Finals: Midterm 1 Content Flashcards

1
Q

pathogen

A
  • a microbe that is capable of causing host damage
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2
Q

pathogenicity

A
  • the ability to produce disease
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3
Q

virulence (2)

A
  • degree of pathogenicity
  • only expressed in a susceptible host
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4
Q

gram negative bacteria (2)

A
  • thin peptidoglycan cell wall, surrounded by outer membrane containing lipopolysaccharide
  • double membrane
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5
Q

gram positive bacteria (2)

A
  • lack an outer membrane
  • surrounded by thick peptidoglycan cell wall
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6
Q

Koch’s Postulate: First Postulate

A
  • the microorganism must be found in abundance in all organisms suffering from the disease, but should not be found in healthy organisms
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7
Q

Koch’s Postulates: Second Postulate

A
  • the microorganism must be isolated from a diseased organism and grown in pure culture
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8
Q

Koch’s Postulate: Third Postulate

A
  • the cultured microorganism should cause disease when introduced into a healthy host
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9
Q

Koch’s Postulates: Fourth Postulate

A
  • microorganism must be re-isolated from the inoculated diseased experimental host and identified as being identical to the specific causative agent
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10
Q

Challenges to Koch’s Postulates: First (3)

A
  • host susceptibility varies in human and animal populations due to a variety of factors
  • opportunistic bacteria can be found in healthy hosts too
  • may be difficult to isolate bacteria from certain areas, such as brain and bones
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11
Q

Challenges to Koch’s Postulates: Second Postulate (2)

A
  • not all bacteria can be cultured in pure form
  • many bacteria undergo genetic and phenotypic changes in culture environments
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12
Q

Challenges to Koch’s Postulates: Third Postulate (2)

A
  • not all bacterial species are equally virulent
  • closely related bacterial strains can be very different
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13
Q

Challenges to Koch’s Postulates: Fourth Postulate

A
  • re-inoculation of a host may not be possible or ethical
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14
Q

Modern Approaches to Satisfy Koch’s Postulates (4)

A
  • PCR
  • immunohistochemistry and immunofluorescence
  • ELISA
  • eliminate the pathogen and prevent/cure the disease
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15
Q

Modern Approaches to Satisfy Koch’s Postulates: PCR

A
  • PCR and 16S sequencing can be used to amplify and detect bacteria during culturing
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16
Q

Modern Approaches to Satisfy Koch’s Postulates: PCR/16S Sequencing steps (2)

A
  • 16S RNA primers are designed to detect conserved regions, with unique regions in-between that act as barcodes to identify the bacteria
  • PCR is done to amplify the bacterial nucleic acid for detection
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17
Q

Modern Approaches to Satisfy Koch’s Postulates: Immunohistochemistry (2)

A
  • microscopic images of immunohistochemical staining can be used to visualize the pathogen in the tissue directly
  • uses of antibodies against the bacteria with attached reporter enzymes
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18
Q

Modern Approaches to Satisfy Koch’s Postulates: Immunofluorescence (2)

A
  • micrographs of cells with fluorescently labelled antibodies against cell markers and bacterial
  • cells infected with bacteria will exhibit colocalization of fluorescent markers, while healthy cells will not
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19
Q

Modern Approaches to Satisfy Koch’s Postulates: ELISA (2)

A
  • use of fluorescently labelled antibodies against bacteria to detect pathogens in infection site without culturing
  • Western Blot/SDS-PAGE can be used subsequently
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20
Q

ELISA methods (3)

A
  • direct ELISA
  • indirect ELISA
  • capture/sandwich ELISA
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21
Q

Modern Approaches to Satisfy Koch’s Postulates: Eliminate the Pathogen and Prevent/Cure the Disease; Why didn’t Koch try this?

A
  • antibiotics did not exist at the time, so treatment was a lot more complicated and time-consuming
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22
Q

virulence factors

A
  • bacterial product or strategy that contributes to the ability of the bacterium to survive in the host/cause infection
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23
Q

Molecular Version of Koch’s Postulates: First Postulate

A
  • gene for virulence should be present in the strain of bacteria that cause disease and absent in avirulent strains
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24
Q

Molecular Version of Koch’s Postulates: Second Postulate

A
  • (i) knocking out or disruption the gene should reduce virulence, and (ii) introduction of the cloned gene into an avirulent strain should render the avirulent strain virulent
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25
Q

Molecular Version of Koch’s Postulates: Third Postulate

A
  • expression of the gene should be demonstrated in human or a relevant model
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26
Q

Molecular Version of Koch’s Postulates: Fourth Postulate

A
  • antibodies or a cell-mediated immune response to a virulence factor should be protective
27
Q

what must a pathogenic microbe do to survive in a host and establish infection (5)

A
  • attach to host cells for colonization
  • evade host’s innate and adaptive immune defenses and persist
  • obtain iron and other nutrients needed to multiply
  • disseminate or spread within a host and to other hosts
  • produce symptoms of disease (to be considered pathogenic)
28
Q

what is adherence mediated by (2)

A
  • bacterial appendages or surface structures
  • interactions can occur directly or indirectly
29
Q

adherence: direct interactions

A
  • “adhesin” on bacterium binds directly to a specific receptor
30
Q

adherence: indirect interactions (2)

A
  • adhesin binding first to the protein in the extracellular matrix (ECM)
  • then, the ECM proteins binds to specific receptors on the host cell, acting as a bridge
31
Q

types of adhesins (2)

A
  • fimbrial adhesins
  • afimbrial adhesins
32
Q

fimbrial adhesins (3)

A
  • fimbriae or pili
  • several types that form polymers
  • Pap pilus is heavily characterized and studied
33
Q

afimbrial adhesins (2)

A
  • several types
  • a single protein
34
Q

why do bacteria use long fragile structures to bind to host cells (3)

A
  • outreach
  • protection and survivability of core bacterial body from host immune system
  • bacterial surface and host cell are negative; allows for connection despite repulsive forces
35
Q

can bacterium have multiple adhesins

A
  • yes, they can be expressed all at once or sequentially during infection at different stages of pathogenesis
36
Q

are all adhesins virulence factors (2)

A
  • as bacteria can have multiple adhesins, it is difficult to prove necessity of any one adhesin
  • not all adhesins are virulence factors
37
Q

host receptors for adhesins (4)

A
  • often carbohydrate-based
  • glycolipids or glycoproteins
  • often integrins
  • receptors determine the tropism of the disease
38
Q

tropism

A
  • tissue specificity
39
Q

pathogen invasion (3)

A
  • pathogens can enter and survive in host cells
  • allows them to breach host epithelial barrier
  • provides them with protected niche for replication and persistence
40
Q

pathogen mechanism of invasion (2)

A
  • can actively direct their entry into host cells
  • usually done by a protein called “invasin”
41
Q

invasins (2)

A
  • activate a receptor
  • leads to a signaling event that enables uptake through cytoskeletal rearrangements
42
Q

mechanisms of particle uptake into cells (2)

A
  • zippers
  • triggers
43
Q

mechanisms of particle update into cells: zippers (3)

A
  • sequential engagement of phagocytic membrane with particle surface
  • pseudopod advances no further than receptor-ligand interaction permits
  • partial engulfment where receptor-ligand interactions are
44
Q

mechanisms of particle update into cells: triggers (2)

A
  • all-or-nothing effect
  • complete phagocytosis
45
Q

zipper uptake steps (4)

A
  1. initial contact (adherence)
  2. receptor clustering
  3. phagocytic cup formation involving actin polymerization and membrane extension
  4. closure of phagocytic cup and retraction involving actin depolymerization
46
Q

what does trigger uptake involve (3)

A
  • mechanism similar to micropinocytosis
  • macropinosomes
  • surface ruffling
47
Q

trigger uptake: mechanism similar to micropinocytosis

A
  • a form of endocytosis that is accompanies by cell surface ruffling
48
Q

trigger uptake: macropinosomes

A
  • large vesicles that serve as a route for cells to take up macromolecules non-selectively
49
Q

trigger uptake: membrane ruffling

A
  • ruffles fall back onto bacteria randomly and this leads to formation of large endosomes, which eventually become smaller
50
Q

trigger: distinct steps (3)

A
  1. filopode formation: membrane ruffles
  2. lamellipode formation: membrane ruffles filled with actin support through polymerization
  3. formation of phagocytic cup made of actin or through adherence
51
Q

zipper vs trigger (2)

A
  • zipper results in localized contact-dependent receptor-medicated particle engulfment
  • trigger results in generalized engulfment event that could include neighbouring particles
52
Q

anti-phagocytosis (2)

A
  • some bacteria actively inhibit their uptake
  • secrete protein YopH through T3SS
53
Q

can one pathogen exhibit both zipper and trigger mediated invasion

A
  • yes, for instance, salmonella can invade host cell using different routes and mechanisms
54
Q

phagocytosis: what is involved in uptake (2)

A
  • microfilament rearrangements
  • lead to formation of a phagosome membrane
55
Q

what does phagocytosis activate

A
  • activation of respiratory/oxidative burst
56
Q

normal endocytic events (4)

A
  • early endosome
  • late endosome
  • fusion with lysosome to create the phagolysosome
  • destruction of the bacteria
57
Q

Rabs

A
  • small GTPases
58
Q

Rab5

A
  • early endosome marker
59
Q

Rab7

A
  • late endosome marker
60
Q

LAMP1 (2)

A
  • lysosomal associated membrane protein
  • phagolysosome fusion marker
61
Q

what tools can we use to see where the phagosome is located in the pathway (3)

A
  • western blot
  • IH
  • IF
62
Q

bacterial phagosome survival mechanisms (4)

A
  • failure to trigger oxidative burst by inhibiting phagosome acidification
  • inhibit/stall the fusion of the phagosome with the lysosome
  • survival within the phagolysosome
  • escape from the phagosome
63
Q

autophagy (2)

A
  • induced when cell undergoes starvation
  • mechanism for recycling proteins