gene Paragraph
• Exon Sentence
• Intron Blank space
• Nucleotides Letters

25
The Two Steps of Protein Synthesis
Graphic Courtesy of TIGR
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Online Transcription/Translation Activity
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DNA Damage

• DNA is damaged by ultraviolet (UV) radiation
• Photodimers are cross-linked bases
• (C-C, T-T) that distort the DNA shape
• Unrepaired DNA damage can lead to DNA mutations
  (altered DNA sequences)
• Mutations in important DNA regions (e.g. p53
  gene) can lead to cancer

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UV Induced DNA Damage
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DNA Repair

• Most DNA damage is successfully repaired
• Nucleotide Excision Repair Mechanism (NER)
• Multiple enzymes remove damaged DNA and fill the
  gap with correct bases
• This occurs on a continuous basis in our cells

http//www.phys.ksu.edu/gene/excisio3.html
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DNA Repair
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UV damage and Skin Cancer

• UV induced DNA damage is the most common cause
  of skin cancer
• Skin cancer is the most common form of cancer in
  the United States (1 million cases diagnosed
  annually)
• One in five Americans will develop skin cancer
  in their lifetime
• UVA and UVB radiation are proven carcinogens
  (i.e. tanning beds are not safe)
• UVB is bigger culprit

Above UV radiation is one part of the suns
electromagnetic spectrum. UVA and UVB radiation
passes through the Earths atmosphere, while UVC
does not. Credits WebShade.com
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UV Sensitive Yeast

• Why Yeast?
• Genome is sequenced
• Eukaryotic (cellular, repair mechanisms similar
  to humans)
• Easy to maintain and fast doubling time (1.5 -2
  hrs _at_30oC)
• UV-sensitive Yeast (rad1 rad18 phr1 mutant)
• Mutation in repair genes inability to repair
  UV-induced DNA damage
• Provides an excellent indicator of the intensity
  of UV radiation as well as the protection factor
  offered by various sunscreens and materials

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Xeroderma Pigmentosum

• XP is a genetic condition
• Mutations in one or more of the genes used for
  DNA repair
• Unrepaired DNA leads to many mutations
• 1,000x more likely to develop skin cancer
• XP individuals must take drastic measures to
  avoid UV exposure

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Pre and Post Lab Activities

• Pre-Lab
• Gene Screen Webquest
• Detecting UV light Using Tonic Water
• UV sensitive beads exposure tally
• Post-Lab
• Class Data Analysis Worksheet
• Exploration Topic projects

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Cells Alive Website
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Learn.Genetics Website
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To the BioBus!
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A Twist of Lyme (grades 8-12)

• Lyme disease is easily detected using a
  scientific technique called an Enzyme-Linked
  Immunosorbent Assay (ELISA). An ELISA is used to
  detect the presence of human antibodies produced
  to combat a foreign substance, in this case, the
  bacteria which cause Lyme disease. BioBus
  scientists perform an ELISA to determine whether
  the symptoms of four different fictitious
  patients are due to Lyme disease.

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Lyme Disease

• First discovered in Lyme, CT in early 1970s
• Transmitted by deer ticks infected with the
  bacteria Borrelia burgdorferi
• Since 1980, over 170,000 cases have been
  reported in the US

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Symptoms

• Bulls eye rash erythema migrans
• Painful and swollen joints
• Nervous system
• Stiff neck, headaches, Bells palsy, numbness,
  pain or weakness in limbs, poor motor
  coordination
• Heart problems, eye inflammation, hepatitis,
  severe fatigue

? Example of erythema migrans, the red bulls
eye rash that may develop with Lyme
disease. Photo Wikipedia Commons
41
Diagnosis of Lyme Disease

• Blood is drawn from patient
• ELISA is used to detect antibodies
• Western blot is used to confirm positive
  results
• PCR of patient samples can be used to detect
  Borrelia burgdorferi

Deer ticks spread Lyme disease in the
northeastern and upper Midwestern U.S. (photo
WebMD)
42
Treatment/Prevention

• Check yourself!
• Bacteria takes 2 days to enter your system
• Antibiotics
• The earlier the better
• After treatment problems may still exist
• Vaccine?
• Limerix pulled off the market, new versions
  underway

Thumb with deer tick nymph (photo FDA.gov)
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Hunting for the Immune Villain

• Human immunodeficiency virus, or HIV, was first
  acknowledged by the CDC in 1983. Today more than
  5 million new infections are expected yearly.
  HIV is easily detected using a scientific
  technique called an Enzyme-Linked Immunosorbent
  Assay (ELISA). An ELISA is used to detect the
  presence of human antibodies produced to combat a
  foreign substance, in this experiment, the virus
  which causes HIV. Here, BioBus scientists
  perform an ELISA to determine whether the
  symptoms of four different fictitious patients
  are due to HIV.

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What is a Virus?

• Submicroscopic particle that infects cells
• Genetic material (DNA or RNA) surrounded by
  protein coat
• Some insert DNA into host cell to make proteins
• Cannot reproduce outside of host
• Alive?

45
Human Immunodeficiency Virus (HIV)

• Retrovirus that infects CD4 T Cells, dendritic
  cells and macrophages (Immune Cells)
• Retrovirus RNA is genetic material
• HIV must make a DNA copy of its RNA before
  insertion into host cell

www.wikipedia.org
DNA
Protein
RNA
46
HIV Life-cycle
http//www.mcb.uct.ac.za/HIVannot.jpg
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HIV Animations
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HIV Timeline

• Early 20th Century Originated from mutation
  in Chimpanzee SIV in Cameroon, Africa
• Early 1980s First US cases were reported
• Homosexual men reporting opportunistic infections
  such as pneumonia and Kaposis sarcoma
• 1983 HIV was identified to be the cause of
  AIDS
• 2007 40 million people worldwide are infected

UNAIDS
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HIV Transmission

• Sexual contact
• Contaminated blood or blood products
• IV drugs users, contaminated transfusions, needle
  sticks
• Mother to child
• in utero, during childbirth, breastfeeding
• Without treatment the transmission rate is 25
• Other
• Present in small concentrations in saliva, tears
  and urine, but not in sweat
• none are transmission routes
• Deep kissing or biting
• very small potential for transmission if open
  sores, bleeding gums or tissue damage are
  involved
• HIV is not transmitted by insects or casual
  contact of any type

50
Signs and Symptoms

• Initial flu or cold-like symptoms followed by
  long, often asymptomatic period (latency)
• Later symptoms fatigue, swollen glands, rapid
  weight loss, fever
• Eventually, T4 helper cell counts drop
  significantly, making one susceptible to
  opportunistic infections
• T4 cell count below 200/ul ? AIDS

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Diagnosis and Treatment

• Diagnosis
• Initial AIDS test or ELISA
• Positive results confirmed with a second test
  (Western Blot)
• Treatment
• Typical treatment is a cocktail of drugs that
  slow HIV replication and infection at different
  points in viral life cycle
• Vaccine is not yet available
• Complicated because virus has very high mutation
  rate

52
Some Vocabulary

• Immunology- The study of the immune system and
  its response to antigens
• Antigen - a substance that elicits an immune
  response and can be bound by an antibody
• environmental agents/infectious agents
• proteins, carbohydrates, or nucleic acids
• part of a virus or bacteria
• Antibodies - proteins produced by white blood
  cells in response to an antigen

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Graphic from accessexcellence.org
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ELISA

• Enzyme-Linked Immunosorbent Assay
• Detects the presence antibodies directed against
  a particular antigen
• Useful for diagnosis of a variety of conditions
• Bacterial infection
• Viral infection
• Pregnancy
• Drug use
• GMO
• Color change indicates positive result

57
Inside the Positive Control Well
Substrate
enzyme
2??
1??
Antigen
Plastic Surface
58
ELISA tutorials
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Pre- and Post- Lab Activities

• Pre-Lab Activity
• Plaster of Paris molds
• Post Lab Activity
• Ouchterlony Test

60
Finding a Perfect Fit

• Materials
• Small disposable containers
• Plaster of Paris
• Small objects

• Procedure
• Cast objects in plaster of paris
• mold antibody
• object antigen
• Mix up antigens and antibodies and look for
  specific fit

61
Wheres the Soy?

• Materials
• Agar plates
• Potassium Phosphate Tribasic
• Calcium Chloride (dihydrate)
• Straws

• Procedure
• Holes are made in agar plates with a straw
• Solutions are dropped into holes and allowed to
  slowly diffuse toward one another
• Interaction between potassium phosphate and
  calcium chloride causes precipitate to form
  between wells
• 3 CaCl2(aq) 2 K3PO4(aq) ? Ca3(PO4)2(s)
  6 KCl(aq)

Key A Soy B Beef C Speedys beef D
Saline E French fries F Soy Antibody
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Ima Munes Story
Ima Mune went to eat a burger at Speedys.
She is very allergic to soy, and knowing that soy
is commonly used as a filler in hamburger meat,
is always cautious when ordering. However,
Speedys restaurant claims to have 100 All-Beef
Burgers. So, being in a ravenous mood for
a burger, she ordered one with lettuce and tomato
and a side of fries. A few minutes after
consuming her burger, Ima Mune started to get
violently ill. She became suspicious that
Speedys claims of all beef burgers was false.
Your job is to determine whether Ima Munes
suspicions are correct.
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Ouchterlony Results
Speedys Beef
Soy
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Back to the BioBus!
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Questions?

• Thank you!