The work I’ve been doing with Dr. Nancy Krucher involves the very invasive disease, Pancreatic cancer. We have worked on a possible solution to the invasive behavior of cancer cells that comprises of knocking down a specific phosphatase subunit and its effect on the Retinoblastoma tumor suppressor protein. We also studied the effects of the drug Erlotinib on cell death. At the start of the year, I worked with 96-well plates for various experiments. Doing this allowed my pipetting skills to improve greatly and I gained more knowledge of knockdowns and cell growth. Some experiments included creating and testing several concentrations of Erlotinib and finding it’s optimal concentration for cell death. Then after finding this concentration, we treated the MiaPaCa-2 pancreatic cancer cells with PNUTS knockdown to inhibit invasion for 48 hours and proceeded with Erlotinib for 24 hours. Afterwards, to count cells we used Cell Titer-Glo spectrophotometer assays to see how effective the combined treatment was in killing off cells. This process took several experiments for me to get the hang of the techniques but the results were great. There were plenty of experiments that were flops, and some that were good but had very big error bars for our data (which isn’t so great). However, multiple experiments of mine showed a decrease in cell count when the cells were treated with both PNUTS knockdown and Erlotinib.
To further look into how PNUTS knockdown would affect the cells, we studied its effects on invasion. I had to perform immunoblotting experiments to see the differences in expression for several EMT (Epithelial-Mesenchymal Transition) markers. Before this experiment, I didn’t know a thing about EMT and it honestly helped me so much in one of my classes afterwards when reviewing other scientific papers. Our main focuses were the markers Zeb, N-cadherin, E-cadherin, and Vimentin. This took me so long to get used to. I had to learn how to lyse cells and extract protein and create samples to run a gel with. The hardest part was finding the concentrations of protein and creating samples properly. If this wasn’t done correctly then my gels were pointless and the bands on my nitrocellulose paper wasn’t reliable. I was able to finally work through my mistakes with some practice and obtained some good results. I found a decrease in both Zeb and N-cadherin expression; which signifies a reduction in invasiveness due to the knockdown. I’m still currently working on experiments like what was just mentioned but with other markers and even touching on different pathways affecting invasion as well.
Throughout this whole experience I’ve realized how great it is to work in the lab and conduct such interesting research. My goal in life is to become a doctor but with my experiences this past year I actually am looking into joining a program to obtain an MD-PHD degree. I want to continue researching in the future. Yes, it gets frustrating when experiments don’t go as planned or when results get messed up, but as Dr. Krucher would always remind me, everything takes time and practice. It is always a learning experience. Even when I thought I knew how to do everything properly there was always something else to be learned and practiced. I am so proud and happy with what I’ve done in this past year and so thankful I’ve been able to do it. I’m very appreciative of the UGR program and Dr. Krucher (for putting up with all my mistakes!). Her guidance has helped me greatly and I hope the project continues to bring positive results!
Extensive research of the lectures of Jacques Lacan has assisted my writing in the discipline of literary criticism in the fragmented representation of addiction in contemporary literature.
Thus far, the most poignant connections between addiction-based lifestyles and Lacan’s psychoanalysis is the idea of identity reformation and recognition coagulating via the unconscious nature that is substance-dependence. A “New Mirror” stage so-to-speak is constructed by the individual’s identity superimposed by the unconscious language of their need for habitual substance abuse.
The “New Mirror” stage (or possibly ‘latent’) allows readers to explore addiction lifestyles as an experience or entity that forces both the addict and the addict’s caregivers to go through identity transformations dictated by the unconscious nature of their relationship.
What is still in need of thorough exploration is how the addict fits into Lacan’s idea of the symbolic order. And how does this relationship to language influence the addict’s desire for wholeness and completion via the impossibility of closure or “recovery.” Scott Heim’s novel We Disappear mobilizes Lacan’s structure of psychoanalysis with the narrator’s relationship to substance abuse and his mother.
UPDATE ON THE INTERVIEW WITH SCOTT HEIM:
Author Scott Heim has written in response to all the questions I asked him and his answers are more than compelling. I am still figuring ways in which to navigate his responses in relation to my research considering I write within the discourse of “death of the author” in which an author releases a text to a public consciousness and therefore is no longer considered whole with the text.
It is even more pressing to figure out how to consider Scott Heim’s responses when he has admitted to me that this novel is infused with autobiographic experience – the addiction.
Use of aromatherapy in freshman college students to decrease test anxiety: A random blinded intervention-placebo study
The experimental portion of this study has ended. Dr. Greenberg and I are now focusing on collecting and analyzing the results of our pre experiment and post-experiment surveys; Pittsburgh Sleep Quality Index, Westside Test Anxiety Scale, and Spielberg State Trait Anxiety inventory.
Dr. Greenberg and I encountered a multitude of challenges in this study;
- We had a large, potential sample size of over 400 freshman students; however, it proved to be an impossible task to recruit many of the students. I sent numerous emails reaching out to the students. University 101 professors posted the consent form for the study to their course’s Blackboard for ease of access for the students. We even offered gift card incentives! We ended with a total sample size of 27 subjects. Due to the difficulty we faced with recruitment, we learned quickly that working with this group of students was going to be a challenge.
- Of the 27 participants, there was huge noncompliance with completing the surveys. Only 4 subjects completed all of the surveys. I even sent mass texts and emails reminding and encouraging the participants to complete the surveys.
- Only 11 subjects actually picked up their inhalers.
- Because of this, our end sample size for the control and intervention groups was limited.
Conversely, despite the challenges faced, we found that utilizing the lavender inhaler a minimum of 3 times per week, in the fall semester, prior to midterm examinations and through final examinations yielded as an effective anxiolytic and improved the subject’s quality of sleep. Specific statistical data is still currently being analyzed and will be discussed in depth on the UG research day that Provost is holding.
I will be graduating this May with a strongly developed skill in evidence-based practice research thanks to Leinhard School of Nursing. The program is strongly invested in teaching nursing students the essentiality of using current, best evidence when making decisions about patient care and utilizing critically appraised, scientifically proven evidence for delivering quality health care. I learned that this is one of my major strengths, and I have become intent on conducting my own research later in my nursing career. When Dr. Greenberg offered me this opportunity to be involved in her research, I was incredibly enthusiastic that she not only recognized my EBP aptitude, but that this opportunity would provide me a foundation on conducting research that I can one day achieve myself. I have become confident in my ability to interpret research data through employing valid statistical tests. Dr. Greenberg and I will be presenting our work on aromatherapy at the Eastern Nursing Research Society’s annual scientific session in Pittsburgh, PA next month. This accomplishment will open a door of opportunities for my career path that I have only begun.
This third blog post features an update on the progress of my project investigating potential experiments for an Inorganic Chemistry laboratory. Since the last blog, a lot of progress has been made. Unfortunately, we have had many bumps in the road of this project but have continued to work around them and make progress. Revisiting the first experiment, the product results were inconclusive and required further testing to confirm the structure. In an effort to move forward, we pushed to our second experiment utilizing the specially-ordered resin and a solvent known as Perchloric Acid.
We had all of our materials ready for this ionic column exchange experiment and were preparing to run an initial test. In science and lab work, it is imperative to know and understand the kind of materials one is going to be working with. Prior to running anything, any risks that could potentially be involved during the course of an experiment should be made well aware to every person working with the materials. In doing our preparative research, we started picking up many warnings regarding the Perchloric Acid. As it would turn out, this material could prove to be highly explosive when it is dried out and can be very caustic to many materials. Upon seeing this, we decided to scrap the experiment in favor of safety. The material has since been properly disposed of.
Following this, we found an alternative experiment that uses the same concepts of the original column experiment with mostly similar materials, aside from the Perchloric Acid. We decided to run this shortly after. The defining concept of this experiment is that it uses a pressurized system of Nitrogen gas in order to push the eluting solvent through the column and resin. This pressure forces the column experiment to proceed at a significantly faster rate and allows each run to be completed quickly. We performed this experiment using regular compressed air instead of Nitrogen due to its ease of access in each of our laboratory rooms. We also looked at an alternative method which we originally felt might be easier to do for an entire lab class full of students. This method used a small syringe and plunger to mimic the column and pressurized system on a much smaller scale and without the use of a compressed air pump. Unfortunately, problems with the syringe resulted in failure of separation of the metals. The procedure could be further adjusted and implemented in to the teaching lab with some more work. Pictures of both the initial set up and the alternative procedure are below.
The most recent experiment we ran as part of this project, is a synthesis between a Cobalt metal and Saccharin molecules. The result that is formed by this demonstrates the unique structures of metal complexes. The experimental procedure here is simple and easy to follow. The reagents involved are all typically non hazardous. The product that was formed through this experiment was then analyzed by ultra-violet visible spectroscopy and confirmed the structure of the product as a successful synthesis. This experiment could be directly implemented in the lab almost as is. A picture of the beautiful crystals that were recovered from this experiment are below.
Recently, Professor Upmacis and I attended the Society of Fellows 2017 meeting at Pace to display the work we’ve accomplished so far. Understanding this post was on the longer side, but it ultimately covered each aspect of the new work my professor and I have completed since the last blog update. Future work includes looking at another experiment to synthesize a metal-quinoline complex utilizing different common transition metals. As an additional note, this project has really been an eye-opening experience for me. It’s demonstrated that it’s a rare occurrence where everything goes perfectly and it’s more likely that something will fail or not work properly. The real world of working on new experiments and researching new procedures or creating a new methods is often full of issues that need to be worked around. A creative head is necessary to succeed and solve various problems!
Thank you for reading this long post!
The research program has expanded since the last blog post we made back in December; we have conducted more research and refined our Qualtrics survey to reflect the most accurate results possible. In addition to displaying biometric facial scanners to the elderly, we have also introduced USB fingerprint scanners to them. Although it is a different kind of technology, it essentially does the same thing; it incorporates a unique feature of each person’s body to authenticate a machine. Fingerprint scans and facial scans can extremely rarely lead to false positives; this is why development is currently leaning towards vein-scanning technology, as it would render less false acceptances into another person’s system (no false positives have been witnessed in the research).
It is actually interesting and exciting to see the elderly actually engaged in using a newer technology; in past cases, older family members often turn away technology but we are seeing an embrace of it at a rapid weight. Some of the elderly people we survey have new iPhone/Android phones that incorporate fingerprint scanners into their hardware; they say they enjoy it because they do not have to remember a password (the less clicks to do something for an elderly person, the better). According to our survey at this point, 76% of elderly people use technology often; with that information, it can be inferred that 100% of that group of people are vulnerable to cyber attacks and need security measures to protect their data and personal information.
We have encountered a challenge; an original goal of this research project was to create an app/software that would improve upon the current biometric softwares provided by Intel and the companies that actually make the hardware, but making one would be almost like reinventing the wheel and would take years of algorithm research and intense development. Rather than create our own version of our vision of these applications, we will forward our research to these companies in hopes that they can make improvements on what we have found to be troublesome aspects. Maneuvering around patents would also make for a huge hassle; it is best if we let the original creators refine their application.
Ever since December, I have been able to make progress with my research. Every single week, I was able to come in and run my four fluoroquinolone drugs. I have been testing them constantly with the same power at 50 and multiplier factor at 50. I’ve tested the samples at 3 different times: 1,3, and 5. Using the scans, I was able to produce spectras for each drug using Igor software. Igor allows me to open up excel files and label the graphs with its peaks accordingly. Since I was able to do run my samples every single week, I took the average of my runs at the different times. This helps to improve the signal to noise ratio and produce accurate results. Seeing all the progress I have made so far makes me feel a lot better about my research because I was struggling a bit in the beginning. I was able to however overcome my issues with the laser Raman and produce results.
From my research so far, I was able to print out a poster of my work and present at the annual Dyson Society of Fellows this past March. It felt good to see my hard work on display for others to understand. I was able to tell those who came up to my poster all of the accomplishments I have achieved thus far. I also was able to answer questions and take in advice that I could use for future work with my fluoroquinolone drugs.
Looking back at the results produced so far, I have been able to obtain accurate spectras for the four fluoroquinolone drugs. Sarafloxacin, norfloxacin, and ciprofloxacin produced peaks using the laser Raman that were similar to its Raman spectras from before. However, enrofloxacin’s spectra produced no peaks that can be used to identify the drug. I find this interesting because the laser Raman was able to produce spectras for the other three types, just not this one. For the rest of my time doing this research, I want to continue running my samples to see if taking the average for all of my trials help my results. I also want to investigate why in particular enrofloxacin only produces peaks with the original Raman, not the laser Raman used.
The two tasks that have been developing are the literature review and data collecting information on Police Shootings with the mentally ill. Paper writing is not my forte. After struggling to write a literature review, something I’ve never done before, I was able to complete it. There still need to be edits and additions but I think I did a good job considering the circumstances.
After that was submitted, I worked on some of the information for the database and researched missing parts of information on a data sheet. Through an excel spread sheet I enter different aspects about many officer involved shootings. The hardest cell to fill was the one that determined mental illness. Many articles made no mention of mental illness or had conflicting views. Some would say mentally ill, and others would say intoxicated or sometimes both. I completed as much of the data sheet as I could in the short amount of time and enjoyed that more than writing the literature review. I want to continue working on it and look more extensively without the time crunch.
Through collecting data I was able to see obvious patterns within geographical areas that struggle the most with police involved shootings. Although I recognized that I was not working on the complete data, just 100 cells, many of the incidents occurred in California. This could be simply because the state is so large, but it could also speak to their policing abilities. The victims were also almost entirely male. Determining the race of a victim was not always easy but there was a high volume of minority victims on my sheet, but again I am aware that it is such a small section of the present data so not to make and final judgments based on that alone.
The progress of this research has been slow and steady. I am currently adding more sources to the literature review. My schedule this semester has been harder than I anticipated, but I will still work on the things Professor Arslan has given me to contribute to the research. The experience of data collecting was so far the best part of this research for me. I hope I do more of it.
March 20th, 2017
Blog # 3
So far only a little more research has happened since the last time a blog post was written. The only field research that has been able to be conducted since the Winter break has been on how the elderly use the mobile application Apple News. This is due to the fact that the researchers had to get seniors to do research with at senior centers. The most common observation of seniors using Apple News is that this application is too difficult for the elderly to use. This is due to how it represents itself, the app opens up to a screen exploding with many different news articles. Which is not bad design by the app, though other functions of the app make this not as wonderful. The seniors were able to click on each article they wanted to read in order to learn more information about an event. The problem occurred when they realized they did not know how to get back to the original screen. This trapped some seniors to only be able to read one article. On a side note other seniors also explored the applications tabs on the bottom of the screens and got lost and were not able to get back to the main page with the news articles.
A simple solution to a problem like this would be to just make the application much simpler. All that would be required of you would be to remove all of the extra functions which may seem important. Perhaps this new version of the app’s only purpose was to provide the news with no special features. This would mean that you could just simply have an article on the page and the user could swipe left or right to the next article and scroll up or down to read the rest of the article. This would follow the golden rule of being simple which allows for a more optimized user interface for senior citizens.
When conducting this research the hardest challenge that has been experienced with doing this research is finding a senior citizen to do research with. This requires the cooperation of a senior center as well as the senior citizen. Then setting up meeting with a senior can take time as well, but once that is all completed conducting the research itself is rather simple. All that is required is for the researcher to observe how the senior citizen reacts to the mobile application, as well as how they use the application. A researcher is able to determine rather quickly if the application is properly optimized for the senior citizen. If it is not then the researcher is able to observe more details of how the senior citizen is using the application and ask them questions to find the reasons for their dissatisfaction with the application. Though this is also done even if the senior citizen likes the application because it will tell the researcher what the mobile application did right.
After doing this research my biggest take away from what I have done would be that more research is always better. The more data that I collect means that I will be able to come to conclusions and to formulate a hypothesis that I am able to back up with research. For example, one of the things that I have realized from all this research that I have been doing is that simplicity is the key for designing mobile applications. It’s also the number one solution to designing a mobile application for the elderly and I have a lot of research that I can use to back this up. Without learning how to collect large amounts of observations I would not know how to go about defending an argument. Therefore I have learned a lot from conducting this research.
The beginning of this semester we continued our work on the C. elegans’ gene T20B12.7 and it was a little shaky due to the snowstorm and school closure. The first RNAi experiment we performed was unsuccessful because we could not get into the lab to record the results. We then performed another RNAi experiment, which show similar results as the previous semester. The RNAi T20B12.7 worms had only a small difference from the L440 worms in unhatched embryos. Another RNAi experiment was performed with rff-3 worms in the hope a more significant difference would be seen. The rff-3 worms are more sensitive to the RNAi treatment. However, the results from the RNAi experiments with rff-3 worms still had an insignificant difference. These results could have been for two different reasons. The first reason was that the RNAi was effecting the worms in another way besides embryo lethality. This was possible, but previous studies showed that this gene influences embryo development so not as likely as the second reason. The second reason for the insignificant difference could have been that something in our RNAi experiment was contaminated or defective. We decided since we were getting deterred by the RNAi experiments for T20B12.7, that we would hold on further experiments for this gene and complete experimentation on another gene, F55A3.3, that had had successful RNAi results.
F55A3.3 is a gene in C. elegans that is involved in embryo development, molting cycle, nucleus organization and reproduction. F55A3.3 is a human ortholog, spt16, that facilitates chromatin remodeling in the FACT complex. Previously performed RNAi experiments in our lab to knockdown F55A3.3 resulted in a sterile phenotype. We hypothesize that the loss of F55A3.3 may be necessary for normal embryo development. Our current experiments will analyze this hypothesis and help further understand the role this gene has in C. elegans. The next step was to use fluorescent microscopy to identify cause of sterility. We have performed two RNAi experiments and used microscopy to image the worms after they were treated. The first experiment we saw several abnormalities in the RNAi treated worms compared to the L44o worms. The abnormalities consisted of changed in the germline shape and irregular dividing in the embryos. The second RNAi experiment was not as successful. The imaging could not be performed properly. We are now in the process of troubleshooting the imaging process to see if the microscope is not working properly or if the worms that are tagged with GFP are contaminated.
Since my last update I have hit a few snags. My samples did not keep as well as I had hoped they would over winter break and I lost the half of the samples that I had not yet identified. I also had some problems getting all of the necessary supplies together, accidentally getting the wrong size pipettes. The good news is that I still have all of the water samples I collected, so it will not be a problem to simply restart that process for those half. This won’t waste much time either, so it isn’t a disaster. I have also gotten the correct pipettes since.
I am currently getting all of my identified samples ready to make frozen stock out of them. Remarkably, all of my already identified bacteria survived the break and are growing wonderfully. (I will attach a picture I took today) This way I will have the samples ready and in the lab to grow whenever I need them. I am about halfway done with identifying the bacteria that I will be using in my experiment. Now that I have all my supplies ready and in the lab, this process should begin moving so much faster.
What I have learned from my project so far is less of a scientific discovery and more of a personal one. I have learned to become adaptive and patient. Working with bacteria, especially those isolated from nature, can be incredibly tiring. It becomes even more complicated when you add work, school, illness, and breaks into the mix. I have had some issues getting my bacteria to grow under lab conditions as they are used to the Hudson supplying their nutrients and growth conditions. The Hudson varies greatly in temperature from season to season, so the bacterial diversity is probably much greater than I am able to see on my simple petri dishes grown in lukewarm conditions. It can be incredibly frustrating when something decides it no longer wants to grow for me, but often times stepping back and giving the organism time or a new growth condition will fix everything.
I have never been a very patient person, but I am slowly getting there with this work. I see things running much more smoothly after from here on out. Hopefully next post I will have much more to tell you!