We open season two of Absolute Gene-ius with two guests sharing their collaborative work in reproductive biology aimed at understanding the role of RNA degradation in oocytes. They share insights on how oocytes are so unique, why they’re so important, and how better understanding them can provide insights to improve women’s health and lead to potential therapies. Join us to meet this interesting couple and hear how they’ve built a marriage in reproductive science.
We are all the product of a reproductive process, yet reproductive biology, or the study of the processes and mechanisms involved in reproduction, is not well understood. Deepening our understanding of reproductive biology is crucial to advancing assistive reproductive technologies (ART) and advancing our collective comprehension of inheritance and evolution.
Our guests for this episode are a couple, and we mean a literal married couple, of reproductive biology experts. Dr. Pavla Brachova and Dr. Nehemiah Alvarez, both working in the Eastern Virginia Medical School’s Department of Physiological Sciences. In their collaborative work they aim to better understand and characterize the role of RNA and cellular events that impact ovarian function in women. We learn about their work with oocytes, which are single cells that grow and mature within the ovary and once fertilized provide the foundations of an embryo capable of maturing to a new individual. They outline how they use digital PCR (dPCR) and other methods to monitor RNA regulation in single cells and how progressing this work and lead to potential RNA-based therapies.
In Cassie’s career corner we hear childhood stories from each guest and learn about their respective career paths, which eventually collided and merged. They share insights on the importance of having mentors experienced in your field, the challenges of shared job searching, and the joys of collaborating as a couple with shared scientific interests.
Visit the Absolute Gene-ius page to learn more about the guests, the hosts, and the Applied Biosystems QuantStudio Absolute Q Digital PCR System.
Nehemiah Seth Alvarez, PhD 00:00
I'm going to scare everyone.
Cassie McCreary00:01
Listen, we're recording this right by Halloween it needs to be scary. So yeah,
Pavla Brachova, PhD 00:05
Yeah, well finding two positions, doesn't get more scary on that.
Cassie McCreary00:09
Yeah, that ups the scare factor, right. Yeah, automatic horror show, right.
Cassie McCreary00:24
Welcome to Absolute Gene-ius, a podcast series from Thermo Fisher Scientific. I'm Cassie McCreary.
Jordan Ruggieri00:30
And I'm Jordan Ruggeri. And folks, the time has come. We are back for season two of our show. And we have got a fantastic slate of Gene-iuses lined up to join you in your ear buds over the next few months. And to kick things off today we have not one but two Gene-iuses with us, Dr. Pavla Brachova and Dr. Nehemiah Alvarez.
Cassie McCreary00:49
Pavla and Nehemiah study reproduction at Eastern Virginia Medical School. Their work focuses on RNA transcription and decay in early embryonic development and the implications this process has for fertility. And they're also married. We had a fascinating conversation with them about their research and their shared life in science. And we're super excited to share it with you.
Jordan Ruggieri01:10
Talk about a plot twist when Cassie and I realized that they were married and studying reproduction and oocyte decay.
Cassie McCreary01:18
Just a show about Gene-iuses hosted by a couple of Gene-iuses.
Jordan Ruggieri01:24
All right. Pavla and Nehemiah thank you so much for joining the Absolute Gene-ius podcast. We are very excited to have you on today. To start off, can you tell us a little bit about yourself and your research.
Pavla Brachova, PhD 01:37
My name is Pavla Brachova, and I am currently an assistant professor at Eastern Virginia Medical School in the Physiological Sciences Department. I went to college and a very small liberal arts college in Iowa. From there, I went to graduate school at the University of Iowa, in molecular biology. I was studying ovarian cancer there. I always wanted to study reproduction and I didn't get that opportunity until my postdoc at the University of Kansas Medical School. I was determined to be there for one year and find the job and start my own scientific career, but I ended up being there for seven years. And then we were able to find two positions here at EVMS. And so that's like my, that's my route that that I took to this point.
Nehemiah Seth Alvarez, PhD 02:32
My name is Nehemiah Alvarez. I'm an assistant professor in Physiological Sciences at Eastern Virginia Medical School. I'm also director of our advanced sequencing program here. I came from the University of Kansas Medical Center. So, I did a postdoc there. I also did my graduate school there. Pavla's postdoc took an extremely long amount of time, but my graduate school took me a long time. So I was, I spent four years in a PhD in one laboratory. And that laboratory subsequently closed down. And then so I had to restart my entire PhD all over again.
Jordan Ruggieri03:13
Oh, man.
Nehemiah Seth Alvarez, PhD 03:13
So initially, I was working in RNA biology and drosophila, and then I had to switch into DNA damage repair in mice. So, it was like, oh, like, you know, up the evolutionary ladder, but a step backwards in transcription, like, you know, I went from post-transcriptional regulation to like, pre-transcriptional regulation. So, I finished my PhD, short postdoc for two years, then I was on research track faculty for like a year and a half, and then we ended up getting both of us ended up getting a position here.
Jordan Ruggieri03:48
Seems like a wild path for sure. Can you tell us a little bit about the research that you're doing now and some of the projects that are happening in your lab?
Pavla Brachova, PhD 03:55
We are interested in female fertility and understanding what makes a good egg. In IVF clinics, when they do in vitro fertilization, the rates of success are fairly low. And even in unassisted fertility when people try to conceive the rates of success are also pretty low. And so, it's very poorly understood what are the requirements within the egg that will allow it to progress into an embryo and then make a successful pregnancy and make a human child. So that's kind of the big picture.
Nehemiah Seth Alvarez, PhD 04:41
So, a lot of what we work on now is trying to understand how in the egg, or the oocyte, what are the mechanisms that regulate RNA stability, and so the reason why we're interested in RNA stability is because the oocytes are very, the egg are very unique in cell type within mammals. And that is where there's no transcription. So, you think about a normal cell, it's making RNA and it's making new RNA about the exact same rate it's decaying the RNA and there's this equilibrium that is occurring. But in the oocyte in early and then during subsequent early embryonic development, there is no transcription. And when it’s going through early embryonic develop, or embryonic development, since with cleavages, it’s still not making any new RNA. And so, the cell has to make do with about 250 picograms of RNA over the span of like, it's like 96 hours without any transcription. Almost like 80 to 90% of the RNA is degraded before the embryo genome turns on to replace that RNA. And what we're focused on is understanding how are they, can be differentially modified and how that leads to that proper degradation. And so, the products that we work in the lab was specifically centered around that. And then one of our other projects is we develop; we work on trying to understand the RNA modifications that are that in relation to like how we can design effective RNA therapies to treat infertility disorders in the context of the oocyte.
Jordan Ruggieri06:22
Very interesting. So, talking again, about that RNA, I mean, the RNA is translated into some type of protein and has some type of, you know, structural function. Are there particular targets or particular pathways that are of critical interest? Are you looking more at just general quantity of RNA?
Pavla Brachova, PhD 06:45
In terms of the RNA modifications, what we've seen is that there are global patterns that happen, but not specific targets that are like super important and critical. It seems to be more of a global pattern and, um...
Nehemiah Seth Alvarez, PhD 07:05
Like a mass action that you need. Like general process, like we look at, we look at like how these RNA modifications could impact. We've talked about stability, but they also impact translation. In oocytes, there's a phenomenon this like translationally-coupled RNA degradation. That means for every RNA, when RNA is translated, it is degraded. It's like, they were like we made the protein we need, now we're going to get rid of that RNA, so that we don't keep making more of it at the wrong time. The RNA modifications can potentially impact how that RNA is translated, if it is translated at all, and it subsequently how it's degraded. That's sort of like we know, as we look at these modification status, and then we look at how that, like feeds into this degradation pathway. And there are other degradation pathways within the oocyte but the main one that were you we may focus on is the translationally-coupled side, because it's a, the tools that we have to study that are better developed than the tools to study like, you know, degradation pathways that only affect like noncoding.
Jordan Ruggieri08:14
Can you give us an overview of maybe what a common workflow looks like in your lab and some of the techniques that you're that you're utilizing? What does that workflow look like as you go from egg to data?
Pavla Brachova, PhD 08:28
For us, we work with single oocytes, and we primarily right now use the mouse model. And so, like a typical experiment, you have to hormonally stimulate mice to get them to be at the proper egg stage. So, we collect, they're called GV oocytes. And then in those, they have the full amount of RNA before the degradation has started. So, we compare those to ovulated eggs after another round of hormone stimulation. And so, we can collect single oocytes or single eggs and compare. Like, for example, we use the digital PCR or RNA sequencing approaches to then quantify the amount of RNA.
Nehemiah Seth Alvarez, PhD 09:16
Yes, that's like the general workflow we use for like our abundance, you know. So, we want to know like, right, we were interested in understanding like the half-life of an RNA, like how does its decay properties change over what does its decay curve look like? In order to measure rate, we need to be as you know, we need to use very like tools that allow us to do like be very quite as quantitative as possible. Pavla mentioned that we do use digital PCR. Sequencing approaches work, but there are some issues associated with those. That's more of like a relative, always at relative abundance. Even the use of UMI approaches within oocytes, just the nature of how they're transcriptionally quiescent leads to, the UMI approach is not a true representation of the molecules, they're just always been like, relative.
Pavla Brachova, PhD 10:08
Can you define UMI?
Nehemiah Seth Alvarez, PhD 10:09
Oh, yeah, so that's like unique molecular identifiers tags. So, these are just random sequences that you can attach to a do library preparations for RNA sequencing. You can attach them to the, there, they can be attached to the reverse transcription stage or after you get the reverse transcription stage. And then you attach the so you're counting what are potentially unique molecules, the UMI counts actually go up. You know, you could think of it as, if you had, if you're a sequencing real estate was a million molecules, and you gave, and you started with a million molecules, you're going to, your sampling of that space, it's going to be, you're going to have a million molecules with sequencing, and you could only get a million molecules output. You're going to get a distribution of those. You know, this is a rate not a random distribution, but you get a distribution that represents that input. So that's why we use the digital PCR approaches, because when we do digital PCR, actually, we actually just count the molecules that are present.
Pavla Brachova, PhD 11:10
And that's incredibly important in our RNA therapeutics studies, because like Nehemiah alluded to this, but we need to, we want to try to replicate the physiological condition as much as possible. And so we are going to be injecting RNA into an oocyte. And we need to know exactly how many copies of that to inject. And then be able to measure how the RNA modification is impacting the stability of that RNA, the translation, and then when it does reach maturity, make sure that it is undergoing the same decay that is happening physiologically. So, we always want to compare to try to, not to try to overload the system. Because we never know even if even if you can, like rescue a fertility defect, you might be altering a lot of RNA binding protein networks within the cells that could have other impacts that you don't even measure at the time.
Jordan Ruggieri12:15
Makes sense. I have a million questions from all of that information. Working with single cells and with RNA can be really complicated, especially when you're looking at degradation. Because you know, you blink at it two instead of one time, or three instead of two times, and the RNA is all of a sudden degraded. Is that something that you run across fairly frequently?
Pavla Brachova, PhD 12:40
Well, we did a lot of experiments when we were postdocs looking at the stability of RNA under different conditions, different incubation times, and it didn't seem to be as impacted by like temperature, for example. We saw that pH was a big issue. We did a lot of these types of like very basic experiments where we looked at RNA quality after certain conditions. And I think that made us feel pretty comfortable handling it. I think what helps us in the lab is that we got digital pipettes. So, they're extremely precise. And from person to person to person, there's very little variability. And so, I think our students also feel really comfortable working with it. But there are times when I am pipetting, one cell into a tube, and it doesn't get in there. I'm not as worried about the RNA as I am about the single cell. We've seen that sometimes there is no cell.
Jordan Ruggieri13:40
Oh, that's crazy. That's just, it's awesome as well. Just really, really cool to think about. Talking a little bit about the RNA therapeutic side of it, what are some of the hurdles when it comes to RNA therapeutics for your particular research? And how are you overcoming those?
Nehemiah Seth Alvarez, PhD 13:59
Just in general, the challenges that would be for the RNA therapeutic in terms of women's health and infertility is that the reason why someone would be infertile is really unclear. That's what makes it the biggest challenge for designing effective RNA therapies. And that's why like initially our primary focus is what is the general features of RNA need to be in order for it to be processed correctly by the oocyte? So, I think that the biggest challenge is figuring out you know, like, is it what would lead to, you know, a situation which we would need to use RNA therapeutics in the IVF.
Pavla Brachova, PhD 14:42
From the literature, it seems to be about one to 3% of women in in the IVF clinics have a genetic component to like their, the oocyte, not being able to mature properly, we're just looking at one very small window of the fertility process and the amount of women impacted is very low.
Nehemiah Seth Alvarez, PhD 15:08
In actuality, like if we had, you know, the budget of like $10 million, we could measure the abundance of every RNA in the oocyte. And then we could go back and say, "Okay, we're just going to make every RNA, we'll just suck out the entire contents of this oocyte and put back all the RNA, and then this and then just…”
Pavla Brachova, PhD 15:32
Total RNA replacement.
Nehemiah Seth Alvarez, PhD 15:34
Total RNA replacement. No, and we joked today,because it is actually it is, like, technically, it's feasible, like, I mean,
Pavla Brachova, PhD 15:40
Theoretically.
Nehemiah Seth Alvarez, PhD 15:41
But budget, budgetarily it's not. The ideal situation would be if we just keep working through this process, you know, we'll try to figure out at the molecular level, you know, how RNA's are processed, we might just, you know, settle on that really drive these oocytes, to get these oocytes kind of over this hurdle of maturation defects, you might just only need, you know, a few genes to do that.
Jordan Ruggieri16:04
Yeah, that's awesome. I want to follow up on the digital PCR elements that you discussed before. Getting that absolute quantification is really important. Are there any other factors that you consider when utilizing digital PCR or any other benefits you get utilizing that technology?
Pavla Brachova, PhD 16:21
In the past I've done a lot of qPCR, in graduate school and in postdoc. And it's always the results and the interpretation of those results. Like what does it actually mean depends on which gene you've chosen, which gene other literature has, like other papers have used to use as your reference. Yeah, to make it relative to that gene. But it's, it's really unknown how that reference gene is changing over time how your experimental conditions are impacted. And so, it makes it more murky, and it complicates the interpretation, and then just the overall entire experiment. So that's another reason why it makes it so much more powerful just to quantify the absolute amount.
Nehemiah Seth Alvarez, PhD 17:13
Just by the nature of what we look at, we use a lot of single cell sequencing technology approaches. And so this is either if you're actually just sequencing the direct, you know, the oocyte itself, or do we know some sort of, you know, using a single cell sequencing platform. So, we can look at potentially what those within the whole tissue, what those oocytes are looking like, transcriptionally, within the tissue, not after we have like, manually taken them out, and you know, and manipulated them in some way. But then the ovaries look at, you know, single cells within that tissue and the oocytes and then we also look at these oocytes in isolation using just a standard sequencing platform. And this gives us a nice large snapshot of what that transcriptional environment looks like and gives us the ability to look at targets that then we can interrogate in much more, you know, much more precise detail using the digital PCR. And so, we could use it for instance to find targets like that we can go after. The nanopore approach requires a huge amount of material and then the digital PCR approach, we just need one cell. And so, all these kinds of work together so we can get, you know, figure out, you know, within oocytes it's what's the modification? What's the spectrum modifications across any given RNA that we can use, like the we can use digital PCR approaches to look at changes in absolute amounts of like the enzymes that are response, potentially responsible for this. Then they can try to draw correlations, like back through the, across our data just to see like, where, you know, where could the defect, you know, if we see changes in RNA modifications in our in our nanopore data, you know, is that correlated with changes in expressions we see across our, you know, our digital PCR data and our sequencing data.
Cassie McCreary19:06
We hope you're enjoying this episode of Absolute Gene-ius. We're huge fans of digital PCR here and want to tell you about Applied Biosystems™ QuantStudio™ Absolute Q™ dPCR system. This instrument enables quantification of your targets without the need for standard curves with excellent precision in only 90 minutes.
Jordan Ruggieri19:24
You can say that again Cassie.
Cassie McCreary19:26
Okay I will! Get dPCR data in only 90 minutes.
Jordan Ruggieri19:30
Unlike other digital PCR systems, the Absolute Q dPCR instrument does not use emulsion or other droplet-based methods to compartmentalize reactions. Rather using distribution channels on a microfluidic array plate, the reagent is consistently delivered to physical micro chambers. The technology utilizes more than 20,000 micro-chambers with the coefficient of variation of less than 1%.
Cassie McCreary19:55
Another feature we're excited about is that the Absolute Q dPCR system analyzes more than 95% of the loaded sample, ensuring highly accurate digital PCR results. No more wasting upwards of 60% of your precious samples when loading, which is especially critical for rare targets.
Jordan Ruggieri20:12
Yes, and Thermo Fisher Scientific has developed a portfolio of robust Absolute Q digital PCR assays, So, if you need precise quantification of gene targets for research applications like AAV viral titer for cell and gene therapy research, eDNA analysis, SARS-CoV-2 wastewater surveillance, or liquid biopsy mutation and monitoring, then you'll want to look into this for sure.
Jordan Ruggieri20:36
You can learn more at www.thermofisher.com/absoluteq or visit the Absolute Gene-ius webpage. Again, that's www.thermofisher.com/absoluteq or visit the Absolute Gene-ius web page. The Applied Biosystems Quant Studio Absolute Q dPCR system is for Research Use Only. Not for use in diagnostic procedures.
Cassie McCreary21:02
Now back to our episode.
Cassie McCreary21:07
Welcome to Cassie's Career Corner, the part of the episode where we all don't have to think so hard.
Nehemiah Seth Alvarez, PhD 21:11
I don't know about that.
Pavla Brachova, PhD 21:13
At least emotionally it might be harder.
Cassie McCreary21:15
This is the part of the episode where I don't have to think so hard. And to our editor, jazzy into music here. So, I wanted to kind of, and I know we touched on this a little bit at the beginning of episode everything, but I kind of wanted to go back to your educational paths and your studies and everything. And I just kind of wanted to get to the root of it, of why you've chosen to kind of go not only just down the PhD route, but study what you have. And then you know, in turn, how you got to focusing in on reproduction and all of this type of thing. Like why is it that you're so passionate about all this? I know Pavla you mentioned in particular, then also Nehemiah, you as well, like how did just how did that all happen, I guess?
Pavla Brachova, PhD 21:55
For me it started actually, when I was eight years old, I clearly remember having this epiphany that if animals don't reproduce, that's just the end. Reproduction is one of the most important things. And so ever since I was eight, I wanted to study reproduction. And I always loved animals and tried to find out as much as possible. There was really no other option for me my whole life.
Cassie McCreary22:19
That's awesome, though. How about you? Nehemiah, how did you end up down this path, really?
Nehemiah Seth Alvarez, PhD 22:24
52You know I had always, I just generally always had a general curiosity of how things work. I just did this like I distinctly remember this, I was five years old. So, live on the cotton farm. And so, in Southern Oklahoma, and I was just pouring just chemicals. I'm pretty sure it's like antifreeze and diesel giving off all these fumes. And I was just thinking, “I just really want to know what's happening here. This is crazy.” I was always like okay, like, you know, how do things work like that. That just sort of kept building as you know as my education, you know, increased. And as I was, you know, exposed more and more different things, you know, that always stuck in the back of my mind. You know, how do things like work in reproduction. Reproduction is an extremely interesting field, you know, you start from one cell, and you end up with organisms that are billions or trillions of cells. You could argue that it's, it's pretty niche. Everyone, you know, it's a very tight knit community. I mean, everybody probably could trace their, like their entire career trip back to one, like, maybe like five people. I mean, it's actually very small. it's a very small field. And so, yours is by the, but it's very exciting. It's very sort of fun.
Cassie McCreary23:40
First of all, it is my personal quest on this show to help solidify the concept that science is the coolest. So, thank you for contributing to that. I said on our very first I think our very first episode I called science the punk rock of academic disciplines. Like its backbone is curiosity. It is the epitome of like, screw around and find out. Science is the coolest. So, we've reinforced that. Now what you're saying about this being a tight knit community, like something that has come up time and time again, in my career corner discussions is kind of like the importance of mentors but also networking and that type of thing. And being in the field that you're in having as kind of a relatively smaller bubble, what is finding a mentor like? How do you hang on to that mentor? And then on the flip side of things I would imagine now being in the positions that you're in, you're starting, you know, you're mentoring people as well. You know, what is all that like?
Nehemiah Seth Alvarez, PhD 24:33
In general, it's really important to find a mentor who's like, who is supportive of your career choice. And I think like, well, so I think that in my observation from my graduate school through postdoc, and even, you know, early. Even my junior faculty right now, is that most trainees in that position, they'll always say, like, you know, they'll be very candid, they'll say, "I don't know what I want to do." Like, you know, that's honest, it's an honest thing. But I think that what really helps, like, where I see people who are, who are very successful, is people who internalize that thought, and instead just say, "This is what I want to do." You are 100% committed. And then it's okay to change your mind. But it's like, it's just like, you know, that your mentor will support you in changing your mind.
Pavla Brachova, PhD 25:30
Yeah, and like how Nehemiah was saying that you just have to go 100% in if you know that's what you want. We were told don't study the same thing. Don't be in the same department. Don't be in the same lab. We had a vision. And that's the only thing that we wanted to do. And so, we just tried as hard as we could to make that happen. And even like, even though we are here, it almost didn't happen. Go after your dreams. I mean, it's corny, but you have to do it. If you know that you want to do it, you have to do it.
Cassie McCreary26:06
Yeah, that's sage advice. I think we only have about 10 minutes left. So, there's one more question I want to make sure we fit in because it's my favorite question. So selfishly, I want to ask it. I ask everybody. What is your best or favorite or proudest lab moment? And what is your biggest, like lab mistake, oops, screw up, oopsy daisy, whatever you want to call it?
Pavla Brachova, PhD 26:34
There's two things that I remember as my biggest oops. One that's most vivid in my mind is the most recent one. In which I was doing an RNA sequencing library preparation. And like we talked about how important it is to have like an internal normalizer. So, in oocytes during the RNA decay process, so that you can quantify a little bit more accurately how many transcripts there are of a certain isoform and, and that's called an ERCC spike in. And so, it's a standard RNA that you can buy from a company, and you have to spike that in at the beginning of your experiment. So, then it undergoes the reverse transcription, the amplifications, everything, just like all the other RNA compare to it. I forgot to put that in there. So that was a whole experiment.
Cassie McCreary27:23
All right.
Pavla Brachova, PhD 27:24
Yeah, I did also miss out the buffers, which Nehemiah had to spend like an hour conversation with the sales rep, or the, the scientists at the company. And we were able to, Nehemiah was able to fix it. And then I also remember in graduate school, I was loading a 384-well plate for qPCR and at the very end, somehow it slipped, and the whole thing just splashed into my face.
Cassie McCreary27:51
Oooh.
Pavla Brachova, PhD 27:52
So that was a day. I sat there, I sat there, and I cried, and I just went home.
Cassie McCreary27:56
That's sad. All right. And on a positive note, then what's your proudest? What's your happiest?
Pavla Brachova, PhD 28:02
When I'm happiest and kind of most excited about the research is when I'm writing grants, and talking about the ideas and developing the projects, that's what makes me the most excited. And that's kind of how we divvy up a lot of the work because we run a joint lab. And so, we have independent offices, independent space, independent funding and everything, but we do everything together and we submit everything together. And we are a joint unit in like, every way possible. That's what is kind of the most enjoyable process to me is developing the ideas and talking with Nehemiah about where we're going to go with our next project.
Cassie McCreary29:05
All right, and Nehemiah when have you screwed up?
Nehemiah Seth Alvarez, PhD 29:17
Either right when I started grad school or when I was a technician, I was doing a lot of, I was in a laboratory where we’re in RNA biology, so we always had to like to make our own antigens and make our own antibodies, purify them and do a lot of protein, not like hardcore biochemistry, but do a lot protein biochemistry. And so, a lot of this involved a lot of cloning or molecular biology. And I remember, I remember the, you know, going through all these the steps of, you know, cloning the gene that we were interested in, and then going through the whole process of cloning into a vector and going through all thing of like, expressing it and E. coli and purifying it. Anyway, this whole thing, and nothing was working, alright, we couldn't get the protein, we couldn't see the protein on the protein gel, you know. Nothing was expressing or changing, like the, the temperature doing all this kind of weird thing. And just like months of like, troubleshooting. You go back and look, and I forgot to add the IRES sequence. So, this is an important ribosomes entry sequence that tells you know, in in eukaryotic organisms to start translation. And all of this was precipitated because of a lack of a control. Right? Like, I didn't load a control on a gel initially. And so that would have told us immediately what was wrong. Instead, we're operating, and I was operating under the assumption that it was expressed that we were just doing something, you know, it was it was expressing in the E. coli, but it was just precipitating. It was insoluble. So, we were doing all these, you know, crazy purification strategies and dialysis and I mean, just, it was just, you know, a colossal waste of time.
Cassie McCreary30:47
How about proudest? Or best?
Nehemiah Seth Alvarez, PhD 30:50
You know think it was just, I think, honestly, it was getting us into a joint position. And I think that was like one of the most, you know, relieving but also exciting, because then we can continue to do research together. It would have been a very, like, different experience than it is in like now. I mean, we're, the only time we actually end up really talking about science is actually at work. You know, our kids are like, you know, our kids are, you know, they we have to, you know, focus on them.
Cassie McCreary31:18
They’re like, “Noooo!”
Nehemiah Seth Alvarez, PhD 31:18
So, like, no, no, no, stop that. Stop talking about that. Stop talking about it. We’re like, “OK, fine.”
Jordan Ruggieri31:24
Pavla, Nehemiah, thank you so much for joining us for Absolute Gene-ius. We really enjoyed having you on.
Pavla Brachova, PhD 31:30
Thanks so much. We also really enjoyed our time with you.
Cassie McCreary31:34
That was Dr. Pavla Brachova and Dr. Nehemiah Alvarez at Eastern Virginia Medical School. Thank you so much for joining us for today's episode of Absolute Gene-ius. We have much more to come this season. This episode was produced by Sarah Briganti, Matt Ferris, and Matthew Stock. Stay curious and we'll see you next time.
Nehemiah Seth Alvarez, PhD 31:53
No, this is this is the same. I just wear a blue shirts, so.
Pavla Brachova, PhD 31:59
Well, Nehemiah has the Obama strategy where you have the same the
Nehemiah Seth Alvarez, PhD 32:03
The same like colored shirt, I just changed my tie.
Cassie McCreary32:07
So, you're like a cartoon character. You open up your closet, there’s just like 40 of the same. Yeah.
Pavla Brachova, PhD 32:11
Yeah, you open his closet and it's like the same exact outfit.
Nehemiah Seth Alvarez, PhD 32:14
I just change the tie. That way you know what day of the week it is because the tie will just change.
Cassie McCreary32:19
There you go.