Level: Intermediate   Steve Figard, Director of Cancer Research Lab, Bob Jones University   Teaching introductory biostatistics to sophomore level biology majors presents some interesting challenges, most notably, the inherent fear of statistics common to those who aren’t really interested in the topic but are being forced to take the course.  Many such students are, at best, uncomfortable with, or worse, totally petrified of, this topic.  Some insights into how to go about this without causing panic attacks in such students will be shared.     Auto-generated transcript...   Speaker Transcript Steve OK, so this is teaching introductory biostatistics using JMP overcoming the fear factor. My name is Steve Figard I'm the director of the Cancer Research Lab at Bob Jones University. I have actually also published a book. Through SAS press biostatistics using jump. Our introduction to biostatistics with Jim, I guess the, the official title. And what we want to cover today is how to teach or some thoughts. On how to teach introductory biostatistics to sophomore level biology majors and part of the reason this is such an interesting topic is there is a problem that We don't always realize, particularly those of us that like to go to discovery conferences, and that's the fact that we are the outliers in this we enjoy statistics. We love doing data analytics and we don't realize that there are that that actually makes this sort of a weird I have a good friend here on faculty who is a medical doctor and he wants saw me with my I love bias. I love statistics will help him on that I got at the Trump discovery conference. And he looked at me and said, you realize that proofs are clinically insane. Or my response back to him was. Yeah, but you know you need people like us, we sometimes forget that there are a lot of people that actually are scared of statistics. And biology majors in particular, particularly those that are starting off identify very much with this quotation from biostatistics text. Let us consider the likely scenario that you are a student of the bio sciences, whether you were at biomedical a physiologist and behavioral ecologist, or whatever. You'd like learning about living things you enjoy learning about the human body bugs and plants. Now, lo and behold, you have been forced to take a course that will make you do things with numbers and tread. Oh, dread even do something with numbers using a computer. You have probably decided that people who are making you do this or mindless status. So I'm those of us those there's actually a large body of people out there. The thing we in fact our mind, let's say to us, or at least clinically insane. And just to prove the point if you don't think that's the case, I start off my course asking my students to provide a one more description of how they feel. One day, one about taking my course. And this is the workflow that I actually created using the text explorer in jump. And you'll notice that The majority tend to be nervous or unsure or afraid and anxious, although we do have some that are excited or intrigued. When I find intriguing that Anyone would use the word a trained to describe about statistics course that's the problem before. So how do we deal with that. So I want to share some insights that I've gained to date using junk. don't require memorization, because in real life statistics are generally accomplished in consultation with others using other resources, besides yourself. You don't have to rely upon your memory. And how can we go about doing that. Well, we have plenty of in class exercises and case studies that are open book and open teacher This allows us to also overcome the initial learning curve for learning how to use jump because these students have never jumped before. So it's not just statistics that they're learning, but they have to actually learn the software package. By working with open book and open teacher, being able to ask questions as they start off, they can Learn these things, not by memorizing them, but just by prep from practice. I encourage collaboration amongst themselves that they don't just rely upon me to teacher to Learn these things that for successful exams that they have to take, but they are encouraged to collaborate with each other and teach help teach each other so that Any teacher will tell you get that you've learned best whenever you actually have to teach someone else. So I encourage collaboration in class. I use take home exams, because again, in real life, you generally don't have one hour to solve the problem, you are Told to go finish working a problem and report back as soon as possible and you generally have more time to Look at other resources and try to figure things out. So I use take home exams, of course, that does Fred provide the potential of cheating on the exam, but Generally, I can tell. You can tell pretty easily if someone's doing if you set up the exam properly, you can tell if they're they're cheating or not. Another point is I provide plenty of information and resources and flow charts and things so that jump is actually not an issue that learning how to use jump. I provide them tools that allow them to and if they figured out that they want to do a T test where in jumped you find the t test. So it jump itself is not the obstacle to learning the by statistics are learning how to Use the tools or to interpret the results they force actually has more stress on how to interpret the results and actually achieve real life solutions, then it does the mechanics of jump. Another point I found that is seems to be helpful is to share your enthusiasm. Whenever you present or teach the course. Make sure that as you're standing in front of your students. They can see that you're having fun up there. You want to approach your class as a mentor. Wanting to help your students not as the grand Grand Poobah and high executioner statistics that they should bow down before Don't be afraid to admit that you don't know something, of course you want to if it's something important that they they need to know and you really should know, then you get back to them and you go look it up yourself. You want to also show your enthusiasm by using examples that show the relevance of biostatistics to what the students actually need. My first lecture on the course. Actually, I showed them literature from the side from the biomedical and biological literature abstracts figure legends and things that incorporate Statistical terminology that if you don't have any clue what that means. You have no idea how to read that paper have no ideas on how to understand that paper so I you want to use examples that are relevant to the different areas of study that the students are actually interested in And last point I would say is employee as much humor as you can muster and your interactions with your students, you want to be relaxed and interact with your students as much as possible. Part of that is you want to begin your collection of means now so that whenever you actually Show things that I mean there's actually a fair amount of very humorous cartoons and things out there related to statistics and to some of the foibles because work via statisticians statisticians in general are still human beings and we still have our foibles and follies so There's More I could say, but we're limited because this is a poster and I will Welcome. Any further comments or thoughts in the future through communications with through sharp and Encourage you shameless plug here to look up my introduction to biostatistics with jump so that you can potentially see whether or not that might be useful in your own efforts to understand statistics and or to actually use it to teach your students.   ... View more

Level: Intermediate   Steve Figard, Director of Cancer Research Lab, Bob Jones University Evan Becker, student, Bob Jones University Luke Brown, student, Bob Jones University Emily Swager, student, Bob Jones University Rachel Westphal, student, Bob Jones University   Colorectal cancer is both the third most common and the third leading cause of deaths associated with cancer.  Previous studies in this lab have demonstrated the in vitro cytotoxic effect of almonds on human GI cancers.  An almond extract was prepared and processed using a pseudo-digestion procedure in order to mimic the effects the extract would have in a physiological system.  This extract demonstrated a dose response in vitro cytotoxicity to human gastric adenocarcinoma and was cytotoxic to a human colorectal adenocarcinoma, but had no effect on a healthy human colon epithelial cell line.  The extract was processed through filters with molecular weight cutoffs of 100,000 Da and 5,000 Da to estimate the size of any anticancer molecules, and it was found that the responsible molecules were less than 5,000 Da in molecular weight.  In addition, a polyphenol extract of the almonds was prepared and shown to have similar effects as the whole almond extract. It was concluded that the anticancer agents are likely polyphenols. Finally, four flavonoids commonly found in almonds were compared to the polyphenol extract, and they showed similar cytotoxicity.     Auto-generated transcript...   Speaker Transcript Evan Becker Hello, this is the 2019 BJU cancer research team. My name is Evan Becker and along with me today are Luke Brown, Emily Swagger, Rachel Westfall and we are all under the direction of Dr. Stephen Figard at the Department of Biology and Bob Jones University. And our paper, our presentation is on the molecular weight characterization of an almond component cytotoxic to gastrointestinal cancer cell lines. All right, for the introduction. Colorectal cancer is currently of great concern in the medical community as it is the third leading cause of cancer deaths for both men and women nationwide. Previous research from the BJU cancer lab has shown promise by demonstrating that almonds have a cytotoxic effect on LoVo colorectal cancer in vitro. A pseudo digestion procedure for an almond extract was also used as a way to mimic how the extract would work in a physiological system. The same almond extract was shown to induce a dose response and the human gastric adenocarcinoma cancer cell line AGS. Also this extract causes no negative effects in the human colon epithelial cell line CCD, indicating that the cytotoxic effects of almonds do not affect normal healthy cells. Passing our almond extract through molecular weight cut off filters of 100,000 Daltons and 5000 Daltons respectively, we were able to determine that the molecules present in the almonds inducing the set of toxic effect must be smaller than 5000 Daltons. As a result, polyphenols were determined to be a possible cause of the cytotoxic effect and the polyphenol extract was conducted on the almonds with this treatment showing very similar cytotoxic effects on the cancer cell lines. I think we're ready. Rachel Westphal To the cell lines that we use included AGS, which is stomach cancer; LoVo, which is colon cancer; and CCD, which was our normal cell line. We used 5-fluorouracil as our positive control for cell death. And we used an in vitro pseudo digestion of the almonds to mimic physiological digest...physiological digestion. WST cell proliferation assay, we utilize that to determine absorbance with a plate reader and use that to calculate percent viability. And then JMP was used for statistical analysis, we used ANOVA analysis, the Tukey-Kramer HSD test and the Wilcox and non parametric comparison. p values less than 0.05 were considered statistically significant. Can go to the next slide. Yeah. Luke Brown All right. The first set of tests I'd like to introduce you to are tests regarding the AGS human cancer cell line and this is a human gastric adino carcinoma. Now JMP was an important tool for us because it helped us to, first of all, determine the standard deviation in a few pilot studies we conducted. This then allowed us to use the software to run a power analysis to determine our sample size. Now looking at some of the data we got here, first of all, you'll see among both Figure 1 and Figure 2 what it has in common is PBS phosphate buffered saline and 5-FU or 5-fluorouracil, which is well documented and established cancer treatment for cancer, such as gastric and colorectal cancers. Now I'd like to direct your attention to Figure 1 here. Previous studies in our lab had already established that it seems almond extract does have some sort of cytotoxic effect that's selective to these cancer lines. Well, we hope to establish in Figure 1 here is, first of all, the dosage effect. And second of all, we wanted to narrow down what molecular weight, we'd be looking at to establish what compound or compounds are responsible for this effect. As you see, moving up from 12% almond extract all the way to 100% almond extract, we do see a dosage response. And we were able to establish this using the Tukey-Kramer HSD test inside JMP to be able to establish that these groups are statistically significantly different. Now you see that both the 100,000 molecular weight cut off filter and less than 5000 molecular weight cut off filter are statistically the same as the 100% almond extract. This led us to believe that whatever compound or compounds is responsible for this effect is going to be relatively small at less than 5,000 Daltons. Given this, we moved on to Figure 2 and we looked at some research showing that flavonoids have been shown to have a similar effect in walnuts. cyanidin, delphinidin, malvidin and petunidin. As you can see both the extract and all these flavonoids were actually more effective than the 5-FU in this treatment. So given this information, I'm going to pass the next slide to Emily here to give you some more information about another set of data. Emily So to make sure that we weren't just looking at results particular to AGS, we also ran another cancer cell line called LoVo. LoVo is a colon cancer cell line. As you can see, we didn't do all of the extensive dosage treatments on this particular one, because we'd kind of shown that with AGS. We were just particularly looking at is this particular to a certain line? So for LoVo, you can see that we don't have as low a value for 5-FU.   ... View more

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Hyohyun Park, Pre-Med Student, Bob Jones University Steve Figard, Director of Cancer Research, Bob Jones University   This poster is about statistical analysis on a cancer research with gastric cancer cells and almond extracts. JMP helped me analyze and understand how significant the differences were among the results of my experimental variables. There is a clear trend of increased cytotoxicity against AGS gastric carcinoma cells when treated with increasing concentrations of each almond extract.    Previous research in this lab showing an apparent cytotoxic effect of an almond extract on gastric cancer cells in the traditional 2D cell culture was expanded into 3D cell culture.   Almond extracts were made with a pseudo-digestion protocol and a new simplified non-enzymatic method.  AGS (ATCC® CRL-1739TM) cells were cultured in Perfecta3D Hanging Drop Plates from 3D Biomatrix until each drop had a cell sphere of approximately 150 µm diameter.  Drops were transferred to Thermo Fisher’s Nunclon™ Sphera 96 well coated plates and treated with increasing concentrations of almond extract for 3 days.  Cell viability was determined using the CellTiter 96® AQueous One Solution Cell Proliferation Assay.  ANOVA analysis of the results used JMP® software (SAS Institute, Cary, NC).   A dose response was observed with both extracts when treating these cells in a 3D culture system, although a statistically significant difference from media alone was not observed until the highest concentration tested (20%) was achieved (p ≤ 0.0013).  Statistically significant differences were also observed between the 10% and 20% extract treatments (p ≤ 0.0239).   These results are consistent with those previously obtained and suggest that almonds contain a natural cytotoxic activity against this line of gastric cancer cells. ... View more

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Partial characterization of the extraction method for and treatment with a potential anti-cancer agent from almonds Machado, Daniel Tuck, Amy Figard, Steve Bob Jones University, Greenville, SC Preliminary data revealed almond extracts created by a mock digestion process were toxic to gastric adenocarcinoma cells in cell culture.  Four variables of the extraction method and treatment were evaluated by DOE methodology to determine their effects on the viability of the AGS gastric adenocarcinoma cell line. Design and analysis were done using JMP ® software. The concentration of the digest added was the most important variable, with higher concentrations reducing viability more (p = 0.0168).  Increased incubation time was also better for killing the cells (p = 0.0250).  As main effects, the grind method and the mock digestion treatment were not statistically significant (p = 0.0546 and 0.4446), but the interaction between the two was (p = 0.0353).  If almonds were prepared by a dry grind, the mock digestion treatment worked better, but when a wet grind was used, the no digestion treatment worked better. It was not surprising to determine that increased concentration and incubation time resulted in reduced cell viability.  The interaction between the grind method and presence/absence of the mock digestion was unexpected.  This suggests that the cytotoxic component requires a liquid milieu to extract it from the almonds, and that it should survive the digestion process. ... View more