30 April 2011

Procrastination Powers Activate!

Back again, hooray sporadic posting!  I hope you all enjoy the Nurburgring webcam. 
This past Thursday at my College’s fencing practice, we had a tournament to determine the first ever Silva Vulcani Champion.  We were fighting outside and towards the end the light was failing fast.  The final three came down to myself, one other older student, and a freshman.  The other older student I know can beat me on a regular basis; he’s got incredibly long arms and is a master of feint maneuvers. We matched up and circled each other for a bit, with a few half-hearted shots here and there to test the waters. We were both losing our game to the failing light, so I took a shot; a deep lunge to his center of mass.  The shot landed dead center, and he was out of the tourney (he had already lost to the freshman). 
Therefore it came down to me and the freshman.  By this point it was dark enough that I could see him and the guard of his sword, but not the blade itself.  I had one large advantage though, I had kept my dagger.  (Participants in the tourney could only fight with weapons they are authorized with; the freshman is only authed with a single sword.)  For a tournament like this, and against a freshman, I wasn’t about to give up that advantage.  If it hadn’t been me in the final, I didn’t want a freshman to win.  I helped start this group three years ago, I wasn’t going to give him any advantage.  Again, we made some passes with nothing landing, we were both playing cautious.  I decided to make a move before my luck ran out (and my light) so I made another lunge; he twisted to the side but not enough to dodge my blade; it drew a wonderful line diagonally across his chest up towards his left shoulder.  I forget if I parried his blade with my dagger or if he just missed, but his sword was nowhere near me.  He called himself dead and I was declared the first Silva Vulcani Champion.  If I make to Pennsic this August, I’ll supposedly have some official duty to do….
            This Sunday will be even more fencing as we’re having a mini-muster.  The focus will be on getting more people authorized with different forms.  I need to get my authorizations with buckler (small shield), cloak (like cloak and dagger), and case of sword.  Case of sword is fighting with two, usually identical swords.  I have very little practice with this form, but the Regional Marshal wants me to have all the forms before making me a marshal as well.  So I’m going to more or less wing the case auth, huzzah!
            My dad came yesterday to tow my dead Pontiac home, a distance of about 300 miles.  Starting the car up to drive onto the two dolly was not fun, it’s most certainly a blown head gasket.  This means the motor will have to pulled….again.  As to whether this means getting another new motor or trying to repair this one, I’m not sure.  I suspect the current one is too far gone.
            In other news, check out XPSMcLaren’s page for some exciting races this weekend.

25 April 2011

Spying on the Germans!

Watch some cool cars and regular commuters from Germany:
Nurburgring.org.uk Webcam

23 April 2011

My Week and Weekend So Far

My it's been a while. Afraid my life isn't terribly exciting.  Anywho, since we last spoke,
     My car, a 2000 Pontiac Grand Prix GT sedan, has blown its 3rd motor.  Yes, 3rd.  The first motor broke a valve spring within a few weeks of buying the vehicle. The 2nd motor's camshaft bearings went bad seven months after installing it. Yes it was a used motor.  Motor 3, the current one, has lasted about eleven months and is now draining coolant into the oil pan. Hooray!  Head gasket? Probably. Top or bottom seal? Hopefully.  Going to attempt fixing it before looking for a replacement. Though any suggestions on a reliable vehicle with a manual trans,  and hopefully a V6 would be nice.
     Last weekend was the Queen's Rapier Championship (yes more sword fighting).  I didn't make as far as I would have liked, I feel I shouldn't have lost to who I did.  Even my friends in the crowd didn't expect me to lose to that particular opponent.  It was a best 2 of 3 deal, she won the first two, barely and questionably, and then we did a third match for fun in which I destroyed her within seconds.  Must have lost my focus or something.
     Today my mom came out to visit and we decided to go hike and bike at Ohiopyle, despite that it was raining in the morning.
We walked in the rain for a little bit along the Youghiogheny River, but then the clouds cleared and the sun came out, making it almost too hot.  

     We continued down a ways past the first falls. We saw some large puddles teeming with tiny tadpoles.  Even saw a salamander.

After the falls we got the bikes out and went for a ride down an old Western Maryland RR trail.  Saw a lot of cool rock formations, including some exposed coal veins.  Saw some chipmunks and even a garter snake.  Anywho, I'm now with a bunch of friends, drinking, and watching Adult Swim. Good times.  Here are some more photos for your general enjoyment.


10 April 2011

Spreading Blades...and Hysteria of Course

     There's a reason I included "Blades" in the name of this blog.  This is because I like sword fighting.  I've been practicing rapier combat through the SCA for about three years now.  I've made a lot friends through it, and the best is, they don't mind when I stab them repeatedly.  Of course, they get to stab me as well, but that's what makes it fun.
     Yesterday, myself and a few others did a fencing demo for whole bunch of BSA Venture Crews.  There were about 400 Scouts at the camp, and we got about sixty of them to play with epees and watch us fight. We had the interested kids write down their names, emails and zip codes so we could help them find local groups to play with.
Sorcha and Adlewolf face off for a group of Venture Scouts.
     I felt pretty good about the whole thing since I'm an Eagle Scout.  I know Venturing works a little different, but I miss the atmosphere of Scout camps...other than the swampiness.  It's a general rule of thumb that land donated to Scouting is almost uninhabitable and useless for anything more productive, but the camps can still be very scenic.
     In other news, there's been much discussion in my SCA "kingdom" about whether or not we need to require "authorization" in epee and foil, also known as "light" blades.  Essentially, in AEthelmearc fencing, you must start with a light blade, and prove that you're safe with it by authorizing.  Once you've done this, you can play with "heavy" blades, which are more true to classical swords in weight, design, and technique.  I found it interesting that at this weekend's demo, people who've never held either type of sword before like the heavy weapons better, stating that while they're heavier, there's "more control" and it "feels better."  I feel this trumps all the current arguments for keeping light blades as a requirement in our kingdom.
     I feel like I had more to discuss, but it's slipped my mind, trying to do too much at once I suppose.  Anywho for anyone who enjoys cars and racing, check out the links below.

Awesome video from this weekend's F1 race in Malaysia:
More details about the race can read at XPSMclaren.

02 April 2011

Going the Distance?

I must say, one of the downsides of attending college several hundred miles from home greatly complicates the aspect of having a relationship.  At one point, there was a girl back home who wanted to date, and I wanted to date her too, but I’d only be able to see her on major holidays.  We eventually did decide to date toward the end of the semester, and it went well for a while.  The relationship fell apart for other reasons.
Now I’m faced with a similar issue. There’s a girl who lives near college who may want to date, but I’m about ready to graduate and may end up being far away for a long time.  We have quite a few common interests, one of which is cuddling.  Yes, I like to cuddle.  Problem with that is, we don’t know if either of us could go those long periods of time not seeing each other, without finding someone else to cuddle.
I’m worried that attempting to date will lead to disaster, but going back to just friends might make it awkward as well.  I’m personally not sure what I want.  Being single, I like that the option to cuddle with any willing girl is there.  At the same time, it’s nice having someone you know is just for you and cares about you and all that good stuff.

01 April 2011

So This is That Paper I've Been Talking About. And yes, it's 4:11AM

A look into the point mutation of c-kit that leads to mastocytosis

Mastocytosis is a disease in which excessive amounts of mast cells build up in bone marrow and skin lesions.  It has been documented as far back as 1869 in the form of a cutaneous disease accompanied by lesions1. The growth of mast cells is controlled by stem cell factor.  This process goes haywire when a point mutation occurs in the proto-oncogene c-kit, specifically an A-T substitution that causes Asp816 to become Val3.  This proto-oncogene codes for a transmembrane tyrosine kinase receptor, which helps mediate the growth and proliferation of these cells3.  Other immune cells, such as T cells and B cells carry this gene but do not express it on their surface, making them less prone to the disease even if they carry the mutation2.  Large advances in understanding the disease and the mutation that causes it have been made in the last twenty years.
Valent1 et al proposed a new classification system for different forms of mastocytosis due to modern advances in understanding different forms of the disease.  They note that patients can shift from one form of mastocytosis to another, each with rather different symptoms, even if they are controlled by the same process. 
This paper while possibly helpful at the doctor and patient level, does little to address the reasons and mechanics behind the mutation of c-kit and development of mastocytosis.  While they attempt to sort varying forms of the disease based on individual patient cases, the paper seems to go in circles and accomplish nothing that can be understood outside the medical community.  The differences pointed out between different cases seem trivial, especially when they are all linked back to the c-kit mutation.
Akin2 et al’s research was meant to find the how the Asp816Val mutation was spread amongst hematopoietic cell lines as well as cells with differentiation markers for myelmonocytic cells such as T and B lymphocytes.  They hypothesize that mutations in the proto-oncogene c-kit, is related to the genesis of Mastocytosis and that it may result from the enhanced growth of mast cells and their precursor cells that express surface c-kit and the Asp816Val mutation.
With blood samples from eighteen patients with Mastocytosis or urticaria pigmentosa, the desired cells were separated out using immunomagnetic beads.  These beads are coated with specific antibodies according to the desired cells.  Once the desired cells were isolated, they were prepared for RNA extraction, as the Asp816Val mutation shows in the RNA.  Using Moloney murine leukemia virus reverse transcriptase, the RNA was transcribed to cDNA.  This DNA in turn was used in a PCR process.  The DNA was then cut with Hae III and Hinf I restriction enzymes and run though a polyacrylamide gel and ethidium bromide staining.  The bands in the gel were then measured for fluorescent intensity.  Peripheral blood was sorted and analyzed with a flow cytometer.
The mutation was detectable in white blood cells, B lymphocytes in one patient, and other monocytes and myeloid cells in another patient.  They determined that it was the use of the specific isolation of certain cell lineages that allowed them to track the mutation.  The mutation was detectable in the peripheral blood of 4 of the patients.  Expression of the mutation was found in bone marrow cells and their lineages.  There was concern that the PCR signal would be false from imperfections from the isolation technique with the immunomagnetic beads, but it was shown that a mutated PCR signal was not detectable under their conditions.   The cytometric data met expectations by showing there is no detectable c-kit membrane expression in peripheral blood cells.
Nagata3 et al, working in 1995, hypothesized that Mastocytosis was linked to a mutation in c-kit.  They noted that stem cells and mast cells express c-kit and that a mutation in c-kit might result in the disease.  One of one patient with Mastocytosis had the corresponding mutation in genomic DNA.  Finding the point mutation that causes the Asp816Val would provide insight into the pathogenesis of the disease. 
Blood samples were obtained from eight patients.  Using RNA extraction techniques and Polymerase Chain Reaction, in fact, the same reverse transcriptase from Moloney murine leukemia virus as Atkin et al used, were employed to isolate the target DNA.  To check for polymorphisms in the cDNA, single-strand conformation polymorphism (SSCP) was performed on the PCR products, any polymorphisms found were sequenced.  These sequences were purified via agarose gel electrophoresis.  These purified samples were then amplified and sequenced.  The restriction enzyme Hinf I was used to find spots where A had been substituted with a T, which creates a restriction site not found in the normal DNA and therefore leads to different bands in a polyacrylamide electrophoresis.
The experiment showed a nucleotide sequence change in six patients.  The A-T substitution was confirmed by the Hinf I digestion of PCR products.  In the four c-kit-mutation positive patients, the bone marrow showed proliferative abnormalities.
The experiment showed that four patients with Mastocytosis and an associated blood disorder carried the A-T substitution.  The overall data did suggest that the Asp816Val point mutation was responsible for the pathogenesis of mastocytosis.  It was found that the majority of cells in blood expressing c-kit are CD34+ cells, which are progenitors of mast cells.  Although the researchers considered it likely that all descendents of these cells would express the mutation, they claimed it was not clear at the time.  The researchers determined that further screening of the patients and follow-ups were necessary to gain a better understanding of c-kit’s relation to mastocytosis.
In summary, mastocytosis is an uncontrolled proliferation of mast cells cause by a point substitution mutation of the c-kit proto-oncogene causing an Aspartic Acid to become a Valine. The various researchers above sought out to explain why mastocytosis occurs at the genetic and cellular levels.  They have pinpointed the location of the genetic error and how it affects the mutated cells.
            Hopefully with this knowledge, new treatments for the disease can be developed at the genetic level and be detected sooner than in the past.  Certainly more research needs to be done, but understanding how some cell lines start uncontrolled reproduction can lead to understanding other forms of uncontrolled cell division.

1. Valent, Peter, Hans P. Horny, Luis Escribano, B. Jack Longley, Chin Y. Li, Lawrence B. Schwatz, Gianni Marone, Rosa Nunez, Cem Akin, Karl Sotlar, Wolfgang R. Sperr, Klaus Wolff, Richard D. Brunning, Reza M. Parwaresch, K. Frank Austen, Karl Lennert, Dean D. Metcalfe, James W. Vardiman, and John M. Bennett. "Diagnostic Criteria and Classifcation of Mastocytosis: a Consensus Proposal." Leukemia Research 25 (2001): 603-25.Www.elsevier.com/locate/leukres. Web. 31 Mar. 2011.
2. Akin, Cem, Arnold S. Kirshenbaum, Tekli Semere, Alexandra S. Worobec, Linda M. Scott, and Dean D. Metcalfe. "Analysis of the Surface Expression of C-kit and Occurrence of the C-kit Asp816Val Activating Mutation in T Cells, B Cells, and Myelomonocytic Cells in Patients with Mastocytosis." Experimental Hematology 28 (2000): 140-47. Elsevier. Web. 31 Mar. 2011. <http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VP8-3YKKBN0-3&_user=3554035&_coverDate=02%2F29%2F2000&_rdoc=1&_fmt=high&_orig=gateway&_origin=gateway&_sort=d&_docanchor=&view=c&_acct=C000060848&_version=1&_urlVersion=0&_userid=3554035&md5=ba092a3e0dac0dd88ac568da5da3228a&searchtype=a>.
3. Nagata, Hiroshi, Alexandra S. Worobec, Chad K. Oh, Badrul A. Chowdhury, Susan Tannenbaum, Yoshifumi Suzuki, and Dean D. Metcalfe. "Identification of a Point Mutation in the Catalytic Domain of the Protooncogene C-kit in Peripheral Blood Mononuclear Cells of Patients Who Have Mastocytosis with an Associated Hematologic Disorder." Proceedings of the National Academy of Sciences 92.23 (1995): 10560-0564. Web. 29 Mar. 2011.