Sunday, April 27, 2014
I have tried many different format in my classes. Most recently I created Facebook pages for my freshmen biology labs and then a group for future neuroscientists. I found, much to my surprise, that many students did not "do" Facebook. So in an attempt to be inclusive and not cause students to participate in something they do not participate in, Biology Research and Interns at CBU will form a blog group. This blog will be about your journey into the research and if applicable internships. In a similar manner to the MHIRT students, this summer you will be required to blog twice. Once concerning your work in the lab experience and once about your journey. Dr. Fitz
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Hey everyone. Just trying this blogging out. I'm still watching my plants grow (or not grow). It won't let me post pictures in comments but I have some of my plants and several of my oh so happy face working in the hot green house with pleanty of Mosquitos.
ReplyDeleteHave a great weekend.
-Cathy
Hello classmates. Hope everyone is enjoying their research so far. I have certainly enjoyed mine so far! I am working on circadian clock research including circadian rhythms influence on diabetes and cardiovascular disease. I am working with Dr. Martin E. Young in the Cardiovascular Department at University of Alabama at Birmingham. During the first week in the lab, I observed the entire biological process of measuring gene expression within a mouse. I was allowed to watch Dr. Young sack mice and remove their hearts which had to be instantly placed in liquid nitrogen (stuff is quite fascinating in the way it evaporates so fast but it doesn't feel quite so fascinating haha). Next, using a mortar and pestle, we crushed the ventricle tissues allowing us to isolate the RNA. This is where my job for the summer comes in. I am running RT PCR (reverse transcription polymerase chain reaction) for numerous genes on all 230 mice samples. I typically run three plates a day and roughly two genes a week. My pipetting skills have improved drastically over the past few weeks. After completing the RT PCR, we must analyze the data. Dr. Young has been teaching me how to enter the data into excel and create the appropriate graphs. All and all it has gone pretty well so far. This week I am actually working in the lab on my own because Dr. Young has gone to an NIH convention at the Arctic Circle for the week (talk about one extreme to another from Birmingham temps to nonstop snow lol). Hopefully I will not make any mistakes while he is gone. I do have the lab assistant Lan (who speaks minimal English) to help me out when needed. Hope this overview was sufficient. Have a great week!
ReplyDelete-Andrew
Hello everyone. Now is as good of a time as any, to do my first post. Currently, I am working on my research in a more indirect way. As I mentioned in class, I am waiting for a method validation for one of the drugs that I am using LEE011 in Ringer's Solution, before I can start my project. I am helping my co-worker Kris with that, as well as doing other lab responsibilities, which as I said before indirectly relate. Since my project will involve a lot of sample running, I am getting practice with that by running Cyclophosphamide patient samples using a solid phase extraction method. (I have had to do a lot of reading, but for the sake of keeping this sort, I will only example when prompted.) Other than my patient samples runs, I have read a lot of journals pertaining to the transporter molecule I will be working with (Beta- cyclodextrin, BCD) as well as articles on pre-clinical operations such as microdialysis and the full plasma pk SOP. I hope to shadow another co-worker, Abi Davis, who will show me the in's and out's of how my samples will be collected and which mice we will be using. Long story short, a lot of reading and even more writing as I am putting together my project write-up and getting ready to present my project as our next lab meeting. I will keep everyone posted.
ReplyDelete-Brooke
Hi guys! So, this week has been a little slow in the lab because of software upgrading. Anyway, for the last two weeks, I have been running different Elisa Assay plates using proteins from hair samples. My very first Elisa was to ran the Cortisol plate (96 well microplate) and be able to understand the significance of the optical density for those proteins. Running the cortisol plate takes about 3 hours, but reading the absorbance of the plate has to be done within 20 minutes. Once the micoplate is in the microplate reader, it gives absorbance reading for each wells and detail statistical analysis of the data. It also constructs a standard curve, which is useful to determine the concentration of the proteins in the sample and whether dilution will be needed. I also ran a serotonin plate which uses a different protocol and the results were a little disappointing because we were only able to detect minute amounts of serotonin per sample. My focus in Dr. Anand's lab in addition to isolation of total protein with measurement of specific proteins of interest according to laboratory specific and established protocols will be to refine and/or develop an optimal ACTH enzyme-linked immunosortant assay (ELISA). Upon completion of the ACTH assay development, CANDLE hair samples will be quantified for ACTH content. The end goal of this particular project is to determine the relationships between ACTH, cortisol and other ACHT targets, and the basic demographic variables of the cohort (e.g., age, sex, race, insurance status).
ReplyDelete-Anqi
Hey Guys. Happy half summer. (Yes, summer is waning away day by day) So Research at UT has been great. The people there are very intellectual. For those who do not know, I am doing Periodontal research (Dentistry). The research is based on finding a good inhibitor of a specific protease, within a certain type of bacteria, porphyromonas gingivalis, which causes periodontal disease. I know you guys know what an inhibitor and protease is! The name of the inhibitor we are testing is E64. We have pretty much been using 96 plate wells to test out what we are researching. We testing to see how much inhibition is caused with increasing amount of this E64 inhibitor we are using. Once we follow the procedures (which you guys probably don't want to read about) we put the plate into an ELISA to test the activity of the cell (how much inhibition occurred).
ReplyDeleteEnjoy your 4th of July Everyone!
-Solomon
Hey guys. So I spent a good majority of last week helping pack up all our lab equipment because we’re moving from a lab at Le’ Bonheur down to one at the University of Tennessee. Tomorrow will be my first day in the new lab and will also be the first day without the pharmacy student there helping me do the experiments. As of right now, we’re running two experiments at the same time. The main one I’m focusing on is called cell proliferation and the other we’re calling caspase. Cell proliferation involves plating our hepatocyte cells and treating them with different concentrations of our omega-3 fatty acid solutions. The current hypothesis we have is that the presence of the omega-3 is somehow induces the cell cycle in the cells which therefore reverses, and may also prevent, the diseases associated with parenteral nutrition.
ReplyDelete-Brent
Hey everyone. I hope everyone is enjoying their summer so far. All of these blogs about research sounds very interesting! My research will be starting in July so I have not collected any data just yet but I will explain the basic outline of my research. I will be measuring contrast sensitivity on low vision patients at the Southern College of Optometry. Low vision patients are patients whose vision CANNOT be corrected with glasses or contacts. These patients include those with ocular diseases including: glaucoma, macular degeneration, optic neuritis, etc. The way that patients will be tested is by reading letters off of a white backgrounded chart. The letters will start out black and the contrast will decrease with each sequential letter. Patients will be asked to read each letter until they cannot distinguish between the letter and the background. After each patient's results have been documented, they will be linked to the intensity of their ocular disease. This study will help provide more information on the effects of each ocular disease that will be tested. My study will be combined and compared to another study that tested non-low vision patients which was performed last summer by Kristen Davis.
ReplyDeleteI hope everyone continues to enjoy their summer and research!
Hey y'all. Just got done checking on my plants which have been over watered due to all the rain this past weekend. I will be transplanting them into 120 individual pots after j take the mcat on Wednesday. As for now nothing exciting is happening in the life of research for me. So sorry this blog post is boring.
ReplyDelete-Cathy Thorn
Hello all (again.) Nothing really exciting has happened to me in the past week. Since I am still waiting for LEE011 to be validated, I am running patient samples for Cyclophosphamide and carboxyethylphosphoramide mustard. Today I ran 95 samples and the whole process of aliquoting, extracting, and processing took about 4 hours. I have also been helping prepare bulk samples for the validation of another cyclo metabolite, 4-hydroxy-cyclophosphamide, which will help me when I am preparing the samples for my validation. If all goes well, the LEE011 will be validated by Wednesday and I will be starting my validation shortly thereafter.
ReplyDelete-Brooke
Hello again to everyone. So the only thing new is I have completed another set of SSR’s via the PCR. After we complete the next set, we will run our check gels on random samples, just to be sure that everything has worked with the PCR. Then we will send all of them to UTHSC to run in their machine and make our SSR’s fluoresce so we can match them up. Dr. Mandel and I are hoping that the variance will be vast enough that it will be unnecessary to finish out the other 300 SSR samples. The chances for that are slim since T. recurvatum is after all a clonal plant, still fingers crossed, because that would mean I can start the data analysis portion of my project in mid July, and possibly be done with that before school starts back up. This project has been tedious and somewhat difficult, but interesting to see how the non-medical plant side of biology takes place. Yet, I have still learned a lot of techniques that are used both in ecology and medical research. So that’s always a plus. I hope y’all’s projects are also going well. Good luck.
ReplyDelete~Colt Terhune
Hi! The lab I work in basically researches the effects of wear particle debris from total joint implants. When a patient with a total joint implant moves, the implant parts move against each other. This creates wear debris particles and sometimes these can cause problems for the patient. For the past two weeks, I've been trying to grow cells (IC-21 mouse cell line) for my experiments. Last week, my cells were growing nicely, but after I replaced my flask with new media to feed them over the weekend, they started dying. I replaced the media again today, so hopefully they start growing again. I'm supposed to start an actual experiment tomorrow, so I'll let you know how that goes. Another CBU student, Karina, is working in my lab as well. She's been growing her cells since I left for California (~a month ago), so we're probably going to use her cells for the experiment. There's also a medical student and lab assistant working in the lab. They're working with THP-1 cells (human cell line). Today, the medical student showed me how to do an ELISA bc I've never done one before. My mentor had me run a couple practice cell proliferation assays to see how well I could pipette. I've also been practicing a bunch of dilution problems, which I have grown an intense hatred for.
ReplyDeleteCharlene
Hi everyone, I haven't started my research yet because its with people its kind of hard to ask them about their business. So ill just inform you on my plans.
ReplyDeleteWellness Care vs, Medical Care
Wellness care seeks to turn on the natural healing ability, not by adding something to the system but by removing anything interfering with normal function. When we think of medical care with think about the medicine doctors prescribe. Most of the time medicine is an on going thing that one would have to continue to take no exactly fixing the problem but sustaining the issue until the next prescription.
Whitney
Hi Again, I also learned something interesting. Any thing as simple as as headache and anything as disgusting as diarrhea could mean that its something dealing with the spinal cord.Spinal Manipulation therapy is very useful for many common body malfunctions. I never really exercised, but taking care of our bodies now would be a lot better than taking different medications when we all get older. I want to research the effectiveness of wellness care with and without medication. I say that wellness care alone is perfectly effective.
ReplyDeleteHello all! I am doing my research at LeBoneur Children's Hospital in the Cardiology Department. I will hopefully begin my research after the 4th of July Holiday. I have received my LeBonheur hospital badge that gives me access to all of the labs and offices in the Cardiology Department. For my research, I will be comparing the development and growth charts of children with Atrial septal defect (ASD), Ventricular septal defect (VSD), and Patent ductus arteriosus (PDA) to the charts of children without these congenital heart defects. ASD and VSD are holes in the heart while PDA is when abnormal blood flow occurs between two of the major arteries connected to the heart. The ages of the children I will be studying range in newborn- early adolescence.
ReplyDeleteSo, I have been running ACTH plates and doing some protein collection from the hair samples. Those proteins collected will be mainly used to quantify cortisol levels. I have learned that getting cortisol from hair samples have many benefits. First, hair is easily obtainable from testing subjects, and usually the more abundant the hair sample, the greater protein concentration. Second, this is a non-invasion method of measuring cortisol level, and this method quantify the amount of stress hormone found in the body over long term. Because cortisol is released in circadian rhythm, varying amount throughout the day, it is difficult to estimate an individual's long-term exposure through blood/saliva/urine test alone. Lastly, quantifying cortisol in hair samples provide an venue for monitoring chronic stress, such as anxiety and depression.
ReplyDeleteHello again! Recently, I have been analyzing some data from my polymerase chain reaction experiments. So as previously mentioned I am studying the circadian clock's role in regulating diabetes and cardiovascular disease. In order to observe this role at a gene level, I am looking at three groups of genes: clock regulated genes (change with presence/absence of circadian clock but may or may not be changed in diabetes), metabolic regulated genes (change with presence/absence of diabetes but may or may not change with presence/absence of circadian clock), and clock component genes (change with presence/absence of circadian clock but not related to diabetes). So we can confer that the last group mentioned (clock component genes) provides a sort of control group to make sure the PCR is working properly and to ensure the mice are genotyped properly. So far I have analyzed one gene (mcd) from the clock regulated gene group and one gene (mcad) from the metabolic regulated group. As might be expected, mcd showed a difference between the wild type mice and the clock mutant mice. Also, mcad showed a difference between the wild type mice and the diabetic mice. However, no difference was seen between mcd levels in diabetic versus wild type mice and no difference was seen between mcad levels in wild type mice and clock mutant mice. Our hope is that eventually, we will find a gene that overlaps between the two groups, or is affected by both diabetes and circadian clockwork at the same time. If such a gene is found, it will become the focus of future studies to see if any beneficial results can be obtained by altering the circadian clock regulation of that gene. Hope this explanation was clear enough! If you have any questions please post them and I will try to answer ASAP.
ReplyDeleteHey, so yesterday Karina and I started our first real experiment (but apparently it was still practice idk?). We put our cells in a 24-well plate in different conditions: cells in medium only (as the control), cells+LPS, cells+Titanium, and cells+Dirty Titanium (LPS+Ti). We also did a standard in a 24 well plate. We had to put the cells in the incubator overnight. Today, we removed the medium from each of the wells. We saved the medium and put each of them into their own tubes to freeze. The medium should have cytokines that we will test for in an ELISA (which we will be doing tomorrow). Then, we set up the cell proliferation assay on the computer. When we actually read the plate, however, our results weren't what we were expecting. I had to leave for work, but Karina told me that I may have set up the computer wrong so that sucks :( oh well...at least it's still practice.
ReplyDeleteFor my research this summer, I am working at UT clinical pharmacy lab. I am working with a fungus called Candida Albicans. I started by making 25 96 well plates, each with increasing concentrations of Amphotericin B (AmB), an anti-fungal drug. Strains of the fungus are grown in YPD from a library of cells and then I streak them onto SD agar plates and incubate them in 30 degrees C for at least 24 hours. Once the cells have grown, I mix them into 2 mls of sterile water and adjust them until the concentration is from 0.109-0.099 when read with a biophotometer. I then dilute the cells twice in RPMI, the media used for the AMb, and plate. I read the plates after they have been incubated in the shaker for 24 and 48 hours and record the first concentration of the drug that has no growth. When I get back from my vacation, I will begin analyzing my data.
ReplyDeleteHey everyone. So carving brains out of mice heads isn't exactly how I would have preferred to have spent the majority of my time in the lab last week, and seeing how the process made me sick to my stomach, I'll spare you most of the highly unpleasant details.
ReplyDeleteIn any case, I'll get down to it, and briefly describe how a perfusion is done: First the mouse is put into a jar of isoflurane which is used as an inhalation anesthesia to paralyze it. The mouse is then further anesthetized with an injection of Avertin. Once it is assured that the mouse feels nothing and is unresponsive to outside stimuli, it is cut open and dies when the diaphragm is cut. After it has perished, the aorta is cut, and saline followed by formalin, is allowed to run through the mouse for about 20 minutes for the purpose of ridding it of the blood in the brain (thankfully, none of this is done by me..). At this point, my task is to remove the brain from the head so that it can be put into a test tube and used for further analysis. I doubt I will ever get used to this...
Hey guys. Today was my first day of research at UTHSC College of Dentistry. My mentor was pretty busy today so I worked with her postdoc. She showed me around and let me culture some cells. All I know so far is that I will be working with SG cells and SCC cells and compare them by doing qPCR. I will probably have a better idea of what I will be doing by the end of the week.
ReplyDeleteHey guys! I hope everyone is having a great summer because I certainly am! Working with human subject is quite a change from working in a lab without human subjects. There are so many regulations that are in place for human subjects and at times it can get a little frustrating, but its rewarding all the same. I'm so much about the clinical and regulatory aspect of research dealing with human subjects. The clinic's specialty is endocrinology and it focuses on Diabetes manly. We have 7 ongoing studies that either deal with DM type 1 or 2. The subjects are mostly older patients who have cardiovascular issues and problems are diabetic complications. A normal research visit includes drawing labs (blood work) and taking a physical assessment of the subject and procedures, such as EKGs. I'm very fortunate to have this position and I am learning so many great things about research. (:
ReplyDeleteHey guys. So far I'm still learning lab techniques and my mentor is still busy closing up some data with the students from last month. She's finally given me my research title: Real Time PCR Analysis of Tumor Suppressor Genes In Oral Cancer. I've learn how to culture cells, split them, harvest them, do RNA extractions and run it on the nanodrop spectrophotometer, and do reverse transcription PCR in order to get cDNA. The last major lab technique that I will learn later this week is Real Time PCR.
ReplyDeleteHello Classmates. So I have finished up the data collection part of my research. Now I am working on my poster presentation that I must complete for my SIBS program this upcoming week. I am going to attach the abstract which I submitted for the program given out at the poster expo here at UAB. Hopefully this is a more concise and clear overview of my research project. Hope everyone is having a great summer so far!
ReplyDelete“Diabetes Attenuates Circadian Clock Output in the Mouse Heart”
Background: Diabetes mellitus affects tens of millions of Americans, and is the primary risk factor for cardiovascular disease development. The cardiomyocyte circadian clock directly regulates a plethora of critical myocardial functions in a time-of-day-dependent manner; following generation of two distinct mouse models wherein the cardiomyocyte circadian clock is genetically disrupted, our laboratory identified Dgat2 (diacylglycerol O-acyltransferase 2), Nampt (nicotinamide phosphoribosyltransferase), and Pik3r1 (phosphatidylinositol 3-kinase regulatory subunit alpha) as novel direct clock output genes. We have also previously reported that this transcriptionally-based molecular mechanism is altered in the rat heart during streptozotocin (STZ)-induced diabetes. Hypothesis: Circadian clock output is disrupted in the heart during diabetes. Methods: STZ was administered to 8-week old FVB/N mice (40mg/Kg body weight/day I.P. for 5 consecutive days); 16 weeks later, hearts were isolated from mice at 4 hour intervals over the course of the day. As a positive control for impaired clock output, cardiomyocyte-specific CLOCK mutant (CCM) mice were utilized. RT-PCR was subsequently performed for gene expression analysis of all hearts. Cosinor analysis was used to test for rhythmicity. Results: As anticipated, STZ treatment significantly decreased circulating insulin levels, and concomitantly increased circulating glucose levels. Similar to previous observations, dgat2, nampt, and pik3r1 expression oscillated in control hearts, and expression was attenuated in CCM hearts. Importantly, we report that oscillations in these clock output genes were absent in STZ-induced diabetic mouse hearts. Conclusions: Diabetes diminishes circadian clock-dependent rhythms in the heart, which could contribute towards cardiovascular disease development.
The past two days, I've had to research articles on hyaluronic acid. I had no clue what that is, but I've found out that it is a carbohydrate polymer and an anionic, nonsulfated gag (glycosaminoglycan). It is naturally present in the body, with highest concentrations in the eyes and joints. HA is a major component of synovial fluid and it increases the viscosity of the fluid, which helps lubricate, cushion and reduce pain in joints. It is present as a coat around each chondrocyte. Its molecular weight decreases with age, but the amount of it increases. A couple articles I read found that proinflammatory or stimulatory effects are associated with low molecular weight HA. One in particular tested the molecular weights of 52, 250, and 970 kDa HA and found that all activated blood phagocytes, however, low MW (52 kDa) had the highest effect, while high MW (970 kDa) had a significantly lower effect.
ReplyDeleteHey guys. Today in the lab we just focused on cutting brain slices with a special machine, that are to be eventually mounted on slides. The machine works by moving the stage on which the brain sits, back and forth, while slowly moving it higher and higher. The slices are very thin cut, and each one is placed into a petri dish well filled with PBS. It is necessary to keep a continuous supply of dry ice around the stage, so that it stays frozen. It is also important to slowly and carefully cut the slices, so that the arcuate nucleus stays in tact.
ReplyDeleteHey.
ReplyDeleteSo I am in New York right now and will be coming home soon. I have had a wonderful experience meeting so many people with Multiple Hereditary Exostoses. Each one of them were also so willing to help me by taking my survey about their GI issues. They were so happy that anyone is taking the time to research more than just the genetics of the disease and showing that there is more to is that growing bone tumors. It really is more than just a bone disease so this research is important to them as well as important to me. They were all great and supportive and will continue to be any help that I may need when it comes to this research. I will analyze all my data this weekend and then move forward from there.
-Cathy
Hey, my mentor has actually changed one of my cell line, so I am working with SCC-25 and GN-23. This whole week has been about culturing cells, splitting them, and doing RNA extractions. It seems like it's harder to get pure RNA for GN-23 but it's not so bad for SCC-25. Hopefully I can get a handful of pure RNA for each cell line by next week so I can get my cDNA from them and then on to qPCR.
ReplyDelete-Vu
I am approved to do research at Methodist! My mentor, Dr. Perez will be out of the office this week and next week for his wedding and honeymoon. I will be working with Jana Hanafin who has access to the cardiology database. She will send me the list of all the surgical and catheter based closures of PDAs, VSDs and ASDs from January 1, 2007- April 30,2014. I will analyze the charts with her until Dr. Perez returns.
ReplyDeleteWe have determined that ACTH was in low abundance in the hair samples. So, for the past 2 weeks, I have been running saliva samples instead of hair sample for ACTH ELISA. I have been using the same protocol for the saliva sample, but I changed the antibody incubation time to 24 hours in 4oC fridge instead of at room temperature for 4 hours with 200rpm rotator. It seemed though, increased incubation time helped with the binding of antibodies to the wells. Saliva samples were collected from my PI's MEG study last year, and those samples were stored in -80oC fridge. The samples were collected in a set of threes. The 1st saliva sample was labeled "AM" because it was collected by the mother in the morning, before tooth brushing or eating. The 2nd one was collected moments before the kid went to sleep according to his/her circadian rhythm, and the 3rd saliva sample was collected right after the lab technician have waken the kid. We had hoped that ACTH will show up with high values for hair samples with high cortisol value, since ACTH is the precursor for cortisol. However, what we have seen is just the opposite, in saliva samples, individuals with high cortisol value actually showed little to no ACTH and those with small cortisol value showed greater ACTH values. I think this reciprocal effect has to do with the negative feedback mechanism of hypothalamic- pituitary- adrenal axis. High levels of cortisol will stimulate the anterior pituitary gland to stop secreting ACTH, and low cortisol level triggers production of CRH and ACTH. This hypothesis seems to hold true for those few saliva samples that I have run so far, but I will have to wait until tomorrow to get all of the results.
ReplyDeleteHey guys
ReplyDeleteSo I've finally finished the portion of my research that deals with the cell proliferation. The results we obtained were not the ones we wanted to see that would've backed up our hypothesis (which was certain concentrations of the omega-3 in the fish oil could induce cell proliferation in our samples).
For some reason, the lower concentrations where we were expecting to see the most cell proliferation gave very different results each time the experiment was ran. And the higher concentrations where we were expecting to see the most apoptosis, or cell death, sometimes showed more proliferation than the lower concentrations. Im not completely sure what direction we will go with this after getting these results but we have a lab meeting next week and I'll know after that.
Hey again
ReplyDeleteThe next direction we're headed is to see by which pathways the omega-3 is working on the cell, whether the pathways deals with cell proliferation or apoptosis. The way we will be doing this is by isolating proteins from the Hep G2 cells after they have been treated with the omega-3 drug solutions and then running a western blot on them. As of right now, two pathways we are looking into are JNK and p53, which both deal with proliferation and apoptosis.
I gathered the proteins and made the solutions this past week. On Wednesday, Dr. Tillman's assistant and I tried to run the solutions on the gel and it messed up for whatever reason; so next week we will try again and hopefully get some results after that
I have finish ELISA for all of the saliva samples, and the next step is to analyze ACTH data sets. Today's plate reading showed the trend that was expected for ACTH values with certain concentration of cortisol. Most of the samples did show the pattern that we anticipated due to the HPA axis negative feedback system. Samples with the highest cortisol values showed lower ACTH values, which corresponds to the pattern that cortisol level peak during the morning hours and decreases throughout the day, and drops during the early phase of sleep. From the data, most samples showed the highest reading for the "post" saliva samples, which indicates lowest cortisol concentration. I think some samples needed to be repeated because the values seems a little off, but I will let my lab manager decide. She will also help me with data interpretation tomorrow.
ReplyDeleteSo I ran a TNF-a ELISA for my hyaluronic acid this week! We did a dose response (50 ug/ml, 25 ug/ml, 12.5 ug/ml, and 6.25 ug/ml) for each of the three molecular weights. We had done practice ELISAs to see if our particles were dirtied with LPS correctly for the controls, and they ended up not being dirtied, so we had to dirty them again. For my ELISA, however, my results showed that LPS had a very large TNF-a release (which it should, as it is the positive control--LPS is pro inflammatory), so we were glad that we dirtied the particles correctly for my experiment. In the articles I read, low MW HA should have had a proinflammatory effect, but my results showed that all of the MW for HA had about the same effect. However, previous studies looked at IL-1a, MCP-1, and IL-6 instead of TNF-a, so maybe TNF-a has different results than the other cytokines. My mentor is on vacation until the middle of August, so I'll have to wait for him to come back to find out if he wants me to do ELISAs for the other cytokines.
ReplyDeleteOn Thursday and Friday, we isolated DNA from mice tails to test for the NPY gene. The mice that don't have the NPY gene cannot be used for the experiment. This genotyping involves suspending each tail in proteinase k, a master mix, and then centrifuging, and placing into a freezer overnight. The next day several more mixtures were put into the test tubes until the DNA was dissolved. The samples were then put into a PCR machine. The final step to this was running the gels. Positive and negative DNA markers were loaded into the machine, and the samples that lined up with the negative marker were negative for the NPY gene. The ones that lined up with the positive marker were positive for it.
ReplyDeleteI tried submitting a post, but it didn't go through so I guess I will retype it. My last summer project has not gone the way that we thought it would. It turns out that the methods we developed are not sound enough at this point to produce reliable data. One of the methods that I was going to use for my project, LEE in Ringer's Solution, seems to have a %CV of 165 when looking at Internal Standard area counts. Since my lab is not sure how long it is going to take to fix this problem, they have assigned me another project. My new project is about determining the difference in Blood Brain Barrier permeability between HbMEC in normal media vs media containing Tumor secreted factors for three different subtypes of Medulloblastoma ( WNT, SHH, and Group 3). The idea is that the reason that drugs are more effective with the WNT subtype could be due to differences in the blood brain barrier caused my some of the different proteins that are being expressed. Our plan is to look at the TEER values as well as do Western blots with our samples in order to determine what tight junction proteins are being expressed. On the upside, since this research has never been pursued before, it means that anything we can discover will be useful as well as publishable and we can choose to look at a lot of different things if I desire to continue working on this project throughout the year. Anyway, I will be able to explain my new project as well as some of the abbreviations I used in this post at our class meeting on Tuesday. Until then Happy Researching!
ReplyDeleteHey guys! Everything is going well here with my research! As I mentioned before, working with human subjects is a bit different working in any other labs. The study I am working on is a 1 year study that involves type 1 and 2 diabetic subjects, testing a diabetic insulin pump. I also mentioned that the reason for the study is to observe A1c deterioration. All the subjects screened have come in for their 1st visit and they are starting to trickle in for the 2nd visits. One subject as come in for their second visit he had his A1c taken.
ReplyDeleteHey everyone, I still haven't gotten to the point of collecting data yet, but I'm getting there! The majority of what I've been doing in lab is the staining of the brain slices and it is a two day process. First a mixture of hydrogen peroxide and PBS is put on the samples for thirty minutes. After the thirty minutes are up, the samples are washed with PBS 3 times for five minutes, then a mixture of goat serum and Tx100 are put on the samples for an hour. After this, the primary antibody is put on and it stays on the shaker overnight. The next day I get the samples and do 5 more washes of PBS. Then the secondary antibody goes on the samples. Finally, I do 5 more washes with PBS and mount the slices on the slides. It is a long process and involves a lot of waiting, but I have come to find that it's my favorite thing to do in the lab.
ReplyDeleteHey guys. Still watering plants while getting eating alive by bugs. The plants are growing at a fast rate. No real updates or complaints. Hope everyone enjoys the rest of their summer!
ReplyDeleteHey guys, so this is the last week of my summer research. So far I am still collecting data. I'm almost at the core of my research project. I'm basically just running PCRs right now.
ReplyDeleteSo, yesterday I finally finished collecting all of my data! I met with my mentor and we looked over all that I have collected these past two months and it turns out, we found a couple of interesting things that he thinks would be worth taking another look at. The rest of this week I'm going to rerun these particular 'interesting' strains and also run them in a different media to see if they react the same way.
ReplyDeleteSo, I finally figured out what questions I am going to ask. We both figured that that wasn't going to be enough data for me. So we decided to ask the medical clinic right next door if I could sit by surveys in his office and he agreed to do so! I am still collecting things from them. Now that school is going to start soon I hope my availablility doesn't conflict with my work.
ReplyDeleteHey again, I forgot to mention that after my research he agreed to hire me as an intern at the office. I am very excited. The patients at the Cordova office and the Whitehaven office has made my experience very interesting. I feel as though my research is going to go by fine.
ReplyDeleteHey everyone! I am a little late blogging, but my research is still going! For everyone that has been in class, I am still testing cerebral arteries from rats for their reaction to ethanol. We are testing which cerebral artery will constrict the most to ethanol between anterior, posterior, basilar, and middle. From our results this far, basilar is the most reactive to ethanol and anterior is the least reactive. This week is our last full week in the lab, I will blog again about our results at the end of this week!
ReplyDeleteThe above comment without a username was done by me, Alex Bickenbach. Sorry about that.
ReplyDeleteOkay, hopefully this username works for Alex Bickenbach!
ReplyDeleteWhile my mentor, Dr. Perez was away on his honeymoon trip, he gave me the contact information to Miss Jane Hanafin at LeBonheur who has access to the database where the data for my research is coming from. I emailed her last week and she sent the charts and data to me on Monday, August 11.
ReplyDeleteHello all, my mentor, Dr. Perez, is back in Memphis from his honeymoon. I spoke to him today and we will be meeting this week to discuss how we will gather and extract the data and going over what things we need to obtain for each patient involved in the research.
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