INTERVIEW:

Q: What led you to pursue this research area and what interests you most about it?

A: My interest in the cardiovascular effects of airborne toxins stemmed from a combination of educational, professional, and personal interests.

While earning a PhD in Environmental Health Sciences, I worked for the EPA, where I analyzed rat ECGs for air pollutant-induced arrhythmias and changes in heart rate variability. Excellent mentors at the EPA, and a strong team of public health researchers focused on environmental determinants of cardiovascular disease during my postdoc at Harvard, furthered my interests in this field.

I also have a vested personal interest in the potential impacts of air pollution being an outdoor enthusiast.

Q: What does the general landscape of this research currently look like?

A: The landscape I’m now most familiar with is largely dictated by funding opportunities and government priorities, which includes characterizing health effects and understanding biological mechanisms of toxicity.

For example, the FDA is more interested in characterization of health effects given their responsibility to evaluate the safety of food and drugs, whereas the NIH is perhaps more interested in work that determines the mechanisms of toxicity.

Attention has recently shifted away from airborne particulate matter (classic air pollutants) to multi-pollutant interactions and volatile organic compounds (VOCs). In inhalation toxicology, emphasis has shifted to some extent to e-cigarette research, given the novelty and our limited understanding of the public health implications of e-cigarettes.

Increasingly sensitive tools are allowing researchers to demonstrate toxicity at lower, more real-world levels. And so, researchers depend all the more on the reliability and sensitivity of their instruments.

Q: What are the real-world implications of your research?

A: My research mostly uses rodents to identify the cardiac risks of inhaled toxins, but it is also designed to clarify whether links between exposures and adverse human health outcomes might causally relate. Right now, there is a proposed menthol ban in combustible cigarettes. This may someday play out for e-cigarettes as well. My research could impact the chosen route forward for regulation of e-cigarettes.

Q: How long have you been an EMKA user?

A: I have used EMKA’s ecgAUTO software for 16 years.

I used it throughout my training at the U.S. EPA and UNC Chapel Hill, as well as Harvard University, and now here at the University of Louisville.

Q: How has using EMKA software helped with improving the translatability and reproducibility of your research?

A: After using ecgAUTO to analyze animal ECGs for over a decade, I began applying it to human data a few years ago. This has enhanced the translatability of my research by allowing me to use the same software and analysis procedures.

The ability to reapply the same configurations across various studies for automated detection of heart beats is a major advantage.

The ability to analyze data from conscious, unrestrained animals ensures a greater level of reproducibility, since anesthetics can interfere with the toxic effects of aerosols and the mechanisms underlying them.

The autonomy to adjust fiduciary points of the library waveforms for pattern recognition in ECG analysis sets ecgAUTO apart from competing software. The software can handle large datasets with multiple physiological signals per file as well.

Q: What were some insights that the EMKA solutions helped you obtain?

A: Throughout my 16 years of working with EMKA, I have truly felt like I am working with the EMKA team. There has been a constant two-way communication to optimize the software and figure out workarounds for various challenges. EMKA has consistently provided me direct, honest, and unfiltered feedback on the limitations and advantages of their products. I really can’t think of a more responsive company.

Q: What features of the equipment or software do you find most useful?

A: The ability of ecgAUTO to handle staggering amounts of data with relative ease has proven highly advantageous.

The customizability of parameters has enabled us to estimate relatively novel endpoints in the field of cardiovascular toxicology. For instance, in rats, we have used negative ST area as a surrogate for ST segment amplitude to indicate myocardial ischemia.

As well, the interlead analysis option has enabled us to estimate excitation-contraction coupling time or electro-mechanical delay when pairing ECG with left ventricular pressure. Spontaneous baroreflex has also been another useful and relatively novel parameter provided in ecgAUTO.

Q: What was the reasoning behind selecting your animal model?

A: I began my research in rats given how conducive their size is to telemetry surgeries as well as their representativeness of human cardiac physiology. However, I made the switch to mice, given the variety of genetic models available for testing biological mechanisms.

Q: What advice do you have for someone starting out in this research area?

A: Glom onto someone knowledgeable in the field and learn everything you can from them. Never stop asking questions, and if your peers and mentors don’t know the answer to the most interesting questions, figure out how to find the answer yourself.

Q: What’s next for your lab and your research?  

A: We have some major publications in the works that examine the ability of e-cigarettes to induce cardiac arrhythmia. I also plan to further enhance the translational aspects of my current research program.

THANK YOU, Dr. CARLL, FOR THIS INTERVIEW!

If you have any questions about the use of ecgAUTO, please Contact Us!

INTERVIEW:

Q: What is your experience with infectious disease research ?
A: I work with different pathogens (viral pathogen (COVID, Flu), bacterial pathogens (anthrax, tularemia), biological pathogen (ricin)) that require testing in a BSL3 facility. Most of these are classified as infectious diseases.

Q: How long have you been doing this type of work ?
A: I have been in CMO/CROs for more than 17 years.

Q: How long has Lovelace Biomedical Research Institute been doing this type of work?
A: Lovelace has been working with infectious disease models over 70 years.

Q: What is your experience with SARS-CoV2 (COVID-19)?
A: I currently work on multiple active SARS-CoV2 (COVID-19) studies on-going in our BSL3 facilities.

Q: How would you describe the current research environment for infectious diseases, for example SARS-CoV2 (COVID-19) which is currently on the rise?
A: Given the current global concern of SARS-CoV2 (COVID-19), it fosters a more collaborative research environment as we are all working toward helping our own communities to find treatment and vaccine options for patients.

Q: What are the clinical signs or endpoints that you are looking for in your SARS-CoV2 (COVID-19) studies and why?
A: We are looking at respiratory outcomes such as respiratory rate (RR), tidal volume (TV), minute volume (MV), and accumulated volume (AV) as well as temperature for onset of fever which are currently used in the clinics to diagnose COVID-19 patients and determine disease severity.

Q:  How are you picking your animal models for the SARS-CoV2 (COVID-19) studies?
A: To begin our SARS-CoV2 (COVID-19) studies we began looking at models that showed promise in previous SARS-CoV and influenza studies. Some examples of currently used subjects in the follow, where each have their own pros and cons for the various research goals each institution is working on.

• Humanized ACE2 transgenic mouse model
• Syrian hamster model
• Ferret
• Common marmoset
• African Green Monkey
• Cynomolgus macaque
• Rhesus macaque

Q: What determines a good subject for SARS-CoV2 (COVID-19) studies?
A: Certain selection criteria are used when looking at any potential subject for inclusion in a research study. In regard to SARS-CoV2 (COVID-19), we are looking at models that show the following:

• ACE2 receptor homology
• ACE2 receptor distribution
• Viral replication similarities
• Clinical signs and disease severity
• Immune response similarities
• etc.

Q: Does Lovelace use large or small animals for their SARS-CoV2 (COVID-19) studies?
A: Both, as both have their own advantages and disadvantages to study the disease in its entirety.

Q: Given the symptoms in the news of COVID-19 patients, how is that like your models?
A: Given the unknowns of COVID-19 along with the wide range of symptoms seen in COVID-19 patients depending on various demographics, model development and symptom assessment are ongoing. Determination of how those symptoms may or may not apply to COVID patients is complicated and requires accurate physiological data collection from animal models.

Q: What diagnostic tools do you use?
A: We use various plethysmography chambers to look at volumetric data. Common respiratory outcomes we measure are respiratory rate (RR), tidal volume (TV), and minute volume (MV). Additionally, we look at temperature changes through telemetry and data logger systems.

Q: Why is RR, TV, MV, and AV important to your SARS-CoV2 (COVID-19) studies?
A:  Physiological data such as RR, TV, and MV may be used to calculate pathogen presentation to a subject or used to assess reaction during and/or after a therapy is presented. Accurate baseline measurements are also critical for comparative analysis of individual subjects.

Q: What type of measurements do you collect post-dosing?
A: Post-exposure we look at behavior, activity, and temperature changes as well as lavage, histology, etc.

Q: Once the subject becomes sick do you look at the changes in TV, MV, RR, temperature, activity, etc.?
A:  Physiological observations are critical to assess reaction to both pathogens and therapies. For example, TV, MV, and RR data may be collected immediately after a therapy is presented, while subjects are periodically monitored for RR, temperature, activity, etc. over a longer period after treatment.

Q: Do you look at higher risk individuals?
A: With some diseases, we will look at additional risk factors. In example, we use genetically modified subjects to better understand and additional risk factors or comorbidities.

Q: Is real-time analysis valuable for your studies?
A: YES. We use real-time accumulated volume (AV) measurements to calculate dosage.

We may also measure RR, TV, and MV during pathogen or treatment exposures. The real-time availability and accuracy of these recordings allows for accurate dosing on a per subject basis.

Additionally, the use of real-time temperature measurements through implanted telemetry, with fever detection, is extremely helpful to research the progression and transmission of different diseases or test articles.

Q: What is your opinion of the pros and cons of implanted verses non-invasive telemetry recording systems?
A:

With large animal subjects, there are more options when working with infectious disease models compared to working with smaller subjects like rodent models. Large animals allow for the use of both implantable and non-invasive telemetry systems, while smaller subjects are restricted to implantable telemetry devices, when working outside the respiratory realm which utilizes plethysmography chambers to measure respiratory volumes.

Being able to use implantable telemetry for chronic studies, or experiments requiring core body temperature, is beneficial where as studies requiring respiratory end points only allow us to utilize non-invasive telemetry options.

The use of non-invasive equipment allows for Lovelace to take advantage of the 3 Rs in research (reduce, reuse, and refine).

In both cases, the advantage of telemetry over logging systems is real-time observation of clinical symptoms, as well as reductions in user time to retrieve and monitor data from BSL-3 facilities throughout the duration of the study.

Q: Is there any specific value to your research from working directly with emka TECHNOLOGIES?
A: One of the emka systems we utilize is the emkaPACK4G respiratory impedance belt (RIP) system, to collect our respiratory volumes. This is critical, as we are using a head-only exposure design, making the RIP system the only method to collect volumetric data during exposure.

Q: Do you find the emka TECHNOLOGIES hardware to easily work across large and small animals?
A: The ability to use the same software set-up and many of the hardware components across study designs and subject models, allow for a more cost-effective core facility to investigate infectious diseases and other applications.

Q: Does emka TECHNOLOGIES specifically add value to your research projects?
A: Personalization of service from emka’s sales and support teams is excellent.

Emka is available for real-time assistance with experiments. The knowledge and ability of each individual emka representative allows for efficient and effective support while working with the same people. This type of service allows us to create a relationship with the emka reps that we learn to trust, when helping with our various research projects and goals. It is very rare that I ask a question and do not receive an answer on the spot, and when the answer is not readily known the response time is still under 24 hours.

THANK YOU, BRENT BARRE, FOR THIS INTERVIEW!