vsv5

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Dr. Alvin Fox

Antibiotics affecting the cell envelope

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Key WordsKey WordsSterilization/disinfection/antisepsis Vancomycin

Antibiotic beta lactam

Selective toxicity Penicillins

Bactericidal Cephalosporins/cephamycins

Bacteriostatic Monobactam

Minimal inhibitory concentration (MIC) Clavulinic acid

Susceptibility testing Polymyxin B

Penicillin binding proteins Isoniazid

Penicillinase/beta lactamase Ethioniamide

Autolysins Ethambutol

Cycloserine Resistance

Bacitracin

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STERILIZATIONSTERILIZATION

• All killedAll killed• non-selective non-selective

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SterilizationSterilization• autoclavingautoclaving

– 121121ooC (heat/pressure)C (heat/pressure)* Heat resistant materialsHeat resistant materials

• ethylene oxideethylene oxide– non heat resistantnon heat resistant– Usually equipmentUsually equipment– being replaced e.g. Hbeing replaced e.g. H22OO22/free radicals/free radicals

• ultra-violet lightultra-violet light– surfaces (e.g operating rooms)surfaces (e.g operating rooms)not totally effectivenot totally effective

• gamma radiationgamma radiation– foodfood– some mailsome mail

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Membrane filters

porespores

bacteriabacteria

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DisinfectionDisinfection

• Liquids that kill bacteriaLiquids that kill bacteria– e.g. phenol basede.g. phenol based– too toxic for skin surfacestoo toxic for skin surfaces

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AntisepticsAntiseptics

• Topical (e.g. skin) Topical (e.g. skin) – e.g. iodine or 70% alcohole.g. iodine or 70% alcohol– “ “reduce” bacterial loadreduce” bacterial load

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ANTIBIOTICSANTIBIOTICS

• Selectively toxic for bacteriaSelectively toxic for bacteria– bactericidal (killing) bactericidal (killing) – bacteriostatic (growth inhibition)bacteriostatic (growth inhibition)

• no harm to patientno harm to patient

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AntibioticsAntibiotics

• destroy structuresdestroy structures– present in bacteriapresent in bacteria

– not present in hostnot present in host

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Antibiotics work togetherAntibiotics work togetherwith immune systemwith immune system

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Minimal inhibitory Minimal inhibitory concentrationconcentration

• lowest level stopping growthlowest level stopping growth

• e. g. zone of inhibition around a disk e. g. zone of inhibition around a disk impregnated with antibioticimpregnated with antibiotic

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• Antibiotics that inhibit cell wall Antibiotics that inhibit cell wall biosynthesis are bactericidalbiosynthesis are bactericidal

• Without cell wall, osmotic pressure Without cell wall, osmotic pressure causes bacteria to burstcauses bacteria to burst

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Cross-linking of peptidoglycanOld New

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Peptidoglycan synthesisPeptidoglycan synthesis

CytoplasmCytoplasm Cell wallCell wall

undecaprenolundecaprenol

sugarsugar

aminoaminoacidacid

Cell MembraneCell Membrane

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CycloserineCycloserine

1.1. alanine (ala) analog alanine (ala) analog 2.2. inhibits conversion of L-ala to D-ala inhibits conversion of L-ala to D-ala 3.3. inhibits formation of D-ala-D-alainhibits formation of D-ala-D-ala

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CycloserineCycloserineAnalog of alanine

XX

CytoplasmCytoplasm

sugarsugar

aminoaminoacidacid

XX

XX

XX

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BacitracinBacitracin

• Inhibits dephosphorylationInhibits dephosphorylation

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TRANSPORT OF TRANSPORT OF PEPTIGOGLYCAN SUBUNIT PEPTIGOGLYCAN SUBUNIT

ACROSS MEMBRANEACROSS MEMBRANECell membraneCell membrane

undecaprenolundecaprenol

PPPP

Cell wallCell wall

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TRANSPORT OF TRANSPORT OF PEPTIGOGLYCAN SUBUNIT PEPTIGOGLYCAN SUBUNIT

ACROSS MEMBRANEACROSS MEMBRANECell membraneCell membrane

undecaprenolundecaprenol

PP

PP

Cell wallCell wall

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TRANSPORT OF TRANSPORT OF PEPTIGOGLYCAN SUBUNIT PEPTIGOGLYCAN SUBUNIT

ACROSS MEMBRANEACROSS MEMBRANECell membraneCell membrane

undecaprenolundecaprenol

PPPP

Cell wallCell wall

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TRANSPORT OF TRANSPORT OF PEPTIGOGLYCAN SUBUNIT PEPTIGOGLYCAN SUBUNIT

ACROSS MEMBRANEACROSS MEMBRANECell membraneCell membrane

undecaprenolundecaprenol

PPPP

Cell wallCell wall

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TRANSPORT OF TRANSPORT OF PEPTIGOGLYCAN SUBUNIT PEPTIGOGLYCAN SUBUNIT

ACROSS MEMBRANEACROSS MEMBRANECell membraneCell membrane

undecaprenolundecaprenol

PPPP

Cell wallCell wall

23MINUSMINUS BACITRACINBACITRACIN

CellCell membranemembrane

undecaprenolundecaprenol

PP

Cell wallCell wall

TRANSPORT OF TRANSPORT OF PEPTIGOGLYCAN SUBUNIT PEPTIGOGLYCAN SUBUNIT ACROSS MEMBRANEACROSS MEMBRANE

24PLUSPLUS BacitracinBacitracin

Cell membraneCell membrane

undecaprenolundecaprenol

PPPP

Cell wallCell wall

TRANSPORT OF TRANSPORT OF PEPTIGOGLYCAN SUBUNIT PEPTIGOGLYCAN SUBUNIT ACROSS MEMBRANEACROSS MEMBRANE

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VancomycinVancomycin

Cell wallCell wall

• binds to D-ala-D-alabinds to D-ala-D-ala• inhibits cross-linkinginhibits cross-linking

Vancomycin

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Beta lactam antibioticsBeta lactam antibiotics

• penicillins penicillins

• Cephalosporins/cephamycins Cephalosporins/cephamycins

• monobactamsmonobactams

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Beta lactamsBeta lactams

CellCell wall wall

Penicillin binding proteinPenicillin binding protein

•inhibit penicillin inhibit penicillin binding proteinsbinding proteins

•stop cross-linkingstop cross-linking

Beta lactam

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C NH CH CH C

O

O C N CH

CH3

CH3

COOH

S

Site of penicillinase action.Breakage of the lactam ring.

STRUCTURE OF PENICILLIN

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Attached to lactam ringAttached to lactam ring

• penicillins penicillins – 5 membered ring 5 membered ring

• Cephalosporins/cephamycins Cephalosporins/cephamycins – 6 membered ring6 membered ring

• monobactams monobactams – no second ringno second ring

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Chemical modifications change Chemical modifications change biological activitybiological activity

• Early lactam antibiotics Early lactam antibiotics

– inactive against Gram negative inactive against Gram negative bacteriabacteria

• no penetration of outer membraneno penetration of outer membrane

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Resistance mechansimsResistance mechansims

• Produce beta lactamase (penicillinase)Produce beta lactamase (penicillinase)

– destroys antibioticdestroys antibiotic

• modified penicillin binding proteins modified penicillin binding proteins

– don’t bind antibioticdon’t bind antibiotic

• modified porinsmodified porins– no internalization of antibioticno internalization of antibiotic

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• beta lactam beta lactam

• binds strongly beta lactamases binds strongly beta lactamases – inhibits activityinhibits activity

Clavulinic acidClavulinic acid

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Polymyxin BPolymyxin B

• bindsbinds– lipid A lipid A – phospholipidsphospholipids

• disrupts outer membrane, Gram negative bacteria disrupts outer membrane, Gram negative bacteria • toxic to human cells toxic to human cells

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Isoniazid, Ethambutol, Ethionamide  

Isoniazid and Ethionamide - chemically related- blocks mycolic acid synthesis

Ethambutol- blocks arabinogalactan synthesis

Anti-tuberculosis drugs