ANTIBIOTICS Antibiotics: - low Molecular weight compounds - either kills bacteria or - inhibit their growth and reproduction.

- primarily produced by bacteria or fungi -Penicillin Penicillium, Mold -Cephalosporins Cephalosporium, Mold - Bacitracin & Polymyxin B Bacillus Bacteria

Bacteria and molds are responsible for antibiotics

Controversy concerning weather physicians should prescribe a broad spectrum verses a specific. Use was narrow initially, but then there was CDIFF.

CDIFF: The illness can cause explosive diarrhea, severe ulcerations of the colon, and death. Several outbreaks in the mid to late 70s among elderly hospitalized patients.

This is a gram (+) bacillus, anaerobic, spore forming, toxin producing son of a gun.

Sudomembroluscholitis is a byproduct of hospitalized patients who were being treated with antibiotics for some other infection.

Antibiotics in question: Clindimycin and Ampacillin.

Wiping out all the good flora in the colon except for about 10-20% of the population that normally has CDIFF, but usually it is such small numbers that is kept in check by the flora in the colon. Wiping out the flora will increase CDIFF. Explosive diarrhea gets into the air and spore formers form, colonizing new patients in the hospital.

When looking for good candidates for antibiotics, want to select a bacterial target site. Ex: DNA Replication, Plasma membrane, Ribosomes, Bacterial cell wall.

Folate Metabolism: Bacteria make their own folic acid, which is necessary for growth, so if we can interfere with folate metabolism, we can control microbial growth.

GROUPS OF ANTIBIOTICS: One of the most commonly prescribed groups of antibiotics are:

THE BETA LACTAM antibiotics

This is because they have minimal side-effects.

4 classes: They all share the Beta Lactam Ring. 1. Penicillin: Ex: Penicillin

2. Cephalosporins: Ex: Keflix- Used against Gram (+) & (-) some are bactericidal, some are bacteriostatic.

3. Carbapenem: Ex: Impenem

4. Monobactum: Ex: Axtreonam

One side effect that can happen is that an allergic reaction can develop because of the production of a Beta Lactam Serum Protein that reacts with the antibiotic. - can occur if one is on a penicillin - if the reaction does happen, that patient will become allergic to cephlasporin and monobactam.

Mechanism of action: They target bacterial cell wall Synthesis. Cell wall consists of peptidoglycan, which consists of alternating units of NAcetyl glucosamine and N-Acetyl Muramic acid (both polysaccharides), linked by glycocidic linkages and peptide linkages. They attack Transpeptidation Bacterial cell wall is impeded, bacterial cell lysis, and bacteria dies. Trans Peptidase enzymes are what are responsible for linkages, sometimes these Trans peptidase enzymes bind to penicillin instead of NAG and NAM

Another mechanism of action: In lag phase when the parent cell is elongating, making more plasma membrane, cytoplasm, ribosome, cell wall Another mechanism of action would be to stimulate Autolysisns to cause bacteria to lysis itself.

GLYCOPEPTIDES As a group, also target bacterial cell wall and interfere with bacterial cell wall synthesis.

In this case they inhibit the second to the last step in bacterial cell wall synthesis: Glycoside linkages: Tranglysylation.

*Bactericydal for Gram (+) bacteria

*Ineffective against Gram (-) bacteria.

Ex. Vancomycin Toxicity issues so it is the drug of last resort.

Ex.Teichoplanin

AMINOGLYCOSIDES Antibiotics that target Bacterial Protein Synthesis:

1. Antibiotic has to get into cytoplasm to the Ribosomes, which means it has to get through cell wall, plasma membrane.

2. Ribosome contains 50s and 30s. To be a functional ribosome, the 50s component has to be bound to the 30 s component. ***They do not inhibit mRNA by binding to the 30s, but they prevent the 30s binding to the 50s component, so the ribosome never becomes active.

****They are BACTERICIDAL

Toxicity issues: hearing loss, kidney damage.

Ex. Kanamycin Gentamycin

TETRACYCLINES Can be taken orally, Had minimal side effects: if given to pregnant women or pre-puberty children, they could cause blackening or darkening of the teeth. In some individuals they would develop nausea, photosensitivity issues. They were over prescribed to treat acne, also given to livestock to prevent shipping sickness.

1. They distort the A site from the EPA which means the anticodon cannot match up with the codon on mRNA and that disrupts bacterial protein synthesis.

****They are BACTERISTATIC

MACROLIDES / LINCOSAMIDES 2 structurally different groups but mode of actions are the same :

They target the 50s component of the bacterial ribosome. 3 steps: a. Innitation b. Elongation- These antibiotics interfere with bacterial Ribosome elongation. c. Termination

Macrolide: Ex. Zithromax, Bioxin

Linosamide: Ex Lincosin

Antibiotics that Target Nucleic Acid synthesis (Both DNA and RNA) QUINOLONES(s) EX. Naladixic Acid DNA-Gyrase: Ability to loosen the super coiled DNA and participate when they reform after DNA replication. Naladixic acid binds to a Beta Subunit of DNA-gyrase and does not allow supercoils to be relaxed or reformed.

2 Groups of Quinolones: 1. Fluroquinolones: Narrow spectrum antibiotics so they dont interfere with the intestinal micro flora and they are extremely effective against enterobacteriasea family: E.coli, clebcella.

EX: Ciprofloxacin Norfloxacin

RIFAMPIN Targets nucleic acid synthesis.

Binds to the Beta Subunit of transcriptase (RNA polymerase) which means no transcription which means no mRNA which means no proteins.

Was used to treat TB cases, now it is used to treat Meningitis as well as for prophylactics when come into contact of Meningitis.

Antibiotics that interfere with Bacterial Folate metabolism: Steps in bacterial folate metabolism PABA Folic AcidDihydrofolic AcidTetrahydrofolic Acid

1. Para Amino Benzoic Acid (PABA): enzymatically converted into Folic acid. Folic acid in turn is enzymatically converted into Dihydrofolic acid which is enzymatically converted into Tertrahydrofolicacid: precursor substrate which is CRITICAL in the carbon transfer reactions that take place during purine and pyrimidine nitrogen base formation and in the synthesis of amino acids. Metabolic inhibitors: They look very similar to the substrates that these enzymes act on; they bind to the antibiotic and do not concert substrate into an end product. 1. Ex.Solfonamides (sulfur based): target the first step; interfere with the conversion of PABA into Folic acid.

2. Ex.Trimethoprins: Interfere with the last step, preventing conversion of Di-hydrofolic acid into Tetra-hydrofolic acid.

Antibiotics that attack Bacterial Cell/plasma Membrane: Ex. Bacitracin Polymixin B

Poke holes in cell membrane, all internal contents spews out into external environment. ONLY FOR TOPICAL USE!!

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Prjessimal Pathogens Becoming resistant to antibiotics that were in use.

Non Pathogens Became more opportunistic, they also were becoming resistant to antibiotics that were in use.

Genetically, how do bacteria become resistant? CHROMOSOMAL MUTATIONS Bacteria acquires a CONJUGATIVE PLASMID Acquiries a CONJUGATIVE TRANSPOSON

Mechanisms Bacteria Possess that make them Resistant to Antibiotics: Gram Negative Bacteria (Fewer layers of Peptidoglycan) but they have: Outer membrane, Surface proteins, LPS components AND PORIN proteins: Act as channels through which substances can enter the Gram Negative bacterial cell, they are non selective AND they give the bacteria about a 10 fold increase in resistance. Beta Lactamases: Ex: Penicillinase: -breaks a bond and converts an active penicillin to an inactive penicillin acid.

Aminoglyosides: -Bacteria codes for genes that have theses modifying enzymes -What they do is attach a chemical group to the enzyme, thereby inactivating it, like a phosphate group. -Effectively inactivate the enzyme.

Enzymatic Inactivation

EFFLUX PUMP: -Microbes acquire a gene that makes an Efflux pump, ATP pump -needs energy, but it will pump out the antibiotic just as fast as it is diffusing into the bacteria -Found in methicillin and tetracycline.

PROTEIN SHIELD: -Bacterium acquires genes to produce a protein. A dome shaped protein, it literally shields the ribosome - tetracycline gets into the cell, but because of the shield it cannot bind to the 30s component and distort the A site.

Another Mechanism of Resistance to Penicillin, affects last step of bacterial wall synthesis, bacteria undergoes a Point Mutation on the enzymes, and Penicillin will no longer bind to them.

Point mutation (single base substitution, modifies the target site.)