Exercise 6: Biomeasures for Stress

Yosup Cho

Professor Pittman

HONORS 231
Investigating the Impact of Caregiving on Grandparent Caregivers through Biomarkers

Grandparent caregivers have a unique role that is both emotionally fulfilling and highly

stressful. Stress can have severe physiological repercussions that impact physical and mental

health. Researchers have looked to biomarkers such as hair cortisol and C-reactive protein (CRP)

as objective indicators of stress and inflammation to comprehend and treat these effects. A

thorough methodological approach addresses the particular difficulties and concerns faced by the

African-American grandparent caregiver group by gathering and evaluating biomarkers such as

hair cortisol and C-reactive protein (CRP).

A long-term measure of cortisol levels, hair cortisol concentration (HCC) represents the

total stress experienced over several months. This biomarker helps evaluate the long-term stress

that grandparents who are caretakers endure, providing information that more transient

measurements like saliva or Cortisol in blood cannot. A 1-6 cm long section cut as close to the

scalp as feasible is ideal for hair sample collection; this is usually from the vertex posterior,

where hair growth rates are most stable. Using this technique, the sample is guaranteed to reflect

cortisol exposure precisely during the given time frame. Gather the hair, give it a good wash to

remove any impurities, and then chop or grind it before using methanol to extract the cortisol.

This systematic procedure makes it possible to measure cortisol levels precisely and establish a

connection between stress levels and caring duties.

Researchers should consider cultural perspectives on hair when obtaining hair samples

from grandparents or other caregivers, particularly within the African American population. For

participant trust and involvement, collection techniques must be courteous and open. Research

shows, like those mentioned by Meyer and Novak, demonstrate the utility of HCC in identifying
elevated stress levels associated with caregiving, highlighting its potential role in creating

targeted interventions to mitigate stress impacts.

Stress can cause both acute and chronic increases in CRP, a biomarker of systemic

inflammation, linking stress to potential health outcomes like cardiovascular disease. Monitoring

CRP can provide insights into the physiological impact of caregiving for grandparents who may

be high-stress caregivers. Saliva samples taken just before bedtime and after the grandparents'

caretakers wake up offer a non-invasive way to measure CRP levels, representing the effects of

inflammation and daily stress. Individuals must adhere to pre-collection protocols, which include

refraining from alcohol, tobacco, and specific foods, to guarantee CRP measurements' accuracy.

The study emphasizes the link between high CRP levels and cardiovascular event risk,

which makes it an essential indicator for grandparents who are already vulnerable to health

inequalities. When gathering and examining CRP data requires careful handling and storage of

samples, it can illuminate the direct effects of caregiving stress on physical health. This

information is critical in creating health promotion strategies to reduce the burden of stress and

inflammation among caregivers.

For grandparent caregivers, hair cortisol and C-reactive protein analysis offer a window

into the unseen toll of caregiving. Researchers can better understand the complex interplay

between stress, health disparities, and caregiving by employing culturally sensitive methods for

collecting and analyzing these biomarkers. Such an approach can inform the development of

targeted interventions to support the health and well-being of grandparent caregivers, ensuring

they have the resources and support needed to thrive in their crucial roles.
Questions

How would it be applied to an African American population?

We need culturally sensitive methods to use hair cortisol concentration (HCC) and

C-reactive protein (CRP) indicators for African American grandparent caregivers.

Acknowledging health inequities and individual stressors while collecting data with dignity and

transparency is imperative. Our methodology uses methods like the "single strand twist and

gentle pull" for hair cortisol to ensure inclusivity and adaptability to various hair types. We must

provide culturally appropriate therapies for CRP, recognizing the stress associated with

caregiving and its health repercussions. To improve health outcomes, involving the community

and adapting support systems to the cultural contexts of African American grandmothers is

essential.

What methodological considerations have to be taken into account? Who should be

excluded from studies using these biomarkers?

When assessing hair cortisol concentration (HCC), researchers must consider several

factors, including variations in hair composition across races and the effects of UV exposure.

African American hair's distinct lipid composition necessitates specific cortisol extraction

techniques, such as chopping hair close to the scalp from the posterior vertex and repeatedly

incubating it in methanol and acetone. These steps are crucial for preserving the integrity of the

HCC measurement, accommodating heredity and variability in hair growth, and accurately

estimating stress levels.

 C-reactive protein (CRP) indicates systemic inflammation influenced by BMI, chronic

illnesses, physical activity, and smoking. Researchers need to control these variables to analyze

caregiving stress's effect on CRP levels accurately. The significant impact of oral contraceptive

pills (OCP) on CRP levels requires consideration in study designs to ensure accurate findings.

Researchers must also account for personal hygiene behaviors that could influence CRP

collection and analysis results.

To maintain data integrity, the study inclusion criteria must exclude participants whose

illnesses or therapies significantly alter cortisol or CRP levels. This exclusion applies to those

with recent acute diseases, chronic inflammatory conditions, and those taking OCPs. Researchers

must also consider variables such as hair treatment and environmental exposure to prevent

altering the cortisol content of hair samples used in cortisol research.

How would the biological material be collected? How does it need to be stored?

To assess hair cortisol concentration (HCC), researchers take hair samples from the

occipital or vertex posterior regions before and after treatments. During collection, they must cut

a 1-6 cm long hair segment—ideally 3 cm—as close to the scalp as possible to avoid

contamination. Researchers need about 10 mg of hair from each month-corresponding section,

and they store the samples at room temperature in clearly labeled plastic vials after thoroughly

cleaning them.

Participants collect saliva samples at bedtime and thirty minutes after waking up to assess

C-reactive protein (CRP) and track daily variations in CRP levels, which indicate stress load.

They allow the saliva to pool in their mouths before transferring 1-2 ml of saliva through a straw

into a collection tube. Researchers recommend that participants avoid alcohol, smoking, and
acidic or high-sugar foods and rinse their mouths with water before collecting their samples.

After collecting the samples, participants stored the saliva samples in a refrigerator and later

transferred them to a freezer maintained at -80 degrees Celsius for long-term preservation.

How would it be analyzed? What lab would we send it to for processing? How much does it

cost?

To assess hair cortisol concentration (HCC), researchers clean, dry, and then mince or

grind the hair samples into powder, using methanol for cortisol extraction. They can analyze

these samples using mass spectrometry or enzyme immunoassay. Labs such as Cortigenix in the

UK and LEAC in São Paulo are potential processing sites. Cortigenix offers a cortisol stress kit

for about USD 126.62, presenting an affordable option for individuals and researchers.

Moreover, organizations like USC provide detailed instructions for collecting cortisol samples,

suggesting the capability to process samples in-house or collaborate with specialized labs.

Labs like Salimetrics LLC supply enzyme immunoassay kits for analyzing salivary

C-reactive protein (CRP) samples. The cost of specific kits, such as the Human CRP ELISA Kit

from Bio-Connect, which costs about USD 574.52 for 96 tests, varies. Additionally, institutions

like the UW Medicine's Department of Laboratory Medicine offer CRP analysis services, though

they might require direct contact for pricing details.

What does it tell us about physiological stress or some other health category?

Research has shown that hair cortisol plays a role in managing and recovering from

stress, linking lower hair cortisol levels to reduced stress and anxiety. Hair cortisol concentration

(HCC) is a marker for chronic stress exposure. High levels of hair cortisol can indicate
individuals at risk for chronic stress-related diseases, as it correlates with poorer recovery

outcomes from treatments and a higher occurrence of cardiovascular diseases. Conditions such

as Cushing's syndrome, PTSD, and other cardiovascular diseases have connections to increased

hair cortisol levels, suggesting a direct link between reduced stress and lower hair cortisol levels.

Beyond its role in cholesterol screening, C-reactive protein (CRP) is a biomarker that

offers predictive insights into cardiovascular risk, metabolic syndrome severity, and systemic

inflammation. Elevated CRP levels among grandmother caregivers highlight the link between

caregiving stress and systemic inflammation, potentially affecting susceptibility to infections and

cardiovascular health. In summary, employing HCC and CRP biomarkers underscores the

importance of culturally sensitive research methods and interventions by providing valuable

insights into the physiological stress and health status of African American grandparent

caregivers.
 Works Cited

Gherardi-Donato, Edilaine Cristina da Silva, Larissa Bessani Hidalgo Gimenez, Maria Neyrian

de Fátima Fernandes, Riccardo Lacchini, Elton Brás Camargo Júnior, Kranya Victoria

Díaz-Serrano, Melissa Melchior, Raquel García Pérez, Jorge Riquelme-Galindo, and

Emilene Reisdorfer. 2023. “Mindfulness Practice Reduces Hair Cortisol, Anxiety and

Perceived Stress in University Workers: Randomized Clinical Trial.” *Healthcare* 11

(21): 2875.

[https://doi.org/10.3390/healthcare11212875](https://doi.org/10.3390/healthcare11212875

).

Iob, Eleonora, and Andrew Steptoe. 2019. “Cardiovascular Disease and Hair Cortisol: A Novel

Biomarker of Chronic Stress.” *Current Cardiology Reports* 21 (10): 116.

[https://doi.org/10.1007/s11886-019-1208-7](https://doi.org/10.1007/s11886-019-1208-7)

Meyer, Jerrold S., and Melinda A. Novak. 2012. “Minireview: Hair Cortisol: A Novel Biomarker

of Hypothalamic-Pituitary-Adrenocortical Activity.” *Endocrinology* 153 (9): 4120–27.

[https://doi.org/10.1210/en.2012-1226](https://doi.org/10.1210/en.2012-1226).

Meyer, J. S., & Novak, M. A. (2012). Minireview: Hair cortisol: a novel biomarker of

hypothalamic-pituitary-adrenocortical activity. Endocrinology, 153(9), 4120–4127.

https://doi.org/10.1210/en.2012-1226

Ridker, Paul M. 2003. “Clinical Application of C-Reactive Protein for Cardiovascular Disease
 Detection and Prevention.” *Circulation* 107 (3): 363–69.

[https://doi.org/10.1161/01.cir.0000053730.47739.3c](https://doi.org/10.1161/01.cir.0000

053730.47739.3c).

Stalder, Tobias, and Clemens Kirschbaum. 2012. “Analysis of Cortisol in Hair – State of the Art

and Future Directions.” *Brain, Behavior, and Immunity* 26 (7): 1019–29.

[https://doi.org/10.1016/j.bbi.2012.02.002](https://doi.org/10.1016/j.bbi.2012.02.002).

Trail Ross, Mary Ellen, Duck-Hee Kang, and Stanley Cron. 2015. “Psychological Profile,

Salivary Cortisol, C-Reactive Protein, and Perceived Health of Grandmothers With

Childrearing Responsibility.” *Journal of Family Issues* 36 (14): 1904–27.

[https://doi.org/10.1177/0192513X14540157](https://doi.org/10.1177/0192513X1454015

7).