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Sayonara!

Well, we leave tomorrow so this will be my last post!  Tonight, we will be attending a “Beer Festival” and will have our last shot at genuine Japanese food and spirits.  It will be located on the NIRS campus and will be with NIRS staff.  We’ve been looking forward to this and it is perfect timing considering the fact that we leave tomorrow.

I want to thank everyone here at NIRS for being so helpful and nice.  Especially, Dr.s Kato, Okayasu, and Fujimori.  Thank you for hosting us, taking us sight seeing, introducing us to new foods, and teaching us the art of radiobiology (cell culture & survival curves)!  It has been a wonderful and unique experience that I am extremely appreciative of.  I have fallen in love with Japanese culture and hope that I will return to this beautiful country.

Sayonara!

-Petra

Radiation Station

Hey all!

So, this is our last week here.  Which means i’ll probably post once more (if that) after this one.  I figured I’d post some good pictures of the equipment and facilities that we’ve gotten to see here.  I can’t tell you how many radiation warning signs-which are very similar to ones in the US- are posted everywhere.  As is Japanese custom, we take off our shoes and step into slippers when entering a person’s home.  Well, for certain rooms and areas at NIRS, we do the same thing.  Notice, the fashionable yellow slippers that we get to wear when in the HIMAC research center (see picture).  I inquired about how they choose which rooms to change shoes and which rooms you don’t and I’ve determined that it’s relatively random.  Almost all of the culturing and lab rooms require that you change slippers…sometimes entire floors or even buildings require it.  However, most offices don’t require that you take your shoes off but some do.  My thoughts on this topic initially would not go away but when I could come up with no answer, I eventually settled for the fact that it’s just random.

Anyway, enough about shoes.  The pictures below show our visit to the HIMAC treatment & research facilities (separate areas) and a few other random pictures of NIRS.

Check ya lata!

-Petra

1,2,3,4 ….ichi, ne, san, chi!

Count, count, count.  I’ve been helping Fujimori Sensei by doing the monotonous and inevitable job that follows cell culture….counting of colonies.  I had a nifty little pen that beeps and counts for you every time you mark a colony but then it dried up.  :(  Which, by the way, I don’t understand since it is sooooo humid here!  Oh well.

Last week I got to help Dr. Allen run an experiment where we irradiated multiple cell lines at different doses with the HIMAC (Heavy Ion Medical Accelerator) and then performed a gratuitous amount of cell culturing till 3AM.  Oi vay. Since then, the cells have been allowed to grow (or not grow) in the incubator.  Some of the dishes already had colonies today so I washed and stained those dishes and will be counting those next week.  To refresh your memory (and mine), most of these projects that I am helping with are all concerned with DNA damage & repair after exposure to radiation (i.e. X-ray or Heavy Ion).

well. that’s it for today!  keepin’ it short!  :)

-Petra

Shockingly Transfected

Hey Hey!  Well, today marks the midpoint of my trip.  In the two weeks I have been here I feel like I have done/seen so much.  It is comforting to know that I have yet another two weeks to do even more!

This week, in the lab, I have been working with Dr. Mori who is a senior scientist at NIRS.  He has been wonderfully helpful and has made a great attempt at explaining things to me in English.  First, he let me practice cell culturing (i.e. washing, trypsinizing, and splitting) with a human colon cancer cell line known as HCT 116.  Later he showed me how to build a DNA vector from E.Coli to be used for transient (as opposed to stable) transfection of a gene.  Apparently, transient transfection allows for a higher quantity of gene expression to occur for a brief period of time (http://en.wikipedia.org/wiki/Transfection).  Today we performed electroporation, a type of transfection process in which foreign DNA is introduced into a eukaryotic cell via formation of transient pores in the cell membrane by high voltage pulses.  In the lab,  cells are placed in suspension in an appropriate buffer (we used K-PBS) and into an electroporation cuvette.  This cuvette is then connected to a power supply and the cells are essentially “shocked” by a high-voltage (today we used 300V & 960µF capacitance) electrical pulse.  After this electrocution 30-50% of the cells actually die and the remainder are what will give us expression of the desired gene.  In this case, we transfected HCT116 cells with a plasmid containing mKate2, a highly fluorescent monomeric fusion protein (http://www.evrogen.com/products/mKate2/mKate2.shtml).

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“Not until we are lost do we begin to understand ourselves.” Henry David Thoreau

I’ve been told that getting lost is the way you learn your way around places.  Yesterday, I got lost not once but two times.  The first time, I was attempting to find Dr. Fujimori’s culture room which happens to be located in an entirely different building from his main office.  I ran up and down multiple floors trying not to look so obviously disoriented but to no avail.  Of course everyone is extremely nice and helpful here but it’s not necessarily any easier to find your way when you don’t understand the language of your helper and vice versa.  Thankfully the Japanese are extremely nice and wonderfully patient and I was eventually found and brought to the correct cell-culturing room that I meant to find in the first place.

Once there, Dr. Fujimori had us distribute multiple cell lines (brain tumour, colon cancer, etc.) into flasks so that they could be irradiated.  After incubation, we exposed them to various doses (2, 4, and 6 Gy) of radiation and then placed them into another incubator.  Later, we fixed the cells onto slides and then treated with goat anti-body.  Apparently we will be assessing DNA Repair characteristics via a protein marker that is specific to these (brain & colon) cancer cell lines.  Of course there is way more to it than what I’ve just described but I will describe more as we progress and when I understand more.

The second time I got lost it was not physically but most definitely intellectually; I attempted to understand a seminar with two presentations in Japanese.  All I could gather was that the first presentation was on miRNA and DNA repair markers having some control over regulation of radio-sensitivity.  The second presentation was given by Hiro (see 7/5 post for picture of him) and was on a HSB 90 (a heat-shock protein) Inhibitor called 17 AGG and its effect on radio-sensitization of mouse tumour cells.  Again, I was saved by a very patient person by the name of Dr. Chris Allen.  Chris is an associate professor in the ERHS department at CSU and is here to use the HIMAC (Heavy Ion Medical Accelerator) for his own research.  Chris’s understanding of the two presentations was just about as limited as mine since he also does not understand Japanese.  HeeHee.  However, he was able to explain some of the DNA repair basics associated with heat-shock proteins to me and my mind did not feel quite so deviated.

Konichiwa!

Konichiwa! My name is Petra and I am another toxicology graduate student (in addition to Chuck) that is studying at the NIRS (National Institute of Radiological Sciences) in Chiba, Japan.  I just arrived along with Lauren (another internee from the toxicology department) a couple of days ago and will be here for the rest of the month so you can expect to see some posts from me over the next 4 weeks.  So far, we have moved into our respective dormitories and have already experienced some REAL Japanese Sushi.  Chuck, Lauren, and I along with Dr.s Ryuichi Okayasu, Fujimori, Kato, and Hiro all went out to dinner the first night we were here (see pic.).  We were exhausted from the traveling but enjoyed the good food and company!

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