Biology 425 - University of Montana

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_____________________________ Biology 425 (BIOB 425) Advanced Cell & Molecular Biology CRN 32222, Spring Term 2014 ______________________________


MWF 1:10-2:00 p.m.; Health Science 411.

Web Page: Course Handouts and Messages will be posted on Moodle. Instructor:

Dr. Jesse C. Hay e-mail: [email protected] Office Hours: Mon. 2-4 p.m. and by appt. Office: Skaggs #390A (New Addition). Research Lab: Skaggs #385

Course Text: Molecular Cell Biology, Seventh Edition, by Harvey Lodish, Arnold Berk, Chris A. Kaiser, Monty Krieger, Anthony Bretscher, Hidde Ploegh, Angelika Amon, and Matthew P. Scott, published by W.H Freeman and Company. Available at the University of Montana Bookstore. Coursepack: will be handed out and posted online in sections as the course progresses.







1/27 1/29 1/31

Introduction to Class, start Methods Methods I: Cells Methods II: Microscopy and Biochemistry

****** 1-22; 397-404; 424-430 93-106; 404-424


2/3 2/5 2/7

Methods III: Genetics and Genomics finish methods lectures Protein Conformation Regulates Cell Events

171-182; 198-206; 212-219; 252-256 ****** 59-92


2/10 2/12 2/14

Protein Conformation (cont.) Membrane Structure and Function Membranes (cont.)

****** 443-470 ******


2/17 2/19 2/21

President's Day, no class Transporters Exam I (65 pts.)

****** 473-494 ******


2/24 2/26 2/28

Regulation of the Cellular Ionic Environment Posttransl. Protein Transport into Organelles Posttransl. Transport (cont.)

495-511 601-614 ******


3/3 3/5 3/7

Transport Across the Nuclear Envelope Cotranslational Protein Transport into the ER Transport into ER (cont.)

615-622 579-594 ******


3/10 3/12 3/14

ER/Golgi Posttranslational Modifications and Quality Control Vesicle Formation & Cargo Sorting

594-601; 644; 651-652 ****** 627-637


3/17 3/19 3/21

Vesicle Targeting and Fusion Exam II (65 pts.) Cytoskeleton I: Actin Dynamics, Muscle

638-650 ****** 773-815


3/24 3/26 3/28

Actin Dynamics (cont.) ****** Cytoskeleton II: Microtubules, Motors, Mitosis 821-859 Cytoskeleton III: Intermediate Filaments 860-867


3/31 4/2 4/4

Spring Break; no class Spring Break; no class Spring Break; no class

****** ****** ******



4/7 4/9 4/11

Cell Signaling Pathways Cell signaling (cont.) Cell Cycle I

673-713; 721-768 ****** 873-889


4/14 4/16 4/18

Cell Cycle II Cell Cycle (cont.) Exam III (65 pts.)

890-913 ****** ******


4/21 4/23 4/25

Apoptosis Mechanisms of Cancer I Mechanisms of Cancer II

1006-1017 1113-1130 1131-1150


4/28 4/30 5/2

Extracellular Matrix and Cell Adhesion I Extracellular Matrix and Cell Adhesion II ECM (cont)

925-945 945-967 ******


5/5 5/7 5/9

Regulated Protein Degradation Nerve Cells and Action Potentials Cellular Basis of Learning and Memory

661-666 1019-1047 ******


5/12-5/16 5/14

Finals Week, no class Final Exam (140 pts); 3:20-5:20 p.m.

****** ******


COURSE POLICIES Course Structure Biol. 425 is designed as an advanced course in Cell Biology for students majoring in the life sciences. To fully absorb the material, it is important that you meet the suggested prerequisites for the course. Some knowledge of both molecular biology and genetics will be assumed. Thus, it is especially important to have previously completed coursework in these areas. It is recommended, but not required, that coursework in biochemistry be taken before or concurrently with this course. Whether you have had the prerequisites or not, if you feel that you are deficient in molecular biology, biochemistry or genetics, be sure you have access to good textbooks to use as background references. Lecture Format There are 3 lectures each week, given on Mon., Wed., Fri. from 1:10-2:00 p.m. in Health Sciences 411. The lectures are mandatory; you must attend the lectures. It is important that you do not miss lectures, since lectures will often contain information or examples that are not covered in the readings. Exam Policy Exam questions will be based primarily on the material covered in lectures. The final exam will contain a section covering the last block of new material in detail, as well as a cumulative section covering the whole semester. Problem Sets There will be a problem set distributed approximately one week in advance of each exam (4 total). These will provide practice for the more difficult types of questions that could appear on an exam, although there is no guarantee that the same precise topics are covered on the problem sets and exams. Students are welcome to work together and to seek the instructor's advice on the problem sets. Problem sets are due at the start of the exam. Writing Assignment This course includes one written paper of at least five pages exclusive of references and figures. The paper will consist of a critical review of current primary research literature and scientific (i.e. not political or social) discoveries/controversies surrounding the cell biology of a disease (human, animal, or plant). A detailed description of the assignment, including content and format requirements, will be distributed at the beginning of the semester. It is hoped that the assignment will help students achieve the following writing goals: 1) identification of sophisticated questions for inquiry; 2) synthesis of information from multiple sources; 3) presentation of multiple perspectives and interpretations; 4) learn appropriate citation and documentation practices for cell biology, and; 5) competence in searching the appropriate electronic databases. A complete draft of the paper, including the appropriate, correctly formatted references, will be due before Spring Break. In addition to the paper, students will turn in results from electronic searches documenting that the relevant literature database(s) was queried with appropriate search terms.


GRADING Problem Sets Written Paper Exam 1 Exam 2 Exam 3 Final Exam TOTAL

80 pts. 85 pts. 65 pts. 65 pts. 65 pts. 140 pts. 500 pts.

4 X 20 pts. (due the lecture preceding each exam) (draft due beginning of Spring Break; final by last day of class) Friday Febr. 21 1:10-2:00 p.m. Wednesday March 19 1:10-2:00 p.m. Friday April 18 1:10-2:00 p.m. Wednesday May 14 3:20-5:20 p.m.

All students are expected to take the exams at the scheduled time. If, however, you feel that you are near death and cannot make an exam, you will be allowed to take a make-up exam ONLY IF YOU CAN PROVIDE A NOTE FROM A CERTIFIED M.D. to that effect. Other emergencies will be considered on a case by case basis. IN ALL CASES, YOU MUST CONTACT DR. HAY IN ADVANCE OF THE EXAM. If you are unable to call, a family member or friend should call instead. If you miss an exam without making any prearrangements, you will receive 0 points for that exam. The dates for all exams are listed above. You have no excuse for not knowing when the exams are scheduled or for leaving before the scheduled final at the end of the semester. To preserve the academic integrity of the course, we reserve the right to alter the content and/or format of the original exam when creating a makeup exam. Make-up final exams will be oral. Individual exam scores will not be curved in this class. Letter grades will be assigned after the semester has been completed, based on the total points accumulated. Adjustments will be made at that time, if necessary. Cutoffs will be no higher than 450 pts. (90%) for A-, 400 pts. (80%) for B-, 350 pts. (70%) for C-, and 300 pts. (60%) for D-. GRADUATE CREDIT Students may receive graduate credit by completing an additional writing assignment. This will involve reading primary research articles, identifying a current gap in our knowledge of cellular processes (or controversial model), and providing a synthesis of recent attempts and future directions to address it experimentally. Detailed instructions will be given for this assignment following Spring Break. Academic Conduct As you proceed through this course you may at times feel the pressure of course work, exams, and assignments. So much to do, and so little time! If you seriously fall behind in your work, you should talk to Dr. Hay. He will try to be sensitive to your individual problems, but to be fair to all students, he will enforce deadlines and grade your performance by standards that apply to everyone. Whatever happens, don't be tempted to cheat; it's not worth it, and the penalties are very severe. If we believe that you have been dishonest on an exam, we reserve the right to award you 0 pts. for that exam. At this stage in your career, suspension and/or failing grades may significantly affect your chances of being able to graduate on schedule. Advice on Mastering the Material in Cell Biology


1. Exam questions are based on the lectures. The readings are intended to supplement your understanding and provide additional examples. However, your lecture notes should always be the priority when studying for an exam. If you want to succeed, you must attend the lectures and take complete notes! 2. Review and outline your notes as soon as possible after lecture. Don’t just read your notes. Cognitive science has shown that writing, for example, outlining your notes, is an important step in the learning process for most people. 3. Keep up with the class. Don’t wait until just before the exam to master all the material! If you find yourself having difficulty with the material DO NOT WAIT until you have problems with the first exam before coming in for help. This is what office hours are for - to clarify material covered in the course. 4. Make use of all the resources available to you. There are many biochemistry, genetics, molecular biology, and other cell biology texts in the library that may serve as good references for you. Make use of office hours. Dr. Hay will also answer concise questions submitted by e mail. 5. It is very helpful for students to form a STUDY GROUP with whom you can meet to discuss the material in the course. Past experience indicates that these groups work best when they are relatively small (2-4 people). This allows everyone in the group to talk about the material. They are most helpful when the group meets on a regular basis (at least once a week - not just before exams


Guidelines for Preparing for the Exams 1. The best way to prepare for the exams is to keep up with the material as we go. Don’t wait until just before the exam to read the textbook and go over your notes!! 2. Discuss the material regularly with a study group of 2-4 classmates. Talk about the information presented in lectures. 3. Make use of office hours to help you understand the difficult topics. If you have brief questions, use e-mail. 4. The exams will test your understanding of the material in three ways: 

Factual Recall: Do you understands the facts presented? These are the “regurgitation” questions - they require relatively straightforward answers.

Concepts: These questions expect you to put several facts together into a concept you have already been introduced to in lecture. They test your understanding of how components fit together. They may be questions about models or mechanisms or processes (simple and complex).

Application: These questions will test your ability to apply the information from lecture to real data examples. These are “story problems” that you will have to reason through. You may be asked to interpret a graph, propose an experiment, or explain some results. In each case, the necessary information will have been presented in lecture or discussion, but you must apply it to this new situation.

5. Concentrate on the lecture notes, since only material covered in lecture will be on exams. Use the textbook or other assigned readings as needed to provide further clarification of the material. Keep in mind that certain information presented in lecture will not be found in the reading. Don’t just memorize the information - make sure you understand it!!


Expected Learning Outcomes for BioB 425 1. Students should understand the experimental basis from which modern cell biological knowledge comes; this includes being able to interpret experiments and evaluate conclusions from studies using: a. Tissue culture (primary and clonal) b. Antibodies as specific probes for cellular components using several distinct protocols c. Recombinant gene expression studies and gene knockout and knockdown approaches d. Light and electron microscopy e. Protein purification and analysis f. Genetic analysis including epistasis experiments for ordering gene pathways g. Basic genomic algorithms and tools for predicting gene function 2. Students should understand the fundamentals of protein structure and how cellular stimuli regulate the activity of proteins. They should be able to describe example mechanisms for how posttranslational events like phosphorylation or GTP binding and hydrolysis translate into altered protein activity. Students should also be able to recognize a number of conserved protein domains and their functions. 3. Students should understand the makeup of cellular membranes throughout the cell; this includes phospholipid asymmetries between cytosolic and extracytosolic leaflets, the abundances of different lipids in different organelles, and the location of synthesis of the major lipid species. Students will also need to understand the different types of membrane proteins, their mechanisms of associations with membranes, and their mechanisms of constraint and microlocalization within membranes. 4. Students should understand how the cell regulates transport across its membranes. This will include knowledge of the major groups and biochemical mechanisms of membrane transporters, ion pumps and channels. It will also include knowing the major ionic equilibria and approximate ion concentrations in cellular compartments. 5. Students should have extensive and detailed knowledge of how proteins become targeted to cellular compartments. This includes mechanisms that occur co-translationally (proteins of the endomembrane system) as well as posttranlationally (mitochondrial, peroxisomal, chloroplast and nuclear proteins). They will also know the primary sequence of localization signals within proteins and how they are interpreted and effected by the targeting machinery. 6. Students should understand the details of protein trafficking within the endomembrane system, once a protein enters this system (see goal 5). This will include understanding and knowing the major examples of transport vesicle coats, cargo recognition and sorting mechanisms, and vesicle targeting and fusion. The students will be able to generalize these mechanisms to specific transport steps of physiological or medical importance such as the uptake of cholesterol via receptor mediated endocytosis, etc. 7. Students should understand the major posttranslational modifications and how protein quality control is maintained. This will involve knowing several pathways, for example the unfolded protein response, autophagy, and ubiquitin-mediated proteasomal activation, for the regulated degradation of inappropriate proteins. 8. Students will know the basic functions, mechanisms of synthesis and regulation, and structural characteristics of cytoskeletal elements, including microfilaments, intermediate filaments and microtubules. This will include understanding at a molecular level the process of treadmilling, branching, dynamic instability and other regulatory phenomena. Students will also know the


major cytoskeletal regulatory proteins and how they control muscle contraction, mitosis, cytokinesis and other cellular transformations. The students will also know the major classes of cytoskeletal motors and which movements and transport events they regulate and how. 9. The students should understand the cell division cycle and its regulation. This will include understanding the historical elucidation of maturation promoting factor and our current molecular understanding of these kinase complexes, their targets, and how they effect cellular events. Students will know about many cell cycle regulators and be able to predict the effects of changes in their activities on rates of cell division and potential roles in cancer. 10. Students will know the major pathways of cell signaling originating from ligand binding by cell surface receptors. This will include tyrosine kinase and G-protein coupled receptors and their entire signaling cascades, second messenger systems, scaffolds and cellular effects. 11. Students will understand the cellular and genetic basis of cancer. This will include understanding the different types of mutations that accumulate in cancer and how they promote cell mitogenesis, tumor production or invasiveness. Common cellular modifications observed in cancer, such as GTPase deficient Ras, should be learned. Students should be able to predict whether a given mutation or modification would have oncogenic vs. tumor suppressor characteristics. 12. Students will learn the major pathways leading to and inhibiting programmed cell death, or apoptosis, and understanding the integration of these pathways with oncogenic and tumor suppressor effects in cancer. 13. Students will know in detail the structure of the extracellular matrix and how it contributes to cell migration, tissue formation, and human disease. 14. Students will know the basic anatomy of neurons, propagation of action potentials, and formation of neuronal signaling circuits. The students should be able to successfully integrate this neuron-specific information into a cutting edge model of learning and memory at the cellular level in Aplysia californica that incorporates many of the cell biological concepts learned throughout the semester. 15. The students should be able to write a paper reviewing current research on the cell biology of a disease, incorporating scientific concepts from class that: a. Identifies a sophisticated question for inquiry b. Synthesizes information from multiple sources c. Presents multiple perspectives and interpretations d. Demonstrates appropriate citation and documentation practices for cell biology e. Demonstrates competence in searching the appropriate electronic databases



Focus and Format Requirements: Choose a disease that has a known or hypothesized cellular/molecular biological basis. Be sure there is little or no overlap with assignments you have done for other classes (this could be considered academic misconduct). The bulk of the paper should deal with basic cell biological research related to your chosen disease. It should not be only a recitation of symptoms or a report of clinical studies, though these things can be briefly summarized in the Introduction section. There should be little or no discussion of patients or patient surveys or human subjects. The main purpose and challenge of this assignment is to dig into and understand the primary literature surrounding the cell biology (ie molecular and cellular mechanism) of the disease. The paper is to be NO SHORTER than 5 pages (1.5 spacing, 11pt. font, 1-inch margins on all sides), excluding figures, references and other literature documentation. It is fine to integrate figures into the text if you want, just make sure the text on it own meets the minimal length. Sections of the Paper: 1. Introduction - this section should include information about the clinical symptoms of the disease and any known epidemiology or related statistics. The purpose here is to tell us, in general terms, about the impact of the disease. Information in this section must cite published journal articles from which you got it, and reviews would probably be the most appropriate type of article to use here (ask a librarian how to limit your literature search to reviews). 2. Cellular/Molecular Basis of the Disease - this section has two parts. a. First, present a model for what causes the disease at the cellular level. This must be clearly related to material presented during lectures or in your textbook, meaning it is put in the context of cell biological knowledge. For example, what cellular regulatory pathways, specific protein structures and functions, membrane/organelle structures and functions, or physiological processes are affected in the disease? How are they affected? The level of detail and focus area of the model presented will need to be sufficient to support part b below. In this context, “model” means current molecular/cellular concepts of how the disease arises, not to be confused with an “animal disease model”, which is an experimental system that replicates a human disease. This section (part a) can cite reviews or primary experimental articles, though reviews should be your starting place for gathering information. You do not have to include a figure with a graphical model, but it would be appropriate; if you borrow one from an article, be sure to properly cite it in the figure legend. b. Second, present three or more specific experiments that were undertaken recently demonstrating how research is evaluating, extending, or changing the model from part a. The experiments discussed should come from at least three separate primary experimental articles. Discuss these three articles plus any others necessary for you to adequately


explain and interpret the experiments. Be sure it is basic science research, not clinical research (i.e., no human subjects, unless patients are used only as a source of cells/tissue for experimentation). As in part a, it is not required to present figures, but it is appropriate and best if it helps explain the conclusions. Figures can be incorporated into text pages or put on separate pages. Avoid exhaustively recapitulating the methods--just briefly explain the rationale for conducting each experiment, the experimental approach, the result and conclusions, and their implications for the model(s) described in part a. You should focus on what you believe is the most significant, conclusive experiment from each of the three experimental articles you are covering. 3. Future Directions - this section could include information on directions of future research and potential therapies to treat/cure the disease that could arise from a better understanding of the cellular basis of the disease. For example, do aspects of the model evaluated in your articles open up new pathways or proteins that could be used as drug targets? This section is more open ended and is mostly your interpretation and opinion. You do not necessarily need to cite more articles unless specific new facts or research is brought up. 4. References - references should be cited parenthetically in the text using the following formats: (Wittman, 2009) for articles with one author, (Cookson and van der Brug, 2008) for articles with two authors, and (Gitler et al., 2008) for articles with three or more authors. At the end of the document, include a separate section titled "References" containing the full citation for all of the articles cited in the text, arranged alphabetically by last name of first author. Each article should appear only once in the reference list, even if you cite it at multiple points in the text for multiple facts or ideas. Include all authors' names (do not use "et al."), year, complete article title, volume and page numbers as shown below (if my examples are not clear please check the reference format in any recent Journal of Cell Biology article). References should be single-spaced with larger spaces between references. Abbreviate the names of journals according to PubMed. Below are examples for journal articles and book chapters (book chapters that comprise primary literature reviews can be cited in parts 1, 2a, or 3 of the paper; they are not suitable for part 2b, which must cite journal articles). Websites are generally not acceptable for citation since they do not represent a permanent, published, information source. Journal article format: Gitler, A. D., B. J. Bevis, J. Shorter, K. E. Strathearn, S. Hamamichi, L. J. Su, K. A. Caldwell, G. A. Caldwell, J.-C. Rochet, J. M. McCaffery, C. Barlowe, and S. Lindquist. 2008. The Parkinson's disease protein alpha-synuclein disrupts cellular Rab homeostasis. Proc Natl Acad Sci USA 105(1):145-50. Book chapter format: Innerarity, T. L., D. Y. Hui, and R. W. Mahley. 1982. Hepatic apoprotein E (remnant) receptor. In Lipoproteins and Coronary Atherosclerosis. G. Noseda, C. Fragiacomo, R. Fumagalli, and R. Paoletti, editors. Elsevier/North Holland, Amsterdam. 173–181. 5. Literature Searches - Every student must also submit documentation of appropriate literature database searches that identify resources used in this assignment. The xi

recommended database for this assignment is PubMed (, but others may be acceptable. Approach a reference librarian at Mansfield Library for help using PubMed or another database if you encounter difficulty or would like some assistance. The Information Center (IC) desk is staffed until 9 p.m. Sunday through Thursday and until 6 p.m. on Friday and Saturday. The documentation should detail your search strategy (database selected, search terms and limits used) and results for at least two searches. The search results do not have to be submitted in any particular format--whatever the database provides is fine. The first database search should be used to identify review articles about the cellular basis of your disease. An example PubMed search to get reviews about synucleinopathies (Parkinson's-related diseases) would be the following: synuclein*[Title] AND review[Publication Type]. When further limited to the last 10 years, this search produces ~175 articles, which is small enough to be perused comprehensively. The other search(es) should be informed by reading one or more of the review articles and use specific proteins, molecular pathways and terms related to your disease to identify recent (late nineties and onward) primary research articles for your paper. For any of your searches, you may have to use more limits to avoid getting hundreds or thousands of clinical reviews and primary papers. One hint would be to combine the disease name with a specific protein name (e.g. hemophilia[Text Word] AND ERGIC-53[Title]). You must turn in sufficiently limited searches—perusing hundreds or thousands of titles is not considered a legitimate search strategy. One downside of being at a small university is that not all research articles will be available to you in full-text. Librarians and the library journal search page (choose the green tab from the library homepage, can help you to identify the articles available in full-text. DO NOT limit your searches to “Free Full Text” on Pubmed. This will cause you to ignore the preeminent journals with the most exciting results. Do not submit the actual full-text articles with your assignment. I require documentation of the searches your performed, not just results. The documentation proves that you performed appropriately crafted, narrowed searches that gave reasonable numbers of results and that you chose to focus on appropriate hits. Submission Instructions A complete draft of the paper is due by 5 a.m. on Tuesday April 1st, 2014. The document should be emailed to [email protected] as an email attachment. I will acknowledge receipt within 24 hours. The email address from which the document is sent will be the same address to which it is returned with a grade and comments. The complete draft, including references, figures, tables, lit. searches etc., must be submitted as a single PDF file of less than 5 MB in size. Word processor files will trigger an automatic 10-pt reduction. Get computer lab help if you have trouble making or sizing your PDF, as this is an important part of the assignment. The database results should be saved as a file (or captured by copying and pasting or screenshots) and incorporated into the submitted PDF. Grading Approximately half of the grade will be determined by writing style and clarity, and half on command of the literature and conceptual framework. The draft will be graded on its merits; this


evaluation will include a list of suggested improvements, including style and content changes. For example, the instructor may require the student to pursue more sophisticated questions about the research, synthesize information from more diverse sources, or present multiple interpretations of the work discussed. In addition, it may be necessary for the student to adapt the paper to include acceptable citation and documentation formats or repeat electronic searches using more sophisticated criteria. Of course, there will also be standard editorial suggestions related to clarity, grammar, organization, and avoidance of redundancy. Standard deductions of 10 pts. will be applied for lateness, missing search documentation or submission of word processor or over-sized files. It will be clearly stated which improvements are considered "minimal" and which are required for a "major" writing overhaul. All of the minimum suggested improvements must be incorporated into the final paper to maintain the same final grade. To get an improved grade on the final paper (10 pts. maximal improvement), all of the stated minimum improvements plus suggested major improvements must be incorporated. The instructor will judge the degree to which suggested improvements were met. Disease Suggestions The disease chosen must have a known or hypothesized cellular basis - this list does not guarantee that all these diseases are appropriate. They are only suggestions, places to start. The disease may affect humans, other animals, microbes, or plants. Suggestions: Tay Sachs muscular dystrophy canine parvo methemoglobinemia cardiomyopathy Huntington’s disease spinobulbar muscular atrophy spinocerebellar ataxia Bannwarth’s Syndrome acrodermatitis chronica atrophicans Kennedy’s disease spinabifida Alzheimer’s multiple sclerosis Friedreich’s ataxia (FRDA) Peutz-Jegher’s syndrome Parkinson’s disease I-cell disease IgA deficiency thalassemia Hodgkin’s disease myasthenia gravis neurofibromatosis

cystinuria Pomp’s disease Shigella - secretory diarrhea (Vibrio cholerae) Zellweger’s syndrome hyperthyroidism pituitary microadrenomas-prolactinoma epilepsy Paget’s disease (increased bone) von Willebrand’s disease (blood clotting) Refsum’s disease hemophilia Gilbert’s syndrome (can’t conjugate bilirubin) Marfan’s syndrome familial polyposis of colon (polyps) Gardner’s disease Zollinger-Ellison syndrome (pancreatic tumors) malaria retinitis pigmentosa measles Polio virus cholera tetanus rabies


fructose intolerance Maple syrup urine disease tyrosinemia Niemann-Pick disease alpha-1 anti-trypsin Osteogenesis Imperfecta phenylketonuria scoliosis homocystinuria Staphylococcal toxic shock syndrome Lyme disease ataxia telangiectasia – IgG erythropoietic protoporphyria myotubule myopia (ryanodine receptor mutations) osteogenesis imperfecta Wolf-Parkinson-White syndrome (cardiac arrhythmias) von Hippel-Lindau (inherited, retinal blastomas) Creutzfeldt-Jakob Syndrome (mad-cow disease) infectious mononucleosis (Epstein-Barr Syndrome) SCIDs - severe combined immunodeficiency (bubble kids) Wiskott-Aldrich syndrome (IgA, IgM, IgG deficiency) spherocytosis - RBCs spherical G6P dehydrogenase deficiency - RBC death McArdle’s disease - no muscle breakdown of glucose polycystic kidney disease primary hyperoxaluria diabetes (specific type) spondyloepiphyseal displasia tarda


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