Notices
Results 1 to 21 of 21

Thread: Postgrad research help...?

  1. #1 Postgrad research help...? 
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    Hi everyone I am new to The Science Forum, I'm doing my Masters degree and I'm starting with new research that I'm not that familiar with... but what better place to get information from than a website full of scientists!!!

    Has anyone ever worked with S9 Fractions or Liver Microsomes?


    Reply With Quote  
     

  2.  
     

  3. #2  
    Forum Professor Zwolver's Avatar
    Join Date
    May 2006
    Location
    Netherlands
    Posts
    1,667
    No, but what is your question?

    Maybe you can explain us what S9 fractions or liver microsomes are.


    Growing up, i marveled at star-trek's science, and ignored the perfect society. Now, i try to ignore their science, and marvel at the society.

    Imagine, being able to create matter out of thin air, and not coming up with using drones for boarding hostile ships. Or using drones to defend your own ship. Heck, using drones to block energy attacks, counterattack or for surveillance. Unless, of course, they are nano-machines in your blood, which is a billion times more complex..
    Reply With Quote  
     

  4. #3  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    First of all this is related to Cell Biology

    Okay in short, I am working with a drug called Sibutramine, I.m doing cytotoxicity and teratogenic tests on it. In an in vivo situation though the sibutramine is metabolized by the liver (CYP2B6 enzymes) so if I use it In vitro (in cell culture) its not metabolised. S9 fraction and liver microsomes are "liver extracts" that contain all the enzymes of liver metabolism. I want to use it to metabolize my drug and then administer the active metabolites to my cells, so I was wondering if anyone has done it before or has worked with liver extracts before so I can pick their brain...
    Reply With Quote  
     

  5. #4  
    Forum Professor Zwolver's Avatar
    Join Date
    May 2006
    Location
    Netherlands
    Posts
    1,667
    Haha, much better. I have worked with the principles, but not with liver extracts in particular, i'm working on the cardiovascular system, cappilaries in particular. So i can't help you.

    What does Sibutramine do? Your testing if it's harmfull for cells, and checking on the cells respons by monitoring these S9 fractions and other liver extracts. Interesting, are there articles about this?
    Growing up, i marveled at star-trek's science, and ignored the perfect society. Now, i try to ignore their science, and marvel at the society.

    Imagine, being able to create matter out of thin air, and not coming up with using drones for boarding hostile ships. Or using drones to defend your own ship. Heck, using drones to block energy attacks, counterattack or for surveillance. Unless, of course, they are nano-machines in your blood, which is a billion times more complex..
    Reply With Quote  
     

  6. #5  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    Sibutramine is a weight-loss drug, but its been banned from most markets as it causes cardiovascular events, some resulting in death. It has an indirect effect though, its metabolized by the liver, then the metabolites (which are much more potent) inhibits the re-uptake of serotonin and nor-epinephrine (nor-adrenaline). And this increases satiety so the subject feels fuller for longer etc. It was a very successful drug.

    Well, basically I did a pilot study with Sibutramine on mouse fibroblast cells and although we could see something happening with the cells the results were inconclusive. So now I want to work wit the metabolites as a certain cell type doesn't have organs or, more specifically, a liver to metabolize the sibutramine. So I want to use the S9 fraction or human microsomes to metabloize the sibutramine and then I give the metabolites to different cell lines ex cardiac, neuronal, renal etc. and then see if it damages the cells in any way.


    When I'm done with that I'll study the teratogenic effects using the in ovo method.

    Biggest problem for me at this stage is to choose between using S9 Fractions or Liver microsomes, and the literature on how to go about doing it is very limited. There are metabolic studies where authors use these liver extracts to study the metabolism of drugs, but none that I have found so far that re-administers the metabolites back into a cell culture or that propose a protocol for doing a study like this.

    It is interesting and very exciting , just wish I could get started now!!! I'm still struggling to get cell lines etc.
    Reply With Quote  
     

  7. #6  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    Last edited by C15ka; May 16th, 2012 at 07:01 AM.
    Reply With Quote  
     

  8. #7  
    Forum Freshman
    Join Date
    May 2012
    Location
    Oklahoma City, USA
    Posts
    14
    I just wanted to wish you the best of luck in your master's research!! I don't have much insight regarding cell lines or your research in particular but, having just completed my master's degree in biology a few weeks ago, I am aware of the adventure you are about to partake. Let me just say it's never too soon to start writing your thesis - getting that introduction/literature review knocked out early will save a lot of headache later on.
    "Don't kick a fresh turd on a hot day." Harry S Truman
    Reply With Quote  
     

  9. #8  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    Thanks and don't I know it!!! But you probably know how many times you change your mind when you do your initial planning, I basically finished everything and then (on recommendation by my supervisors) everything changed, just last week, so I have to start all over again, but thank you for your encouraging words. What did you do your MSc about (if I may ask...)?
    Reply With Quote  
     

  10. #9  
    Forum Freshman
    Join Date
    May 2012
    Location
    Oklahoma City, USA
    Posts
    14
    I screened a population of white-throated woodrats from Arizona for genetic variation using major histocompatibility complex (Mhc) loci to try to find an explanation as to why this species is naturally associated with at least 3 viruses. My data weren't all the spectacular but I was able to provide some preliminary results that others may use and build upon. My biggest challenge was using single-strand conformational polymorphism (SSCP) analysis to detect alleles -- it ended up being a huge pain in the ass!
    "Don't kick a fresh turd on a hot day." Harry S Truman
    Reply With Quote  
     

  11. #10  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    There's always something that you have to struggle with, I've been doing cytotoxicity assays from the beginning of the year at least every two weeks under the exact same conditions, and my results were different every time, so now I have to review the whole assay protocol, an assay we've been using in the labs for forever!!! so have to sort out that first, and I cant find fertilized chicken eggs, just found a place though and they are SOOO expensive, but I guess anything regarding research in South Africa is expensive

    Sorry, I'm obviously frustrated...
    Reply With Quote  
     

  12. #11  
    Forum Freshman
    Join Date
    May 2012
    Location
    Oklahoma City, USA
    Posts
    14
    Dude, it's good - feel free to vent your frustrations! If it was easy, it wouldn't be science. And I know things get expensive, especially lab work. Sometimes protocols just stop working for whatever reason -- maybe there is some contamination going on or your chemicals have expired. I have been there.

    Hang in there!!!!!!
    "Don't kick a fresh turd on a hot day." Harry S Truman
    Reply With Quote  
     

  13. #12  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    So what are your plans for the future? career options for me is very limited, especially here and especially in research, so I'm looking into other options, maybe starting over as a first year next year...

    (Just ignore me if I'm asking too much too soon, just trying to get an idea of what research entails in other parts of the world)
    Reply With Quote  
     

  14. #13  
    Forum Freshman
    Join Date
    May 2012
    Location
    Oklahoma City, USA
    Posts
    14
    I applied to a PhD program but did not get accepted due to budget cuts -- seems to be the major problem right now in academia in the US. I'm going to apply locally to research labs (as a research assistant), colleges to be a part-time (adjunct) instructor, and other state-funded industries, such as the water treatment facilities or the health department. I'd really like to continue with research but because those jobs are inherently tied to available funding, they are limited.
    "Don't kick a fresh turd on a hot day." Harry S Truman
    Reply With Quote  
     

  15. #14  
    Forum Freshman
    Join Date
    May 2012
    Location
    Oklahoma City, USA
    Posts
    14
    What types of future endeavors would you like to pursue? Now, I'm curious.
    "Don't kick a fresh turd on a hot day." Harry S Truman
    Reply With Quote  
     

  16. #15  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    Well, in a perfect world, I'd do my Phd, work at an Academic Research Institution, get a BBIIGG salary and become a world renowned scientist for curing cancer or Aids or something... and just be happy with that...

    But I've applied for veterinary sciences, and I'm considering applying for medicine, I'm not sure if I really have a future in research in SA
    Reply With Quote  
     

  17. #16  
    Forum Freshman
    Join Date
    May 2012
    Location
    Oklahoma City, USA
    Posts
    14
    I totally agree with your view of the perfect world!
    "Don't kick a fresh turd on a hot day." Harry S Truman
    Reply With Quote  
     

  18. #17  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    Have you considered doing your Phd abroad? Or what are you doing to kill time?
    Reply With Quote  
     

  19. #18  
    Forum Freshman
    Join Date
    May 2012
    Location
    Oklahoma City, USA
    Posts
    14
    Well, my husband has a good job here so it wouldn't be wise for us just to pick up and move for the hell of it. For the moment, I'm just enjoying my time off. How about you?
    "Don't kick a fresh turd on a hot day." Harry S Truman
    Reply With Quote  
     

  20. #19  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    I haven't really thought about it...
    Reply With Quote  
     

  21. #20  
    Forum Bachelors Degree
    Join Date
    Jul 2008
    Posts
    420
    Hi C15ka,

    I don't have any experience working with liver microsomes but some of my colleagues in the Lab carry out liver microsomal prep's on a regular basis. I could ask them about it, if you provide specific questions?

    Also - why don't you start your toxicity studies in a mouse model first?

    Best wishes,

    Tridimity
    Reply With Quote  
     

  22. #21  
    Forum Freshman
    Join Date
    May 2012
    Location
    South Africa
    Posts
    17
    Hi Tridimity, thanks for your reply

    From my previous posts I'm sure you now have some idea of what I am trying, I've found literature on studies where people have metabolized sibutramine , but no studies where the metabolite has been re-administered back into a cell culture, I'm still doing some research on this side and as soon as I have all my questions I'll start posting them. But I haven't used liver microsomes before so any usefull tips are welcome

    The reason why I'm not using a animal model is because we did the animal study last year, and we're still processing data, however there are very little in vitro studies on sibutramine and I have done a pilot study where I can definitely see a change in the cells but the assays tell a different story. So I want to study its mechanism in vitro.

    However, I will be using chick embryos for my teratogenic studies.
    Reply With Quote  
     

Similar Threads

  1. Research Help
    By bobsanchez in forum Environmental Issues
    Replies: 0
    Last Post: October 12th, 2010, 02:29 PM
  2. Research
    By paulmichelnovo in forum Philosophy
    Replies: 2
    Last Post: September 29th, 2010, 09:13 AM
  3. New Research Software
    By Dekstrus in forum Links
    Replies: 2
    Last Post: January 23rd, 2008, 03:35 PM
  4. Methylbenzoate research
    By krystelkaos in forum Chemistry
    Replies: 2
    Last Post: March 7th, 2007, 07:32 PM
  5. Question about research
    By mental in forum General Discussion
    Replies: 0
    Last Post: November 6th, 2006, 02:47 PM
Bookmarks
Bookmarks
Posting Permissions
  • You may not post new threads
  • You may not post replies
  • You may not post attachments
  • You may not edit your posts
  •