Proteins: Bacterial Infection Flashcards

1
Q

What is bacteria?

A

Diverse and ubiquitous group of small, unicellular prokaryotes.

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2
Q

What’s the difference between good ‘friendly’ bacteria and bad bacteria?

A

Good is useful for the ecosystem and food production as a source of useful compounds whereas bad is disease causing, causes food spoilage and bio fouling/corrosion.

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3
Q

What are the four Koch’s postulates?

A
  1. The microorganism must be present in every case of the disease (not in healthy animals)
  2. The microorganism must be isolated from the diseases host and grown in pure culture
  3. The specific disease must be isolated when a pure culture of the microorganism is inoculated into a health susceptible host
  4. The microorganism must be recoverable from the experimentally infected host and shown to be identical to the original causative agent.
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4
Q

What are Koch’s postulates?

A

Guidelines to what a bacterium is, although not all are met all of the time.

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5
Q

What’s a) pathogenic, b) opportunistic/ conditionally pathogenic and c) non-pathogenic in terms of bacteria?

A

Pathogenic: disease causing
Opportunistic/conditionally pathogenic: sometimes disease causing
Non-pathogenic: harmless

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6
Q

How do diseases come about?

A

Some bacteria are entirely pathogenic, some bacteria acquire extra virulence factors which make them pathogenic, some bacteria from normal flora can cause disease if they gain access to deep tissue (especially if associated with a foreign body) or in immunocompromised patients many free-living bacteria and components of the normal flora can cause disease.

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7
Q

What is the normal flora?

A

All body surfaces possess a rich normal flora, the nose, mouth, GI, urogenital tract and skin in particular. It provides colonisation resistance and includes numerous conditional pathogens.

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8
Q

What type of infection can normal flora cause?

A

Endogenous infection

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9
Q

What is zoonosis?

A

An infectious disease that can be transmitted from animal to human, which may or may not cause disease.

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10
Q

What percentage of known human pathogens are zoonotic?

A

~60%

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11
Q

What is disease invasion?

A

The movement of a pathogen into a new host

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12
Q

Discuss tuberculosis?

A

Caused by mycobacterium tuberculosis, an infection of the lungs. Causes a cough, chest pain, fever, weight loss, fatigue etc. 1/3 of the worlds population is latently infected, 1/10 of latent infections become active diseases. Untreated it kills 50% infected.

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12
Q

Describe pneumonia?

A

Most commonly caused by streptococcus pneumoniae but also caused by other bacteria. Symptoms include cough, fever and chest pains due to compromised oxygen transport through the alveoli due to bacteria nucleophiles and fluid from damage. Major cause of death in young and old.

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13
Q

What does synergy between viral and bacterial diseases mean?

A

When they both infect together and become lethal. Eg. Flu and pneumonia or HIV and tuberculosis

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14
Q

How much more likely are you to develop active tuberculosis if you have HIV?

A

x800

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15
Q

What is a major cause of food poisoning?

A

Bacteria, 1 million cases per year and ~ 500 deaths.

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16
Q

What does the term exotoxin- mediated disease mean?

A

Caused by bacterial toxins grown outside the body but ingested.

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17
Q

Describe cholera?

A

An exotoxin-mediated disease, caused by vibrio cholerae. Causes rapid dehydration and electrolyte imbalance which causes rice-water diarrhoea and vomiting. Around 5 million cases per year and 100 000 deaths. Treatment is rehydration therapy-fluid and electrolytes.

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18
Q

What is the mechanism by which Choleragen works?

A

The toxin binds to GM1 ganglioside recepto tom enterocytes via b subunits. The a subunit undergoes receptor mediated endocytosis and catalyses ADP-ribosylation of the Gs protein. This stimulate adenylate cyclase and overproduction of cAMP which in turn stimulates the secretion of ions from cells to lumen and a large scale movement of water into the lumen.

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19
Q

What is septic shock?

A

Lipopolysaccharides (contains lipid and sugar portions) are endotoxins which cause septic shock. The symptoms are raised temperature, fever, hypotension, coagulation and leads to multiple organ failure and death in over 50% of cases.

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20
Q

What are some methods of preventing bacterial disease?

A

Disinfection/sterilisation, aseptic technique, decolonisation and prophylactic antibacterials and vaccination.

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21
Q

How are bacterial diseases treated?

A

Antibacterial chemotherapy, anti-toxin, surgery, electrolyte replacement etc.

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22
Q

What are cell walls common to?

A

Plants, bacteria and fungi but not found in protozoa and animals

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23
Q

How do cell walls vary?

A

Different biological kingdoms have difference cells walls which are comprised of polymers of repeating subunits.

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24
Q

What are the repeating units that make cells walls in plants?

A

Cellulose (a glucose polymer)

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25
Q

What are the repeating units that make cells walls in bacteria?

A

Peptidoglycan (a polymer of two sugars cross-linked by peptides)

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26
Q

What are cell walls required for?

A

Structural protection from osmotic pressure, enables cells to survive hypotonic environments, a scaffold for anchoring extra cellular proteins, roles in virulence, immune system recognition, major antibiotic targets and fundamental biological interest.

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27
Q

What is the structure of peptidoglycan?

A

Made of N-acetyl muramic acid and N-acetyl glucosamine linked by peptide bridges.

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28
Q

What are other terms for a) N-acetyl muramic acid and B) N-acetyl glucosamine?

A

A) NAM or MurNAc

B) NAG or GlcNAc

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29
Q

How does he peptidoglycan vary?

A

Many variations in the stem peptide (originally 5 amino acids long but cleavage between 4 and 5 is required for energy) region. Position 4 is always D-alanine.
In E.coli meso-diaminopimelate (an amino acids related to Lysine) is at position 3.
In S.aureus L-Lysine is present at position 3.

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30
Q

What are the sources of peptidoglycan precursors?

A

D-alanyl-D-alanine: alanine racemase, D-ala-D-ala ligase

D-glutamate: glutamate racemase

31
Q

How is peptidoglycan synthesised?

A

The UDP-sugar pentapeptide in the cytoplasm is synthesised, MurA and MurB join then C, D, E and F do sequentially. A lipid carrier, a carbohydrate and additional cross linking amino acids are added and MraY is changed to MurG. Polymerisation and cross linking of pentapeptide and carbohydrate occurs to yield the final polymer.

32
Q

How are the peptidoglycan polymers joint?

A

Transglycosylation involves the polymerisation of the chains and transpeptidation involves crosslinking between glycan strands.

33
Q

What is transglycoslation and transpeptidation catalysed by?

A

By penicillin binding proteins which are bifunctional and membrane associated enzymes.

34
Q

What is the difference between gram-negative and gram-positive bacteria?

A

Gram-negative have an additional outer membrane, as a dedicated diffusion barrier to avoid toxic substances, therefore the peptide is between the membranes in a single, 8nm layer. In gram-positive the peptide is on the outside as a thick mesh of many layers, ~80 nm, which acts as a tethering point.

35
Q

What other constituents of a cell wall to gram-positive bacteria have?

A

Additional teichoic and lipoteichoic acid which are polymers of glycerol phosphate.

36
Q

What other constituents of a cell wall to gram-negative bacteria have?

A

Lipoproteins (Brauns), 7.2 kDa in size which have a hydrophobic head gorup buried in the outer membrane and the other end forms a peptide bond with D-amino pamelic acid to tether the outer membrane to the cell wall.

37
Q

Describe some unusual cell walls?

A

Archae lack peptidoglycan and have an S layer instead. This is a thick later (25 nm) of proteins or glycoprotein which acts in the same way.
And some have different forms of peptidoglycan (eg. Pseudo). Chlamydiae have a P layer which is composed of large cysteine rich proteins.

38
Q

What is the chlamydiae paradox?

A

Chlamydiae has most of the cellular machinery to make peptidoglycan but does not. New evidence that they do make some which is for cell division not cell walls.

39
Q

How does the mammalian immune system recognise peptidoglycan?

A

As it has an antigenic structure that is uniquely bacteria. Recognised extracellularly by membrane-bound CD14 and TLR2 which triggers signalling cascade via cytokine release. Intracellular recognition is mediated by two members of NOD-like receptor proteins. NOD-1 recognise gram-negative bacteria and NOD-2 recognises muramyl di peptide. Also soluble recognition molecules which activate complement cascade and lysosome that degrades peptidoglycan.

40
Q

How do lysosome act?

A

Cleaves the cell wall of bacteria and fungi.

41
Q

What is lysostaphin?

A

A metalloendopeptidase capable of destroying peptidoglycan which is produced by staphylococcus stimulants.

42
Q

How do you treat bacteria infections?

A

By targeting cell wall synthesis using B-lactams, vancomycin, bacitracin, fosfomycin, D-cycloserine.

43
Q

How do B-lactams work?

A

B-lactam ring is homologous to the D-alanyl-D-alanine so competes for its position in the cell wall. If incorporated it is irreversibly weakened and leads to rupturing.

44
Q

How does vancomycin work?

A

It binds to the substrate of the penicillin binding protein and clas them to prevent them being used.

45
Q

Define an antibiotic?

A

A natural substance produced by one micro-organism that kills or inhibits the growth of another.

46
Q

Define an antibacterial agent/antibacterials?

A

Compounds capable of killing or inhibiting the growth of bacteria, including semi-synthetic or synthetic compounds.

47
Q

Define disinfectants/antiseptics?

A

Compounds that kill microorganisms but are two toxic for internal use in human patients.

48
Q

Define antibacterial drugs?

A

Compounds that shoe selective toxicity against bacterial cells versus mammalian cells and can be used by patients.

49
Q

Define antibacterial chemotherapy?

A

The use of antibacterial drugs to treat and cure bacterial infection of animals and humans.

50
Q

What is selective toxicity?

A

A central concept in antibacterial drug action, when the growth of an infecting organism is selectively inhibited or killed without damage to the host cells. Based on exploiting differences in the structure or biochemistry of infecting agent and host, eg. the agent cannot enter mammalian cells as easily as it would bacterial, the agent target processes/structures only present in bacteria, or agents are pro drugs only activated in bacteria. Ideally would have no adverse effect but would be lethal to the organism.

51
Q

What is the difference between bactriostatic and bacteriocidal?

A

Bacteriostatic halts the growth whereas bacteriocidal kills.

52
Q

How are antibiotics biosynthesised?

A

They are secondary metabolites produced by bacteria or fungi. Some organisms make over 25 types. Synthetic antibiotics have been synthesised with the same action but they are less complex compared to their natural analogues.

53
Q

Why is antibacterial chemotherapy important?

A

Underpins modern economics, huge benefits for individuals and societies health (survival, longevity, quality of life and productivity), modern medicine relies on it.

54
Q

What are the targets for antibacterials?

A

Cell wall biosynthesis, the membrane, nucleotide metabolism, DNA, transcription, protein synthesis.

55
Q

How do antibacterials target cell wall biosynthesis?

A

Both B-lactams and glycopeptides (including vancomycin and teicoplanin), interfere with the final stages of cell wall biosynthesis. And isoniazid and ethionamide are pro drugs that block mycobacterial my colic acid synthesis.

56
Q

What antibacterials target the membrane?

A

Daptomycin and polymyxin B and E (colistin).

57
Q

What antibacterials target nucleotide metabolism?

A

Sulfamethoxazole and trimethoprim (which can be used together as co-trimoxazole). Sulfa drugs are competitive inhibitors and alternative substrates for DHPS.

58
Q

What antibacterials target DNA?

A

Synthetic antibacterials; quinolones and fluoroquinolone which target DNA gyrase and Topoisomerase 4.

59
Q

What antibacterial targets transcription and what is its mode of action?

A

Rifampicin, binds to the beta subunits of prokaryotic RNA polymerase and interferes with transcription initiation.

60
Q

What antibacterials effect protein synthesis?

A

Large number, some effect the small ribosomal subunit and some the large. Most act directly and are bacteriostatic. Eg. Linezolid

61
Q

Define antibacterial drug resistance?

A

The ability for bacteria to survive and grown in the presence of an antibiotic at a connect ration that can be safely achieved in patients at site of infection.

62
Q

What does antibacterial drug resistance imply?

A

That an antibiotic will not assist in resolution of the bacterial infection, i.e. That it is no longer effective for treatment.

63
Q

What’s the difference between intrinsic and acquired resistance?

A

Intrinsic results from inherent features of a particular bacterial population, eg. E.coli is intrinsically resistant to Rifampicin and mupirocin because they can’t get through the outer membrane.
Acquired is when a previously-antibiotic susceptible bacterial population becomes resistant.

64
Q

What are the types of acquired resistance?

A

Spontaneous mutations, when various mutations can give rise to reduced antibiotic susceptibility and horizontal acquisition, where horizontal gene transfer from one to another occurs.

65
Q

What are the negative consequences of antibacterial drug resistance?

A

Increased mortality, morbidity and cost.

66
Q

What drives antibacterial drug resistance?

A

The use of antibiotics due to evolution, selective pressure and survival of the fittest.

67
Q

What are the mechanisms of antibacterial drug resistance?

A

Altered target site, decreased uptake, inactivation/modification and bypass.

68
Q

How does alteration of the target site occur and cause antibacterial drug resistance?

A

The target can be mutated, for example amino acid substitutions reduce the binding and cause a reduction in Rifampicin activity. The target can be modified, for examples methylations of ribosomal RNA or vancomycin resistance in enterococci. Or the target can be over expressed and therefore require more drug for action, eg. Vancomycin

69
Q

How can decreases uptake cause antibacterial drug resistance?

A

Reduced permeability to the antibiotic (uncommon), efflux (more common) where toxic compounds are pumped from the cell by efflux pumps which require a proton motive force of ATP, resistances arises by up-regulation of acquisition of a new pumps, removed the drug before it can act.

70
Q

How can inactivation cause antibacterial resistance?

A

Destruction of the antibiotic, for example B-lactam rings can be targeted and hydrolysed leaving the inactive.

71
Q

How can modification cause antibacterial drug resistance?

A

For example amino glycosides can be adenylated by adenylate transferase, acetylated by acetyltransferase or phosphorylated by phosphotransferase.

72
Q

How does bypass cause antibacterial drug resistance?

A

By acquisition of a new target, foe example methicillin resistance in S.aureus.

73
Q

Can multiple methods of antibacterial drug resistance work together?

A

Yes

74
Q

How can the problem of antibacterial drug resistance be addressed?

A

Better training for prescribers, coordination of surveillance of resistance, better guidelines for therapy, possible cycling of antibiotics to reduce the selective pressure and restriction of use as growth promoters, improved infection control process and the development of new, novel antibacterials.