Lecture 1 (Ch. 1): 1. Microorganisms are all around us. They are a major source of disease in the world but also have many beneficial roles both in the environment and in/on our bodies. 2. We use microbes in the lab all the time. Pasteur’s Swan neck flask experiment used microbes to prove that cells come from existing cells, and not spontaneous generation. He also surmised that it was these microbes that were the cause of many illnesses. Koch formulated his postulates to formalize how we determine if a specific microbe causes a specific illness (also, there are limits to these postulates that you should now be able to articulate). 3. We use microscopes to look at bacteria, viruses are too small. Microscopy can tell us a lot about a bacteria (arrangement, shape, size, Gram designation). Lecture 2 (Ch. 3) 1. There are many cellular structures we learned about. Some of the most important ones come back again and again, usually because they have some role in disease, either by helping bacteria to colonize, avoid the immune system, etc. or as targets for antimicrobials. Think: capsule (and biofilms), cell wall, flagella, outer membrane (LPS/endotoxin), 30S ribosome. 2. Transport in and out of the cell: it’s important to understand that it takes energy to move something UP its concentration gradient but it does NOT to move something DOWN its concentration gradient. Also, we talk about the electron transport chain/hydrogen pumps a lot in Ch. 6. 3. Chemotaxis is a type of cellular movement that directs movement towards or away from a stimulus. This should make sense, especially since we have spent some time talking about how a cell senses the outside world and then responds to it (think lac operon). Chemotaxis is simply a way bacteria (or other cells, like macrophages and neutrophils) sense the outside world and then respond via movement. Lecture 3 (Chs. 3 and 4) 1. Eukaryotic cells are characterized by the presence of a nucleus and compartments that have specialized functions. Eukaryotic cells have different ways of taking in materials, and phagocytosis is a specialized kind of endocytosis. 2. Mitochondria and chloroplasts are particularly interesting organelles because they have many traits making them resemble bacteria (endosymbiotic theory). 3. Prokaryotes divide by binary fission and have varying generation times which reflect how quickly they can grow (including during infections). Some grow in biofilms, which offers protection from the immune system and the outside world. 4. It’s important to understand that bacteria have different nutritional and environmental needs. Remember, some bacteria require NO growth factors (E. coli) and some require many (N. gonorrhea). Bacteria have different oxygen requirements, as well (you should know the terminology by now). You should be comfortable with terms describing where bacteria get their carbon and energy, as well. 5. At this point you are all probably experts in the idea of selective and differential media. Make sure you understand how these might be used to enrich for a particular bacteria. Lecture 4 (Ch. 6) 1. All cells undergo glycolysis. Some then stop (fermentation), while others can undergo a transition step to generate acetyl-coA from pyruvate. This enters the TCA cycle. The reducing power (NADH, FADH2) can then be used to take excited electrons to the electron transport chain. These electrons then gradually lose their energy as they are shuttled from one membrane protein to the next and as they do this, this energy is used to pump hydrogen ions UP their concentration gradient. As the hydrogen ions enter back into the cell via ATPases, ATP is generated in large amounts. This process is cellular respiration (and usually the terminal electron acceptor is oxygen). You should know what the major products of each step are. 2. You should understand that these processes are often conducted by enzymes, which bind to their substrates and allow some kind of chemical change to occur. Enzymes are regulated by either competitive inhibition (competitor binds to the active site) or by non-competitive inhibition (molecule binds to an allosteric site, a site other than the active site). 3. Photosynthesis uses photons to directly excite electrons found in the chloroplasts. These electrons can either be recycled (no terminal electron acceptor, they return to Photosystem I and are excited by photons again) or can instead be used to generate NADPH, which provides reducing power that the cell needs during biosynthesis. 4. All this energy is used to either fix carbon (in the case of autotrophs) or fuel growth, movement, reproduction, etc. The pathways that result in the generation of new cellular structures is called anabolic and is essentially the opposite of catabolic pathways. Lecture 5 (Ch. 5) 1. Different settings require different levels of microbial control. It is especially important in the medical setting that pathogens be either reduced in number (like on the skin before surgery) or completely absent (like on a scalpel). People in hospitals are especially vulnerable to infection because of open wounds/incisions and potentially being immunocompromised. 2. The bacteria that cause nosocomial infections are generally ubiquitous (found everywhere), but they usually only cause problems in the special setting of a hospital. They can be extremely drug resistant. 3. Sterilization will kill everything, including endospores (one of the hardiest forms of life on the planet). Disinfection aims to reduce or eliminate the majority of bacteria and viruses, but some small number may still remain. You should be familiar with the constraints of different kinds of sterilization and disinfection methods (i.e. when would it be appropriate to use an autoclave versus a filter?). Lecture 6 (Ch. 7) 1. The central dogma of biology is that DNA is transcribed into RNA which is then translated into proteins. DNA is made up of 4 nucleic acids: A, T, C, G which base pair (A-T, C-G). mRNA is RNA that is transcribed from a gene in the DNA. Many copies of mRNA can be made for the same gene and generally the amount of mRNA is related to the amount of protein to be made. It is important that mRNA is unstable so that the cell can quickly stop making a protein if necessary. 2. Three nucleotides of mRNA make up a codon. Each codon corresponds to one amino acid. The mRNA is translated into amino acid chains (peptides, proteins) by ribosomes which are composed of protein and rRNA. These are responsible for the catalysis of the addition of new amino acids to the growing polypeptide chain. tRNA is responsible for “reading” the codons and bringing the correct amino acid to the ribosome for incorporation into the new protein. 3. The process of transcription is tightly regulated so that the cell only makes proteins necessary for its environmental conditions. This is why sensor proteins help to communicate what is going on and this signal then travels through the cell and to the DNA. Transcription can be regulated by the presence of a repressor protein which must bind an inducer in order to fall off the gene and allow the RNA polymerase to transcribe. Alternatively, often a corepressor will need to bind to the repressor in order to allow the repressor to bind to the DNA. In some cases, an activator is required to turn a gene on and this activator must be bound by an inducer. In all these cases, often the presence or absence of an inducer or corepressor is dictated by what the sensor protein is sensing. The Lac Operon was a complex example of this. Lecture 7 (Ch. 8) 1. Bacteria can evolve via spontaneous mutation or horizontal gene transfer. Spontaneous mutation occurs rarely and can have different outcomes depending on the exact location of the mutation (silent, missense or nonsense mutations). Deletions or insertions can be very destructive since they cause frameshifts to occur. 2. Transposons are mobile genetic elements that can jump around the genome and plasmids of a cell. Transposons can carry genes. 3. Mutation can be induced by introducing chemicals that can either modify or take the place of the normal nucleic acids. Usually, this causes a mistake in base-pairing. Intercalating agents can cause insertions or deletions. 4. DNA can be repaired by either light-dependent or light-independent methods. Usually, the cell will excise a small piece of the problematic DNA (identified as the unmethylated new strand) and then use the good DNA as a template for the repair. In the case of UV light, thymine dimers form and some bacteria can use visible light to break these bonds. If not, the cell will often start the SOS response (leading to the use of a polymerase that makes many mistakes—this is how thymine dimers indirectly lead to mutation). 5. We can select for mutants that have gained an ability (like resistance to antibiotics) by plating them on selective media. We can select for bacteria that have lost an ability (like making the amino acid histidine) by replica plating on a minimal media and a rich media and looking for colonies that can no longer grow on the minimal media. 6. You should understand conjugation, transduction, transformation in terms of how they occur, what must happen to the DNA for it to become permanent and what kinds of genes that we care about might be transferred in this way. Chapter 12 1. How does “zombification” of ants help parasitic fungi to spread? 2. Why can eurkaryotic infections be harder to treat (why don’t we have any many drugs to treat them)? 3. What diseases do protozoans cause? Lecture 8 (Ch. 20) 1. Antimicrobial targets are generally something unique to bacteria. You should know the general targets and be familiar with how different drugs target different parts of cell wall synthesis. Also understand that pencillin (and all antimicrobials) must be able to access their target or they cannot work. 2. While you don’t need to memorize all the different kinds of antibiotics again, you should be familiar with the targets: 30S, 50S ribosome, nucleic acid synthesis, folate synthesis. Lecture 10 (Bacterial and Viral Pathogens) 1. Diarrheal diseases are spread via the fecal-oral route (water, food, your hands can be contaminated). It makes sense that the bacteria would cause lots of diarrhea as the bacteria is shed that way. For respiratory organisms, they cause coughs in order to be transmitted to the lungs of the next host. 2. TB is a slow-growing bacteria that will slowly cause granuloma formation in the lungs. 3. Legionella is interesting because it infects free-living cells that resemble our macrophages. 4. Staphyloccocus aureus and Streptococcus pyogenes both cause a variety of infections depending on where in the body the infection is located (the skin, the throat, in the blood, under the skin in the fascia [necrotizing fasciitis]). Alternative treatments are important for MRSA because we are running out of options.