MIcro Exam II – Lectures 10-12 – Flashcards

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question
Is a virus alive?
answer
No, it can direct life, but has no metabolism, no cytoplasm, organelles.
question
T/F Bacteria outnumber viruses.
answer
False - viruses outnumber 10:1.
question
What is a capsid?
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Surrounds the genome in a virus.
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Can a virus have both RNA and DNA?
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No - mutually exclusive.
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What is a naked virus?
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No viral envelope, which is stolen from the host cell.
question
Describe the virus contents.
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DNA xor RNA, a few viral proteins for replication, and a protein shell.
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What is a virion?
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Extracellular form of a virus - also called virus particle.
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What are the contents of a nucleocapsid?
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DNA xor RNA, structural proteins/protein shell, enzymes and nucleic acid binding proteins, and Nucleocapsid may be the outer structure (if it's naked).
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What is the envelope made of?
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Lipid bilayer.
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Which is hardier - naked or not naked virus?
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Naked - resistant to the environment. The lipid bilayer of envelope makes it more "fussy".
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How does our I/S recognize viruses?
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The viral proteins that project out of the membrane for attachment - peplomers, spikes, VAPs).
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What is a capsomere?
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A viral nucleocapsid structure - can be icosahedral - 20 sides, or alpha helical shaped (coiled).
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Name nucleocapsid structures?
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Capsomeres, helical(rod) or icosahedral, bacteriophage (spaceship).
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How do we get rid of viruses?
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Antibody response can kill it, but not clear it. Need CMI.
question
Name the 2 bacteriophage lifecycles.
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Lytic, Lysogenic.
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What are the details of the lytic lifecycle?
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attach, penetrate, inject (no uncoating), biosynthesis, maturation (assembly of new virus), release (lysis of the host cell).
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What is the process of DNA integrating into the chromosome, and the virus sitting dormant in the cell?
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Lysogenic life cycle of bacteriophage.
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Where does an envelope come from?
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Host cell membrane.
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What is a VAP?
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Viral attachment protein.
question
What does influenza have that helps it attach to its host cell?
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Hemaglutinin.
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How do enveloped viruses spread?
answer
Needs moisture - airborne droplets, secretions, etc... Fragile b/c of membrane lipids. Also has VAPS, spikes, peplomers, etc...
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What happens in the Attachment phase of viral replication?
answer
Uses specific receptors, bacteriophages use tail fibers.
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What happens in the Penetration phase of viral replication?
answer
Animal viruses are taken up by endocytosis, whole virus inside the cell. Bacteriophages attach and inject their material. Some bacteria have a primitive I/S to chop up foreign DNA to protect against bacteriophages.
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What happens in the Synthesis phase of viral replication?
answer
Bacteriophages - all steps in the cytoplasm. Animal viruses - diff for DNA, RNA.
question
How does a DNA virus replicate itself?
answer
Replicate in the nucles and use ribosomes in the cytoplasm.
question
How does positive sense RNA replicate?
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They use negative sense RNA to transcribe mRNA in the nucleus, then the proteins are made in the cytoplasm.
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How does negative sense RNA replicate?
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Converted to +sense, then the positive sense RNA is used as mRNA to directly make proteins.
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What happens in the Assembly phase of viral replication - where does it occur?
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Cytoplasm of the host cell, some viruses need envelope from the host cell.
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What happens during the Release phase of viral replication?
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Lytic cycle - host cell is killed when virus lyses. After lying dormant in lysogenic cycle, it can enter lytic cycle an lyse. In eukaryotic viruses, infection leads to lysis or budding.
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How are viruses classified?
answer
Size, morphology, nucleic acid content, structure, diseases causes, means of transmission, host cell.
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What is the Baltimore System?
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Viral classification system. Most used currently. 7 families that considers structure, biochemical characteristics, genome structure, and mode of replication.
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What is tropism?
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Virus host cell specificity - determined by VAPs on the surface. Ex. Hep C is hepatotropic. Some have a wide range, some very narrow.
question
Describe a typical viral growth curve.
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Wish I could copy the chart in here. On Section 10 handout.
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What is the viral burst size in a viral growth curve?
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The number of virions produced when a cell lyses. Ranges from a few to 1000s.
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What is the replication cycle in a viral growth curve.
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Process that starts when a virion enters a cell and ends when virions are released.
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T/F A typical human virus replication cycle is 48-72 hrs?
answer
False - 8-40 hrs.
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What is the eclipse period of a viral growth curve?
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The period of time between infection by a virus and the appearance of the mature virus within the cell. If you're exposed during this time you won't get infected.
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What are Early Proteins?
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Biosynthesis of viruses happens in steps. Proteins needed initially for replication are Early Proteins.
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What are Late Proteins (examples)?
answer
Proteins needed by a virus to assemble virions. Include capsomeres and capsids.
question
Name the 7 Baltimore classification categories.
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dsDNA, ssDNA, dsRNA, +sense ssRNA, -sense ssRNA, RNA reverse transcribing viruses, DNA reverse transcribing viruses.
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Describe the replication of dsDNA viruses.
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host cell needs to transcribe the DNA->RNA->protein. Host cell already knows how to do this.
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Describe the replication of ssDNA viruses.
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Translocated to the nucleus. Synthesizes into dsDNA. This is how our DNA is replicated, so machinery is already in place to do that.
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Describe the replication of dsRNA viruses.
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Viruses have an enzyme to make mRNA. Can then translate this to a protein. Viral RNA polymerase.
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Describe the replication of +sense ssRNA viruses.
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Could be directly used as mRNA to make a protein. But making more copies is a problem. A viral enzyme is needed to copy.
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Describe the replication of -sense ssRNA viruses.
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This must be converted to +sense. Can then be used as mRNA.
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Describe the replication of RNA reverse transcribing viruses.
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RNA->DNA. Virus needs to bring an enzyme to do that.
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Name the DNA viruses.
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Hepadnaviridae, Herpesviridae, Adenoviridae, Poxviridae, Papillomaviridae, Parvoviridae, Polyoma.
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Name the ssDNA viruses.
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Only one - Parvoviridae.
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Name the naked DNA viruses.
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Pre-PAP... Polyoma, Papilloma, Adenovirus, Parvovirus (Not Pox).
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Name the reverse DNA virus.
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Hepadnaviridae - Hepatic DNA virus.
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Name the positive ss RNA viruses.
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A RETRO TOGA party with FLAVorful PICO de gaillo and CORONAs.
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Name the negative ss RNA viruses.
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Paramyxo, Orthomyxo, Rhabdo, Filo, Bunva, Arena.
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Name the dsRNA.
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Reoviridae.
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Name the Reverse RNA.
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Easy - Retroviridae - HIV.
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What unique problem is presented to viruses using eukaryotic ribosomes?
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Eukaryotic cells generally uses monocistronic (one protein for one mRNA) and the virus has polycistronic mRNA.
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How does HIV adapt to the monocistronic/polycistronic problem?
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The virus makes monocistronic mRNA, the host cleaves the viral mRNA into monocistronic, a long polyprotein is made then cut, viral mRNA has sites for ribosomes to internally AND at 5' end.
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List virus virulence factors.
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Attachment proteins, enzyme.
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What is an attenuated virus?
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Lost its virulence factors.
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What is viremia?
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Viruses in the blood. Can then spread to secondary site.
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What is significant about your I/S fighting a viral infection.
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Must kill your own cell since it's intracellular. Cytokines cause dmg. Immune complexes and complement also cause damage.
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What is viral incubation period? How is it measured?
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Time between exposure and symptoms. Usually measured in days to years.
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What is the significance of a latent viral infection - how does this relate to a chronic infection?
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Latent - viral genome present, but shed virus is NOT DETECTABLE. Chronic - small amt of virus is present.
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How are viruses spread?
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Inhalation, fecal/oral, blood.
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What is syncytium?
answer
Cell to cell transfer w/out being exposed to your I/S.
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How can viruses cause cancer?
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Provide/Activate growth stimulating genes, Remove stops for DNA synthesis and growth, Prevent apoptosis, Integrate into host and turn on neighboring genes, Cause cells to divide faster and get more mutations.
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How are viruses grown?
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Tissue culture or live animals.
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What is CPE?
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Cytopathic effect of the virus - what you see. CMV - large cells, inclusion bodies - negri bodies, rabies in brain, HSV, Varicella-Zoster, etc....
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How are viruses diagnosed?
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Serology, detection of viral components, symptoms, history, isolation and growth, electron microscope.
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How are viruses treated?
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Antivirals, Disrupt the virion (envelope, stop infection), Block attachment, Block penetration and uncoating, Block RNA synthesis/transcription, Blcok genome replication, Protein Synthesis, Virion Assembly and Release, Stimulate Host I/S.
question
What is the only known human virus of the Polyoma family?
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SV-40.
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Is SV-40 oncogenic?
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Yes - although we don't know why. Rarely causes disease in monkeys unless infected w/ SIV. It does bind MHC I, so the I/S doesn't see the cell.
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What is the shape of the capsid and nucleic acid of HPV?
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Naked nucleocapsid, icosahedral capsule.
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What virus causes cervical cancer?
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HPV (HSV).
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What is Gardasil a vaccine for?
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HPV (HSV) - combines 16, 18, 6, and 11 capsid proteins.
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Where does HPV replicate?
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Squamous cell epithelium. Becomes warts. Causes cell proliferation in mucosa/cutaneous. Inhibits tumor suppressor, so unchecked growth.
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What are some disease caused by adenoviruses?
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Resp. infections, Conjunctivits, Pharyngoconjunctival fever in infants, Gastroenteritis,
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How is adenovirus transmitted?
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Droplet or fecal/oral.
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Describe the Adenovirus.
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DS DNA naked nucleocapsid icosahedral virus.
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What virus is used as a vector for gene therapy?
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Adenovirus.
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Describe the herpes virus.
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Enveloped DS DNA virus w/ icosahedral capsidds. Can cause latent and recurrent infections.
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What is the only virus family w/ a nuclear envelope (instead of an outer cell membrane)?
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Herpes.
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Name some diseases caused by Herpes.
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HSV 1, HSV2, Varicella Zoster, Cytomegalovirus, EBV, Herpes Virus-8.
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What does HSV 1 infect?
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Herpes Simplex Virus 1 infects the lips/face/skin/oral cavity.
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How is HSV 1 treated?
answer
Acyclovir.
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Who is at risk with HSV 1/2?
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Immunosuppressed, newborns, can spread to brain and be fatal.
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How are HSV-1/HSV-2 acuired?
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Cuts, infection. HSV-2 is an STD.
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What does HSV-2 infect?
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Mucoepithelial cells. Latent in neurons.
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What does Varicella Zoster cause?
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Chicken pox/shingles. Infects mucoepithelial cells, latent in neurons, resp. and close contact transmission.
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Where does Zoster lie dormant?
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Neurons - they'll follow the nerve track.
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What does Varicella/Zoster spread thru?
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Macs, so it spreads quickly.
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Which Herpes disease do we have a vaccine for? What type of vaccine?
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Live attenuated for Varicella/Zoster.
question
What cells does CMV infect?
answer
monocytes, lymphocytes and epithelial cells, latent in monocytes and lymphocytes
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How is CMV spread?
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close contact, transfusions, tissues, congenital.
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What has caused CMV to be worse?
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More prevalent due to HIV - immunosuppressed. Also pregos at risk.
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T/F Most of us have had CMV.
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True - just not serious to us.
question
What caused Jimmer Fredette to miss a critical stretch of last season?
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Mono-EBV.
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What cells does EBV infect?
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B-cells - Jimmer played for BYU. Also epithelial cells.
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How is EBV transmitted?
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Spit - the kissing disease.
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How long does mono last?
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2-3 weeks, no vaccine or treatment. That's why Jimmer took a month to get back to normal.
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How is mono diagnosed?
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Mono spot. Your serum + sheep RBCs -> clumping.
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What is associated with Burkitts lymphoma?
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EBV and co-infection w/ malaria.
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What causes Kaposi's Sarcoma?
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Herpes 8 - Type of skin cancer.
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Why do AIDS pts get Kaposi's Sarcoma?
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Type of skin cancer, need T cells to clear it.
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Describe the pox virus.
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DS DNA enveloped virus.
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What is unique about the Pox replication cycle?
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Replicates in the cytoplasm - brings in enzymes to copy its DNA.
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What does Pox cause?
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Variola - one of these is small pox. Also Molluscum contagiosu (warts - only spread by close contact, sex and fomites), vaccinia virus (cowbox), monkeypox (milder, 1-10% fatal).
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Talk about the small pox virus.
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Spread by inhalation, travels in blood, fatal, acute, highly infections. BIG CANDIDATE FOR BIOTERRORISM. No treatment.
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Describe the Parvo Virus family.
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SS naked DNA (only SS one).
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Name the SS naked DNA virus?
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Parvovirus. This is a small virus - needs growing cells to replicate.
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What disease does Parvovirus cause?
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Strain B19 causes Fifths disease. Slapped cheek, Fever, problem for anemic kids.
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What causes slapped cheek syndrome?
answer
Parvovirus, B19, Fifths Disease.
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What are the characteristics of the Picorna virus family?
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Small, SS + strand naked RNA, icosahedral capsid.
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Which Picornavirus that is not stable in acid (the rest are stable)?
answer
Rhinovirus.
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T/F Picorna virus doesn't need to bring in its own enzymes.
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False - brings in proteases and RNA dependent RNA polymerase.
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Name the 3 important genera of Picornaviruses.
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Enterovirus, Rhinovirus, Hepatovirus.
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T/F Picornavirus is cytolytic.
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True. Repicates and spreads quickly as well.
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What do polio and coxsackie viruses belong to (family/genus)?
answer
Picornavirus family, Enterovirus genus.
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What makes Enteroviruses resistant to harsh conditions (GI tract and sewage)?
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Their capsid.
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Which enterovirus has a vaccine?
answer
Polio.
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How is Polio spread?
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Fecal contamination of water.
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Describe the lifecycle of the polio virus?
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Replicates in pharynx and GI tract, enters bloodstream, then goes to the CNS.
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Why does polio cause paralysis?
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It's cytolytic for motor neurons.
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Name the vector/reservoir for polio.
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Us (so no vector). So this can be eliminated.
question
Herpangina is caused by what?
answer
Coxsackievirus - Group A - not related to Herpes virus - hand/foot/mouth. Also causes conjunctivities.
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What causes Pleurodynia?
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Coxsackievirus - Group B. Myocardial/pericardial infections.
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How are enteroviruses treated?
answer
Antiviral pleoconaril - prevents penetration into the cell. Must be given early.
question
T/F Sabin is inactivated polio vaccine.
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False - Sabin is Oral - live attenuated.
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What does Rhinovirus belong to?
answer
Picornavirus family.
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T/F There just a few serotypes of Rhinovirus.
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False - over 100.
question
How is Rhinovirus treated?
answer
Nasal vasoconstrictors, antiviral pleocarnil for asthmatics.
question
Describe the calci virus family.
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Postive sense SS RNA naked capsid icosahedral.
question
What causes Norwalk virus?
answer
Calcivirus. This is the prototype.
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What causes 50% of gastroenteritis?
answer
Norwalk virus.
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How is Norwalk virus treated?
answer
Prevention and rehydration. Must use bleach or steam to prevent.
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T/F the infections dose is very high for Norwalk virus.
answer
False - very low b/c it' stable in acid, bleach. Need fresh conc. bleach or steam.
question
What is the only +SS RNA enveloped virus?
answer
Coronavirus.
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How does Coronavirus survive in the GI tract?
answer
Surface projections.
question
What is the 2nd most prevalent cause of the common cold?
answer
Coronavirus. Also found in GI tract.
question
What caused SARS?
answer
Coronavirus.
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What are the characteristics of the Paramyxovirus?
answer
-Sense, SS RNA - enveloped, helical nucleocapsid.
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How can you tell family members of the Paramyxofamily apart?
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By their VAPs.
question
T/F Paramyxoviruses induce cell fusion?
answer
True - they induce cell fusion - giant multi-nucleated cells.
question
What causes measles/rubeola?
answer
Paramyxovirus, morbillivirus genus. Highly contagious - droplet transmission - not prevalent due to vaccines.
question
What causes Koplik's spots?
answer
Paramyxovirus, on the buccal mucosa.
question
What family/genus is parainfluenza part of?
answer
Paramyxovirus/paramyxovirus.
question
What kind of vaccine is the rubella vaccine?
answer
Live attenuated.
question
What is infected in rubella?
answer
Testes/CNS - due to viremia.
question
What do Nipah and Hendra belong to?
answer
Paramyxovirus - unassigned genus. Broad host range - accidental host causes DEADLY OUTCOME.
question
What causes RSV?
answer
Paramyxovirus Pneuovirus genus.
question
How are infants w/ RSV treated?
answer
Ribavarin - inhaled. Also Ig shots - anti-RSV - somebody else's antibodies.
question
What causes Influenza A/B/C?
answer
Orthomyxovirus.
question
T/F Orthomyxovirus is hardier than most enveloped viruses.
answer
True - can live on fomites for a day.
question
Name the VAP that elicits antibody response on orthomyxoviruses.
answer
Hemaglulutinin.
question
What is neurominidase?
answer
Enzyme important for release in orthomyxovirus. Target for drug therapy.
question
Describe the orthomyxovirus.
answer
SS -sense RNA enveloped helical virus.
question
T/F vaccines are available for orthomyxovirus.
answer
Of course - flu vaccine.
question
How are orthomyxos treated?
answer
Antivirals - stop attachment and uncoating.
question
How are influenzas classified?
answer
Time/Place of orig isolation/Antigen HA and NA.
question
What disease is associated with Reye's syndrome?
answer
Influenza B and chick pox and aspirin. Anyone <19 shouldn't be given aspirin.
question
Describe the Rhabdovirus family.
answer
Enveloped -sense SS helical RNA virus.
question
What causes rabies?
answer
Rhabdovirus.
question
T/F Rabies has never been fatal.
answer
False - before vaccine it was 100% fatal. Replicates for days/months at site of bite/inhalation, goes to CNS then brain.
question
How is rabies treated?
answer
Vaccine/antibodies. Takes a long time to replicate. Ab can block spread during incubation period.
question
How is rabies diagnosed?
answer
Negri bodies in neurons. CPE in neurons.
question
What is the reservoir for rabies?
answer
Dogs.
question
What are the important diseases caused by Filovirus family?
answer
Ebola and Genus Marburg virus. Hemorrhagic fevers.
question
Where is Filovirus endemic?
answer
Africa - spread by bats/monkeys/humans/syringes.
question
How is Filovirus treated?
answer
Anti-serum, quarantine, killing infected animals.
question
Describe the Reovirus family.
answer
DS + RNA - segmented genome.
question
T/F Reovirus is enveloped.
answer
False - double layered icosahedral protein capsids.
question
T/F Reovirus is stable in acids.
answer
True - also airborne transmission.
question
What types of illnesses are caused by Orthoreovirus.
answer
Not treated, not surveyed. Mostly asymptomatic in humans, mild upper resp/ GI, biliary atresial (Gall bladder blockage).
question
Where is Ortheovirus genus found?
answer
Rivers/water.
question
What family is Ortheovirus in?
answer
Reovirus.
question
What family is Rotavirus in?
answer
Reovirus.
question
What types of illnesses are caused by rotavirus?
answer
GI tract.
question
How is Rotavirus transmitted?
answer
Human fecal/oral.
question
Who does Rotavirus infect?
answer
Birds/mammals/humans.
question
T/F Rotavirus is hardy.
answer
True - stable in env., may resist handwashing, survives stomach acid.
question
T/F Rotavirus does not have a vaccine.
answer
False - live attenuated - oral - so you're trying to give local immuninity.
question
What causes 50% of all diarrhea in kids < 2 years old?
answer
Rotavirus.
question
What types of illnesses are caused by coltiviruses?
answer
Colorado Tick Fever - CO and West US, Canada.
question
What are the reservoir/vector of coltvirus?
answer
Rodents (reservoir) Ticks (Vector).
question
How do you treat coltvirus (CO tick fever)?
answer
It resolves itself. Fever, headache, myalgia, rash. Not reportable so no good stats.
question
How is coltvirus (CO tick fever) diagnosed?
answer
Blood serology, rule out RMSF - which is bacterial, treated w/ abx).
question
Describe Togavirus.
answer
+ SS RNA icosahedral. Enveloped.
question
Name 2 generas of togavirus.
answer
Alphavirus (not important human pathogen) and Rubivirus.
question
What is unique about rubivirus?
answer
Spread by resp. droplets
question
What does rubivirus cause?
answer
Rubella (German Measles)
question
What are Forchhiemer's Spots associated with?
answer
Rubivirus (rubella).
question
Can rubella cause abortion?
answer
Yes.
question
What is the vaccine for Rubivirus?
answer
MMR - live attenuated.
question
Describe Flaviviridae.
answer
Enveloped + SS RNA.
question
How are arboviruses (Flaviviridae) transmitted?
answer
Mosquitoes - think Dengue/Yellow Fever.
question
What causes West Nile/Dengue/Yellow Fever?
answer
Arboviruses/Flaviviridae.
question
Describe the Bunyaviridae family.
answer
Enveloped segmented - ssRNA.
question
What causes Hantavirus?
answer
Bunyaviridae. Rodent (urine) spread - lethal - pulmonary disease.
question
What causes B. Rift valley fever and California?
answer
Bunyaviridae - though this is mosquito born - disease of livestock.
question
What organ do hepatoviruses target?
answer
Liver - jaundice, dark urine, lymphadenopathy and muscle pain.
question
What family does Hep A belong to?
answer
Picornavirus - genus Heparnavirus. Generally mild.
question
T/F Hep A is stable.
answer
True - Inactivated by detergents, salt water, drying, autoclave, Cl-, etc...
question
How is Hep A spread?
answer
Fecal/oral, water, shellfish, or FOOD PREP WORKERS.
question
T/F Hep A causes chronic disease.
answer
False.
question
T/F Hep A damange is due to the Immune Response.
answer
True.
question
How is Hep A treated?
answer
Antibody (artificial passive) for contacts.
question
What vaccine is available for Hep A?
answer
Killed vaccine here (age 2), live in China.
question
Which Hep Virus is a DNA Virus?
answer
B
question
What causes Hep B?
answer
Hepadnavirus.
question
Described Hepadnavirus.
answer
enveloped partially ds cirucular DNA genome, icosahedral.
question
T/F Hep B can cause cancer.
answer
True - integrates into host genome.
question
What is an infections Hep B particle called?
answer
Dane particle.
question
How is Hep B spread?
answer
needles, sex, perinatally.
question
What is serum hepatitis?
answer
Hep B - 60-90 days incubation. seroconversion for needle stick so universal precautions taken.
question
How is a Hep B needle stick treated?
answer
HB Ig.
question
What percentage of the world population is infected w/ Hep B?
answer
1/3 - that can't be right - double check this. 5% of US - 25% of those have symptoms.
question
What does the T-cell response from Hep B cause? What kind of immunopathology is this?
answer
Liver damage. Your T cells kill your liver cells. Type II response.
question
What kind of immunopathology results from Hep B?
answer
Type II (vasculities and renal dmg) and Type III.
question
How is Hep B treated?
answer
Antivirals aimed at polymerase.
question
How is Hep B diagnosed?
answer
Hep B surface antigen - HBsAg - 1st antigen to appear.
question
T/F If you're seropositive for HBcAg Ab, you could have been vaccinated.
answer
False - core antigen - host makes IgM to this when you clear the virus, so NOT present from vaccine.
question
What comes after HBsAg?
answer
HBeAg - comes after HBsAg and this means the virus is very infective, active.
question
What causes Hep C virus?
answer
Flavivirus - RNA genome. Hepacivurs + sense RNA - enveloped icosahedral.
question
How is Hep C spread?
answer
blood, needles, sex, perinatally.
question
T/F Hep C doesn't cause cancer.
answer
False - Increases liver cancer - high cell turnover.
question
Who is infected by Hep C?
answer
Humans/chimps - 170 million carriers.
question
Flavivirus causes which Hepatatises?
answer
C and G.
question
T/F Hep G is more serious than Hep C and related?
answer
False - related yes, but less virulent. Unlikely a major human pathogen.
question
How is Hep C diagnosed?
answer
ELISA.
question
How is Hep C treated?
answer
IFN-alpha or ribavirin - 50% recovery.
question
Describe Hep E virus.
answer
Resembles Calcivirus.
question
Describe Hep D.
answer
-Sense SS RNA.
question
T/F Hep D acts alone.
answer
False - needs HBV as a helper - uses HBV cell to produce protein - viral parasite. SUPERINFECTION TO HBV. Makes it worse.
question
Describe Hep E.
answer
non-enveloped +sense ss RNA.
question
T/F Hep E disease resembles calcivirus.
answer
False - DISEASE resembles Hep A, and the VIRUS resembles Calcivirus.
question
Describe a prion.
answer
Proeinaceous infectious particle.
question
What does a prion cause?
answer
encephalopathies -
question
How do prions act?
answer
Binds to your proteins - causes re-forlding.
question
T/F prions are not antigenic, inflammatory or immunogenic.
answer
True - to all 3.
question
T/F Prions are resistant to heat and radiation.
answer
True on both accounts.
question
How are prions transmitted?
answer
Injection, transplantation, contact w/ medical equipment, food, genetics for humal familial
question
How are prions treated?
answer
No serotests, prevention is key. Need to treat w/ NaOH for autoclave - 30 mins, 1 hour in hour and autoclave, then 1 hour rinse in water.
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