So it has now been five weeks, eight spawning trials and at least 20 different fish, and we have still been unable to get our fish to spawn. Because we are now coming down to the last two and a half weeks in Xiamen, and I need to give a presentation on the results of my project to a group of professors here in Xiamen, I am now starting an entirely new experiment on Monday. Although this is frustrating, I am just looking at this as an opportunity to learn new skills that I would not have learned otherwise. I will now be using high performance liquid chromatography to try to separate and identify the different sex pheromones in the testes, ovaries and urinary bladder of the Chinese black sleeper. I truly hope that this experiment will be more successful than my previous efforts.
I am continuing to have a great time in Xiamen, and I really wish I had more time left in college because I would love to spend an entire semester here. The few things that seemed strange or I didn’t like when I first came here I have grown to live with and even enjoy some of them. For instance, my journey through the world of Chinese cuisine really got off to a slow start, and I thought I wasn’t going to make it 8 weeks eating Chinese food. I have now tried food I would have never imagined eating, and other than the occasional pig intestine or bee in the chicken, I really love most of the food I have eaten.
Language wise, I still struggle most of the time communicating with people outside of my lab. For the most part I am able to get through situations just by pointing and looking confused, but it was very frustrating when we were at the club the other night and there was a beautiful girl that I asked to dance with me, but of course she had no idea what I was trying to say so I just had to slowly walk away.
A little side story… for the most part we have done very well with the public transportation using the buses only requires memorizing a few characters and the cab drivers can always get you back to the university, but the other night we truly took a bus trip for the ages. So before we even get on the old 48 bus, the driver manages to hit a man driving on a moped and although he wasn’t injured, it was still very shocking to see. So despite our best judgments we still get on the bus and as we are going we realize the bus driver is an absolute maniac. He was using all four lanes on the road, two of which were full of cars heading in the opposite direction. So as we are coming into a stop our bus legit crashes into another bus. Instead of stopping and seeing the damage, the lunatic shoots a dirty look at the other bus driver and speeds off as if trying to make up time. We then pull up to a traffic light and as it turns green, the driver looks out the window and as a cab starts to pass us, the bus driver guns it and forces the cab into oncoming traffic. By the time we finally got off this hellish bus trip I was truly thankful for my life, and I can guarantee I will never ride the 48 bus ever again.
This past week I took a trip to Shanghai to see the World Expo with two of my lab mates and Cathy, another Eckerd student who is currently attending Xiamen University for a study abroad program. The theme of this year’s World Expo was “Better City, Better Life,” and one of the most interesting parts of the expo was learning about the ecological problems in other countries. Many of the pavilions explained major issues in their cities, suburban areas, and rural regions and explained their efforts to solve those problems. For example, Saudi Arabia presented their new technology that gives hundreds of thousands of people access to clean water at the annual Muslim gathering at Mecca. Each country also presented cultural aspects in the form of art, dance performances, food samples, and videos. It was interesting to see how cultural ideals influence how ecological problems are addressed in each country. For example, the Australian pavilion focused on the challenge of balancing life, work, and family. I was able to read short encouraging stories about people who understand the importance of taking time for themselves and their family, but are also extremely successful and have a significant influence on the decisions made in their country.
We spent three full days at the expo visiting pavilions for different countries and interacting with other tourists. While the World expo attracts tourists from across the globe, Cathy and I still found ourselves to be two of the very few international tourists. With the combination of my height and Cathy’s dark skin, we clearly stood out in the vast crowds. The great thing about this was that we had the opportunity to talk with hundreds of people who were curious about where we came from and why we look the way we do (and when I say hundreds of people, I am not exaggerating!). Whether we were standing in lines, exploring the pavilions, eating meals, or watching performances, curious people were never hesitant to ask questions or take pictures of us. Back in the United States it would be odd for a stranger to ask to take a picture with us, so this unusual type of attention really made me think about the differences between American and Chinese culture.
In the United States, midwest, southern, east coast and west coast states have certain cultural aspects that make them different from one another, yet most Americans speak the same language and have major cultural aspects that bring people together. American students from Eckerd College come from all over the U.S., but can easily communicate and generally eat the same foods and interact with others in similar ways. Chinese provinces have very different cultures that make them distinguishable, much more so than regions of the United States. Throughout China there are many different dialects that might as well be considered separate languages. People who speak different dialects cannot understand one another, which makes communication very difficult. Food preferences are also very different throughout China, so people from the different provinces may have completely different dishes and diets. Provinces might also have their own ways of interacting with others, for example, beginning conversations with strangers or playing with strangers’ children might be encouraged in some provinces and thought to be rude in others. After learning about so many cultural differences throughout China, I thought about how unique our American culture is. People in the United States are often brought together by current cultural similarities like foods, language, music, and by their ideas for a better future. Since Chinese people currently participate in such different cultures, everyone is brought together by their common past. America’s history is relatively young and since our American culture formed from pieces of very different cultural backgrounds, it seems that we are brought together by our current culture and our plans for America’s future. I had an interesting conversation with the group in Shanghai about this difference, and one of my lab mates said that young people in China have an obligation to carry China’s past with them as they grow older. As Americans, I think we consider ourselves more as a part of the future than as a part of the past.
As you may recall, last week was a very successful week in terms of progress in my research. More specifically, I got some results from my cDNA samples by performing real-time PCR. So we kicked this week off with some analysis! First, we had to convert the cycle numbers of the specific transporters we tested into their relative expression levels. After this was done we could than compare the relative expression levels of specific transporters within the cell for the different conditions (salinities) and organisms (freshwater environment eel and saltwater environment eel). With the data we collected we think that there may be a difference in the relative expression level of two of the transporters, sodium hydrogen exchanger 1 and 3 (NHE-1 and NHE-3), between the freshwater and saltwater eel within the same salinity treatment! However, before we get too excited we have to realize that it is difficult to say for certain because we only treated our cells for six hours so they had low expression levels. So in order to test this hypothesis we need to treat our samples for a longer time frame in order to get a better grasp on what is happening within the cell, which means that we must start the process from the beginning.
So this week we dissected again and collected the gills (from both freshwater and saltwater environment eels), performed primary gill cell culture and treated the cells with a smaller range of salinities (because we realized that the extreme salinities were causing too much cell death) for a longer time frame (6 and 24 hours). After the treatments, the idea was that we would perform RNA extraction and convert our RNA samples into cDNA so that way we could re-test the expression levels of NHE-1 and NHE-3 along with other transporters. The only obstacle that stood in the way of this was bacteria!
Yes, unfortunately 2 of the 3 freshwater samples somehow became contaminated and bacteria made its home right smack dab in the middle of all of our beautiful samples. When I saw that all of our samples had turned red in color (because we treat our samples with a pH color sensitive solution, so they normally are a wonderful array colors from yellow to purple) I went from happy and excited to sad and discouraged. But Oscar and Professor Wong were very encouraging and even told me some stories of their own in which bacteria had overtaken their samples too. So I just decided to take Oscar’s usual advice and to not worry about it.
But thankfully the freshwater sample that we prepared for the MTT assay and all three saltwater samples were unaffected! So we were able to continue on with the protocol for the saltwater samples but this week we did not have time to test the transporters. We also got to perform the MTT assay but once again we did not have time to interpret our results (yet). So next week will be another busy week for us in the lab! So overall, I would like to think that this last week as another success. Plus as Professor Wong and Oscar would say that anything that is not considered a “success” is considered experience… so this last week I would like to think that I had both success and experience! And I would like to keep this in mind for the rest of my time here, so that no matter how I get accomplished here in terms of results and in term of progress that I am definitely getting some good experience!!!
The internet refers to my situation as the “Pilot error hypothesis”; I call it sabotage. When a young scientist initially begins doing PCR on their own, it can often go wrong. This hypothesis states that it is a common issue simply due to inexperience. The first two weeks of PCR were excellent (Under lab-mate Yanan’s supervision of course), it was looking like I could finish my project on time. However, the past week has surely put me behind. For a reason unknown to me, my DNA never comes out on my PCR.
This is an issue. I have tried: changing the amount of DNA in the initial mixture, creating a new DNA template, checking DNA concentrations in the template, changing the PCR program temperature, using brand new buffers, new primers, new polymerase, changing PCR machines, changing electrophoresis machines, increasing/decreasing electrophoresis voltage, everything I can possibly think of… but nothing has given me the correct results.
It takes nearly the entire day to learn the results; there is no DNA in my agarose gel. Then I do it again. I have been working 10-6, a couple times coming back at 8:30 pm. It is really frustrating to work all day, and then finish with no results.
Last Thursday was even more frustrating. At 2 pm, after waiting all day for the PCR to finish, I found the machine had been turned off! As if my week was not discouraging enough, someone ruined my samples! I had to completely re-start the day’s work.
Today, I booked the PCR machine in advance. (THE PROPER THING TO DO) Then, when I went back downstairs with my samples, someone had put their samples in the machine and had started running their PCR! This is a 2.5 hour procedure, so obviously I was annoyed.
Having 10 failed attempts at PCR is clearly upsetting. Today, (otherwise known as attempt #11) I am trying to use a new gene, with a new primer. If this does not work, then I will have to think of a new method for #12… While reading a National Geographic (I am finding myself with a lot of free time while PCRs are running…) I read the phrase “Persistence commands success”. Generally I do not give inspirational quotes a second glance, however I find this exceptionally relevant. I kept it in the back of my mind for the past 5 or so PCR/electrophoresis attempts, and I think it is an appropriate summation of the past week.
Hopefully next time I report back it will be with positive news!
Week three turned out to be quite exciting/time consuming. I began the week by receiving two projects for the week from my professor. First i had to prepare 700 sample bottles, and second i had to clean 200 PUF’s (small filters used to remove large particles from seawater) for use on the Arctic cruise i mentioned last week. I bet this all sounds quite boring and lets be honest it was not the most exciting work. However it provided me with a wonderful opportunity to get to know my lab mates (since i was stuck in a small lab room with them for 10 hours a day). This turned out to be great because to pass the time we would do Karaoke, watch sports, and play some fun games to pass the time. In the end i believe my lab mates and i really turned a corner last week and it should definitely make my last few weeks in Xiamen unforgettable.
On top of the bonding experience i had two major scares while working in the lab both involving toxic chemicals. The first occurred Monday morning when I was washing bottles with what i thought was distilled water (that is until some of this mystery liquid spilled on my shirt). Within about 30 seconds the skin on my stomach started to burn and my shirt sprung a few holes. What i thought was water actually turned out to be a seven molar HCl (Hydrochloric Acid) solution. I blame my clogged nose for not allowing me to smell the solution, and my lab mates for not telling me what i was working with haha. As i am writing this blog today you can see that i survived the situation, however my shirt did not haha.
On to scare number two. On Thursday morning i was cleaning some PUF’s in a Soxhlet extractor with toluene, a fairly toxic organic solvent. Out of nowhere one of the beakers holding the toluene broke when i picked it up. Good thing one of my lab mates was working with me, because the beaker literally shattered in my hands when i picked it up, i have never seen anything like it before and who would have believed me on that one. Well that is not where the fun stops because when i began to clean up the mess i noticed that the toluene had partially dissolved the latex gloves i was wearing. so not only was i cleaning up a broken beaker, but not my hands were now covered with a toxic chemical. I should have known the toluene would dissolve latex since it is often used as a paint thinner to dissolve latex based paints. O well i just washed my hands thoroughly and put on non latex based gloves to finish the cleanup process. I will say that watching your gloves dissolve while your handling a toxic chemical is not the most reassuring sight.
As i said before they experience of working with my lab mates in such close quarters provided me with the best opportunity yet to get to know my lab mates (especially the ones who’s English is not the strongest). By the end of the week i had gotten even my shyest lab mate Lin Jing to say a few words in English. Those few words were her asking me for help in preparing for her big English language exam she and another one of my mates Sunny ( Shi Xianming) has in three weeks. We have done two sessions so far and it is amazing how much they have both improved, i assume it is because they can both write very well in English, it is just that their verbal skills have never received any use prior to my arrival. Well i believe that is it for week three, stay tuned for next weeks edition of Anthony Reporting from the Orient!!!!!!!!!!!!!!!!!!!!!!!
So I know it has been a while since my last blog, but fear not, I am back and more mediocre than ever. The long period between fascinating blog posts has been due to a drastic increase in workload for my project. When I last wrote, my project really hadn’t gotten started and I was getting a bit anxious to get underway. Luckily I got my wish, but even more than I asked for. It is kind of like walking into a Chinese McDonalds and ordering a #2 with large fries, but instead getting two #2’s and two medium fries. Both of these can be a little frustrating, but only one results in an extra burger and fries. While the workload has been pretty heavy, I am still having a great time and getting a lot done. For the past two weeks, my days have basically consisted of getting to the lab at about 9:00 in the morning and setting up the scanning electron microscope. Then I essentially process samples straight through the day until 5 or 6 at night. Each sample can take anywhere from an hour and a half to three hours, depending on the levels of pyrite in each one. I am usually pretty dead by the end of the day, so you can all imagine why it has been a little hard to convince myself to run back to my room and blog. I also just learned that instead of processing samples from just one core, which takes quite a bit of time, I will actually be processing three separate cores, so I will certainly have a full plate of work in the coming weeks. Even though the work, at times, has been a bit overwhelming, I am getting a lot of very good data and should be able to pull together a pretty solid final product. However, I have said that many times in my life and am still waiting to develop anything of substance, so we will see….
For the sake of filling up a little more space, I will tell a couple of quick food-related stories. Ever since we arrived here, we have constantly been searching for a big plate of Pad Thai. Sadly, even our best efforts came up empty and often resulted in ordering something that looked like Pad Thai in the picture, but was something completely different that usually tasted pretty terrible. After crying myself to sleep every night for the first week because of the lack of Pad Thai, I gave up my search. Then, out of the jaws of defeat rose a new hope not seen since the Obama campaign. A restaurant was spotted advertizing Thai food. Was this, at last, the location of our much sought after noodle dish? Well it was worth a shot. So we bused our way over there and we entered a rather fancy restaurant. a little too fancy, if you ask me. Upon searching through the oversized menu, it was not until the final page that our illusive Pad Thai dish was listed. Just below it was a picture of some tasty looking shrimp fried rice. When it came time to order, I asked for the Pad Thai and the rice, but to my horror and shock, the waitress looked me in the eyes and told me that it was too much food. She literally would not let me order both, and claimed, foolishly, that the Pad Thai alone would feed three people. I did not come all the way there to eat rice, so sadly it had to go. Once the moderately sized portion of Pad Thai arrived, I ate it all, like I anticipated, and still had some room for more. I didn’t order more because I wasn’t allowed to the first time and I didn’t want any trouble. The food was good so I was happy to leave knowing I could have eaten more, but was still pleasantly full. As a side story, while we walked around the area surrounding the restaurant, we witnessed an elderly man find a half eaten ice cream on the ground, and proceed to pick off some dirt and eat the rest. It was horrifying…
Another short food related story occurred last week. There was a café located next to a restaurant we had tried earlier in the trip, so we thought it would be a good idea to go there for dinner. Poor decision. What looked like a delightful little establishment was far from it. I figured that the grilled chicken with orange sauce was a safe bet, but once again I was wrong. After waiting for fifty minutes for my chicken dish, it finally arrived but included a little something extra. When I turned the piece over, I noticed that a decently sized bee had been cooked into my chicken. Not wanting to seem like an ignorant American, I was hesitant to alert our waiter because I didn’t know if it was supposed to be there or not. As gross as that sounds, we had already encountered other types of foods that incorporated bees and other bugs, so for all I knew it might have just been part of the sauce. Finally, it was decided that this dead bee was not intentionally included, and when it was brought to their attention, we were given all of our food for free, even though we were too grossed out and opted instead to just leave, not risking another encounter with something else in our food.
So I would love to talk about all my new developments on my project but due to fish that refuse to spawn and a national holiday for most of the past week, I really haven’t gotten much further on my project. We are now trying to use a ceramic nest instead of the plastic one in hopes of the fish spawning. Also next week, we will try to go to the market with my professor in hopes of getting fish that are ready to spawn.
Outside of some celibate fish, I am having an amazing time. As I said, last week was a holiday in China; therefore I went with five of my lab mates to one of their family’s apartment. We had an excellent lunch of mudskippers, pig stomach, and chicken feet. Although it sounds like an unusual lunch, I absolutely loved it, and I had a great time spending time with my labmates family and becoming ‘Uncle Bryan’ to my labmates nephew. Although my actual experiment is developing slowly, my time spent with my labmates has truly been the most fun part of being here.
I am still having a great time exploring around Xiamen. Last weekend, we went to see the earthen houses in Tulou. These houses are like giant villages that are contained within a circular or square house that are made entirely from dirt, clay, and bamboo. Some of these houses are so big, that around 400 people lived in one at one time. For our trip, we had an awesome tour guide who spoke great English and even brought us to her home village where we had tea with her and her mother. It was an excellent day that ended with our guide serenading me with a beautiful Chinese love song.
This past week, I kept very busy going out to Gulangyu Island, climbing the mountain behind Xiamen University, and trying to go see the dragon boat races which we ended up missing because we went on the wrong day. I have also been playing basketball just about every day of the week. It is a lot of fun just going down to the courts and finding random people to play with.
Hopefully by next week, I will be able to talk about all my fresh accomplishments in lab! Until next time…
This last Friday was the moment of truth…
The reason why I say this is because all of the hard work, time, and effort that I had put into my research this past week all came down to Friday. My goal was to perform the entire protocol for RNA extraction to cDNA, so this means that I started off this past week from scratch.
So on Monday, I dissected an eel, extracted its gills, and performed the protocol for primary gill cell culture. On Tuesday, I treated the cells with the specific salinities that I wanted to test. I decided to do five different treatments (5pH, 6pH, 7pH, 8pH and 9pH), excluding the control. After treating the cells for 6 hours, I was then able to perform the RNA extraction. Then on Wednesday, we got a break :) The reason is because it was a public holiday! The public holiday is known as the Tuen Ng (Cantonese name) Festival and is most commonly known for its dragon boat races! Gretchen and I were able to actually go watch the dragon boat races at Stanley which was a really neat experience!
But we were back to work on Thursday… So I had to determine the concentration of the RNA for each of my samples which is very nerve wracking! The reason why I say this is because if the concentration isn’t high enough than you can no longer proceed with the procedure and have to start from the beginning with the dissection, so you really only have one shot at this. But luck was on my side and my samples had a high enough concentration! However this is not the end of it yet. On Friday, I then put my samples into the thermocycler to synthesize cDNA from my RNA samples. Then to make sure that everything up to this point has been done correctly and that everything is still good, I had to test the quality of my cDNA by preparing them with a “housekeeping” gene and loading them into the real-time PCR machine. Then after the real-time PCR machine has performed its cycles you can retrieve the cycle number of each of your samples which you can compare with your control to make sure your quality is good enough to continue on with the procedure (plus the cycle number is very important because we will use the cycle number to calculate the level of expression of that specific gene/transporter within the cell which is what we are really interested in).
And this is where the moment of truth comes in….
The moment where you will see if all of the work for the entire week has been done sufficiently enough to actually start the testing (I am pretty sure I had my fingers crossed for the entire PCR cycle which is approximately 2 1/2 hours long! It was especially hard because we were at lunch during this time and eating with chopsticks while your fingers are crossed is not an easy task!) On a side note, the students in my lab think it is actually very funny that we cross our fingers. I did not realize that they do not do it here but I think I may have convinced them to start because… Once again luck was on my side!!! That’s right, my quality of cDNA was good enough to continue on! I cannot even describe how excited I was!! I think Oscar, the graduate student I work with, even commented on how big my smile was!
But this really was fantastic news because I was able to test two different transporters on Friday alone! Plus I am very excited because the amount of cDNA needed to run the test is very a small amount so I still have enough cDNA to run even more tests this week!!! After work I was so happy and everything that I even bought a celebratory ice cream from the local student store for my walk back to the dorms! haha So hopefully (knock on wood), this week will pan out just like last week! Cross your fingers ;)
In my lab, we have been researching the embryonic development of the shark Scoliodon laticaudus, which is commonly fished as a food source in the Fujian province. We buy pregnant female sharks from a street market vendor in Xiamen and dissect them in the lab in order to remove the embryos from the mother’s uterus. The embryos range in size, anywhere from 1 - 30 cm. The main purpose of the research is to determine where the gonads begin to develop in embryos and to observe how this growth progresses throughout the stages of embryonic development.
One of the most memorable cultural experiences for me was taking the opportunity to go berry picking with my lab mates. The English translation of these berries was “wax berries,” a dark red or purple fruit that does not grow in the United States. I had tasted these berries once before and I absolutely loved them, so I was excited to venture out to pick them by hand. Before leaving for the day long trip, I had pictured myself picking berries at an American style orchard, but when we arrived I was surprised to find out that the wax berries, which I had been practically drooling over the entire car ride, were located at the very top of a mountain! After about an hour of maneuvering through trees and ducking under low branches, we reached the top of the mountain. After picking our basket of berries, the entire group took a long rest at the top of the mountain, eating our wax berries as we looked out over the ocean.
After slipping in and sliding all the way back down the mountain, we stumbled across a local seafood restaurant for dinner. The building was set up on some sort of floating device so it stood alone out on the water. There was a man there who paddled customers from the land to the restaurant on a small wooden ferry, as simple as a large plank of wood. As a group we ordered every kind of seafood imaginable. The meal was delicious! Everything about that day was completely unexpected, but I wouldn’t have had it any other way.
Well where should i begin, with the mundane week or the magical weekend… I think i will make you suffer first!! haha. Well the second week in lab sure was a bore, my project all but derailed within the first few minutes Monday morning. My Professor arrived back from Hong Kong and informed me that my project was being put on hold for up to two weeks!!!!!!!!!! There were two reasons he gave for this, first one of our lab mates was giving his masters thesis defense on Tuesday morning, and second two of my other lab mates are leaving for a three month Arctic research cruise on June 18th. Thus all lab equipment must be devoted to helping those going on the cruise prepare. So in effect i gather that i will not be starting my research in earnest until the week of June 21. This will leave me under three weeks to complete my project which is quite a tight schedule. Other then that the lab has been fun, my lab mates are great and im sad to see two of them leave in just under a week (who am i going to play Badminton with now haha).
Well lets move on to the good stuff!! On this past Saturday all the Eckerd students and Professor Duncan went to go see a tǔlóu or 土樓 in simplified Chinese characters. The translation from Chinese is earthen building. They are extraordinary structures constructed from the 15th century up until the 1950’s by the Hakka people of Fujian Province. On July 7, 2008 UNESCO added the Tulou’s of Fujian Province to the World Heritage List under the moniker Hakka Houses. These structures are breathtaking they are basically an entire village enclosed into an area smaller then a football pitch. Some of the larger tulou’s contained 800 or more people at some point during there existence.
We had an amazing tour guide her name was Miss Zhang she took us to many different sites and even took us to her home town, which was a little town which did not pander to tourists like some of the other villages. Her town used the tourism dollars it earned to enhance the experience not hinder it like some of the other Hakka villages. We all sat in her mothers little tea shop drank tea, and talked for about it an hour it was truly a wonderful experience. She was also a wonderful tour guide as she took us to four different Tulou sites and gave us great insight at each location.
The tulou’s were an interesting mix of old and new. We went to one in which no commercialization had occurred and we went to others that were basically tourist traps in old buildings. It definitely shows how China’s rush to modernize has been both good an bad. I hope these communities are able to survive as more then tourist destinations. The Hakka People have a wonderful and unique culture and they should preserve as much of it as they can!!!!!!!
Now the final touch to an amazing weekend occured sunday morning around 4 A.M. Beijing time. THE UNITED STATES TIED ENGLAND IN THEIR FIRST GROUP STAGE MATCH. I stayed up to watch the game and was elated by the result. England has arguable the best team in the world cup, and the US a team full of no names tied them. I bet most Americans can name three players at best (Tim Howard, Clint Dempsey, and Landon Donnovan). Well as a huge football fan (yes football not soccer) i could not be happier i wish the US team my best and hope they can advance far beyond the Group Stages. Well that’s it for this week keep tuning in for more up dates!!!!!!!!!!!!!!