Living the Bucket List

I feel like I was a bird flying high dropping tears like rain over the earth. I witnessed sad tale above sad tale of women abused, the religious forming dischord, and the diverse peoples of the world fighting over money. As a child the news was never my primary choice scene to marvel. However now I have realized I have changed. I have changed and my life has changed. My environment changes and I am forced to change with it at speeds I am not comfortable. I break in the strong winds of change and learn that if desired I can lessen the impacts of the winds of change or exacerbate the situation. It's a learning process by which failure is the vehicle.
Soaring over the world taking a sample via CNN I see the pain others in worse positions go through to acheive their living. While I am fully equipped with a living, I still manage to put myself in a hell of suffering on par with those who truly suffer. I constantly beat myself emotionally falling short to realize the true beauties of my existence and environment. These abused women finding work have nothing and do not know any better. Children sold into slavery do not know any better and suffer so much more than I have, yet they know who they are. Even they have vision, so why not me?
Vision is something I struggle with. A healthy non-idealistic sense of vision is a trait I wish to have and learn to cultivate. I've wasted away enough of my life trying to jump out of my skin and into others. It's time to wrap myself tight and see where I end up.
I may not be like the people on the news--those on the Next Line. But I am an individual with individual dreams and will power. Will power is something I am discovering. Innovators inspire me because despite the odds against them they still plow on and maintain that determination and vision creating something beautiful. They realize their potential and take the risks to make it possible. Maintain the course in the midst of uncertainty. I admire that in others and hope someday very soon to see this play in my life as well.
What I'm getting at is seizing opportunity. Living an enriched life. Feeling spiritual again and therefore knowing myself.
Be the shark. Keep moving or you'll die. And yes, you will die from the inside out and be left wondering what happened to the world, but really it's all you. Live today like your last day. Live to be inspired. Live. Laugh. and Love

Dreams...

Why is it the only thing that gets me excited is thinking about ways I can get out of the country? I wonder who I know in other parts of the world, where I can go and just exist. What value do I think that will have? Don't I know I'll be neglecting everything I have here? A place to live, computers, books, clothes, friends and family nearby, hygeine, source of income? Why do I think making my life so difficult would be worth it when I can't even get by as I am now?
Perhaps the desire comes from wanting the extraordinary and keeping myself from connecting the dots. The extraordinary requires what I think would be an extermination of fear. The extraordinary would be the experience of a lifetime. The extraordinary takes a lot of money.
I think of small scale extraordinary adventures and I can see how I can do this. I vaguely have an idea of places I could go. Maybe India, most likely Argentina, perhaps California. if i split them evenly across the year it would mean 4 months in each place. It's nice to dream.
Money is a bit of a necessity haha.

And this is where the cosmic dancer comes back to earth. I have a lot to be thankful for here. Many opportunities and comforts, still a variety of cultures if I just look. I look everywhere but my own hands instead of tending to my own grass to make it greener. It's so much easier to wish for something you don't have. But alas, it's to work, build, acheive, and dream. Setting acheivable goals. That one up there is not acheivable. But what is acheivable is a smaller adventure. And I pretty much just recently got back from a trip to California that a salesman reminded me of. I'm clearly not happy with myself. I just want to reach this point where I'm not running anywhere. Where I have a bit of a steadier more settled mentality about myself. At times yes I dream and dream big. I think of the small group of people I know and point to them wanting to try them on. But I'm not like everyone else. I never will be. I'm not sure if that will click in my head anytime soon.

I haven't realized until today where my actual unhappiness comes from. It comes from big ideas implanted in me and ideals influencing me from home. Ideas I adopt because I am told, but have neglected to challenge them for a while. I also can't seem to understand that an acheivable goal means setting standards in place to reach them. I will always fall short without the proper discipline. I am very naive and idealistic about things which keep me from experiencing for what they are. Make the most of life and each moment we are given. I am so pissed with myself for letting anxiety take over my life for 2 months. It was a waste of my life and I'll never get those moments back. Moments when I could have experienced full range of my emotions I know I am capable of won't come back. So everyday I need to consciously remind myself to live each moment to the fullest since it doesn't come naturally for me yet. It's why my zest has left me. It's why my passions have faded. It's why I see myself as less than others because I know I am less than myself.

If I knew I couldn't get a moment back, what would I do?

Discipline is the key. Maybe military service would be good for me...

Confidence

Lately I've been going through a rough time, when it finally dawned on me. Life is not absent of the tough times, and the easier cheery times are not such intrinsically. It's all about your outlook and your ability to take mountains for mountains and molehills for molehills. It's less about strength and more about calm and focus. I question myself of late, and so have delt my self confidence a heavy blow.

Confidence I've read, in it's Latin root, means "with trust" and so I can say I have lost trust in myself. The idea was posed that we lack this self confidence when there was an advantage to someone else to make us feel this way. I see clearly as to how my childhood has raised me to be this way. Taking several steps backwards to discover myself as I live on my own and stand on my own two feet I see just how shaky I've been raised to be. Crippled to remain so close to home and maimed with blurred vision to keep my dreams from me. I even wonder if I know what my dreams mean, but that's just it. Trust myself. That I'm taking positive steps and will always be on a journey of self discovery no matter where I am.

World travel. It sounds wonderful but is it a goal of my life? If it was truly a goal wouldn't I have worked that into my career plan? Is it really something I want since I covet it so much or is it just my displeasure with the way things are here. It may be another dream I need to fulfill. Which will be outweighed? This is something I need soon: to travel the world or a small portion I wish to see. I can immediately see not getting into school next year as a huge blessing. This means I can have a chance to see the world before long and again be encouraged. Pave the way for myself. Maybe that's the best idea.

More dreams, dreams of rainy days in Seattle or flowers in crisp summer weather in the midwest. I will go places and not be afraid to step outside. I will have a dynamic and adventerous life. Let no one take that away from me. These are my dreams to have for as long as I want them to be my own.

And in light of my recent love affair, why can't I connect the dots with that one. Somehow it's connected to confidence. Maybe I felt I could trust myself only if I could corner myself with him? Maybe I needed more of a go getter? I don't know, but I lost myself somehow.

More later...
musing

Pursuit of happiness

There's an anathema after me. It's come to a point I only remember once every year. I've been either hiding or running away and I'm hanging on a cliff all daggers out come to make me pay for my choices. My choices are my own, and they are what have put me in this awful position. A position where I am everyone's enemy. I can consider the situation from all sides. I can say why am I so bull headed with family and not with anyone else. Why do I take offense with my stupid teenage sister when I can't say if I was any different at her age. In truth I probably wasn't. I know many times I'd keep her out of my life not treating her with compassion. I was so stressed I wouldn't care to entertain a stupid middle school child. I remember my mother used to encourage this by comforting Sonia and telling her Ali is being mean and it's okay don't worry about her Mommy's here. And so I began the vicious cycle that has now come to bite me. My relatives do their best to haunt me while they are still alive, about their God who helps them through everything, "they turn to God" and put their trust in him. I know better. You do you're best with what you have and where you are. The world turns because of the motions set in place and the choices of the people living on earth. These choices affect our own and thus we call it fate...God...Allah...karma. Humans out of their own insecurities feel an innate need to judge others' decisions. And so we hinder each others' free will and understanding. We do not 'get what we deserve'. It's just human nature to want to get even. There is cause and effect and choices. Whether they are right or wrong is up for opinion. The truth is what keeps our world going is cause and effect.
The cause was my own hatred and stupidity when I was a teenager. And now I am seeing the effects of that. If you sow a seed into the ground, you will reap the benefits. Or in this case, detriment. I don't believe a God smiting humans and all creation. I don't see us paying for consequences. It is simply a cause and an effect.
I can spend a week under a tree meditating on what went wrong with the cause or I can choose to move forward and live.
I obviously cannot learn to get along with my family. I have developed my own way of thinking but cannot quite come to terms with living in a judgemental environment with people whom I've been told are to provide you with love and warmth. I always made the mistake of telling my parents my plans. I've made the mistake of trusting them with my relationships. I've made the mistake of misunderstanding their convoluted versions of generosity. I've learned a lot.
I've learned to take gifts even when you don't want them. I've learned that anything you say will never be taken the way you mean. I learned to step back from attempting to raise my own bratty sister.
What I didn't learn was to keep family problems to themselves. I've never once been truly happy since living at home. All my happiest times no matter how disgusting were outside the home. In some eyes I've sinned and disrespected. But merely cause and effect has played out. A cause I knew better than to set into motion and have now become more miserable because of it. I would call that a bad choice.
And so, I am sorry to myself for that. I am sorry I have a will and mind of my own. I am sorry I still cannot tolerate only critical and negative comments from my parents especially my mother. I'm sorry I have learned to be almost as cold and heartless as my younger blood relative. I am sorry I try to be everything you want me to be. I am sorry we don't get along. I am sorry I stayed out of mercenary means. I am sorry I was so foolish to think we could actually live in peace.

So now I hope we can accept what we've needed to all along. I'm sorry Daddy that you're hope of a real family is probably never going to happen, but I love you dearly for being so strong to put up with what I cannot. I am sorry you cannot know who I really am, but I must protect myself and pursue the right this wonderful country has given me, the pursuit of happiness whether I may find it or not.

I love you

First lab meeting

Today Nahir and Oleg gave presentations on their biochemical projects involving AdoMetDC, which is a pro-enzyme for which T. brucei needs in order to synthesize necessary amino acids. The sequence of this pro-protein at the N terminushas been concerved across several species excluding mammalian species including humans. This makes it a viable protein for drug targeting. Nahir's project involves creating a better way to produce more of this target protein in order to purify the protein in it's entirity for crystallography purposes. It seems her first problem was mainly getting the N-His tag to come off the protein without disrupting it's enzymatic activity--you need to be able to prove that and therefore you should be able to first cleve the N-His tag. She had problems doing that. So she cloned the gene into a vector in a new way that included a novel product known as th e SUMO fusion sequence. This sequence assists the protein in it's cloning and facilitates folding, therefore increasing the amount of viable protein. She first designed her primers and also designed primers to cleave portions of the N terminus off the gene, later testing for enzymatic activity. How to test for enyzmatic activity and not just folding? Immuno-assay. or is it co-immuno precipitation assay?

So to ensure the gene was in the new vector she ran a gel. Showing the reaction inserted. She may also may have had to do some mini preps. She then digested the vector to show that she could isolate the gene and put it back inside. Isolating the gene meant she needed to sequence it.

confirming the correct gene she then began the process of large scale bacterial cultures. She'd lyse the cells and then purify the protein with the Ni2+. Through a 2 step process of running the solutions through the column on a gradient, she was able to isolate the protein without the His-tag. She would run gels using affinity chromatography which elutes the proteins based on size. In some mutants with a more upstream N-terminus deletion she lost the pro-enzyme associated with AdoMet. Ideally you want a clean protein for crystallization purposes.

Oleg was having trouble eluting enough of his protein. Seemed that in purification processes he would lose more than half of his protein. It seemed that there was too much protein and the column was too old to handle becoming overloaded. It was recommended to run more batches but diluted and with a better column.