Challenges in research and the defective DNA tubes

We have a research project which is ongoing up here. There are two parts to the study. One is similar to the prior study I did in 2008 with ibuprofen as an alternative treatment for high altitude headache. This time we are also comparing it to another medication. The second part of the study involves DNA sequencing of people who do and don't have symptoms of altitude illness. Both of these have had some challenges.

First, the clinical trial (headache treatment part of the study) requires that our subjects have a headache at that moment which they are enrolled and that they are willing to track their headache for the next two hours. Initially, we had planned to keep the subjects in the clinic, but cold temperatures and space made this difficult. Also, we are having trouble recruiting subjects. Patients who come to the clinic have often already taken a variety of medications for their headache which makes interpretation difficult. We tried recruiting from the tea houses at dinner, breakfast, and even mid-afternoon with limited results. This technique had worked for the previous study I was involved in, but was not very productive this time.

Second the DNA blood collection has been difficult. First, many people do not want to give up their blood. Although we are not taking much, they prefer to retain all the red blood cells they can for transporting oxygen. I admit that I do understand their concern, but the blood is a very small amount and shouldn't matter in their performance. Second, the blood is difficult to collect. The clinic is near freezing most of the day which makes veins constrict and drawing blood a challenge. In addition, the DNA collection tubes are malfunctioning. These blood tubes are very special in that they do not have to be kept at a certain temperature and the DNA is still good. They are fairly new and although skeptical, it will be awesome if they work. I had called the company before leaving Boston to ask if their tubes would still function after flying around the globe on four flights, being transported to 14,000 feet on a yak, multiple freeze-thaw cycles over several months, and then the whole thing in reverse. The company assured me this would not affect the DNA stability and that they would still function at altitude. They were wrong. In order to collect blood, most blood tubes have a vacuum inside which helps them fill even though they are sealed. Initially, we tried several methods of venipuncture thinking the butterfly needles were the problem with the convicted veins which resulted in Chris and Penny getting covered in blood during one collection gone awry. However, it finally dawned on us that with the much lower atmospheric pressure here, the tubes did not have enough of a lower pressure to create a decent vacuum. By drawing 20mL of air out of the tubes before using, they then worked. Hopefully, this does not alter anything significantly upon descent or cause the tubes to break when repressurized.

There are also two things which are also out of our hands that have decreased our potential subjects. First, there are fewer people trekking in part from fewer flights from Lukla due to colder, snowy weather and new weight and passenger restrictions on flights. Second, the large Extreme Everest group is doing their own research and so none of their subjects are allowed to participate. They are taking up over half of the biggest tea house for over eight weeks and so many other trekkers are staying in Dingboche instead.

These challenges aside, we are hoping for more research subjects so that we do not have to continue the study next season.