Name: A Method for Manipulating Blood Glucose and Measuring Resulting Changes in Cognitive Accessibility of Target Stimuli (Video) | JoVE Journal
Uploaded: 2016-08-12T13:00:00.000+00:00
Duration: 8 min 1 s
Description: Learn to manipulate blood glucose and measure changes in implicit cognition using a lexical decision task. This cost-effective method helps researchers study energy needs and psychological behavior.

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5 minutes

This research was conducted with grant funding from the Anthony M. Marchionne Foundation (70256-23284) and TCU IS. The authors thank Danielle DelPriore, Amanda Morin, and Christopher Rodeheffer for their helpful contributions towards shaping these methods.&#160;The authors would also like to thank Hannah Bradshaw and Randi Proffitt Leyva for their assistance with filming this protocol.

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The authors have nothing to disclose.

This manuscript outlines a simple, inexpensive procedure for manipulating blood glucose, as well as a procedure for measuring resulting changes in the cognitive accessibility of target words and concepts. The above outlined methods can be applied to a wide range of research areas including social psychology, cognitive psychology, and nutrition sciences. Given that this method may be used by people in multiple research areas (some that do not typically use survey data), this section will present some tips on critical step...

Researchers in cognitive psychology and neuroscience have long studied the impact of blood glucose fluctuations on the brain, cognition, and behavior1. For example, researchers have found that fluctuations in blood glucose predict differences in memory (verbal, digit span, working, and episodic)1-5, attention6,7, puzzle solving8, and performance on tasks of varying levels of cognitive demand2,9,10. This research finds that increasing blood glucose enhances attention and memory, and that these effects are strongest when working on cognitively demanding tasks, and in those of older age1,3.
In recent decades, research on the impact of blood glucose fluctuations on human psychology has also become a topic of interest in the field of social psychology11-13. Fluctuations in blood glucose have been linked to changes in mood14, motivation15, self-control11-13, attitudes16,17, prejudice18, and to changes in decision-making in consumer19, eating20,21, and financial domains22. Because these results transcend a wide variety of dependent variables, the message about the role of blood glucose in decision-making is clear: blood glucose is the critical form of energy to the brain, and depletions in blood glucose lead to important changes in cognition and self regulation11,13,15.
Given the important role that blood glucose plays in various psychological and behavioral outcomes, the current manuscript offers a simple method for manipulating blood glucose that researchers can use for testing a range of cognitions and behaviors. Additionally, the current manuscript provides a method for assessing the cognitive accessibility of target words or concepts that may change in response to fluctuations in blood glucose using a lexical decision task. The efficiency of the present methods make them ideal for experimental research, as they effectively manipulate blood glucose and measure cognitive changes using relatively inexpensive, commercially-available products. The affordability of the present blood glucose manipulation makes it easier to run the large numbers of participants needed to obtain the adequate power needed to detect statistically significant differences in behavior caused by any manipulated changes in blood glucose23. Additionally, the lexical decision task is useful in that it can reveal implicit changes in cognition not easily accessed through more explicit measures such as self-report surveys24.
The described blood glucose manipulation task involves an 8 hour fasting period followed by the directed consumption of a caloric or a calorie-free drink22. After fasting for eight hours, participants are randomly assigned to consume either a sugar sweetened beverage or water. This method also allows for the testing of any unique effects produced by non-caloric sweeteners (e.g. aspartame, sucralose), which taste sweet like traditional caloric sweeteners, but do not contain the sugars needed to raise blood glucose levels20,22,25. If interested in the effects of non-calorically sweetened beverages, this task includes an optional, 3rd between-subjects condition in which participants consume a &#34;zero-calorie&#34; drink. The current method has been applied successfully in multiple social psychological paradigms12,18,22, and offers a shorter way to test for the effects of changes in blood glucose upon self-regulation than previously used measures like glucose tolerance tests26.
The authors also provide a procedure for assessing the cognitive accessibility of target words, or concepts, using a lexical decision task27,28. In this task, letter strings and words are presented via computer in rapid-fire format, and typically fall into a few distinct conceptual categories (in the described research, the categories were: high fat food words, low fat food words, non-words). Participants are asked to quickly identify (typically via keystroke) whether a string of letters (e.g. fhens; pizza) is a word or a non-word. The reaction speed in which participants correctly identify letter strings in each conceptual category measures the relative ease with which the participant is able to mentally process words in that concept category. Quicker reactions mean that a concept is more mentally accessible, and thus, &#34;on someone&#39;s mind&#34;24,29,30. As a consequence, lexical decision tasks make useful dependent variables, and can also be used as effective manipulation checks to measure if a recently primed concept (see reference30 for more information on priming) is indeed subsequently more cognitively accessible than it would be in the absence of a prime. Although the lexical decision task is a task that has been used in social psychology for decades29, the current manuscript presents some minor variations to the procedure, a link to an editable program template27, and an empirically validated list of words from the authors&#39; previous work that measures the cognitive accessibility of healthy versus non healthy foods20. It is the authors&#39; hope that the further adaptation of these two procedures will help lead to innovations in the fields of social psychological research, in eating behavior and energy regulation research, and will facilitate further comparative research that combines the perspectives and methodology of&#160;multiple disciplines.

<table><tbody><tr><td>Survey Building Software</td><td>Qualtrics</td><td>Qualtrics Research Suite</td><td>Alternative survey building softwares/applications include Survey Monkey, Google Forms, Media Lab, and Inquisit software.</td></tr><tr><td>Behavioral Task Software (for lexical decision task)</td><td>Inquisit</td><td>Inquisit 4 Lab (4.0.8.0)</td><td>Alternative behavioral task softwares/applications include Media Lab/Direct RT or programming the task into an internet browser using a programming language of your choice (such as java).</td></tr><tr><td>Batch File&amp;nbsp;</td><td>Microsoft&amp;nbsp;</td><td>Microsoft Notepad; Windows</td><td></td></tr><tr><td>Lexical Decision Task Template</td><td>Millisecond</td><td>Millisecond survey library, cited template author is linked on page</td><td>Can build a lexical decision task by hand in other behavioral task softwares.</td></tr><tr><td>Participant Scheduling and Compensation Software</td><td>SONA systems</td><td>SONA systems scheduling software</td><td>Appointments can be arranged manually, too.</td></tr><tr><td>Statistical Analysis Software</td><td>IBM</td><td>IBM SPSS Statistics Standard, 22</td><td>Alternate softwares include SAS and R.</td></tr><tr><td>Blood glucose manipulating beverege paradigm</td><td>Coca Cola&amp;nbsp;</td><td>Sprite, Sprite Zero, Sparkling water</td><td>Can use any store brand sugary beverage, non calorically sweetened beverage, and sparkling water beverage, as long as beverages are not easily discernable from each other by sight.&amp;nbsp;</td></tr><tr><td>Lancet</td><td>Assure</td><td>Assure Lanets 23 gauge</td><td>Many brands of testing lancets available both online and at local pharmacies.</td></tr><tr><td>Blood glucose testing meter</td><td>Bayer&#039;s</td><td>Breeze 2</td><td>Many brands of testing meters available both online and at local pharmacies.</td></tr><tr><td>Blood glucose testing strips</td><td>Bayer&#039;s</td><td>Breeze 2</td><td>Many brands of testing strips available both online and at local pharmacies, but they must be compatible with your chosen meter.</td></tr><tr><td>&amp;nbsp;Nitrile exam gloves (400 count)</td><td>Kirkland</td><td>Kirkland Signature Nitrile Exam Gloves</td><td>Any medical grade exam glove that provides sufficient protection from blood exposure can be used.</td></tr><tr><td>Disinfecting wipes</td><td>Lysol</td><td>Lysol Disinfecting Wipes Lemon Scent</td><td>Any wipe that can kill off any bloodborne or contact born contaminants.</td></tr></tbody></table>

a method for manipulating blood glucose and measuring resulting changes in cognitive accessibility of target stimuli

A Method for Manipulating Blood Glucose and Measuring Resulting Changes in Cognitive Accessibility of Target Stimuli

Much research in social psychology has investigated the impact of bodily energy need on cognition and decision-making. As such, blood glucose, the body's primary energy source, has been of special interest to researchers for years. Fluctuations in blood glucose have been linked to a variety of changes in cognitive and behavioral processes, such as self-control, political attitudes, and eating behavior. To help meet growing interest in the links between bodily energy need and these processes, this manuscript offers a simple methodology to experimentally manipulate blood glucose using a fasting procedure followed by administration of a sugar-sweetened, unsweetened, or artificially-sweetened beverage. This is followed by presentation of a method for measuring resulting changes in implicit cognition using a lexical decision-task. In this task, participants are asked to identify whether strings of letters are words or non-words and response latencies are recorded. Sample results from a recent publication are presented as an example of the applications for the experimental manipulation of blood glucose and the lexical decision task measures.

Participants
The above methods were implemented in a study run by Hill and colleagues20 at a midsize, private university in the southern United States. The undergraduate population at the university provided the subject sample and participants received partial course credit as compensation for study participation. Using the exact methods outlined in the current manuscript protocol, the authors ran participants through...

The authors introduce a method for manipulating blood glucose and measuring resulting changes in cognitive accessibility of target words using a lexical decision task.

Much research in social psychology has investigated the impact of bodily energy need on cognition and decision-making.  As such, blood glucose, the body's ...

The overall goal of this methodology is to experimentally manipulate blood glucose using a fasting procedure followed by administration of a beverage. This is followed by the presentation of a method for measuring changes in implicit cognition using a lexical decision task. This method will show researchers how to experimentally manipulate participants'blood glucose levels and then program a task to measure changes in implicit cognition that results.The main advantage of this technique is that it's relatively inexpensive and easy to perform. Although this method can be used to measure the impact of energy need on changes in implicit cognition, it can also be used to measure the impact of energy need on other psychological processes and behavior. To begin, set up a two part survey using survey building software.In addition to the research task, make sure that the survey begins with a consent form and ends with a debriefing statement. Next, send out a recruitment email and select a pool of participants based on the inclusion and exclusion factors described in the accompanying text protocol. Ask them to refrain from eating or drinking any non water beverages for eight hours prior to their participation session.It is critical that you instruct participants to fast for a minimum of eight hours prior to the experimental session. Any participant that violates these instructions, needs to be excluded from data analysis. Before participants arrive in the laboratory, assign each participant a unique ID number and randomly assign them a beverage type using a participant log.Then, write each participant ID number being used in a session on an individual sticky note. Pre-pour the participant's chilled drink into an opaque disposable cup labeled with that participant's participant ID number. It's important that participants'ID numbers are correctly entered into the experimental log and their testing computer so that data can be linked at the end of the study.On the computer, open the study folder, double click on the Batch file icon, and select Run to open the survey. Bring participants into the lab and ask if they have met the pre-agreed fasting requirements. Have the participant digitally sign the consent form before starting the experiment.Following this, ask the participants a few short questions that will help to maintain the integrity of the cover story. When the computer screen indicates a stopping point, guide the participant to a sterile spot away from the survey to take a pre-manipulation blood glucose reading. Clean the participant's finger with a sterile wipe and then use a lancet to gently prick the side of the participant's index finger.Squeeze a small amount of blood onto a blood testing strip, and insert the strip into a glucometer. Write down the corresponding blood glucose reading and the time that the blood glucose reading was taken in the participant log. Then, wipe down the area with a sterilizing wipe to prepare it for the next participant.Offer the participant some hand sanitizer and a tissue to clean their finger. Also, provide the participant with a small adhesive bandage if they desire to use one to cover their finger. Following the glucose test, direct the participant to take a seat at the computer terminal.At this point, check that the cup matches the assigned condition and participant number recorded on the participant log and hand the participant the cup containing their assigned drink. Instruct the participant that the next task will involve making a series of consumer evaluations and that they should inform the experimenter when they reach the stop message at the end of the timed beverage consumption task. Next, instruct the participant that the task requires that they consume the entire beverage in two minutes or less and that the computer will time them.When the participant begins to drink the beverage, click the Next arrow on the survey page to begin the timed beverage consumption task. After the two minute period has elapsed, the survey will automatically continue to a message that prompts the participant to fill out the beverage evaluation questions. It will then automatically follow these questions with a filler task.While the participant completes the task, collect the used beverage cup from the participant's work space but do not throw it away. Mark down whether or not the participant finished their entire beverage in the experimental log next to the participant's ID number. The filler task will last for 10 minutes to allow the changes in blood glucose level to occur.Upon completion of the 10 minute filler task, a Stop message will appear. At this point the participant should submit and close the survey. Then, immediately take another blood glucose reading and proceed to the lexical decision task.To begin the lexical decision task, double click and open the software. Enter the participant's ID number into the prompt and initiate the programmed lexical decision task by clicking the Run button on the prompt window. At this point, briefly describe the lexical decision task to the participant and instruct them to notify the researcher when the task is finished.Then, allow the participant time to complete the lexical decision task on the computer. After finishing the task, close the lexical decision task out fully by hitting the space bar. Next, ensure that the batch file opens the final survey portion, which contains the socio-demographic questions and a debriefing statement as described in the accompanying text protocol.At the conclusion of the entire study, debrief, compensate, and dismiss the participant. After the participant leaves, flag the data of any participant that did not meet fasting requirements, whose fasting blood glucose read higher than 100 milligrams per deciliter, or that did not finish their drink. Results of a study based on this methodology revealed that participants who drank non calorically sweetened beverages responded more quickly to high calorie words than participants who drank sugar sweetened or unsweetened beverages.Results also revealed no differences in mean reaction time to these same high calorie words between participants who drank sugar sweetened or unsweetened beverages. In addition, there were no effects of drink type on participants'reaction times to low calorie food words or non words. After watching this video, you should have a good idea of how to experimentally manipulate participants'blood glucose levels and then measure resulting changes in implicit cognition.These procedures can be used either together or separately and can be done in under one hour if performed properly. Don't forget that working with human blood, even in small amounts, can be extremely hazardous. Always take proper precautions such as wearing rubber gloves and using appropriate sanitation techniques when performing this procedure.

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Research

JoVE Journal

Behavior

The hyperinsulinemic-euglycemic clamp is the "gold standard" for the assessment of insulin action. Insulin is infused at a constant rate stimulating glucose uptake. The amount of exogenous glucose infused to counter this drop is indicative of insulin sensitivity. Here the procedure is performed on a conscious, unrestrained rat.

Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat

The hyperinsulinemic-euglycemic clamp, or insulin clamp, is the gold standard for assessing insulin action in vivo.  A method for performing insulin clamps in mice is described.  This includes a method for arterial catheterization that permits experiments to be performed in conscious, unrestrained mice with minimal stress.

Hyperinsulinemic-euglycemic Clamps in Conscious, Unrestrained Mice

Modulation of hippocampally-dependent spatial working memory by direct intrahippocampal microinjection, accompanied and followed by in vivo microdialysis for metabolites in conscious, behaving animals.

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

This video demonstrates the effect of continuous drug infusion into the mouse brain on blood glucose levels. Glucose metabolism is assessed by collecting blood from a fasted mouse's tail and measuring glucose levels before and after oral glucose administration. Insulin tolerance is evaluated by injecting insulin in the mouse, with glucose levels recorded at specific intervals.

Evaluating the Effect of Continuous Drug Infusion into the Mouse Brain on Peripheral Glucose Levels

Microfluidic oxygen control confers more than just convenience and speed over hypoxic chambers for biological experiments. Especially when implemented via diffusion through a membrane, microfluidic oxygen can provide simultaneous liquid and gas phase modulations at the microscale-level. This technique enables dynamic multi-parametric experiments critical for studying islet pathophysiology.

Quantitative and Temporal Control of Oxygen Microenvironment at the Single Islet Level

The activity of single neurons from adult-aged mice can be studied by dissociating neurons from specific brain regions and using fluorescent membrane potential dye imaging. By testing responses to changes in glucose, this technique can be used to study the glucose sensitivity of adult ventromedial hypothalamic neurons.

Membrane Potential Dye Imaging of Ventromedial Hypothalamus Neurons From Adult Mice to Study Glucose Sensing

This protocol studies the role of chemokine (C-C motif) ligand 5 (CCL5) in the hypothalamus by delivering an antagonist, MetCCL5, into the mouse brain using a micro-osmotic pump brain infusion system. This transient inhibition of CCL5 activity interrupted hypothalamic insulin signaling, leading to glucose intolerance and peripheral systemic insulin sensitivity.

Studying the Hypothalamic Insulin Signal to Peripheral Glucose Intolerance with a Continuous Drug Infusion System into the Mouse Brain

Extracellular depletion of fluorescently labeled glucose correlates with glucose uptake and could be used for high-throughput screening of glucose uptake in excised organs and cell cultures.

Extracellular Glucose Depletion as an Indirect Measure of Glucose Uptake in Cells and Tissues Ex Vivo

This article describes a detailed protocol to increase glucose concentration in the cerebrospinal fluid (CSF) of mice. This approach can be useful for studying the effects of high CSF glucose on neurodegeneration, cognition, and peripheral glucose metabolism in mice.

Osmotic Minipump Implantation for Increasing Glucose Concentration in Mouse Cerebrospinal Fluid

A hyperglycemic clamp is used for measuring insulin release with a maintained higher blood glucose concentration. A hypoglycemic clamp is for measuring glucose production induced by counter-regulatory responses. Both methods use the same surgical procedure. Here, we present a clamp technique to assess systemic glucose metabolism.

Hyperglycemic Clamp and Hypoglycemic Clamp in Conscious Mice

Learn to manipulate blood glucose and measure changes in implicit cognition using a lexical decision task. This cost-effective method helps researchers study energy needs and psychological behavior.

A Method for Manipulating Blood Glucose and Measuring Resulting Changes in Cognitive Accessibility of Target Stimuli

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