LTU3IND Placement Logbook - Sara Radovanovic by

DAY 1: Safety induction and Tour (21/08; 5hrs)

21069622 — 2024-08-28 00:24:53 UTC

Today was my first day at the Centre of Cardiovascular Biology and Disease Research (CCBDR). I was nervous and didn't know what to expect as I had never been to a research facility before, but the fact that it was located at La Trobe University, my nerves slightly settled. I met up with Vivian, my secondary supervisor at 9 am and she showed me around the PhD student office and their laboratory; where I got my laboratory safety induction completed by my primary supervisor, Maria. Vivian then took me to La Trobe's animal house where the dissections and direct animal work happens for the PhD student's research. This is where I met another lovely PhD student, Buddhila, in the middle of a cull (killing animals for dissection). Vivian and I ended up staying with Buddhila for the rest of the day where I weighed the kidneys and spleens of her test mice. I was slightly overwhelmed since it was my first day. Nonetheless, I was grateful for the push into practical work on the first day and can not wait for the days to come!

DAY 3: Weekly Team Meeting (23/08; 2hrs)

21069622 — 2024-08-28 00:24:53 UTC

Today I attended the weekly meeting where Maria's Honours student, Roberto, explained his current research so far. Prior to the meeting commencement, Vivian and I helped media representatives produce videos that showcase the key facilities and collaborations associated with the CCBDR for advertisement purposes to industry partners. During the meeting, Roberto explained the research he has collated so far on the causal relationship between Gamma Delta T-cells and Abdominal Aortic Aneurysms. To work on my oral communication goal, today I asked 1 question.

QUESTION: What is the mortality rate of abdominal aortic aneurysms and are their any current treatments?

ANSWER: "AAAs are almost immediately fatal with a mortality rate 90%, and there are no pharmacological treatments, only surgical interventions."

DAY 4: Heart Camp Day (26/08; 6.5hrs)

21069622 — 2024-08-28 00:24:53 UTC

Today was Heart Camp Day!! Heart Camp is an event that the CCBDR runs as an engaging and informative way for Primary-Secondary schools to learn about the function of the heart and the importance of keeping it healthy. At the beginning of the day, there were 4 speakers who each talked about different parts of heart health including; nutrition, brain, kidneys and the heart itself. The kids then rotated between 'Science Stations', which included playing around with anatomy games using VR goggles, seeing the difference in their heart rate and blood pressure before and after exercise, and looking at the Zebrafish anatomy under the microscope. After lunch, Vivian planned an 'amazing race' type challenge where the children went around the university and completed challenges at each checkpoint to receive a ticket stating the location of their next challenge. I volunteered to host one of the challenges to exercise my public speaking and leadership goals. The challenge involved reassembling a model of the brain, and my role was to explain it to the kids and assist and give them hints as they performed the challenge. I also helped in the blood pressure measuring science station as well. This experience put me out of my comfort zone as I've never done a role like this before, however, as the day progressed, I became more confident and comfortable with speaking and directing the kids through the challenge/station and I am grateful for the experience it gave me.

DAY 2: SABE Seminar (22/08; 1.5hr)

21069622 — 2024-08-28 00:24:53 UTC

Today, I attended a SABE seminar at La Trobe University, where researchers are invited to talk about their projects and spread awareness of their findings. I had never been to a research seminar before and one of the speakers was one of my supervisors, Vivian, so I was pretty excited but also nervous. As I am still new to the research team, I am not yet familiar with many students or researchers, so I had no one to go and sit with, but I still wanted to come and support my supervisor regardless! The first speaker explained the health and environmental benefits that legumes provide, highlighting how special bacteria within the nodules of legume plants undergo nitrogen fixation, which converts atmospheric nitrogen into a form that enriches our soil. Vivian spoke second and her performance was nothing short of amazing! Since I already knew her project, I felt much more comfortable being a part of the audience than I did during the first speaker. Overall, I really enjoyed that day and may start eating more legumes!

DAY 6: Weekly Team Meeting (30/08; 2hrs)

21069622 — 2024-08-28 00:24:53 UTC

Today was lab meeting day again! It was Bill's (the other lab head of the research division) turn to host the meeting. He decided to have a relaxed meeting today and he kindly shouted us coffee, and then we sat in the agora. As we sat and sipped on our coffees, the team discussed housekeeping and lab maintenance before letting the conversion drift into more casual topics. The homely and comforting environment the research team creates together took me by surprise, but a pleasant one! It was at this moment when I realised my perception on the research field and the environment I would endure (if I completed my honours or a PhD) was completely inaccurate!

DAY 5: Lab Day w/ Vivian! (27/08; 3hrs)

21069622 — 2024-08-28 00:24:53 UTC

Today was mostly spent in the lab with Vivian! She taught me what a Haematotoxylin & Eosin (H&E) stain is and allowed me to perform the procedure, while she guided me through the steps. This was a big moment that helped me with my leadership skills, as later that day, I explained the procedure to another PhD student to confirm what I learnt. I noted the procedure for the H&E stain, as this was going towards my evidence at the end of the semester. We then began preparing for other stains required for her PhD, including Oil-Red-O staining. This involved filtering diluted isopropanol through Oil-Red-O powder and filter paper. This stain is used to visualise fat/lipids in tissue samples; which can be helpful in pathological studies. This day was very productive as I recorded a lot of laboratory experience which can be appealing for future jobs in research and laboratories. Overall, I had a lot of fun!

DAY 7: Lab Day w/ Buddhila! (02/09; 4.5hrs)

21069622 — 2024-08-28 00:24:53 UTC

Today was another lab day! Vivian and I assisted another PhD student, Buddhila, with flow cytometry and flow processing of different organs, including mice kidneys, spleens, and aortas. I have minimal experience with handling organ tissue, so this was very exciting! My part in the flow cytometry process was to 'mash' and strain the organs into falcon tubes to make them as broken up as possible, ready for the individual cells to be separated from each other. I also weighed mice hearts which wasn't as easy as it sounds! It requires a lot of focus and a delicate hand to make sure the tweezers don't drop the heart but don't squish it either. Today was very lab-heavy. I felt some stress since I was careful to avoid mistakes that might impact Buddhila's research experiment! But gaining more experience with handling mice organs was still very exciting!

DAY 10: Team Meeting (06/09; 1.5hrs)

21069622 — 2024-08-28 00:24:53 UTC

During today's meeting, Herika, a post-doc student, presented how her research is going. Herika's project involves analysing sex-related differences in different types of B-cells. Based on my prior knowledge in immunology, I understood that B-cells were a part of the adaptive immune response and when activated, they undergo clonal expansion. However, I learnt today that the type B1a-cells don't undergo clonal expansion! I also learnt that females have a quicker and more robust inflammatory response in chronic diseases than males. This explains why more male mice die or have a more severe prognosis as their immune system acts later.

My question for this weeks meeting is below!

Q: Why don't B1a cell undergo clonal expansion?

A: "unlike B2 cells, B1a cells undergo self-renewal processes and enter a state of 'functional unresponsiveness' which important so the cells don't make antibodies against self-antigens (attacking itself), which can potentially result in an autoimmune disease".

DAY 8: World Heart Day Preparation + Lab Errands (03/09; 7hrs)

21069622 — 2024-08-28 00:24:53 UTC

For the first couple hours of my shift today, I assisted Vivian with editing the informative posters for World Heart Day which is coming up in a few weeks. Vivian then had a meeting at 1:00 pm, and she allowed me to do any notes or assessments from my university subjects, which I gladly did while she was away. When she returned, we ran a few errands at La Trobe animal house and then picked up mail from the postal/storage area on level the bottom level of the TLC building at La Trobe. Once we returned to the office, Vivian told me about her plans for World Heart Day, which will involve a 2km walk and a fruit bar brunch for everyone involved!

[Relatively busy day today; forgot to take a photo!]

DAY 9: SABE Seminar and Lab Day w/ Vivian + Jake! (05/09; 7.5hrs)

21069622 — 2024-08-28 00:24:53 UTC

Today, Vivian and I started in the laboratory! We spent the morning (~3hrs) going through the Oil Red-O staining procedure again, as the results from the first time we did it together, didn't come out too well. But this time, Jake, one of the PhD students, joined us!

At 12:00pm, Vivian and I attended another SABE seminar. The researcher who spoke, Elaine, discussed her project which was all about the microbiome. I learnt that our microbiome has a major influence on illnesses, including cardiovascular diseases, mental health issues (like depression) and various cancers!! After this seminar, I am now going to be more mindful of my food and drink choices!

We then returned to the office, where I helped Vivian prepare for World Heart Day. Specifically, I helped her edit the posters that will be on display during the event. The posters explain topics on heart attacks, kidney health, nutritional health and cardiovascular health!

DAY 11: Team Meeting (09/09; 1.5hrs)

21069622 — 2024-08-28 00:24:54 UTC

In this week's meeting, it was the amazing Vivian's turn to present!! Vivian's project focuses on determining the sex-related differences in metabolic syndrome (MetS). With her project, Vivian aims to bridge the gap between the sex differences present in pathophysiology, so patients can be medically attended to appropriately and prescribed the correct pharmacological treatments. She explains that after 4 years of research (which she thought felt much longer!) she was able to identify biological differences between male and female mice with MetS. Most MetS studies involve inducing the disease in mice by using diet models such as a high fat, salt or sugar diet. What makes Vivian's project so special is that she uses a high fat + salt + sugar diet model, which provides more information than a diet just on one of these factors. It was very motivating watching Vivian express the powerful passion she has for her research and inspires me to be just as passionate about my hard work one day!

Q: What is the FEAST trial? (she mentions it in her research)

A: "FEAST is short for the 'Effects of Anti-Inflammatory diet for Knee Osteoarthritis'; this trial compares the efficacy of two diets on improving factors such as pain and function in patients with knee osteoarthritis"

DAY 12: H+E Stain Imaging (10/09; 4hrs)

21069622 — 2024-10-06 11:20:51 UTC

For most of the morning today, I tagged alongside Vivian with running laboratory and office errands around the university. We visited the animal house to discuss clean-up dates and other housekeeping updates.

We then headed back into the laboratory to take snapshots of some of the liver stains she had performed previously. The cross-sections were from various mice she studied, including healthy males, males with Metabolic syndrome (MetS), healthy females and females with MetS. It was very interesting analysing the differences between healthy and diseased tissues, but very difficult! Diseased tissue would consist of cell 'ballooning' (swelling) and have high immune cell infiltration.

At 12:30 pm, Vivian spent lunch watching a seminar, where a foot podiatrist spoke about her work and different types of foot conditions. Unfortunately, I had to leave at 1:00 pm because I had class, but overall, today felt very productive!

DAY 13: Errands + Histological Analysis (11/09; 3.5hrs)

21069622 — 2024-10-06 11:21:06 UTC

Since Vivian's role in the CCBDR is very important, she is a very busy lady! So most of the shift today, like yesterday, consisted of running a lot of errands for the first part. We visited the upstairs office (the microbiology research department) to pick up some posters and other things Vivian was looking for, for World Heart Day.

To finish off the day, Vivian showed me some ultrasound images of mice aortas she took last year as part of her research. She taught me how to identify differences between healthy and diseased aortas, and the general process of taking ultrasound images. Mice aortas are extremely small, being only a couple of millimetres in diameter, and 2 or so centimetres in length; it was mindblowing to understand how researchers can get such accurate images on something so small!

Before we were finished for the day, Vivian wanted to head into the laboratory so I could practice analysing healthy and diseased tissues, but this time, we didn't just look at liver cross-sections; we also analysed cross-sections of mice kidneys, aorta and heart under the microscope!

I forgot to take photos of the histology samples or ultrasound images (I was too distracted by their beauty!), so here's a photo I took, while we were running errands, of one of the PhD students, Tayla, picking up mice aorta's from storage in the liquid nitrogen tank!

DAY 14: First Yr Fest (12/09; 5hrs)

21069622 — 2024-10-06 11:21:19 UTC

Today was a HUGE day for me; I co-chaired a university seminar for the first time! I cannot form into words how nervous I was for this moment, but I knew this would be a major moment of growth, especially in my oral communication skills and confidence. The seminar was a part of the First Year Fest, which involved 1st year PhD students and 2nd year Masters students from La Trobe sharing the research they've collated so far, and the aims of their research for the future. I co-chaired the first session of the day with Tayla and I'm very grateful I did so. Tayla's confidence and way with words were so impressive, I couldn't have done it with anyone better! As part of the job of a chairman, we have to be ready to ask the speaker a question if the crowd doesn't have any questions after their talk. This was a struggle and very stressful for me because most of the research being presented was very advanced, so listening and paying attention to the speaker was very critical to formulate a question. Throughout the seminar, Tayla gave me tips about speaking, as well as what types of questions to ask. Luckily the audience was very inquisitive today, but I was determined to ask at least one question before our session ended; so after the last student of our session finished discussing their research on lymphodaemas using zebrafish, I asked my question!

Q: Do you plan on looking into whether there are sex-related differences in the response to different lymphoedema treatments?

A: "At the moment, my research uses zebrafish that are not yet fully matured into adulthood, and not until adulthood, zebrafish aren't sexually dimorphic, meaning there are no systematic differences between males and females; therefore I wouldn't be able to test for sex-related differences".

DAY 15: Weekly Team Meeting (16/09; 2hr)

21069622 — 2024-10-06 11:21:31 UTC

In today's weekly meeting, Buddhila was allocated to present and update us on her research so far. Buddhila's research looks at determining the role of the cytokine IL-1RA (Interleukin-1 Receptor Agonist) in kidney inflammation and kidney fibrosis. She mentioned that as part of her PhD research, she had to measure the blood pressure of mice using a method called the 'Tail-Cuff method' - so for my question for this meeting, I asked Buddhila this:

Q: Is measuring BP using the tail-cuff method reliable? and are there alternative ways to measure BP in mice?

A: "No, it isn't overly reliable as the tail is relatively far from the heart. The closer the measurement is taken to the heart, the more reliable the measurement will be; but, the tail-cuff method is much cheaper and more convenient to perform".

"the golden standard for measuring BP in mice is called radiotelemetry; which is generally used to confirm tail-cuff readings"

DAY 16: Liver Scoring in Lab (19/09; 3hrs)

21069622 — 2024-10-06 11:21:38 UTC

Today, I came into placement to assist Vivian with scoring liver tissue sample images she took of her mice. Liver scoring consists of analysing stained liver cross-sections and giving them ratings on the level of steatosis (fat build-up), lobular inflammation and hepatocyte (liver cell) ballooning. Firstly, Vivian and I went through a couple of images together so she could teach me how to rate the tissues, but since liver scoring is an independent and subjective task, for the rest of the images we individually rated them based on what we thought. I found this difficult to do because I don't have much experience in histological analysis (and to me, all the images looked almost identical!). It took me much longer to rate each liver image compared to Vivian and this reminded me of how talented researchers are due to the enormous amounts of effort and time they put into their work - Vivian was a pro at scoring liver! This was a long process but very interesting and will be very beneficial for when I head into the clinical scene!

DAY 17: Oil Red-O Staining day (20/09; 3.5hrs)

21069622 — 2024-10-06 11:21:45 UTC

Today, Vivian and I spent the day in the laboratory staining another set of liver cross-sections using the same method we have used previously; Oil Red-O staining. The past couple of times Vivian and I have performed this stain, the results don't come out as clean and crisp as what Vivian would like them to. Each time we've performed the stain, Vivian has tweaked certain steps. This time round, she left it submerged in the dye for 5 minutes longer, so hopefully the results will come out much better this time! This stain is an overnight process so we won't know until the next time we come back into the lab! Today highlighted that experiments don't always work out perfectly (not even if you stick strictly to the protocol given by the company!), so it's very important to stay persistent and motivated in a field such as research.

DAY 18: More Staining + Team Meeting (23/09; 4.5hrs)

21069622 — 2024-10-06 11:21:55 UTC

Today was a very, very fun day. Firstly, Vivian explained how our recent Oil Red-O staining session turned out better, but not yet up to her standards. So for most of the morning, Vivian and I prepared another round of Oil Red-O stain solution and scavenged some old cross-section sample slides to use for the stain. Once again, Vivian tweaked a different aspect of the protocol (fingers crossed this is the last tweak we will have to make!). After leaving the tissues to stain, Vivian and I tidied our laboratory bench and made our way to have lunch with the rest of the team at 12:00 pm until our team meeting at 1:00 pm.

(Now for the most fun part) In today's team meeting... I was allocated to present! I was lucky enough to take my other supervisor's (Maria) place and present why I'm here, what I've achieved so far and what I plan to do for the rest of my placement. I spoke about my degree and the subjects I've so far completed, and my current career aspirations and future workplaces of interest. I also explained the goals I made as part of LTU3IND, and what I've done, and intend to do, to improve them. Finally, I ended my presentation with a Kahoot quiz based on my presentation; but I also added in questions tailored for each member of the research team (so at least everyone got one question right!). The response from my presentation (mostly regarding the fun quiz) was filled with so much positivity, with the PhD students saying they loved it and had so much fun! It was so overwhelming and made me feel so happy that I was able to bring a little fun to the workplace, like a research lab, which can become quite stressful at times.

Today was definitely one of my most memorable days at placement so far!

DAY 19: Peer-reviewing w/ Vivian! (25/09; 8.5hrs)

21069622 — 2024-10-06 11:22:04 UTC

Today was a very long day at placement, but very fascinating. Today, I assisted Vivian in peer-reviewing a PhD thesis written by a student from the UK. Since Vivian has a stronger background in pathophysiology, she was able to identify mistakes in the procedures and technical issues with the analyses the student had performed. Although I have never written or read a thesis before, Vivian mentioned that my input would be valuable because I lack specific background knowledge on this research area. This allows me to assess whether the student has explained the concepts enough for someone unfamiliar with the topic. I also picked up any spelling and grammar mistakes. I was not expecting this task to take too long, but (believe it or not) it took the whole day to go through the thesis and properly note the errors and critiques. However, I learned a lot about peer-reviewing and what it entails, which was very interesting!

DAY 20: Team Meeting (30/09; 1hr)

21069622 — 2024-10-06 11:22:11 UTC

In this week's team meeting, Jake was allocated to present. Jake's PhD is on coronary artery disease and analysing the prevalence of "SMuRFs" (Standard Modifiable Risk Factors) which include: smoking, hypertension, hypercholesterolemia and diabetes. He discussed the difficulties in understanding why some patients who develop cardiovascular disease (CVD) do not present with SMuRFs; and this is what led to the term "SMuRF-less", meaning they suffer from CVD without the traditional risk factors that typically increase susceptibility. Jake also mentioned that most "SMuRF-less" cases are women, which led me to wonder whether ethnicity might also contribute to CVD, even when standard risk factors are absent.

My question for this week's meeting:

Q: Does ethnicity play a in increasing someone's risk of developing CVD, as a "SMuRF-less" patient?

A: "Yes! factors such as ethnicity as well as genetics and socioeconomic status can majorly influence someone's risk of CVD, even when they don't present any SMuRFs - examples include Indigenous cultures due to deficient health literacy, however, SMuRF-less cases are observed at lower rates in Asian countries when compared to Western countries; this could be due to the more sedentary lifestyle and differences in diet observed in Western cultures".

DAY 21: World Heart Day! (01/10; 4hrs)

21069622 — 2024-10-06 11:22:18 UTC

Today is World Heart Day and for placement, I helped out with the World Heart Day event that the CCBDR held. Before the event started at 11:00 am, Vivian and I set up the brunch area by hanging up the heart and health informative posters and laying out the fruit and snack platters for the participants after the 2km walk! I volunteered to undertake a job for the day, and I got given the part of a traffic/road-crossing marshall. This meant I, unfortunately, couldn't participate in the walk; instead, I stood at the crossing point where I was marshalling, until those walking reached my point. I felt honoured to be given such an important role, but I was a little nervous at the same time as I was ultimately in charge of everyone's safety (even though it was only during the time they crossed the road which was for about 30-60s). At 1:00 pm, Vivian, the other PhD students and I went to a live seminar where a successful physics engineer talked about failure and success, using his life as an example to inspire the up-and-coming researchers of La Trobe.

It was motivating and I'm glad that I was able to attend the seminar before I went to class at 2:00 pm.

DAY 22: Team Meeting (07/10; 2hrs)

21069622 — 2024-10-06 11:22:23 UTC

In this week's team meeting, the rotation restarted and the honours student, Roberto, presented again. But first, Bill, the other lab head, addressed any housekeeping notices. But for this week's meeting, I made chocolate rum balls for the team because it's my birthday!!

Roberto has his honours oral due next week, so as a group, we went through his plan and presentation slides and provided suggestions on how to improve the aesthetic and the flow of his presentation. While the advice was for Roberto, I took note of the tips and tricks they provided him as I have two oral presentations coming up myself!

DAY 23: Errands + Cleaning (09/10; 7hrs)

21069622 — 2024-10-06 11:22:37 UTC

Today was a full-on day in the lab. Firstly, Vivian and I headed into the microscope room where we imaged/took snapshots of healthy and diseased mice aortas. Since I've watched her do this before, today she allowed me to do it myself!

Vivian and I then met with Roberto to clean the ultrasound machine and other pieces of equipment which took a couple of hours. During this time, Vivian also taught how the ultrasound machine is used, which was interesting!

We then returned to the office to analyse the images we took this morning; then finished the day with running errands around the laboratory!

DAY 24: Blood Pressuring Mice w/ Buddhila! (14/10; 5hrs)

21069622 — 2024-10-06 11:22:42 UTC

This week's team meeting was cancelled, so today, I assisted Buddhila with blood-pressuring her mice! This was such a fun experience. I have always wondered how the tail-cuffing method of measuring BP in mice worked, so I was really excited for her to show me. It works similarly to human BP arm cuffs but their tails are placed in an inflatable cuff instead, and a special light sensor is focused on the tail. This light measures and monitors the changes in blood flow, before and after the cuff inflates; then the machine use these values and calculates the BP of the mice.

This task took the majority of the evening. Once we finished, Buddhila showed me her other mice and we did their evening check-ups. I was not sure how I was going to feel about being around live mice (because so far I've mostly been around dead mice), but it was way better than I thought, besides the fact they smell and poop everywhere! Overall, a very fun evening in the animal house.

DAY 25: Flow Cytometry w/ Buddhila! (16/10; 4hrs)

21069622 — 2024-10-06 11:22:47 UTC

Today was another lab day with Buddhila! She was performing flow cytometry today, which involved separating cells into a single-cellular solution, so they are ready to be quantified using a cell counting machine. After cell separation, a dye called Typran blue dye is added to the cells which distinguishes dead from live cells so the cell machine only counts the live cells. After the counting process, we were glad to see that the cell solutions were successfully separated and each had between 1-2 million live cells!

After we had lunch, Buddhila showed me some immunofluorescence images of nephrons in mice kidneys, that she had taken throughout her PhD. One of the images she showed me can be seen above!

DAY 26: Honours Oral Presentations (18/10; 2hrs)

21069622 — 2024-10-06 11:22:52 UTC

Today was a very emotional day as it was my last day at placement! I spent my final shift watching the Honours oral presentations. This was very insightful as it gave me an insider view of what I would expect if I were to complete my Honours next year. The Honours student who is a part of the research division I've been doing my placement at, Roberto, presented his oral presentation as well today. It was nice seeing how far he has come, and expressing how much he enjoyed the process and relationships he has made.

Today, as well as my whole placement experience, has made completing Honours and going into research such an appealing pathway to pursue. I will definitely be considering this path after I graduate!