Nitya Talreja #2: Border Patrol Tests

This week at the Penn Working Dog Center, I got to see a 5 week old puppy in Delaware. Beginning at 4 weeks old, Border Patrol Tests are conducted on donated puppies to test their confidence, drive, and ability to succeed as working dogs. The puppy’s name is Rigo and he is a black lab, and not to mention a singleton. The term singleton is used to describe a puppy that was the only puppy in its litter. Due to Rigo’s circumstances, as a fetus he received the nutrients equivalent to that of 6 puppies, hence he was twice the weight he was supposed to be, but nonetheless adorable J.

Border Patrol Test stimuli include keys, toys, and unstable surfaces. Rigo’s reaction to each stimulus is rated on a scale of 0 – 6. 0 being the least driven or lacking confidence, and 6 being the most driven or reactive to the stimulus. It was quite difficult to keep Rigo’s attention and he did not seem very interested in most of the toys for the test, however, the keys did seem to scare him. He did not seemed bothered by the unstable surface we had placed him on, which was a good quality for a working dog. Otherwise, his reaction to most of the objects were accurate with that of a 5 week old puppy.

Because Rigo was extremely obese, we often called him the meatloaf because there was barely any room between him and the ground :-). 

The other puppy at the PWDC was named Lucas and he was an 8 week old lab when I first arrived at the center. Lucas’s border patrol (BP) tests were also conducted on a weekly basis. Lucas has excellent drive and confidence for a puppy of his age, and Dr. Lorenzo has great confidence in his success as a working dog. Even at his young age, Lucas expressed strong vocalizations and was not intimidated by any of the stimuli. On his 8 week BP test, Lucas scored a 6 in all of the categories! And his strong drive did not waiver on subsequent BP tests (he was so excited one time that he actually bit my hand instead of the toy).

*One interesting fact I learned this week was that Labradors make such excellent working dogs because they lack the gene that makes them feel full after a meal, which is why they are often overweight and always food-motivated to complete tasks. Not to mention that they are retrievers by nature, which makes the toy-motivated as well. 

             

 

Nitya #1: Weeks 1-4, Getting Lost, and Lots of Puppies

Out of all the struggles I expected to encounter this summer, little did I know lunches would be at the top of that list. After not having to make my lunch for the last 5 years of my life, I found it to be a great struggle to start making my own lunch again. First week in, coming into my lab, I was at first intimidated by the location of the lab (35th and Gray's Ferry Avenue--not the safest of places), but it does make sense to not be in a very public place and surrounded by abandoned buildings to successfully train working dogs for a wide array of purposes. Thankfully, since I was commuting to my lab everyday, my PI was very flexible with my hours (usually from 9am-3pm everyday). I usually began my day with shadowing a trainer on a walk by video recording or collecting data on fitness and husbandry related tasks, then I would assist in the cancer trials by either helping clean the materials between every session, collecting data, randomizing the order of the samples, or by clicking when the dog alerts on the correct sample port on the wheel. I would assist in the breakdown of the study as well.

 

 

There were three dogs that were classified as "cancer detection dogs" which belonged to the facility for research purposes and would not be sold. There was one German Shepard (Tsunami), a spaniel mix (McBaine), and a yellow labrador (Ffoster). Collectively, all the dogs had a 98% success rate on which they alerted at the malignant cancer sample. The dogs were tested on a wheel on which there were 12 ports. The ports consisted of plasma samples from three different people. One person had normal blood plasma, another had benign plasma for ovarian cancer, and the third had malignant ovarian cancer. All the samples were donated to Penn for research purposes and provided to the PWDC. The 9 other ports on the wheel consisted of blanks or controls. 

The people at my lab were mostly Drexel co-op interns and Penn Vet students. My PI  (Dr. Lorenzo Ramirez) was a post-doc employed by the Penn Working Dog Center. 

 

 

My first week commuting to Philly I made every mistake in the book. I somehow managed to be bitten by an adorable 8-week old lab with razor sharp teeth, I forgot my wallet at home (thank god for uber!), got on the wring train (and ended up in North Philly!), fell asleep on the train and missed my stop, and somehow managed to let my phone die on the way back home. But somehow, despite these occurrences, I survived.

Alex Larson, Entry #5, Coming Full Circle (Cause we're working with Plasmids get it)

Well if everything goes as planned, I should be finishing up my portion of the project really soon. Now that the finish line is in sight I can finally see how everything I did led up to our ultimate goal, which was something that seemed so abstract to me even just a couple weeks ago. In short, our Lab does a lot of mRNA analysis to see the various types of proteins that are being made during periods of influenza infection and periods without infection. To do this, people have to do a lot of RNA extraction, which is almost too good at getting RNA. I say it’s too good because upwards of 90% of the extracted RNA is ribosomal RNA, or rRNA, which will be there regardless of infection, person, or even species (Every living organism uses ribosomes). I did the extraction of the 45S precursor gene for rRNA. I created primers flanking the coding regions within the 45S precursor gene. I then amplified these regions through PCR, which actually was the hardest part of everything, and then sent off the successful amplicons for sequencing. Then we plan on using a cloning protocol to put these amplicons into separate plasmids and grow them in E. Coli. This is done so we don’t have to continuously run PCR to get more rRNA sequences. Right now I am working on the online tool, Benchling, once again, to make specific sequences for single-guide RNA, also known as the part of CRISPR/CAS9 that binds to the appropriate sequence of DNA that is subject to cleaving. Adam is going to stay in touch with me for the duration of the project and will inform me how successful DASH is when we put our specialized CRISPR/CAS9 into transcriptomic samples.

            Being in the Ghedin Lab for seven and a half weeks has really opened my eyes up to both the frustrations and elations of doing hands on research and pushing the limits of human scientific knowledge. I always knew that I liked science and I think that this experience has really cemented into my mind that biological sciences is the field for me. I’d like to thank everyone in my lab, specifically Adam, Nick, Tim, Alan, Mathilde, Alex and everyone else who I’ve gotten to know so well over the past two months. I would extend a special thanks to Dr. Ghedin for putting up with me for the past several weeks and always being so helpful and open. I’m especially thankful to have that sodium borate jug in the back named in my honor ;).

            Outside of the lab setting, just being in New York City and commuting and seeing the people has also been very eye-opening. Just being surrounded by so many different people of so many different backgrounds has no only changed the way I see New York, it has also changed the way I behave (I’m certain my people skills and ‘street smarts’ have improved this summer). So as I finish up my last few days in the lab and prepare to return to the boring summer life I’ve known since I was little, I really only have one question that I can’t for the life of me figure out. How does everyone have pictures of themselves to doing work? I feel like when I do bench work, I’m out there with gloves on, and so is everyone else. So where people in these blog posts get their pictures from is beyond me. But I suppose there will be plenty of time for lab pictures throughout higher-education anyway, so of all the problems that I could have encountered this summer, I can’t say that I’m too upset about this one. All in all, this has been one of the best summers yet. 

Gonna miss the ol' bench