Rare things

I'm trying to get my hands on a copy of a very rare animation from 1975 that I watched in Psychology class senior year of high school. I just recently remembered it and how it made me feel and looked it up. The title is "Everybody Rides the Carousel" and it is a cartoon consisting of different watercolor techniques (from what I remember) based on psychologist Erik Eriksson's idea of the Eight Stages of Man. The cartoon features a jester who is like God taking you along a carousel that stops at each stage of development of a normal human being from birth until death to give you a peak at what life is like. The jester narrates it with a calming, nascent voice.. the kind of voice you'd use to whisper into the ear of a new-born baby or someone on their deathbed. It is very stimulating, reminds me of how it feels to wake up bewildered in a late afternoon during the end of summer after a long strange, dream-filled nap and I am sweating. Some of the images still haunt me.
Another rare video that I am glad I actually own (back in Illinois) is "The Snowman," the short animation we used to watch as a family and often with the Gulezians around every Christmas time. We have another VHS copy than the one we used to. The old one was slightly shakey since it was a TV recording. It had an antiquated feel to it, even then. The video started off with a real-life man walking through snow near the edge of the woods, and it always reminded me of the back field behind our house. The man stops and his British voice comes on as a narration. He says: "I remember that winter because it had brought the heaviest snows I had ever seen. Snow had fallen steadily all night long and in the morning I woke in a room filled with light and silence, the world seemed to be held in a dream-like stillness. It was a magical day... and it was on that day I made the snowman." And then the video turns into a cartoon and this really dramatic piano song comes on. That video (and its contents) is an important thing from my childhood. Thinking of it now brings me back to a nostalgia of Christmas in Chicago. Everything from picking out the Christmas tree with my mom and decorating it with my brother(s). Sometimes snow would fall on the Day, and sometimes the Eve which we spent with my dad and went out to eat at Grand Mandarin. And it makes me think of the cookies I'd put out for Santa, the following morning reduced to crumbs and a thank-you note scribbled in my mom's hand-writing.

Fragmented thoughts at 2 am

I'm rolling into the finish line of my undergraduate degree. Exactly two weeks from tomorrow at 10 am I will be done with school having accomplished a biology degree at the University of Oregon, so one cannot blame me for my erratic sleep patterns, my difficulty focusing on school, and my present blogging moment.
I finally set up my desktop computer today because my life was nearly without music and that has been greatly disturbing me for awhile. So for a couple hours now I have i-tunes set to shuffle and it feels great. I thought I'd scribble a few notes down before bed because sometimes it can be therapeutic and help me get to bed. I like to write sometimes because it reminds me that I am human; I remember riding in the passenger seat of my friend Simon Kwan's rice-burner Honda while we were driving all over the Bay area... night had approached and he was telling me about his internship where at the end of everyday he feels less of a person after a day of mind-numbing busywork. Writing keeps me from that feeling sometimes. I remember the car ride well, as it was a very happy moment for me. Simon is a friend I really value, absolutely one-of-a-kind.
That's not what I wanted to go into though... there were a few other unrelated thoughts. I may as well just throw them out there and let the audience organize them.
Often times as I lay in bed at night, trying desperately to fall asleep with no luck in sight, I carry on imaginary phone conversations with my dad in Chinese. I guess it is a good way of practicing my Chinese nowadays.
I want to star in a horror movie, preferably a nice, stylish one but any horror film would surely be fun to participate in.
I have few friends in Oregon anymore and I really don't care. Most of the people I have met here I got bored of really quickly. I know exactly what I look for in people, but I don't know how to define it. My friends are as follows: The Architect, The Historian, The Anthropologist, and The Economist/Biologist. One of them is my roommate. Two of them are my coworkers in the organic chemistry lab; one of them is Nigerian. Two of them I have dinner with almost every Wednesday evening. One of the two I have dinner with I do not meet with otherwise. One of the coworkers I do not meet with outside of work.
Sometimes I miss my bed in Nanjing. I liked sleeping in on the weekends with the curtains pulled shut, it cast the room in a nice light conducive to rest and incubation. I also miss the hardness of the bed contrasted with the softness of the comforter.
When I was in Thailand I hitched a ride with a tuk-tuk driver who told me he would take me to all these wonderful temples as long as I go to a tailor shop first, all for a low low price. I agreed and jumped on and he zipped through horrific traffic to a tailor shop, stopped, forced me inside where I was shuffled into a back room and brought face-to-face with a Burmese man where we had a private chat. He asked me where I was from and why I was in Thailand. I told him about my study abroad going on in China. He mentioned he used to have Chinese friends. Then he said, "So would you like a suit?" The tuk-tuk driver was waiting for me outside in his colorful ride and then he took me to some really pretty temples. After the second I went out to meet him and he was gone. I waited for the longest time and even walked around the temple to see if I could find him. I had never paid him, but I needed to eat and get to the boxing stadium which was still a ways off. So I hailed another driver hoping karma nor the original driver would ever catch up to me for not paying. He knew I was going to the boxing stadium too.

Saurabh, Redux

Methinks it is nearly time to pull “Saurabh’s Darkest Hour” out of the old vaults, a short-story of which I have been continuously conscious and editing over the course of probably a decade. Julian first introduced to me the ill-fated story of Saurabh, and around that time (4th grade) I wrote the very first draft of “Saurabh’s Darkest Hour,” which underwent several revisions for years to come. The last draft to be written was in 2005 when I was a senior in high school and enrolled in a creative writing class. At that time, I submitted “Saurabh” as an open-ended short-story piece and got positive feedback from my teacher. As the years went by, the Saurabh character became more developed, his story more existential and persistent. I stayed quite true to the original in its core, though this of course was a loose adaptation of the “true” story Julian had told me when he was 16 and I was 10. That last version of 2005 was filed away indefinitely, crystallized in an adolescent form, replete with angst and yearning. Indeed, the first “Saurabh” was very well suited for a pre-teenage mentality: action-packed, irrevocably goofy. Then, the early teen phases displayed more of an undeveloped sense of desperation and anxiety with a self-conscious approach, which finally gave way for the no-holds-barred I-don’t-give-a-shit-if-my-parents-read-this reworking that revolutionized the story and gave it a significantly deeper message. The version that exists today is flawed. I think it will be forever flawed, but I can at least rework it to the best of my means, like a physicist breaking down particles faced with the obstacle of a limitless smallness of things. If I do revise it with much sweat and tears (quite possibly bloodshed too), it will not be much different. Saurabh will stay the 18-year-old me, not the 22-year old (as of now) me. But the betterment of my writing since 4 years ago, and a little bit more common sense as to what works and what doesn’t may be poured into it to allow for a more solid piece. “Saurabh” has become, after all, a story of adolescence and therefore must stay in that moment of time. Any changes I must make will be purely cosmetic; although maybe some subtle changes that affect the message will be undertaken as well. Stay posted…

hookah

So languidly does the smoke rise from my contorted mouth after a big inhale. The freedom and pace the smoke embodies is what I wanted in certain moments. When I let it all out, it would stream through my lips like a river, shaping itself around the fine patterns of air-flow as the water does to its scattered rock bed. Watching smoke unfold over and again is like a sped-up rendition of a flower in bloom minus the pretty colors and instead of decay it simply disappears from sight.

"all that we see or seem...""

Before I forget completely, I should mention a dream I had not long ago. I'm not trying to transform this blog into a dream journal, but while I am on the subject and all...
I dreampt I was walking in a forest and I found a corpse with its severed head laying next to it. I was frightened, but I reached through some branches of a fallen tree next to him and pulled his body out, found a place to dig a hole and buried him. A week later some guiding force within me told me to unbury him, take his remains to another spot for proper burial. So I dug him up and he was in a big gray plastic container where people usually store sports equipment or toys and I carried him through a marsh (very reminiscent of the marsh in our yard in Downers Grove). While wading through the reeds and water I turned into Leonardo DiCaprio. That is all I remember.
My ability to remember dreams vividly is getting better after a long absence. A life without dreams feels like a life without music in some ways. I think of a Tom Waits song, a certain line that goes: "Life is so much different than it is in your dreams," and while I understand the metaphorical context of that saying, I always take it quite literally. Example: This morning I awoke from a dream in which I was flying above a colorful little town and caught on to a sky-tram. Delightful as it was, I woke up feeling sad that I could not fly and immediately I thought of the line from that song. Last night I was hit with a whole assortment of dreams. In one of them I invented a viral strain... maybe it was of Ebola? Yikes...
Last week I was taking a nap between classes and dreampt I had a bacterial infection and I was performing a series of diagnostic tests (much like I have been doing in microbiology with a set of unknown bacterial types) to identify what it was. I narrowed it down to two species and was set to finalize my tests when I awoke from an incoming phone call. It was my friend Simon who goes to UC-Davis. He told me in his Cantonese-twangy voice that my dreams of bacteria were "crazy" and he ought to let me get back to them. Was it methicillin-resistant Staphylococcus aureus (MRSA)? Yikes...

Maggot Therapy

The other night I read about maggot therapy right before going to sleep. Maggot therapy is the use of maggots (fly larvae) in certain wounds as a debriding and antimicrobial agent. The maggots selectively eat away necrotic tissue leaving live tissue unharmed while giving off antibacterial secretions to which many skin populations of bacteria are not immune. Perhaps because the generational cycles of flies are rather quick, I wonder if the maggots’ antibacterial properties are able to keep up with the resistance that bacteria can develop in just a few generations (?). At any rate, I really like the idea of maggot therapy actually. But then that next morning about half an hour before my alarm went off I had a dream that I woke up at 8:48 am, just 12 minutes before class was to begin. Panicking to get ready and run out the door, I went to the bathroom to freshen up and low and behold half of my face and right shoulder and pec region were decaying and yellow puss was oozing out. Meanwhile, packs of maggots were in bundles on my skin and I could feel them writhing. When I woke up the thought of maggot therapy wasn’t as appealing as the night before.

Shiga Toxin

Shiga toxin (STx) is encoded and expressed by the bacterium Shigella dysenteriae and largely contributes to the pathogenesis of shigellosis, otherwise known as bacillary dysentery [1]. Among the three species of Shigella that are capable of causing the condition, S. dysenteriae is regarded as the most worrisome owing particularly to its expression of Shiga toxin. The disease shigellosis is characterized by severe bloody, mucosal diarrhea and is endemic to developing nations with overcrowding and little or no sanitation. Correspondingly, the amplified bacteria are transmitted through fecal contamination of water or food, although only a remarkably small cell number is required for infection. Recent estimates indicate that Shigellosis is responsible for over 100,000 deaths each year, predominately among children in the developing world [2].
Irrespective of Shiga toxin, Shigella bacteria invade an infected host’s colonic epithelial cells by means of surreptitious M-cell transcytosis, followed by macrophage lytic activity. After successful entry into the cells, the bacteria proliferate and spread to adjacent cells, often killing them and inducing inflammatory responses [1]. The effect of such activity assists in the further invasion and ulceration of the intestinal mucosa; instances of low-volume diarrhea associated with dysentery are an attribute of the bacteria-induced inflammatory response. In spite of this toxin-independent aspect of shigellosis pathogenesis, the Shiga toxin can ultimately be held accountable for much of the devastating effects of Shigella dysenteriae infection [3].
The Shiga toxin is composed of an A subunit connected to a pentameric B moiety. The B subunit acts as a binding domain by preferentially binding the cell-surface ganglioside globotriaosylceramide (Gb3) [4]. Gb3 is most prevalent in a select few epithelial and endothelial cell types, which will become important later on in the discussion of STx use in research. Once bound to the Gb3 receptor, the toxin-receptor complex enters the cell via either clathrin-dependent or calveolae-dependent (lipid raft) endocytosis, the precise mechanisms of which are still not entirely known. The early endosome-encased toxin is then described as being shuttled
directly to the trans Golgi network (TGN), circumventing the late endosome pathway that leads to degradation in a lysosome. During its transport in the early endosome, the A subunit of STx undergoes proteolytic cleavage by furan, a crucial step in activating toxicity. In this manner the A subunit is split into two components (A1 and A2) held together by an internal disulfide bond. The endosome is transported in retrograde fashion beginning with the TGN through the Golgi apparatus and subsequently on to the ER. In the ER, the A1 component of the STx A subunit is cleaved and translocated into the cytosol by what is considered to be STx interactions with
and manipulations of the sec61 membrane protein translocator in the ER. As strongly evidenced, the movement of Shiga toxin through a cell is quite valuable for the study of retrograde trafficking of intracellular materials from the endosome to the Golgi, ER, and cytosol [5],[6].
Essentially tantamount to the toxic mechanism of ricin, the enzymatically active A1 subunit of Shiga is an N-glycosidase that acts by removing adenine 4324 from the 28s rRNA. This blocks binding of elongation factor 1-dependent aminoacyl-tRNA to the 60s ribosomal subunit, leading to inhibition of protein synthesis and ultimately cell death. In relation to shigellosis, Shiga toxin can rapidly induce cell death in the intestinal epithelium, which is peppered with Gb3 [5]. STx therefore participates in the apoptotic death of intestinal cells,
while also possessing the ability to affect the vascular endothelium of capillaries in the gut leading to hemorrhaging [6]. STx has furthermore been discovered to act systemically in a variety of ways that makes Shigella dysenteriae infection even more complicated and dangerous than other non-Shiga expressing Shigellae [7].
Studies have shown that red blood cells, platelets, neutrophils, and monocytes all carry Gb3 receptors for the Shiga toxin, however, they are much less sensitive to its toxic nature. These cell types are proposed to bind the toxin and transport it to other regions of the body, in particular the renal glomerular endothelium and tubular epithelium, where Gb3 can be found in abundance. The irreversible ribosome-inhibiting action of STx can then work its course, inducing apoptosis and inflammation of the kidney and its microvascular network. As
a result, Shiga toxin can cause hemolytic-uremic syndrome (HUS), most notably in children who have higher Gb3 receptors in these tissues [7]. HUS is often characterized by acute renal failure, to which the severity of Shigella dysenterae infection is attributed [2].
Shiga and analogous protein toxins such as the plant-produced ricin toxin have been studied vigorously in attempts to understand retrograde vesicular transport. In fact, STx is apparently the first molecule noted to be transported from the cell surface through the Golgi to the ER. Not only have toxins played a crucial role in elucidating many gray areas of cell biology, but they have also been used in medical applications such as engineering of immunotoxins to help combat various cancers and HIV [8]. In the case of Shiga toxin, recent efforts are being made to manipulate the B subunit alone as a means of targeting tumors for imaging purposes. By removing the toxic component of STx, researchers have been exploring the potential uses of the B subunit. In one study by Janssen et al [9], the markedly high proportion of Gb3 expression in tumors of the digestive tract prompted the researchers to consider the STx B subunit (STxB) as a valuable tool for not only imaging/detection purposes, but chemotherapeutic agents as well. STx is especially viable for these purposes on account of its propensity to move along a retrograde path and thus avoid recycling or degradation in the cell.
In order to test the efficacy of STxB for tumor targeting, Janssen et al developed several transgenic mouse models to which they subjected STxB with a fluorescent probe, either orally or intravenously. An initial Gb3 analysis indicated a stark contrast between high levels of Gb3 expression in tumor cells compared to relatively low levels in normal cells. In the orally-fed mouse assay, fluorophore-labeled STxB was found to significantly accumulate in adenocarcinomas of the digestive tract, whereas the STxB marker was not strongly
persistent in most normal cells, nearly disappearing within 3 hours of those few normal cells it did happen to target. Moreover, STxB was found in tumor cells for up to 24 hours and in some cases even longer. Immunofluorescent techniques affirmed that STxB traveled in a retrograde manner through the cells, providing an explanation for its persistence. PET imaging was utilized to monitor STxB coupled with another contrast agent in its ability to target carcinoma cells, and an equivalent set of results was turned over.
Viel et al. [10] conducted a similar experiment in 2008 using two mouse models with human colorectal carcinoma xenografts. STxB was found to accumulate in these high Gb3-expressing tumors, and in monocytes and macrophages. In contrast to the previous study mentioned, Viel et al found intravenous injection much more successful than oral administration of STxB. They also discovered a heterogeneous distribution of Gb3 in tumor cells, however, this did not affect the affinity of these cells for STxB adhesion. Again, STxB was markedly localized to grafted human colorectal carcinoma cells as opposed to normal cells in the mice models. Ultimately both of these in vivo research attempts have elucidated the potential use for STx B
subunit as drug vector for various cancers, in particular colorectal cancers that are known to express the STx receptor in high degrees. Accumulation of STxB in immune cells furthermore suggests a possibility for immunotherapy drug targeting. At the very least, STxB may be developed for use as an efficient tumor imagingdevice which would be crucial for early diagnosis.
Further research into the efficacy of Shiga toxin B subunit for the purposes of early tumor detection and tumor cell-specific drug delivery may well have promising discoveries. This has been evidenced by Shiga toxin’s ability to evade intracellular degradation and recycling, along with its specificity for Gb3-overexpressing carcinomas.


References
[1] Ryan, Kenneth J., and C. G. Ray. Sherris Medical Microbiology: An Introduction to
Infectious Diseases. 4th ed. McGraw-Hill Medical, 2003.
[2] "Shigellosis." World Health Organization. 21 Apr. 2009.
http://www.who.int/vaccine_research/diseases/diarrhoeal/en/index6.html
[3] Hecht, Gail A. Microbial Pathogenesis and the Intestinal Epithelial Cell. New York: ASM P,
2003.
[4] Sandvig, K. Shiga toxins. Toxicon. 2001; 39: 1629-1635.
[5] Tarrago-Trani, M.T., Storrie, B. Alternate routes for drug delivery to the cell interior:
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[6] Cherla, R.P. et al. Shiga toxins and apoptosis. FEMS Microbiology Letters. 2003; 228: 159-
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[7] Ray, P.E., Liu, X. Pathogenesis of Shiga toxin-induced hemolytic uremic syndrome. Pediatr
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[8] Washbourne, Philip. "Diphtheria/Ricin Toxins." University of Oregon, Eugene, OR. 14 Apr.
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[10] Viel, T. et al. In vivo tumor targeting by the B-subunit of Shiga toxin. Molecular Imaging.
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