outsourcing this post

Last weekend I was hanging out with my brothers. Justin was good enough to spend some of his precious leave time with me before he heads to Wisconson then Iraq. We had some fun together up at Sunny side park. Go here to see. Tim (my brother in law) and I hung out by taking the tubes up (by car), then down (by water), the Selway river. We hit some nice rapids, hit some not so nice rocks, took in the silence, and watched fish swim around. Go here and scroll down a bit.

Thanks guys! It was a lot of fun and I enjoyed it all immensely.

Edit: We seem to be popping up all over the blogs. Click here for proof that we went to grandpa’s in Port Orchard and hung out with the Deffenbaugh’s.

at the farm

Thank you Dad and Laurie for having us and our friends over last weekend. I realize the fortune of having a loving family who appreciate the company of me, my family, and friends. Daniel, Jen, Marian and I all felt at home and there wasn’t a moment when we wished to be somewhere else.  I am grateful how you share your time, hospitality, and food without reservation. But most importantly, I am greatful of the faith we share in Jesus Christ who provides rescue from death. I love you both.

Here are some of the pictures we took before the camera quit working:

Marian very very happy

Jen and Shadow meeting each other

Alice realizing the farm is wonderful place filled with interesting smells. She carries her prized oyster shell.

Wiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiii golf

Alice finds commonality

Dad likes to cook good things. Tonight for dinner is halibut wrapped in bacon, twice baked potatoes, corn, and tomato slices. good good good.

No trip to the farm is complete without several pinochle games with grandma Carol.

Photogenic

A dentist friend came over to my paramedic father to get fixed up after he puts a knife through his finger.

The pictures don’t show all that we did, but on the way home everyone felt like Alice looked.

Marston/Gibbs/Laughery family vacation photos

We took a little weekend trip to a nice bread and breakfast that sits on the Clearwater river. Here are some of the pictures we took of our excursions. I think it will be more fun if you insert your own commentary.

*Thanks to Paula for taking most of the pictures.*

weekend schedule for the rest of our summer

July 12-13

• Work, work, work, bbq, work, then sleep.

July 19-20

• Marston/Gibbs/Laughery family vacation. Includes bed and breakfast, hiking, and maybe a little Rook

July 26-27

• Possible trip to the Laughery farm with a couple friends. Expect to see if the waves are any good for surfing, do a little horsing around, have a totem milkshake, and maybe a little Rook.

August 2-3

• Meet Dad at uncle Pat’s for dinner.
• Possibly go mountain biking with wife and dog.

August 9-10 [Edit]:

• Hang out with Tim.

August 16-17 [Edit]:

• Hang out with the Deffenbaugh cousins in Port Orchard!
• Go to a bbq in Bremerton at a friends new house.

August 23-24

• Gear up for the WSU fall semester starting the following Monday.
• Summer is unofficially over. Visit our offices if you ever want to see our faces.

we celebrate two years!

And give divorce the finger.

I ride a motorcycle

I caught the motorcycling bug early in life. My dad had a street bike for a little while when I was around four or six years old. He would sit me on his lap and take me for a little ride around the neighborhood. I don’t remember too much about the bike or the time, but I do remember feeling privileged whenever I could take a ride. Later, my parents let me spend three summers’ worth of saved up lawn mowing allowance on a used dirt bike. When the bike wasn’t broken down, or when my motorcycle privileges weren’t rebuked because of unrelated disobedience, I had a lot of fun on it. After it finally died I didn’t ride until just after I was married. Marian and I (emphasis on the I) wanted a second vehicle when schedules conflicted. An old motorcycle seemed like the most affordable option, and it was an old standard that could barely reach 55mph which became the solution. The season that this bike ran was all that was needed for my motorcycling bug to resurface. Since then I have learned much, and tested many bikes. After owning another that was really too shiny, I finally found this: 

It is a 2003 Suzuki SV1000N. For some reason Suzuki only made the N (naked) model for one year but kept producing the S (sport) model, which makes this bike hard to find.

There are two initial responses whenever someone learns that I ride a motorcycle. One is “cool, what kind?” even though they have no idea even when I do tell them, and the second is “be careful” or similarly “don’t kill yourself” to which normally I respond only with a smile, but say in my mind “you’re the one driving the 4000lb car; you don’t kill me, please”.

I don’t deny that motorcycling is dangerous; I have read the statistics. In 2001 the fatality rate per 100,000 registered motorcycles was 88, and the fatality rate per 100,000 register passenger cars is only 15.72. This means 0.88% of all the motorcycles of 2001 in the US resulted in the death of it’s driver, and a motorcyclist is five times more likely than the driver of a passenger car to have a fatal crash. However, if you take a closer look at the statistics you can start finding ways to beat the statistical odds (become non-normal). Of all the motorcycle fatalities roughly 30% of them involved the driver having a BAC of 0.08% or higher. So, by not drinking and driving, riders will reduce their statistical chances of a motorcycling fatality from 0.88% to 0.62%. There are many other factors that may reduce the chances of crash fatality, such as proper gear, like a helmet and armored clothing, riding during favorable weather conditions, riding during favorable traffic conditions, and so on. There are a lot of riders that don’t take these precautions who, if they had, would still be riding. These stats only pertain to when a motorcyclist gets into an accident, but let me tell you why motorcycles are better equipped than cars to keep you out of an accident.

Awareness:Obviously, in a car or on a bike, being alert and aware of your surroundings keeps you out of accidents. However, on a bike the level of awareness is naturally increased. When riding a motorcycle you are out in the open where the senses are stimulated, and this helps influence safer diving directly and indirectly. For example, when the temperature is below 32F it is very uncomfortable to ride the motorcycle, and I take the car instead, thus keeping my two wheels off of the icy roads. Likewise, my senses alert me when riding on windy, wet, gravely, or oily roads, whereas in a car, the outside conditions are muted to my senses. The other way a motorcycle increases awareness is the nature of the controls. All my limbs are used in operating the bike, and my body weight is constantly being used to balance and steer. Driving most motorcycles (large cruisers potentially exempt) there is no convenient opportunity to chat on the cell phone, mess with the stereo, eat a fast food dinner, converse with a passenger, or any other thing that my distract from what is going on the road. In fact, as a motorcyclist, I always am aware of, and increase the distance between, me and the driver of the passenger car that is talking on the cell phone (blue tooth or not). 

Deceleration: My 500lb motorcycle is equipped with three disk breaks. Two in front with 310mm rotors and four-piston calipers, and one in back with a 220mm rotor and duel-piston calipers. I don’t have the stats for stopping distance, but an instructor at a motorcycle training school once said that a novice rider can stop his bike just as quickly as a car, and an experienced rider can stop much quicker. On a motorcycle, you know when you can’t stop any quicker when the rear tire starts leaving the road. 

 

 

 

 

 

Acceleration: Yes, acceleration is used for safety. The most dangerous place to be on the road is around other cars. So, if you put them behind you, the chances of one of those cars not seeing you is reduced. Most motorcycles are naturally quicker than cars, which allows the rider to get ahead without putting much strain on the bike. If fact, my motorcycle is much happier accelerating from 4500-9000rpm than it is lugging along at 3000rpm. This safety practice must be utilized in a safe and respectful manner. Novice riders may get in trouble by using too much throttle for their experience, and, even towards the experienced riders, this practice is often looked at with contempt by other motorists who do not understand the reason a motorcyclist may jet out in front of them.

 

 Size: Being small on the road can make you less visible to other drivers, which is not a good thing, however there are benefits when finding yourself in a tight spot. Along with quick acceleration and deceleration, the small size of a motorcycle leaves many more options of escape over most passenger cars. If a car cuts into my lane I can easily use whatever shoulder there is to escape from being hit. If no shoulder, I can nearly fit in the same lane as the car cutting me off until I can get away. Also, I have many options of lane position. Normally I use the lane position that keeps me most visible, but if there are any obstacles in the road it is no problem to miss that item without slamming on my breaks creating potential for being rear ended.

 

 

Rider Education and Training: In Washington state it is mandatory for a rider operating a motorcycle with an engine greater than 50cc to have a valid driver’s licence and pass a motorcycle written and skill test. This is minimal, but because motorcycling for many is more of a hobby than a necessity, most motorcyclist keep reading up, and practice how to drive more efficiently and effectively. There are a number of books and online resources that talk about safe riding. Many motorcyclist often practice safety maneuvers on their bike on a regular basis. There are many opportunities for advanced motorcycle classes for any one with a valid endorsement and a bit of cash.  For sport bike riders there are track days where many go to keep the fast riding off the public roads and on a safe course where falling off your bike at 100mph is survivable without need for a hospital trip. These practices will make motorcyclists better drivers whether riding their bike or driving a car, decreasing the potential for accidents.

————————————————————————————————————————————–

 

At the beginning of the month I met up with my dad and brother in Ephrata, and we all went for a little ride together. Riding in a group is very enjoyable especially when the group is your family. I hadn’t ridden with my dad since last year when we took a trip to Oregon to see my grandparents, and my brother is taking a one year tour in Iraq with his squad this coming August. It was good to hang out with both of them. I don’t have too many pictures to show, because, well, we were riding.

 

 

 

 

Some river we used to cool off a bit.

 

 

Grand Coulee Dam being released.

summer snippet

immunofluorecence assay (IFA)

The term antibody (Ab) is pretty self-explanatory: anti “against”, body “body”.  Ideally, the “body” that the antibody is “against” is a foreign substance that breaks through your first line of defense (skin, hair, mucus) into your… body.  How the antibody works has a very complex answer. However, let me give a general explanation of how our immune system creates them.  In the body, our immune system is made up of a very diverse infantry of cells. When a foreign substance (bacteria, parasite, virus, transplant organ, another’s’ blood, or baby) enters our body the immune system doesn’t just start shooting random Ab missiles at the intruder. If it did, those random Ab may miss the target and start hitting “self”, and, almost always, shooting missiles at yourself is not a good thing.  Instead, our immune system has several fail-safes to keep the Ab specific for intruders only. It all starts with the T cell…well, it actually starts in the bone marrow where T cells differentiate from stem cells. Naive (inactivated) T-cells travel to the Thymus where they are put through an immuno obstacle course to determine if they will recognize and attack self. If a T-cell is unfortunate enough to recognize “self” that T-cell is executed immediately, but if not, then it is free to float around the body looking for foreign substance, or we will now call it, antigen (Ag). Now, when the T-cell finds a foreign Ag it takes a chunck of it and travels to where the B-cells hang out, the lymph nodes. Unique to individual B-cells are receptors attached to its outside surface with a unique combination for recognizing, or not, Ag presented to it by the T-cell. When a B-cell has the right combination for the Ag it will send signal to the T-cell and wait for the T-cell to signal back. Only after the T-cell “pings” back will the B-cell differentiat into a plasma cell and start making antibodies with specific targets against the antigen.

All this to say, the antibody is very specific for a particular antigen. Not only is it specific to the Ag, but it is also known that the Ab is only specific to a certain part of an Ag called an epitope. Having something that is so specific is not only helpful to fight off the right pathogens in our body, but can also be a helpful tool in the lab. Immunofluorecence is one way we utilize the specificity of antibodies to determine and localize a particular antigen. 

I will show you how to do an immunofluorecence assay just incase you need to determine an Ag at home.

Here is my antigen I will be testing. These two wells contain an intraerythrocytic (inside red blood cell) parasite called Babesia bovis. By growing these parasites in culture I am able to have immediate access for use in my experiments.

To conduct my assay I make a blood smear from my culture on to a glass slide.

I am fortunate enough to work in a lab that uses very expensive and elaborate instrumentation, but sometimes you just go with what works the best.  Fingernail polish does the trick here.

With the fingernail polish I fence in a section of my Ag that I will use against several different Ab.

Antibodies for laboratory use are created by inoculating a bunch of known antigen into an animal (mouse, rabbit, goat, whatever) in order for it to launch an immune response against it. Then, when the immune system of the animal is at its peak, the specific Ab for the known Ag can be isolated. Here I have several Ab that I want to test against my Ag; most of them are for negative and positive controls. Not only do I have different Ab, but I also use different dilutions to aid in a sort of pseudo quantification.

Here I spot the Ab on the fenced in Ag, referring to my napkin map along the way.

Let the samples incubate to allow the Ab to find and bind the Ag.

Rinse and…

wash.

Now my Ab is bound to the specific Ag that my parasites may or may not have. But now what? How am I supposed to see it? This is where the “fluorescence” meets the “immuno” in my IFA. Antibodies look like a capital Y with the specific Ag binding part on the top two ends, and antigen looks like spaghetti. So, you can imagine a bunch of Y tails sticking up on one particular area on a noodle in a pile of spaghetti. It is only the Ag binding part of the Ab that is specific, and the tail is common to almost all types of Ab. We can create a second Ab that will recognize and bind to the tail of the first. Not only that, but we can put a “tag” on the second antibody that we will be able to visualize. I used FITC for my tag in this experiment, and it goes on the same way as the primary Ab.

After my primary and secondary antibodies have been fixed I just go look at the slide with a scope. This scope has a camera mounted on it so I can have a live video feed and take pictures. It also is able to shine different light spectrums at the sample, which is fortunate since my tag fluoresces at 532nm when a blue laser diode hits it.

Here is a negative control that informs me of the background level.

 

Each green spot is where a parasite has the peptide specific for the Ab I used. I can conclude that my parasites express a gene we call RAP1 (Rhoptry Associated Protein One), which is known to aid in cellular invasion. 

“normal life” for Tim

fairy rings

Mushrooms are beginning to become a bit of a hobby of mine. They are unlike any other plant or animal, and I’m always curius when I find them outside or pass them in the grocer. I remember one of my first mushroom hunts with my dad as a small boy of 7 or 9 years old. On a whim we stopped the car on the shoulder of the highway that ran from our house to the nearest town, and hiked straight into the forest. I remember walking through the giant wall of trees that the utility workers cut back far enough from the power lines to keep stray branches from potential power outages. Once through the wall we passed into a soft, misty land where the traffic of the highway was muffled and the silence of the forest was noticeable. In there I was taught to speak with a hush, and to not step on sticks that may snap in order to keep the woods quiet and alert. My dad and I were searching the moss covered forest floor, and discriminating against the many inedible mushroom varieties.  I don’t remember the variety we were after, maybe the Chanterelles or Corals, but I do remember we happened upon quite a score. The most distinct memory I have is that we had no bag to fill or sack to hold our find, and it was too much to carry out in our hands. Without much choice my dad opened his jacket to reveal a deep inside pocket. We filled my fathers pocket with the fungi we had collected.  Once home, I remember that there was quite a bit of dirt forgotten inside the pocket my father’s jacket in the excitement of preparing those mushrooms.

After the snow melted yesterday and the rain stopped this morning, I came home to find a fresh ring of mushrooms in the yard. I had noticed this ring before, but not payed much attention until the mushrooms were old and dried up. I did however look them up in our trusty “Audubon Field Guide to Mushrooms” that I believe Tim gave Marian or I as a gift. It turns out that the ring of mushrooms were an edible variety called marasmius oreades “fairy ring mushrooms”. The book comments: “It dries readily and becomes easily overlooked in grassy areas: then, when it rains, it revives and appears to have sprung up overnight. In early times, the rings were thought to be magical places where fairies bounced in a circle on moonlit nights or gnomes buried their treasure.” Sounds like a cool idea for a water painting if you ask me.

 

These are the ones in my yard. The rings increase in size each year as the mycelium expands out to find new nutrients. 

 

 

 

 

 

 

 

 

 

 

 

Here I have them drying out a little on my table. Unlike the Morel which has a “foresty” taste and a meaty texture, these mushrooms have a “woody” taste and a more fibrous texture.

 

You will probably see more posts about mushroom in the future, and don’t be surprised when my laboratory skills start playing a part.