Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

Study Questions for Pathogenic Microbiology (Bio321)

Epidemiology Define Etiology and Epidemiology. What are some of the key factors in the occurrence and spread of disease? What are some strategies used to trace the cause of a disease outbreak? Why are cities referred to as disease magnets? What are some additional modern "conveniences" that have led to the spread of disease? Body defense mechanisms What are the differences between constitutive and induced defense mechanisms? What is the main goal of surface defense mechanisms? What are some of the defense mechanisms of the skin and mucosal surfaces? How and why do these defense mechanisms differ? How do the MALT and SALT systems work? Compare some of the defense systems of the various specific mucosal surfaces. Mouth and respiratory tract, eyes, urinary tract, intestinal tract etc. Defenses of tissue and blood What are the functions of transferrin, ferritin What are the major types of phagocytic cells and what are some of their functions. What are some of the strategies phagocytic cells use to kill bacteria? What does it mean to say phagocytic cells are sloppy feeders? What are the advantages and disadvantages of sloppy feeding? What are the functions of the complement system What are the differences and similarities between the alternative and classical pathways for complement activation What is the role of serum proteins H and I? What are the roles of C5a and C5b Why do some bacteria incorporate sialic acid or hyaluronic acid into their capsules?

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Study Questions for Bio321

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What are the major parts of antibody molecules? What are the three major groups of antibodies and how do they differ? How do the differences between the major types of antibodies affect their function? How do host defenses respond differently to extracellular versus intracellular bacteria? Experimental systems Why is selecting an appropriate bacterial strain and species important for setting up a model experimental system? What are some of the factors to consider when selecting a model pathogen? What are some of the factors to consider when selecting an animal model? What are the advantages and disadvantages of Animal models in general Infant or immunocompromised animals Transgenic animals Germfree (gnotobiotic) mouse systems Cultured cell lines Organ culture What are some of the common transgenic model animals available today? What are some of the common measurement of virulence? Do these measurements accurately reflect pathogenicity? Why or why not? What factors would influence the accuracy of virulence measurements? What are some of the strategies used to identify virulence factors? What are the advantages and disadvantages of using a biochemical approach? What are the advantages and disadvantages of using a genetic approach How would you screen/select for virulence genes using an avirulent strain of E. coli? How can transposons be used to identify virulence genes? How are reporter genes used to identify virulence factors? Under what circumstances would you use an in vitro constructed gene fusion or use a transposon to construct a gene fusion? How can reporter gene fusions be used to determine if a gene is controlled by a repressor or an activator protein? How are PhoA fusions used to identify virulence factors? Why are PhoA fusions such a powerful tool for identifying virulence factors?

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

Nucleic acid detection systems Describe the various nucleic acid based techniques that can be used to detect pathogens. How is a southern blot performed? What are some of the possible sources of DNA probes What types of labels are available for detecting DNA-DNA hybridization? How can the washing step be used to modify the specificity of DNA-DNA hybridization? What are the advantages and disadvantages of PCR and LCR as amplification techniques? What types of adherence mechanisms can pathogens use for colonization? What are pili and what is their function? What is the structure of a pilus? How is a pilus assembled? What are the roles of the signal peptide, chaperones, termination peptide What are afimbrial adhesins? How do pathogens avoid phagocytosis Once phagocytized how can bacteria escape the phagosome? What are invasins? How are capsules used to evade complement? How do host defenses compete against capsules? Intracellular pathogens - Salmonella, Shigella and Listeria Compare and contrast the diseases caused by all three organisms i.e. - transmission, ID50, incubation period, reservoirs, invasion mechanism etc. How are Salmonella and Shigella distinguished from other bacteria and from each other What are the three major antigen types for Salmonella spp. In a swine model, S. typhimurium is found within absorptive mucosal cells - not M cells or Peyers patches but S.choleraesuis is found in M - cells but not mucosal absorptive cells. What is the significance of this observation? In stained cells and colony morphology L. monocytogenes looks like what other type of organism. How is L. monocytogenes distinguished from this organism?

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

What is the role of actin rearrangement in bacterial invasion? How were the inv genes cloned from Salmonella? What is the significance of the relatively low % G + C of the inv genes relative to other genes? How does ruffling, induced by Salmonella, compare to ruffling that occurs in response to other factors? What is the evidence that a specific receptor is involved in triggering Salmonella invasion? What is the eventual end result of the Salmonella induced phosphorylation cascade that apparently results in membrane ruffling? What happens if you add calcium chelators during Salmonella invasion? What are the 2 possible outcomes of phospholipase A2 activity that could lead to Ca++ influx? How was the hil locus identified? Why do transposon mutants in hil behave differently from deletion mutants of hil? What is Rck and how does it protect cells from complement? What effect does Cytochalisin B and Cytochalisin D (which block actin polymerization) have on Shigella invasion? Would you predict the same effect on Salmonella invasion? What are the roles of IpaB and IpaC in invasion by Shigella? What do the mxi gene products do? What is the significance of the observation that the ipa gene products seem to bind to integrins What is the evidence that flagellar motility is not important for L. monocytogenes invasion. How was it shown that InlA alone (which codes for an 80 kDa membrane protein) is required for entry into host cells? What are the roles of Listeriolysin O, Phospholipase C and Lecithinase in entry into host cells? How was it shown that the hly gene product is involved in escape from phagosomes? Only bacteria from inside macrophages can demonstrate actin based motility in vitro. Why? What host factor(s) are required for actin based motility? List some of the strategies bacteria can use for survival in vesicles. What is the significance of the observation that invading bacteria are often not surrounded by "true" phagolysosome? What is some of the evidence that invading bacteria do not enter through a "true" phagolysosome? How might bacteria that invade macrophages trigger apoptosis? Observations relating to bacterial invasion triggering apoptosis.

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

Bacteria unable to enter epithelial cells do not induce apoptosis in macrophages Degree of apoptosis correlates with level of activation of macrophages Only entry of bacteria induces apoptosis - entry without replication still induces apop. Any gene that interrupts Salmonella invasion interrupts apoptosis What mechanism can bacteria use as a defense against bactericidal cationic peptides? What is the evidence that filament formation by Salmonella inside a host cell is correlated with survival and multiplication What is the significance of the observation that there are 2 populations of Salmonella in macrophages? How might intracellular pathogens influence the lysosomal events inside the host cell? What type of mechanisms might intracellular bacteria use to obtain nutrients from a host cell even though they are encased inside a membranous vesicle? Toxins What are toxins? What are the basic features or exotoxins? In what ways are toxins like enzymes? What makes a toxin specific? What are some of the general names for toxins related to their specificity? Name some examples of toxins with narrow specificity and some with broad specificity. Why are toxins highly antigenic? What is an antitoxin? Why are antitoxins often more effective in vivo compared to in vitro? What are toxoids and how can they be made? What are the 3 major types of exotoxins and describe each type? What are the differences between simple and compound A-B toxins? What are the two possible pathways a toxin can take into a cell? Which part of the A-B toxin typically enters the cell and what are the possible mechanisms it can use to kill the target cell? How do membrane-disrupting toxins work? What are some of the other general names used for membrane-disrupting toxins?

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

What are some of the possible mechanisms for insertion of pore forming proteins into the membrane? What are amphipathic helices? What device can be used to depict amphilicity? What are some of the basic properties of thiol-activated cytolysins? What are superantigens and how do they work? What are some of the compounds released in response to superantigens? Name some of the diseases associated with superantigens. What is an endotoxin How do endotoxins differ from exotoxins? What are some of the symptoms of endotoxin diseases? What are some of the accessory indicators to look for in order to suspect endotoxin desease? What are the steps leading to septic shock? What are the likely receptors involved in endotoxin binding to target cells? Which cytokines are released in response to endotoxin What are the best forms of treatment for endotoxin disease? What are the advantages/disadvantages of each? Which part of LPS is the actual toxin portion? Vibrio cholera What are the two main strains of Vibrio cholera that are responsible for the majority of the serious epidemics worldwide? What is the general effect of Cholera toxin and what is the prescribed treatment? What is the role of the TCP pili in Cholera infection? How were the accessory colonization factor (acf) genes indeitifies? What is the role of detachment in the spread of Cholera? Describe Cholera toxin (i.e. type of toxin, genes, subunit composition, structure, secretion, activity) What is the host cell receptor for Cholera toxin and what is its basic composition? What is the role of neuraminidase (sialidase) in toxin activity? What is the enzymatic activity of Cholera toxin and what is it's target? How does this enzyme activity lead to the symptoms of the disease?
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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

Where are the key targets of Cholera toxin localized in the host cell? What impact does that have on the course of events leading to Cholera? What does it mean when we say that the Cholera toxin genes are located on a virulence cassette? What are the implications associated with the ctx genes being on a virulence cassette What are the regulatory signals for Cholera toxin expression? How was the toxR - gene cloned? What other genes are involved in ctx expression? How do ToxR and ToxT differ? Corynebacterium diphtheria What is Diphtheria? Describe the causative agent of diphtheria- Corynebacterium diphtheria What are the characteristic features of diphtheria and how is it identified? What are the 3 colony types of C. diphtheria? How can you confirm that a C. diphtheria strain is toxigenic? Describe the basic properties of Diphtheria toxin What are the three functional regions of Diphtheria toxin? How can genes encoding toxin receptor proteins be identified? How is Diphtheria toxin translocated into the cell? What is the target of Diphtheria toxin and how does it kill the host cell? What is diphthamide? How is the dtx gene regulated? Toxinoses What are toxinoses? Name some organisms that cause toxinoses. What is the causative agent of botulism? What are the symptoms of botulism? How do infant botulism and wound botulism differ from food borne botulism? What are the 3 main C. botulinum toxins and how do they differ?

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

What are the target cells of botulinum toxin? How does botulinal toxin inhibit host cells? What is the structure of botulinal toxin and how does it differ from most otherfood borne pathogens? Compare and contrast botulinal toxin and tetanus toxin. Yersinia What are the 3 species of Yersinia that cause disease in humans? What are the differences in the disease caused by each of these three Yersinia sp. How are Y. pseudotuberculosis and Y. enterocolitica infections acquired? What are the target cells for Y. pseudotuberculosis and Y. enterocolitica. What is the animal reservoir for Y. pestis? What are the two modes of transmission of Y. pestis and how do the resulting diseases differ. What strategies do the bacteria use to aid intransmission? What are Buboes? Which of the three strains are most closely related? Describe the three plasmids found in Y. pestis - including genes located on each? Which plasmid is shared by all three pathogenic Yersinia sp.? What are the functions of Inv and Ail in Yersinia infections? How were the Inv and Ail genes cloned and how are they regulated? Name the adhesins found in Yersinia sp. What are Yops and what are their functions? What gene products are required for Yop secretion? What are the conditions required for Yop expression and secretion? What are the roles of YmoA and VirF in Yop expression a secretion? What are the roles of YopN, LcrH and LcrQ in the calcium response? What is the response of Yersinia Yop expression and secretion on contact with a host cell. Pseudomonas aeruginosa Describe Pseudomonas aeruginosa

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

Why is P. aeruginosa a true opportunistic pathogen? What are some of the defenses of the respiratory tract? What is the impact of cystic fibrosis on these defenses? What are some of the defenses of the skin? What impact do burns have on these defenses? What impact do burns have on the LD50 for P. aeruginosa What are the main virulence factors for P. aeruginosa What is the host receptor for P. aeruginosa pilin? Nonpilus adhesins of P. aeruginosa are involved in binding what types of cells or molecules? How is Pilin expression regulated? What is the role of P. aeruginosa exoenzyme S in pathogenesis How does P. aeruginosa exotoxin A compare to diphtheria toxin? What is the role of P. aeruginosa elastases in pathogenesis? Describe LasA and LasB and how they act on elastin and tissue defenses? How are lasA and lasB regulated? What is the composition of alginate? Why is alginate production highly regulated? Why is the ability to produce alginate easily lost? What is AlgD and what does it do? What are the regulatory signals for alginate production? What other factors are similarly regulated? What are the three levels at which alginate production is regulated? What proteins are involved in phase variation (genetic switch) What are the two forms of LPS in P. aeruginosa? What proteins are involved in transcriptional regulation of alginate production? What is the role of AlgB in alginate production? What are the pigments produced by P. aeruginosa and what is their role in pathogenesis? Why is P. aeruginosa resistant to so many antibiotics? What are some of the mechanisms of P. aeruginosa antibiotic sensitivity and resistance? Quorum sensing

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

What is quorum sensing and what role might it have in virulence? Why was quorum sensing first called autoinduction? What is some of the evidence for a quorum sensing system in P. aeruginosa? Describe the quorum sensing system in P. aeruginosa. What is the effect of a lasR knock-out mutation? What are the two main structural domains of LasR as well as other LuxR-like proteins. Why does adding lasR in trans to a lasR mutant not restore expression to 100% of wild-type How could you test for production of a diffusible molecule? What is the basic structure of most autoinducer molecules? Can the P. aeruginosa and V. fischeri quorum sensing systems be interchanged? General mechanisms for regulation List the possible mechanisms for regulation of genes and gene products. Give one specific example of each. Bordetella pertussis How do transcriptional activators differ from transcriptional repressors? Where does Bordetella pertussis typically colonize? What is the typical course of infection by B. pertussis? What factors are involved in B. pertussis attachment ? Describe Pertussin toxin. Compare Pertussin toxin to Cholera toxin a B. pertussis Invasive Adenylate Cyclase Describe the BvgA and BvgS two-component regulatory system. Include the key structural features of each protein and the phosphate relay system. Iron acquisition Why is iron acquisition and regulation important for virulence? Why do pathogens often have multiple iron acquisition mechanism? What is Fur? How do bacteria acquire iron for nutrition but not so much that it becomes toxix?

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

Describe the possible mechanisms for iron acquisition. How do bacteria utilize siderophores. What is the role of TonB and ExbB/D in siderophore uptake? Heme utilization is best studied in what bacterial system. What are transferrin, lactoferrin and ferritin and how do their uptake systems differ? Antigenic variation What is Antigenic variation as defined for N. gonorrhoeae? What effect does antigenic variation have on immunity and the course of infection? What is the basic structure of the N. gonorrhoeae pilin protein How do pilE and pilS differ? How do so few genes result in such a high variation in pilin structure? How does antigenic variation relate to phase variation in N. gonorrhoeae? What are the two probable mechanisms for exchange of pilE and pilS genes? What is some of the evidence in favor of each mechanism? Antibiotics - mechanisms What are some of the factors to consider when selecting an antibiotic? What are the differences between bactericidal and bacteriostatic antibiotics? Be prepared to recognize representatives of each class of antibiotics. Describe the target and mechanism for each class of antibiotics. Antibiotics - resistance List the possible mechanisms of antibiotic resistance? Give a specific example of each mechanism of antibiotic resistance. If you were given unlimited funding to find a new antibiotic to cure any bacterial disease you wanted to cure what disease would you select and how might you go about finding or creating that antibiotic? Vaccines What is the difference between active and passive immunity? What is the difference between natural or artificial immunity?

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Study Questions for Bio321

http://web.mst.edu/~microbio/BIO321_1998/Bio321studyQ.html

What are some of the desired properties of vaccines? Why are polysaccharides poor vaccines? How can you improve the effectiveness of a polysaccharide vaccine? Explain why it is important to design a vaccine that stimulates the correct immune response. Give an example of how this might be accomplished. What are some of the safety concerns to consider when designing vaccines? What are the advantages or disadvantages of using whole cell, live or acelluar vaccines? Successful whole cell vaccines for cholera include the B subunit of cholera toxin. Why? Salmonella typhimurium is a popular choice for constructing live vaccines against a number of intestinal pathogens. Why? There appears to be little hope for designing a successful vaccine against Neisseria gonorrhoeae. Why? Current, vaccines against Streptococcus pneumoniae are not very effective for infants and elderly patients, which are also at the highest risk for S. pneumoniae infections. Why are the current vaccines ineffective and what can be done to improve them? E. coli diseases How can E. coli cause such a wide assortment of infections, anything from diarrhea to dysentery, hemolytic uremic syndrome, bladder and kidney infections, septicemia, pneumonia, meningitis? What are the methods currently used to distinguish different strains of E. coli? What are the 5 major virotypes of E. coli that cause gastrointestinal infections? Enterotoxigenic E. coli causes a disease that is similar to what other major gastrointestinal pathogen? How does enterotoxigenic E. coli infection differ from the more severe disease? How do the LT and ST toxins of ETEC differ from each other? How does enteropathogenic E. coli (EPEC) gastrointestinal disease compare to ETEC? What are the 3 stages of EPEC invasion? How is enterohemorragic E. coli infection similar to Shigella infection? What is the difference between enteroinvasive E. coli dysentery and Shigella dysentery?

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Study Questions for Bio321

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This Document is maintained by djwesten@ umr.edu Last Modified: Thursday, 10-Dec-1998 23:00:00 CST

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