User:Z5129708

From CellBiology

Welcome to Cell Biology 2017!

Lab 1 Assessment

Lab 2 Assessment

  1. Identify a chemical SDS and the risks and hazards of that chemical in text. Add a link to the original SDS
  2. Select 4 reference papers papers related to your selected group project topic sub-section. Read the research papers and write a brief description of their findings and relevance to the selected topic sub-section. The reference along with your description should then be pasted on both your group discussion page and your own personal page.

Lab 3 Assessment - Endo/Exo worksheet questions.

Lab 4 Assessment

  1. Identify a cytoskeletal antibody.
  2. Identify the species deriving the antibody.
  3. Identify the working concentration for the antibody.
  4. Identify a secondary antibody that could be used with this antibody.
  5. Identify a paper that has used this antibody.

This assessment will be due by the next lab (Lab 5).

Lab 7 Assessment

The following peer assessment exercise should be completed before next lab (Lab 8 - 2 May) as your individual assessment for this week (lab missed due to public holiday).

Your answer should be pasted in 2 places

  1. onto each project discussion page (Note you should add anonymously to the discussion page)
  2. your own individual student page for my assessment.

Each individual will provide a brief assessment of the other groups projects. This should take the form of a brief critical (balanced) assessment identifying both the positive (good) and negative (bad) aspects of the project page as it currently exists online.

You may if you choose, use the final project assessment criteria as a guide. Though you are also welcome to use your own criteria.

Group Assessment Criteria

  1. The key points relating to the topic that your group allocated are clearly described.
  2. The choice of content, headings and sub-headings, diagrams, tables, graphs show a good understanding of the topic area.
  3. Content is correctly cited and referenced.
  4. The wiki has an element of teaching at a peer level using the student's own innovative diagrams, tables or figures and/or using interesting examples or explanations.
  5. Evidence of significant research relating to basic and applied sciences that goes beyond the formal teaching activities.
  6. Relates the topic and content of the Wiki entry to learning aims of cell biology.
  7. Clearly reflects on editing/feedback from group peers and articulates how the Wiki could be improved (or not) based on peer comments/feedback. Demonstrates an ability to review own work when criticised in an open edited wiki format. Reflects on what was learned from the process of editing a peer's wiki.
  8. Evaluates own performance and that of group peers to give a rounded summary of this wiki process in terms of group effort and achievement.
  9. The content of the wiki should demonstrate to the reader that your group has researched adequately on this topic and covered the key areas necessary to inform your peers in their learning.
  10. Develops and edits the wiki entries in accordance with the above guidelines.

Attendance

Z5129708 (talk) 16:05, 7 March 2017 (AEDT)

Z5129708 (talk) 16:10, 14 March 2017 (AEDT)

Z5129708 (talk) 15:14, 21 March 2017 (AEDT)

Z5129708 (talk) 15:58, 28 March 2017 (AEDT)

Z5129708 (talk) 15:05, 4 April 2017 (AEST)

Z5129708 (talk) 15:00, 16 May 2017 (AEST)

Tutorial

Student Image Template

Note - This image was originally uploaded as part of a student project and may contain inaccuracies in either description or acknowledgements. Please contact the site coordinator if the uploaded content does not meet the original copyright permission or requirements, for immediate removal.

Referencing

PubMed

Search Plasma Membrane

  • No Link - PMID: 28257417
  • link - PMID 28257417

Javier U Chicote, Rob DeSalle, Antonio García-España Phosphotyrosine phosphatase R3 receptors: Origin, evolution and structural diversification. PLoS ONE: 2017, 12(3);e0172887 PubMed 28257417


Human R3 RPTP members.png

Lab 1 Assessment

Model for rapid modulation of the Plasma Membrane.png

A Ca 2+ dependent model for repair of the Plasma Membrane [1]

When the plasma membrane of a cell is damaged, it needs to be repaired in order to maintain homeostasis in the cell, the image shows a model for how the membrane is repaired. When the plasma membrane is broken there is an influx of Ca 2+ ions, which subsequently causes exocytosis of lysosomes. This paper shows that the proteases present in the lysosomes play a major role in the regulation of the repair of the membrane. An example of these proteases are Cathepsin B, L and D. Cathepsin B and L are released because of the exocytosis of the lysosomes, this release is shown by the green curve on the graph in the figure, the release of Cathepsin D is delayed and released around 30 seconds after the wounding of the cell membrane, this is shown by the red curve on the graph. After their release, Cathepsin B and L alter the extracellular matrix of the cell by cleaving the proteins allowing better access to the surface of the outer leaf of the membrane, this makes it easier for Acid Sphingomyelinases to hydrolyse the sphingomyelin which results in the release of ceramide and promotes the resealing of the plasma membrane. As a result of this release Cathepsin D’s activity is unregulated which down regulates the activity of Acid sphingomyelinase and finishes the process, leaving a repaired plasma membrane.


Mark Hill (talk) 14:24, 28 March 2017 (AEDT) Good with the image upload and associated information summary. Additional information required is with the image file. Try not to use sub-headings for image titles, I have adjusted. (5/5)

Lab 2 Exercise

Picric Acid

  • Molecular Weight = 229.10 g/mol
  • Molecular Formula = C6H3N3O7
  • Explosive when dry (moistened with water to make sure the compound does not become explosive)
  • Dangerous

Find full safety information at Picric Acid Sigma Aldrich [2]

Pancreatic Ductal Cells Research

Dynamic regulation of CFTR bicarbonate permeability by [Cl-]i and its role in pancreatic bicarbonate secretion.

The pancreatic ductal cells secrete HCO3-, which provides an environment in the duodenum which is optimum for the function of digestive enzymes. This paper explores how the cells secrete the HCO3- using concentration of Cl- ions. To do this they use human pancreatic cells and guinea pig pancreatic tissue and analyse using approaches such as imunnoblotting and electrophysiology. The results of the study suggested that the pathway, WNK-OSR1/SPAK pathway, is sensitive to the intracellular concentration of Cl- and this acts as a switch which causes the cells to secrete HCO3-.

[3]

ATP release, generation and hydrolysis in exocrine pancreatic duct cells.

This study explores whether pancreatic duct cells release ATP in order to regulate the ductal function, and the factors that are important in the control of this process. A human pancreatic cell line Capan-1 was used, as these cells are known epithelial cells and have ion channels and transporters in their membrane used for secretion. The conditions around the cells were changed, for example changes to the pH, and the fast ATP release from the cells was detected using an online luminescence measurement. [4]

Mechanisms of KGF Mediated Signaling in Pancreatic Duct Cell Proliferation and Differentiation

The aim of this paper is to explore how pancreatic ductal cells proliferate and differentiate as they can be used as an alternative source of insulin producing beta cells, this is important as it could be an opportunity for new therapies of cell replacement in Type 1 diabetic patients. The study investigates the effects of Keratinocyte Growth factor (KGF), which activates Fibroblast growth factor (FGFs) receptors which are expressed on the surface of the ductal cells. To do this they used a BrdU assay to measure the cell proliferation of diabetic rat cells, the results measured using immunolocalization. The results of these in vivo and in vitro experiments show that the KGF promotes duct cell proliferation by inducing PDX1 pathway and the regenesis of beta cells. When the pathways involved were investigated they found that the MEK-ERK1/2 pathway is responsible for duct cell proliferation and the PI3K/AKT pathway is unregulated by KGF and mediates cell differentiation to beta cells. The findings from this study is important as it could be used in regenerative medicine for diabetes. [5]

Inactivation of TGFβ receptor II signalling in pancreatic epithelial cells promotes acinar cell proliferation, acinar-to-ductal metaplasia and fibrosis during pancreatitis.

Pancreatitis is a disease in the pancreatic cells where digestive enzymes attack the cells as they are prematurely activated. After pancreatitis the tissue needs to repair itself, Transforming growth factor-β (TGFβ)’s experession is unregulated during this repair. TGFβ is involved in activation of transcription genes and activating smads which activate growth. The aim of this study was to explore how the signalling of TGFβ in pancreatic duct cells is in involved in pancreatitis.The study used transgenic mice models, that had a knockout allele of the TGFβ-RII receptor, allowing them to analyse the signal involved in each key part of the disease. To study the cells, pancreatitis was induced in the mice using injections, then cultures of their acing cells were taken and incubated with the TGFβ inhibitor. The cells were analysed by using antibodies to label the proteins of interest and quantify and then RNA was then extracted from the cells and analysed. The results of the experiments suggested that TGFβ-RII signalling is not involved in the damage observed from pancreatitis, it also increases the expression of kinase inhibitors and the lack of TGFβ-RII in the cells causes a increased number of inflammatory cells present in pancreatitis. The results of this study are important as they show that TGFβ can act as a tumour supressor and is involved in pancreatic tissue regeneration. [6]



Mark Hill (talk) 19:20, 24 April 2017 (AEST) Very good for papers could have improved for SDS (7/10) .

For the SDS you have not identified the hazards and therefore not completely correct even without the full statement could have included the following codes and information:

  • Symbol - GHS02, GHS06
  • Hazard statements - H228-H301 + H311-H332
  • Precautionary statements - P210-P280-P301 + P310-P312
  • Supplemental Hazard Statements - Explosive when dry.
  • Personal Protective Equipment - Eyeshields, Faceshields, Gloves, type P2 (EN 143) respirator cartridges

These are the Globally Harmonized System as described in the practical class. The 4 papers are relevant, hopefully they can be used in the final group project.

Lab 4 Exercise

Antibodies

Actin Monoclonal Antibody (ACTNO5 (C4))

  • Host of antibody is mouse
  • Anitbody reconises Actin in Dog, Chicken, Human, Mouse, Non Human primate
  • Monoclonal Antibody
  • IgG 1
  • Unconjugated Antibody

ThermoFisher Actin Antibody [7]

Anti-Mouse Antibody

  • Conjuagted with a Fluorescent dye - Alexa Fluor 488
  • Host of anti-antibody is Goat
  • Poly Clonal

ThermoFisher IgG1 Anti-antibody [8]

Needed to identify a paper using this antibody. (3.5/5)

Lab 5 Exercise

Cytoskeleton Exercise

Comparison of B35 Cells with 'normal' and overexpressed TM4.png

A graph showing the comparison of two B35 cell groups and their percentage of each phenotype shown. Group A has an overexpressed TM4 and Group B acts as a control.


Mark Hill (talk) 20:51, 24 April 2017 (AEST) This lab assessment will be marked by the guest presenter and the mark added here when I have received.

Lab 7 Assessment - Peer Assessment

Group 1 - Delta Cells

On first look at this Group project you can see that you have an extensive content of headings and sub headings which are good, although these are slightly repetitive and could be cut down slightly. So far, there is good use of videos from YouTube and 1 image which is useful and easy to read and there seems to be significant research into function and cell matrix interactions. It could be worth considering re-ordering your subheadings, for example putting signalling and cell matrix interactions before pathology as this knowledge may give a better understanding of why diseases arise? More images could be added and adding a few images made by you for the signalling would be a good way to have images and relay information. Sub-headings such as Structure and current research could do with some more research and although the diseases have videos there needs to be more information here – but I’m sure this is just not been done yet. The background/introduction could have some additional information, at the moment I don’t think it gives a an easy to read introduction to Delta Cells. Overall, I think this is a good start to the project as there is a lot of information.

Group 3 - Beta Cells

There is a good amount of sub headings – although these could have subheadings within, in order to make it easier for the reader to find information? The images that have been used so far are good; I especially like the Beta-Cell destruction image. The introduction is a helpful overview of the islets of Langerhans although could introduce the Beta cells in slightly more detail? I think the information in the ‘Role in Disease’ sub topic is good and you have outlined Type 1 Diabetes and Type 2 Diabetes well, one way in which you could improve this section is you could make an image of the different stages of Type 2 Diabetes meaning less text and an easy to understand/view image. Function, Development and signalling could do with more information but I’m sure this is just due to time, although the points that you have outlined in each sub heading here is good. A suggestion for a sub topic to add is current research? You could link this to the treatments. Overall though, each sub topic seems to have some information and images used so far are good.

Group 4 - Alpha Cells

There has been a good start to this page, I especially like the table in 'History' and the images used so far - although these images don't seem to be referenced properly. The sub headings used so far are useful, although these could be linked better - for example why endocrine cells link to development. Introduction and structure need more adding - although you have lots of time to do this. Overall referencing needs to be looked at but the information down so far is helpful. I also like the use of videos on this page. Some sections are still in bullet points - these could be combined into sentences - also with some of the signalling processes in this part you could make images to make them easier to follow? Overall, a good start though.


Very good feedback includes banned assessment comments. (8/10)

References

  1. Thiago Castro-Gomes, Matthias Corrotte, Christina Tam, Norma W Andrews Plasma Membrane Repair Is Regulated Extracellularly by Proteases Released from Lysosomes. PLoS ONE: 2016, 11(3);e0152583 PubMed 27028538
  2. Picric Acid Sigma Aldrich
  3. Hyun Woo Park, Joo Hyun Nam, Joo Young Kim, Wan Namkung, Jae Seok Yoon, Jung-Soo Lee, Kyung Sik Kim, Viktoria Venglovecz, Michael A Gray, Kyung Hwan Kim, Min Goo Lee Dynamic regulation of CFTR bicarbonate permeability by [Cl-]i and its role in pancreatic bicarbonate secretion. Gastroenterology: 2010, 139(2);620-31 PubMed 20398666
  4. J M Kowal, G G Yegutkin, I Novak ATP release, generation and hydrolysis in exocrine pancreatic duct cells. Purinergic Signal.: 2015, 11(4);533-50 PubMed 26431833
  5. Benjamin Uzan, Florence Figeac, Bernard Portha, Jamileh Movassat Mechanisms of KGF mediated signaling in pancreatic duct cell proliferation and differentiation. PLoS ONE: 2009, 4(3);e4734 PubMed 19266047
  6. Kamile Grabliauskaite, Enrica Saponara, Theresia Reding, Marta Bombardo, Gitta M Seleznik, Ermanno Malagola, Anja Zabel, Carmen Faso, Sabrina Sonda, Rolf Graf Inactivation of TGFβ receptor II signalling in pancreatic epithelial cells promotes acinar cell proliferation, acinar-to-ductal metaplasia and fibrosis during pancreatitis. J. Pathol.: 2016, 238(3);434-45 PubMed 26510396
  7. ThermoFisher Actin Antibody
  8. ThermoFisher IgG1 Anti-antibody


2017 Course Content

Moodle

Lectures: Cell Biology Introduction | Cells Eukaryotes and Prokaryotes | Cell Membranes and Compartments | Cell Nucleus | Cell Export - Exocytosis | Cell Import - Endocytosis | Cytoskeleton Introduction | Cytoskeleton - Microfilaments | Cytoskeleton - Microtubules | Cytoskeleton - Intermediate Filaments | Cell Mitochondria | Cell Junctions | Extracellular Matrix 1 | Extracellular Matrix 2 | Cell Cycle | Cell Division | Cell Death 1 | Cell Death 2 | Signal 1 | Signal 2 | Stem Cells 1 | Stem Cells 2 | Development | 2017 Revision

2017 Laboratories: Introduction to Lab | Fixation and Staining |


2017 Projects: Group 1 - Delta | Group 2 - Duct | Group 3 - Beta | Group 4 - Alpha

Dr Mark Hill 2015, UNSW Cell Biology - UNSW CRICOS Provider Code No. 00098G