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The Journal of Nutrition Nutrition and Disease
Bioactives in Blueberries Improve Insulin Sensitivity in Obese,
Insulin-Resistant Men and Women1–4
April J. Stull, Katherine C. Cash, William D. Johnson,
Catherine M. Champagne, and William T. Cefalu*
Center for the Study of Botanicals and Metabolic Syndrome,
Pennington Biomedical Research Center, Louisiana State
University System, Baton Rouge, LA 70808
Abstract
Dietary supplementation with whole blueberries in a preclinical
study resulted in a reduction in glucose concentrations over
time. We sought to evaluate the effect of daily dietary
supplementation with bioactives from blueberries on whole-
body insulin sensitivity in men and women. A double-blinded,
randomized, and placebo-controlled clinical study design was
used. After screening to resolve study eligibility, baseline (wk
0) insulin sensitivity was measured on 32 obese, nondiabetic,
and insulin-resistant subjects using a high-dose
hyperinsulinemic-euglycemic clamp (insulin infusion of 120
mU(861 pmol)×m22×min21). Serum inﬂammatory biomarkers
and adiposity were measured at baseline. At the end of the
study, insulin sensitivity, inﬂammatory biomarkers, and
adiposity were reassessed. Participants were randomized to
consume either a smoothie containing 22.5 g blueberry
bioactives (blueberry group, n = 15) or a smoothie of equal
nutritional value without added blueberry bioactives (placebo
group, n = 17) twice daily for 6 wk. Both groups were
instructed to maintain their body weight by reducing ad libitum
intake by an amount equal to the energy intake of the
smoothies. Participants’ body weights were evaluated weekly
and 3-d food records were collected at baseline, the middle, and
end of the study. The mean change in insulin sensitivity
improved more in the blueberry group (1.7 6 0.5 mg×kg
FFM21×min21) than in the placebo group (0.4 6 0.4 mg×kg

FFM21×min21)(P = 0.04). Insulin sensitivity was enhanced in
the blueberry group at the end of the study without significant
changes in adiposity, energy intake, and inflammatory
biomarkers. In conclusion, daily dietary supplementation with
bioactives from whole blueberries improved insulin sensitivity
in obese, nondiabetic, and insulin-resistant participants. J. Nutr.
140: 1764–1768, 2010.
Introduction
Increased consumption of berries has been shown to improve
cognitive function, risk of cardiovascular disease, and cancer
(1,2). Studies have also reported that specific berries, i.e.,
blueberries, have antidiabetic effects. Specifically, a study
performed in mice (3) found that supplementation with
whole blueberries reduced the blood glucose area under the
curve (AUC)5 in vivo and cell culture studies (4,5)
demonstrated increased glucose uptake in vitro (6). In addition,
inflammatory genes have been reduced in mice after consuming
blueberry bioactives, which suggests an antiinflammatory
response (3). The purported health benefits from blueberries
have been attributed to their phenolic bioactive compounds,
such as anthocyanins, which also have antioxidant properties
(6–8). Given the concern regarding the ability to greatly
increase and maintain an individual’s fruit and vegetable
consumption over a long-term period (9), the role of dietary
supplementation with bioactive components in blueberries
remains a very attractive and feasible daily dietary intervention.
To the best of our knowledge, there is no human research that
has reported on the efficacy of increased blueberry bioactive
consumption on insulin sensitivity by using the
hyperinsulinemic-euglycemic clamp technique (10), which is
the gold standard for measuring in
1 Supported in part by the NIH training grant T32 AT004094
(supporting A.J.S.), by the United States Highbush Blueberry
Council, and P50AT002776-01 from the National Center for
Complementary and Alternative Medicine and the Office of
Dietary Supplements (W.T.C.), which funds the Botanical

Research Center of Pennington Biomedical Research Center and
The Biotech Center of Rutgers University. This project used
facilities that are supported in part by Centers of Biomedical
Research Excellence (NIH P20-RR021945) and Clinical
Nutrition Research Unit (NIH 1P30-DK072476) center grants
from the NIH. 2 Author disclosures: A.J.Stull,K.C. Cash,W. D.
Johnson, andC. M.Champagne, no conﬂicts of interest. W. T.
Cefalu received research funds from the United States Highbush
Blueberry Council. 3 This trial was registered at
clinicaltrials.gov as NCT01005420. 4 Supplemental Figure 1
and Table 1 are available with the online posting of this paper
at jn.nutrition.org. * To whom correspondence should be
addressed. E-mail:
[email protected]
edu.
5 Abbreviations used: AUC, area under the curve; hsCRP, high
sensitivity C-reactive protein; MCP-1, monocyte
chemoattractant protein 1; TNFa, tumor necrosis factor-a.
ã 2010 American Society for Nutrition. 1764 Manuscript
received April 13, 2010. Initial review completed May 18, 2010.
Revision accepted July 12, 2010. First published online August
19, 2010; doi:10.3945/jn.110.125336.
by guest on January 11, 2014jn.nutrition.orgDownloaded from
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Supplemental Material can be found at:
vivo insulin action. Therefore, this project’s overall objective
was to examine the role of dietary supplementation with
bioactives in freeze-dried whole blueberry powder on insulin
action in vivo with the use of hyperinsulinemic-euglycemic
clamps in individuals who were obese, nondiabetic, and insulin
resistant. We hypothesized that increased daily consumption of
blueberry bioactives, based on preclinical data, would be
effective in increasing insulin action in vivo and ultimately
result in improved insulin sensitivity in a human population at
high risk for type 2 diabetes.

Subjects and Methods
Subjects. Participants inthe study were recruited fromthe
Greater Baton Rouge area. A total of 32 men and women
completed all evaluations (Supplemental Fig. 1). Those included
were adults ($20 y old), obese (BMI between 32 and 45 kg/m2),
and insulin resistant (nondiabetic). The exclusion criteria
included: 1) diabetes; no diabetes status was confirmed by a 2-h
oral glucose tolerance test; 2) medications known to affect
glucose metabolism; 3) untreated thyroid or chronic liver, renal,
or cardiovascular disease; 4) a history of drug and/or alcohol
abuse, or psychiatric disease prohibiting adherence to study
protocol; 5) history of allergic reactions to blueberries; 6)
consuming berries, grapes, and wine .3 times/wk; and 7)
fluctuation in body weight . 5% in the preceding 2 mo. The
Institutional Review Board for human subjects at Pennington
Biomedical Research Center reviewed and approved the study
protocol. All participants gave written consent prior to starting
the study.
Study design.This study design was double blinded, placebo-
controlled, and randomized. All study evaluations and
measurementswere performed onparticipantsthat had fasted for
10h. Aweekwasdefined as7 d (6 2 d).
Clinical intervention and source of whole blueberry bioactives.
The freeze-dried whole blueberry powder was prepared by the
United States Highbush Blueberry Council (USDA oversight).
The whole blueberry powder was made from a 50/50 mixture of
2 varieties of highbush blueberries, Tifblue (Vaccinium ashei)
and Rubel (Vaccinium corymbosum). The whole blueberries
were freeze-dried, milled, and stored in aluminum cans under
nitrogen. Based on the compositional analysis, the 45 g of
blueberry powder contained 1462 mg of total phenolics, 668 mg
of anthocyanins, and 16.02 mmol TE of antioxidants (oxygen
radical absorbance capacity). Also, the 45 g of blueberry
powder that was provided to the participants equated to an
amount of bioactives in ~2 cups of fresh whole blueberries.
After the participants were assessed as being insulin resistant

(glucose disposal rate # 650 mg/min), they were randomized to
receive twice daily a smoothie with blueberry bioactives added
or an identical smoothie without blueberry bioactives (i.e.,
placebo) (Supplemental Table 1). The participants were
instructed to consume 1 smoothie at breakfast meals and the
other smoothie at dinner meals (at least 6 h apart). The
smoothies were prepared in the metabolic kitchen and a week’s
supply of frozen smoothies was provided in a cooler for the
participants to pick up at each weekly visit. Participants were
instructed to keep the smoothies frozen, thaw them in the
refrigerator, avoid exposing them to direct heat, and avoid
adding any other ingredients to them. For study compliance, the
participants verbally reported their smoothie consumption to the
dietitian at each visit. A compliance of .75% was mandatory for
continued participation in the study.
Physiologic assessments. Hyperinsulinemic-euglycemic clamps
(10) were performed to assess insulin sensitivity after a 10-h
fast. Participants were admitted into the inpatient research unit
the evening prior to their insulin sensitivity testing day and
consumed a eucaloric standardized meal (50% carbohydrates,
35% fat, and 15% protein). The next morning, an i.v. catheter
was placed in an antecubital vein for infusion of insulin and
glucose. A second catheter was inserted in a dorsal vein of the
contralateral arm for blood withdrawal. The hand was placed
between a
heating pad for arterialization of venous blood sampling. During
the 45 min prior to the clamp, bloodsamples were collected
every 15 min for glucose and insulin. Then insulin was
administered at a primedcontinuous infusion rate of 120
mU(861 pmol)×m22×min21 for 2 h and blood samples were
collected every 5 min for glucose and every 15 min for insulin
during this period. Serum insulin was measured by a Siemens
Immulite 2000 using immunoassay with chemiluminescent. A
variable infusionof dextrose (20% solution) was given to
maintainserum glucose concentrations at ;5.6 mmol/L (100
mg/dL). Arterialized serum glucose was measured using a YSI

2300 Stat Plus glucose analyzer (model no. 2300 STAT Plus D)
and Beckman Coulter DXC600. During the steady state (last 30
min of clamp), the mean rate of exogenous glucose infusion was
corrected for changes in glycemia and divided by fat-free mass
to assess insulin sensitivity.
Body weight/fat distribution. Fat-free mass, fat mass, and body
fat percentage were measured by dual-energy X-ray
absorptiometry with CV for measurements assessed at 0.6, 1.1,
and 1.1%, respectively. Overall, biologic, instrument, and
reader variability was assessed at ~10%.
Serum inflammatory biomarkers and lipids. During the baseline
of the clamp, blood was collected for measuring serum
inflammatory biomarkers, including high sensitivityC-reactive
protein (hsCRP), tumor necrosis factor-a (TNFa), and monocyte
chemoattractant protein 1 (MCP-1). TNFa and MCP-1 were
measured on a Luminex system using kits from Millipore. High
sensitivity C-reactive protein was measured by automated
immunoassay as assessed on a Siemens 2000 instrument. In
addition, the serum lipid profile was measured (triglycerides,
total cholesterol, LDL-cholesterol, and HDL-cholesterol).
Triglycerides and total cholesterol were measured by using a
Beckman Coulter DXC600 and HDL-cholesterol was measured
by using a Trinity DXC600. LDL-cholesterol was based on a
calculation [cholesterol 2 (1/ 5 triglycerides) – HDL].
Food records and questionnaires. At the screening visits, a
registered dietitian instructed participants to record a detailed
3-d food record (i.e., 2 weekdays and 1 weekend day).
Participants were asked to provide labels and/or recipes for
accuracy of the food records. The dietitian reviewed the food
records for accuracy and completeness. Based on their eating
patterns and usual intake, participants were counseled by the
dietitian on ways to remove ~2000 kJ/d (500 kcal/d) from their
daily intake to compensate for the energy consumed in the
blueberry and placebo smoothies. Food records were also
administered at the midpoint and end of the study. The food
records were analyzed using the Pennington Biomedical

Research Center’s Food Diary Program (Pennington Biomedical
Research Foundation). Participants were asked to maintain their
current body weight and physical activity or they would be
eliminated from the study. The participants’ body weights were
measured weekly to monitor weight maintenance. A change of
$1 kg of body weight was addressed by the dietitian and proper
counseling was provided. They also reported adverse events and
changes in medication during the study. The smoothie rating
and fruit/wine questionnaires were also used in the study.
Before starting the study, participants were given the
opportunity to taste the smoothie for acceptability. The
fruit/wine questionnaire was administered at each visit as a
reminder to abstain from berries, grapes, juices that contained
berries and grapes, and wine throughout the study. The rationale
for these questionnaires was to eliminate consumption of
anthocyanin-containing foods and drinks.
Statistical analysis. All analyseswere performedusing SAS
version 9.2. Repeated-measures ANOVA with week as the
repeated factor was used to compare the blueberry with placebo
groups. Differences between the blueberry and placebo baseline
characteristics were analyzed by a 2-sample t test (continuous
data) and within groups analyzed by a paired t-test. Categorical
data were summarized as counts and analyzed by chisquare
tests. Nutritional value of food intake was analyzed by
mixedmodel ANOVA. P # 0.05 indicated a significant difference
between the groups. Data were expressed as means 6 SEM.
Effect of blueberries on insulin sensitivity 1765
Results
At baseline, the groups did not differ in age, body composition,
lipid profile, blood pressure, and inflammatory biomarkers
(Table 1).
Energy intake, body composition, and metabolic variables.
Throughout the study, the groups did not differ in energy and
macronutrient (protein, carbohydrate, and fat) consumption
(data not shown) or in body weight or adiposity (Table 1). In

addition, the inflammatory biomarkers, lipid profile, and blood
pressure did not differ between the study groups from the
beginning to the end of the study (Table 1). None of these
variables changed within each group during the treatment period
(Table 1).
Insulin sensitivity. When evaluating the percent change of
insulin sensitivity, 67% of the participants (10 of 15)
randomized to the blueberry group had at least a 10% or greater
favorable change in insulin sensitivity, whereas only 41% of the
placebo participants (7 of 17) demonstrated this change (Fig. 1).
Themeanchangeininsulin sensitivitywasimprovedsignificantly
more in the blueberry group compared to the placebo group
(Fig. 2). Also, the percent change in insulin sensitivity was
greater in the blueberry group (22.2 65.8%) than in the placebo
group (4.9 6 4.5%) (P = 0.02).
Discussion
To our knowledge, this is the first reported human study that
evaluated the effect of daily dietary supplementation with
bioactives in blueberries on whole-body insulin sensitivity in
obese, nondiabetic, and insulin-resistant men and women. The
uniqueness of this study relates to the design, which was
randomized, double blinded, and placebo controlled. By design,
the blueberry and placebo smoothies were identical in physical
appearance and macronutrient content with the exception of
adding the blueberry bioactives to the blueberry smoothie.
Another strength of the study was the use of the most precise
metabolic technique for assessing whole-body insulin
sensitivity, i.e.,hyperinsulinemic-euglycemic clamps.The
majorfindingwas
TABLE 1 Anthropometrics and serum biochemistry of obese,
insulin-resistant participants before (pre) and after (post) the
blueberry and placebo treatments1
Variables
Blueberry Placebo Pre Post Pre Post
Race (African American/Caucasian), n/n 8/7 — 8/9 — Gender
(male/female), n/n 2/13 — 3/14 — Age, y 54 6 3 — 49 6 3—

Body weight, kg 98.7 6 3.1 99.1 6 3.1 102.9 6 3.4 103.4 6 3.5
BMI, kg/m2 36.8 6 0.9 37.0 6 0.9 38.0 6 0.9 38.2 6 1.0 Body
fat, % 40.9 6 1.3 40.9 6 1.3 42.5 6 1.4 42.8 6 1.4 Fat mass, kg
40.8 6 2.0 40.8 6 2.0 44.2 6 2.3 44.7 6 2.3 Lean mass, kg 58.7 6
2.1 58.7 6 2.1 59.2 6 2.0 59.4 6 2.1 Systolic blood pressure, mm
Hg 116.9 6 3.2 115.2 6 3.2 122.6 6 3.7 118.5 6 3.2 Diastolic
blood pressure, mm Hg 73.5 6 2.3 73.2 6 1.9 75.7 6 1.9 76.6 6
2.1 Serum biochemistry2 Glucose, mmol/L 5.7 6 0.1 5.7 6 0.1
5.9 6 0.1 5.9 6 0.1 Insulin, pmol/L 132 6 15 140 6 17 142 6 15
148 6 16 Triglycerides, mmol/L 1.53 6 0.18 1.66 6 0.17 1.44 6
0.21 1.67 6 0.26 Cholesterol, mmol/L 5.34 6 0.21 4.76 6 0.24
5.18 6 0.19 4.65 6 0.18 LDL cholesterol, mmol/L 3.28 6 0.21
2.88 6 0.19 3.22 6 0.18 2.84 6 0.17 HDL cholesterol, mmol/L
1.35 6 0.08 1.12 6 0.06 1.30 6 0.07 1.05 6 0.06 C-reactive
protein, mg/L 5.3 6 1.3 6.9 6 1.8 6.9 6 1.1 8.5 6 1.9 TNFa, ng/L
7.4 6 1.5 6.2 6 1.0 11.5 6 4.3 6.5 6 0.5 MCP-1, ng/L 358 6 37
377 6 44 401 6 58 396 6 38
1 Values are means 6 SEM, n = 15 (blueberry) or 17 (placebo)
except TNFa, where n = 11 or 13, respectively. 2 Blood was
drawn from participants after a 10-h fast.
FIGURE 1 Percent change in insulin sensitivity in individual
obese, insulin-resistant men and women who consumed the
blueberry (black bars) or placebo (white bars) smoothies for 6
wk. % D = [(postintervention 2
preintervention)/preintervention] 3 100. Values are means 6
SEM, n = 15 (blueberry) or 17 (placebo).
1766 Stull et al.
that daily consumption of whole blueberry bioactives for 6 wk
improved insulin sensitivity in a population at high risk for type
2 diabetes compared with ad libitum dietary intake alone.
Consumptionofsmoothies(in the caseofthisstudy, bioactives in
blueberries) may be a more attractive and convenient dietary
approach for those adults who do not consume the recommended
daily amounts of fruits and vegetables. In the current study, we
made sure that the energy in the smoothies did not contribute to

any body weight gain. Specifically, our study dietitian worked
with the participants during the weekly visits to eliminate 2000
kJ/d (1000 kJ/smoothie) from their diets to compensate for the
energy provided by the smoothies. As such, the participants
were able to maintain a constant body weight throughout the
study. The observation that insulin sensitivity increased
withouta change inbodyweight suggests thatthe
blueberrybioactives had a direct effect on increasing whole-
body insulin action. The current study evaluated the synergistic
effect of all the bioactive compounds in blueberries. Limited
data exist on using whole blueberries as the intervention. In a
previous preclinical study, DeFuria et al. (3) used a comparable
dose of an identical freeze-dried whole blueberry powder and
observed similar health effects to the current clinical trial. The
study showed that mice who consumed a high-fat diet with
blueberries for 8 wk had a lower plasma glucose AUC during a
90-min intraperitoneal insulin tolerance test compared with the
mice fed the highfat diet alone. Plasma insulin concentrations
were unchanged. These results suggest that blueberries
improved the high-fat diet–induced hyperglycemia. However,
Prior et al. (11) found that freeze-dried whole blueberry powder
did not affect the plasma glucose AUC during a 120-min
intraperitoneal glucose tolerance test in high-fat diet–induced
obese mice. Perhaps the null finding was due to the type of
freeze-dried blueberry powder usedin the experiment, whichwas
different from thecurrent and previous (3) studies or the specific
technique used could have potentially lacked the precision to
adequately assess carbohydrate metabolism. It is well
established that any change in adiposity can greatly alter whole-
body insulin sensitivity (12). In the current study, body weight
was kept constant throughout the study, so that it would not be a
confounding factor that contributed to the improved insulin
sensitivity. Furthermore, participants were instructed not to
alter their physical activity during the study. Even after
controlling for certain variables, as expected for human studies,
there was variability in insulin sensitivity values for both

treatment groups. However, compared with the placebo
group overall, insulin sensitivity improved significantly more in
the blueberry group without any changes in body weight,
adiposity, or energy intake. Also, no changes in body
composition were observed in diet-induced obese mice fed
whole blueberries (3). Another study (11) found the opposite in
that whole blueberry supplementation increased body weight
and adiposity in mice that were fed a high-fat diet with added
blueberries compared with mice fed only a high-fat diet. The
increase in the body weight and adiposity of the mice
throughout the study could have potentially affected the
outcome of unobserved improvements in glucose tolerance with
whole blueberry supplementation, as discussed previously.
Emerging data have clearly linked inflammation to adiposity
with significant reports on the mechanisms by which
inflammation at a whole-body level attenuates insulin action
(13). Specifically, DeFuria et al. (3) found that supplementing
obese mice with blueberries reduced the gene expression for
inflammatory biomarkers TNFa and interleukin-10.
Unfortunately, significant changes were not observed in all the
measured inflammatory biomarkers (MCP-1, interleukin-6, and
inducible nitric oxide synthase). In the current study,
consumption of the daily dose of bioactives in blueberries did
not alter the participants’ inflammatory biomarker profile, which
consisted of hsCRP, TNFa, and MCP-1. The previous study (3)
and current study cannot be compared because of the different
research species and evaluations of inflammatory biomarkers
[gene expression (3) vs. serum (current study)]. Given the
enhanced insulin sensitivity in the group randomized to the
blueberry bioactives, a determination of insulindependent or -
independent signaling pathways in muscle would provide a
cellular basis contributing to the understanding of the clinical
effect. However, muscle biopsies were not obtained in the
current study and cellular mechanisms were not evaluated.
Some may view this as a study limitation, but we did evaluate
whole-body insulin sensitivity, which is a critical step before

evaluating cellular mechanisms. Furthermore, an in vitro study
showed (4) that 21-h incubation of the blueberry extract in
muscle cells enhanced glucose uptake only in the presence of
insulin. Another study (5) found that 6-h treatment of fermented
blueberry juice with and without insulin increased glucose
uptake into the muscle and adipocyte cells. However, the
nonfermented blueberry juice had no effect on glucose uptake.
The fermented blueberry juice also increased the
phosphorylation/activation of proteins in the insulin-
independent pathway (i.e., AMP-activated protein kinase) and
did not phosphorylate/ activate proteins in the insulin-dependent
pathway (i.e., AKT and ERK1/2). These results suggest that the
addition of fermented blueberry bioactives increased glucose
uptake into the cells in an insulin-independent mechanism.
More cellular mechanistic studies are warranted to elucidate the
speciﬁc cellular pathway involved in the improvement of insulin
sensitivity that was observed when blueberries were consumed
in our study. In conclusion, our double-blinded and placebo-
controlled study showed that daily dietary supplementation of
bioactives in freeze-dried whole blueberry powder improved
insulin sensitivity over 6 wk in obese, nondiabetic, and insulin-
resistant participants. The bioactives in blueberries enhanced
insulin sensitivity independent of any changes in inﬂammatory
biomarkers or adiposity. This study is not conclusive, but it
strongly suggests a need to further explore the cellular
mechanism for the effect. In addition, our study suggests the
need for studies of longer duration that will evaluate blueberries
and their potential role in improving insulin sensitivity in an
insulin-resistant human population.
FIGURE 2 Mean change in insulin sensitivity in the obese,
insulinresistant men and women who consumed either the
blueberry or placebo smoothies for 6 wk. D = postintervention 2
preintervention. Values are means 6 SEM, n = 15 (blueberry) or
17 (placebo).

Acknowledgments A.J.S. designed research, conducted research,
collected and analyzed the data, and wrote the manuscript;
C.M.C. and K.C.C. designed dietary research, conducted dietary
research, and collected and analyzed dietary data; W.D.J.
performed statistical analysis; and W.T.C. was the principal
investigator who designed research and had primary
responsibility for final content. All authors read and approved
the final manuscript.
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Disease
Bioactives in Blueberries Improve Insulin Sensitivity in Obese,
Insulin-Resistant Men and Women1–4
April J. Stull, Katherine C. Cash, William D. Johnson,
Catherine M. Champagne, and William T. Cefalu*
Center for the Study of Botanicals and Metabolic Syndrome,
Pennington Biomedical Research Center, Louisiana State
University System, Baton Rouge, LA 70808
Abstract
Dietary supplementation with whole blueberries in a preclinical
study resulted in a reduction in glucose concentrations over
time. We sought to evaluate the effect of daily dietary
supplementation with bioactives from blueberries on whole-
body insulin sensitivity in men and women. A double-blinded,
randomized, and placebo-controlled clinical study design was
used. After screening to resolve study eligibility, baseline (wk
0) insulin sensitivity was measured on 32 obese, nondiabetic,

and insulin-resistant subjects using a high-dose
hyperinsulinemic-euglycemic clamp (insulin infusion of 120
mU(861 pmol)×m22×min21). Serum inflammatory biomarkers
and adiposity were measured at baseline. At the end of the
study, insulin sensitivity, inflammatory biomarkers, and
adiposity were reassessed. Participants were randomized to
consume either a smoothie containing 22.5 g blueberry
bioactives (blueberry group, n = 15) or a smoothie of equal
nutritional value without added blueberry bioactives (placebo
group, n = 17) twice daily for 6 wk. Both groups were
instructed to maintain their body weight by reducing ad libitum
intake by an amount equal to the energy intake of the
smoothies. Participants’ body weights were evaluated weekly
and 3-d food records were collected at baseline, the middle, and
end of the study. The mean change in insulin sensitivity
improved more in the blueberry group (1.7 6 0.5 mg×kg
FFM21×min21) than in the placebo group (0.4 6 0.4 mg×kg
FFM21×min21)(P = 0.04). Insulin sensitivity was enhanced in
the blueberry group at the end of the study without significant
changes in adiposity, energy intake, and inflammatory
biomarkers. In conclusion, daily dietary supplementation with
bioactives from whole blueberries improved insulin sensitivity
in obese, nondiabetic, and insulin-resistant participants. J. Nutr.
140: 1764–1768, 2010.
Introduction
Increased consumption of berries has been shown to improve
cognitive function, risk of cardiovascular disease, and cancer
(1,2). Studies have also reported that specific berries, i.e.,
blueberries, have antidiabetic effects. Specifically, a study
performed in mice (3) found that supplementation with
whole blueberries reduced the blood glucose area under the
curve (AUC)5 in vivo and cell culture studies (4,5)
demonstrated increased glucose uptake in vitro (6). In addition,
inflammatory genes have been reduced in mice after consuming
blueberry bioactives, which suggests an antiinflammatory
response (3). The purported health benefits from blueberries

have been attributed to their phenolic bioactive compounds,
such as anthocyanins, which also have antioxidant properties
(6–8). Given the concern regarding the ability to greatly
increase and maintain an individual’s fruit and vegetable
consumption over a long-term period (9), the role of dietary
supplementation with bioactive components in blueberries
remains a very attractive and feasible daily dietary intervention.
To the best of our knowledge, there is no human research that
has reported on the efficacy of increased blueberry bioactive
consumption on insulin sensitivity by using the
hyperinsulinemic-euglycemic clamp technique (10), which is
the gold standard for measuring in
1 Supported in part by the NIH training grant T32 AT004094
(supporting A.J.S.), by the United States Highbush Blueberry
Council, and P50AT002776-01 from the National Center for
Complementary and Alternative Medicine and the Office of
Dietary Supplements (W.T.C.), which funds the Botanical
Research Center of Pennington Biomedical Research Center and
The Biotech Center of Rutgers University. This project used
facilities that are supported in part by Centers of Biomedical
Research Excellence (NIH P20-RR021945) and Clinical
Nutrition Research Unit (NIH 1P30-DK072476) center grants
from the NIH. 2 Author disclosures: A.J.Stull,K.C. Cash,W. D.
Johnson, andC. M.Champagne, no conflicts of interest. W. T.
Cefalu received research funds from the United States Highbush
Blueberry Council. 3 This trial was registered at
clinicaltrials.gov as NCT01005420. 4 Supplemental Figure 1
and Table 1 are available with the online posting of this paper
at jn.nutrition.org. * To whom correspondence should be
addressed. E-mail:
[email protected]
edu.
5 Abbreviations used: AUC, area under the curve; hsCRP, high
sensitivity C-reactive protein; MCP-1, monocyte
chemoattractant protein 1; TNFa, tumor necrosis factor-a.
ã 2010 American Society for Nutrition. 1764 Manuscript

received April 13, 2010. Initial review completed May 18, 2010.
Revision accepted July 12, 2010. First published online August
19, 2010; doi:10.3945/jn.110.125336.
6.DC1.html
http://jn.nutrition.org/content/suppl/2010/09/20/jn.110.12533
Supplemental Material can be found at:
vivo insulin action. Therefore, this project’s overall objective
was to examine the role of dietary supplementation with
bioactives in freeze-dried whole blueberry powder on insulin
action in vivo with the use of hyperinsulinemic-euglycemic
clamps in individuals who were obese, nondiabetic, and insulin
resistant. We hypothesized that increased daily consumption of
blueberry bioactives, based on preclinical data, would be
effective in increasing insulin action in vivo and ultimately
result in improved insulin sensitivity in a human population at
high risk for type 2 diabetes.
Subjects and Methods
Subjects. Participants inthe study were recruited fromthe
Greater Baton Rouge area. A total of 32 men and women
completed all evaluations (Supplemental Fig. 1). Those included
were adults ($20 y old), obese (BMI between 32 and 45 kg/m2),
and insulin resistant (nondiabetic). The exclusion criteria
included: 1) diabetes; no diabetes status was conﬁrmed by a 2-h
oral glucose tolerance test; 2) medications known to affect
glucose metabolism; 3) untreated thyroid or chronic liver, renal,
or cardiovascular disease; 4) a history of drug and/or alcohol
abuse, or psychiatric disease prohibiting adherence to study
protocol; 5) history of allergic reactions to blueberries; 6)
consuming berries, grapes, and wine .3 times/wk; and 7)
ﬂuctuation in body weight . 5% in the preceding 2 mo. The
Institutional Review Board for human subjects at Pennington
Biomedical Research Center reviewed and approved the study
protocol. All participants gave written consent prior to starting
the study.
Study design.This study design was double blinded, placebo-

controlled, and randomized. All study evaluations and
measurementswere performed onparticipantsthat had fasted for
10h. Aweekwasdeﬁned as7 d (6 2 d).
Clinical intervention and source of whole blueberry bioactives.
The freeze-dried whole blueberry powder was prepared by the
United States Highbush Blueberry Council (USDA oversight).
The whole blueberry powder was made from a 50/50 mixture of
2 varieties of highbush blueberries, Tifblue (Vaccinium ashei)
and Rubel (Vaccinium corymbosum). The whole blueberries
were freeze-dried, milled, and stored in aluminum cans under
nitrogen. Based on the compositional analysis, the 45 g of
blueberry powder contained 1462 mg of total phenolics, 668 mg
of anthocyanins, and 16.02 mmol TE of antioxidants (oxygen
radical absorbance capacity). Also, the 45 g of blueberry
powder that was provided to the participants equated to an
amount of bioactives in ~2 cups of fresh whole blueberries.
After the participants were assessed as being insulin resistant
(glucose disposal rate # 650 mg/min), they were randomized to
receive twice daily a smoothie with blueberry bioactives added
or an identical smoothie without blueberry bioactives (i.e.,
placebo) (Supplemental Table 1). The participants were
instructed to consume 1 smoothie at breakfast meals and the
other smoothie at dinner meals (at least 6 h apart). The
smoothies were prepared in the metabolic kitchen and a week’s
supply of frozen smoothies was provided in a cooler for the
participants to pick up at each weekly visit. Participants were
instructed to keep the smoothies frozen, thaw them in the
refrigerator, avoid exposing them to direct heat, and avoid
adding any other ingredients to them. For study compliance, the
participants verbally reported their smoothie consumption to the
dietitian at each visit. A compliance of .75% was mandatory for
continued participation in the study.
Physiologic assessments. Hyperinsulinemic-euglycemic clamps
(10) were performed to assess insulin sensitivity after a 10-h
fast. Participants were admitted into the inpatient research unit
the evening prior to their insulin sensitivity testing day and

consumed a eucaloric standardized meal (50% carbohydrates,
35% fat, and 15% protein). The next morning, an i.v. catheter
was placed in an antecubital vein for infusion of insulin and
glucose. A second catheter was inserted in a dorsal vein of the
contralateral arm for blood withdrawal. The hand was placed
between a
heating pad for arterialization of venous blood sampling. During
the 45 min prior to the clamp, bloodsamples were collected
every 15 min for glucose and insulin. Then insulin was
administered at a primedcontinuous infusion rate of 120
mU(861 pmol)×m22×min21 for 2 h and blood samples were
collected every 5 min for glucose and every 15 min for insulin
during this period. Serum insulin was measured by a Siemens
Immulite 2000 using immunoassay with chemiluminescent. A
variable infusionof dextrose (20% solution) was given to
maintainserum glucose concentrations at ;5.6 mmol/L (100
mg/dL). Arterialized serum glucose was measured using a YSI
2300 Stat Plus glucose analyzer (model no. 2300 STAT Plus D)
and Beckman Coulter DXC600. During the steady state (last 30
min of clamp), the mean rate of exogenous glucose infusion was
corrected for changes in glycemia and divided by fat-free mass
to assess insulin sensitivity.
Body weight/fat distribution. Fat-free mass, fat mass, and body
fat percentage were measured by dual-energy X-ray
absorptiometry with CV for measurements assessed at 0.6, 1.1,
and 1.1%, respectively. Overall, biologic, instrument, and
reader variability was assessed at ~10%.
Serum inflammatory biomarkers and lipids. During the baseline
of the clamp, blood was collected for measuring serum
inflammatory biomarkers, including high sensitivityC-reactive
protein (hsCRP), tumor necrosis factor-a (TNFa), and monocyte
chemoattractant protein 1 (MCP-1). TNFa and MCP-1 were
measured on a Luminex system using kits from Millipore. High
sensitivity C-reactive protein was measured by automated
immunoassay as assessed on a Siemens 2000 instrument. In
addition, the serum lipid profile was measured (triglycerides,

total cholesterol, LDL-cholesterol, and HDL-cholesterol).
Triglycerides and total cholesterol were measured by using a
Beckman Coulter DXC600 and HDL-cholesterol was measured
by using a Trinity DXC600. LDL-cholesterol was based on a
calculation [cholesterol 2 (1/ 5 triglycerides) – HDL].
Food records and questionnaires. At the screening visits, a
registered dietitian instructed participants to record a detailed
3-d food record (i.e., 2 weekdays and 1 weekend day).
Participants were asked to provide labels and/or recipes for
accuracy of the food records. The dietitian reviewed the food
records for accuracy and completeness. Based on their eating
patterns and usual intake, participants were counseled by the
dietitian on ways to remove ~2000 kJ/d (500 kcal/d) from their
daily intake to compensate for the energy consumed in the
blueberry and placebo smoothies. Food records were also
administered at the midpoint and end of the study. The food
records were analyzed using the Pennington Biomedical
Research Center’s Food Diary Program (Pennington Biomedical
Research Foundation). Participants were asked to maintain their
current body weight and physical activity or they would be
eliminated from the study. The participants’ body weights were
measured weekly to monitor weight maintenance. A change of
$1 kg of body weight was addressed by the dietitian and proper
counseling was provided. They also reported adverse events and
changes in medication during the study. The smoothie rating
and fruit/wine questionnaires were also used in the study.
Before starting the study, participants were given the
opportunity to taste the smoothie for acceptability. The
fruit/wine questionnaire was administered at each visit as a
reminder to abstain from berries, grapes, juices that contained
berries and grapes, and wine throughout the study. The rationale
for these questionnaires was to eliminate consumption of
anthocyanin-containing foods and drinks.
Statistical analysis. All analyseswere performedusing SAS
version 9.2. Repeated-measures ANOVA with week as the
repeated factor was used to compare the blueberry with placebo

groups. Differences between the blueberry and placebo baseline
characteristics were analyzed by a 2-sample t test (continuous
data) and within groups analyzed by a paired t-test. Categorical
data were summarized as counts and analyzed by chisquare
tests. Nutritional value of food intake was analyzed by
mixedmodel ANOVA. P # 0.05 indicated a significant difference
between the groups. Data were expressed as means 6 SEM.
Results
At baseline, the groups did not differ in age, body composition,
lipid profile, blood pressure, and inflammatory biomarkers
(Table 1).
Energy intake, body composition, and metabolic variables.
Throughout the study, the groups did not differ in energy and
macronutrient (protein, carbohydrate, and fat) consumption
(data not shown) or in body weight or adiposity (Table 1). In
addition, the inflammatory biomarkers, lipid profile, and blood
pressure did not differ between the study groups from the
beginning to the end of the study (Table 1). None of these
variables changed within each group during the treatment period
(Table 1).
Insulin sensitivity. When evaluating the percent change of
insulin sensitivity, 67% of the participants (10 of 15)
randomized to the blueberry group had at least a 10% or greater
favorable change in insulin sensitivity, whereas only 41% of the
placebo participants (7 of 17) demonstrated this change (Fig. 1).
Themeanchangeininsulin sensitivitywasimprovedsignificantly
more in the blueberry group compared to the placebo group
(Fig. 2). Also, the percent change in insulin sensitivity was
greater in the blueberry group (22.2 65.8%) than in the placebo
group (4.9 6 4.5%) (P = 0.02).
Discussion
To our knowledge, this is the first reported human study that
evaluated the effect of daily dietary supplementation with
bioactives in blueberries on whole-body insulin sensitivity in

obese, nondiabetic, and insulin-resistant men and women. The
uniqueness of this study relates to the design, which was
randomized, double blinded, and placebo controlled. By design,
the blueberry and placebo smoothies were identical in physical
appearance and macronutrient content with the exception of
adding the blueberry bioactives to the blueberry smoothie.
Another strength of the study was the use of the most precise
metabolic technique for assessing whole-body insulin
sensitivity, i.e.,hyperinsulinemic-euglycemic clamps.The
majorﬁndingwas
TABLE 1 Anthropometrics and serum biochemistry of obese,
insulin-resistant participants before (pre) and after (post) the
blueberry and placebo treatments1
Variables
Blueberry Placebo Pre Post Pre Post
Race (African American/Caucasian), n/n 8/7 — 8/9 — Gender
(male/female), n/n 2/13 — 3/14 — Age, y 54 6 3 — 49 6 3—
Body weight, kg 98.7 6 3.1 99.1 6 3.1 102.9 6 3.4 103.4 6 3.5
BMI, kg/m2 36.8 6 0.9 37.0 6 0.9 38.0 6 0.9 38.2 6 1.0 Body
fat, % 40.9 6 1.3 40.9 6 1.3 42.5 6 1.4 42.8 6 1.4 Fat mass, kg
40.8 6 2.0 40.8 6 2.0 44.2 6 2.3 44.7 6 2.3 Lean mass, kg 58.7 6
2.1 58.7 6 2.1 59.2 6 2.0 59.4 6 2.1 Systolic blood pressure, mm
Hg 116.9 6 3.2 115.2 6 3.2 122.6 6 3.7 118.5 6 3.2 Diastolic
blood pressure, mm Hg 73.5 6 2.3 73.2 6 1.9 75.7 6 1.9 76.6 6
2.1 Serum biochemistry2 Glucose, mmol/L 5.7 6 0.1 5.7 6 0.1
5.9 6 0.1 5.9 6 0.1 Insulin, pmol/L 132 6 15 140 6 17 142 6 15
148 6 16 Triglycerides, mmol/L 1.53 6 0.18 1.66 6 0.17 1.44 6
0.21 1.67 6 0.26 Cholesterol, mmol/L 5.34 6 0.21 4.76 6 0.24
5.18 6 0.19 4.65 6 0.18 LDL cholesterol, mmol/L 3.28 6 0.21
2.88 6 0.19 3.22 6 0.18 2.84 6 0.17 HDL cholesterol, mmol/L
1.35 6 0.08 1.12 6 0.06 1.30 6 0.07 1.05 6 0.06 C-reactive
protein, mg/L 5.3 6 1.3 6.9 6 1.8 6.9 6 1.1 8.5 6 1.9 TNFa, ng/L
7.4 6 1.5 6.2 6 1.0 11.5 6 4.3 6.5 6 0.5 MCP-1, ng/L 358 6 37
377 6 44 401 6 58 396 6 38
1 Values are means 6 SEM, n = 15 (blueberry) or 17 (placebo)
except TNFa, where n = 11 or 13, respectively. 2 Blood was

drawn from participants after a 10-h fast.
FIGURE 1 Percent change in insulin sensitivity in individual
obese, insulin-resistant men and women who consumed the
blueberry (black bars) or placebo (white bars) smoothies for 6
wk. % D = [(postintervention 2
preintervention)/preintervention] 3 100. Values are means 6
SEM, n = 15 (blueberry) or 17 (placebo).
1766 Stull et al.
that daily consumption of whole blueberry bioactives for 6 wk
improved insulin sensitivity in a population at high risk for type
2 diabetes compared with ad libitum dietary intake alone.
Consumptionofsmoothies(in the caseofthisstudy, bioactives in
blueberries) may be a more attractive and convenient dietary
approach for those adults who do not consume the recommended
daily amounts of fruits and vegetables. In the current study, we
made sure that the energy in the smoothies did not contribute to
any body weight gain. Speciﬁcally, our study dietitian worked
with the participants during the weekly visits to eliminate 2000
kJ/d (1000 kJ/smoothie) from their diets to compensate for the
energy provided by the smoothies. As such, the participants
were able to maintain a constant body weight throughout the
study. The observation that insulin sensitivity increased
withouta change inbodyweight suggests thatthe
blueberrybioactives had a direct effect on increasing whole-
body insulin action. The current study evaluated the synergistic
effect of all the bioactive compounds in blueberries. Limited
data exist on using whole blueberries as the intervention. In a
previous preclinical study, DeFuria et al. (3) used a comparable
dose of an identical freeze-dried whole blueberry powder and
observed similar health effects to the current clinical trial. The
study showed that mice who consumed a high-fat diet with
blueberries for 8 wk had a lower plasma glucose AUC during a
90-min intraperitoneal insulin tolerance test compared with the
mice fed the highfat diet alone. Plasma insulin concentrations
were unchanged. These results suggest that blueberries

improved the high-fat diet–induced hyperglycemia. However,
Prior et al. (11) found that freeze-dried whole blueberry powder
did not affect the plasma glucose AUC during a 120-min
intraperitoneal glucose tolerance test in high-fat diet–induced
obese mice. Perhaps the null finding was due to the type of
freeze-dried blueberry powder usedin the experiment, whichwas
different from thecurrent and previous (3) studies or the specific
technique used could have potentially lacked the precision to
adequately assess carbohydrate metabolism. It is well
established that any change in adiposity can greatly alter whole-
body insulin sensitivity (12). In the current study, body weight
was kept constant throughout the study, so that it would not be a
confounding factor that contributed to the improved insulin
sensitivity. Furthermore, participants were instructed not to
alter their physical activity during the study. Even after
controlling for certain variables, as expected for human studies,
there was variability in insulin sensitivity values for both
treatment groups. However, compared with the placebo
group overall, insulin sensitivity improved significantly more in
the blueberry group without any changes in body weight,
adiposity, or energy intake. Also, no changes in body
composition were observed in diet-induced obese mice fed
whole blueberries (3). Another study (11) found the opposite in
that whole blueberry supplementation increased body weight
and adiposity in mice that were fed a high-fat diet with added
blueberries compared with mice fed only a high-fat diet. The
increase in the body weight and adiposity of the mice
throughout the study could have potentially affected the
outcome of unobserved improvements in glucose tolerance with
whole blueberry supplementation, as discussed previously.
Emerging data have clearly linked inflammation to adiposity
with significant reports on the mechanisms by which
inflammation at a whole-body level attenuates insulin action
(13). Specifically, DeFuria et al. (3) found that supplementing
obese mice with blueberries reduced the gene expression for
inflammatory biomarkers TNFa and interleukin-10.

Unfortunately, significant changes were not observed in all the
measured inflammatory biomarkers (MCP-1, interleukin-6, and
inducible nitric oxide synthase). In the current study,
consumption of the daily dose of bioactives in blueberries did
not alter the participants’ inflammatory biomarker profile, which
consisted of hsCRP, TNFa, and MCP-1. The previous study (3)
and current study cannot be compared because of the different
research species and evaluations of inflammatory biomarkers
[gene expression (3) vs. serum (current study)]. Given the
enhanced insulin sensitivity in the group randomized to the
blueberry bioactives, a determination of insulindependent or -
independent signaling pathways in muscle would provide a
cellular basis contributing to the understanding of the clinical
effect. However, muscle biopsies were not obtained in the
current study and cellular mechanisms were not evaluated.
Some may view this as a study limitation, but we did evaluate
whole-body insulin sensitivity, which is a critical step before
evaluating cellular mechanisms. Furthermore, an in vitro study
showed (4) that 21-h incubation of the blueberry extract in
muscle cells enhanced glucose uptake only in the presence of
insulin. Another study (5) found that 6-h treatment of fermented
blueberry juice with and without insulin increased glucose
uptake into the muscle and adipocyte cells. However, the
nonfermented blueberry juice had no effect on glucose uptake.
The fermented blueberry juice also increased the
phosphorylation/activation of proteins in the insulin-
independent pathway (i.e., AMP-activated protein kinase) and
did not phosphorylate/ activate proteins in the insulin-dependent
pathway (i.e., AKT and ERK1/2). These results suggest that the
addition of fermented blueberry bioactives increased glucose
uptake into the cells in an insulin-independent mechanism.
More cellular mechanistic studies are warranted to elucidate the
specific cellular pathway involved in the improvement of insulin
sensitivity that was observed when blueberries were consumed
in our study. In conclusion, our double-blinded and placebo-
controlled study showed that daily dietary supplementation of

bioactives in freeze-dried whole blueberry powder improved
insulin sensitivity over 6 wk in obese, nondiabetic, and insulin-
resistant participants. The bioactives in blueberries enhanced
insulin sensitivity independent of any changes in inflammatory
biomarkers or adiposity. This study is not conclusive, but it
strongly suggests a need to further explore the cellular
mechanism for the effect. In addition, our study suggests the
need for studies of longer duration that will evaluate blueberries
and their potential role in improving insulin sensitivity in an
insulin-resistant human population.
FIGURE 2 Mean change in insulin sensitivity in the obese,
insulinresistant men and women who consumed either the
blueberry or placebo smoothies for 6 wk. D = postintervention 2
preintervention. Values are means 6 SEM, n = 15 (blueberry) or
17 (placebo).
Acknowledgments A.J.S. designed research, conducted research,
collected and analyzed the data, and wrote the manuscript;
C.M.C. and K.C.C. designed dietary research, conducted dietary
research, and collected and analyzed dietary data; W.D.J.
performed statistical analysis; and W.T.C. was the principal
investigator who designed research and had primary
responsibility for final content. All authors read and approved
the final manuscript.
Literature Cited
1. Bagchi D, Sen CK, Bagchi M, Atalay M. Anti-angiogenic,
antioxidant, and anti-carcinogenic properties of a novel
anthocyanin-rich berry extract formula. Biochemistry (Mosc).
2004;69:75–80, 1 p preceding 75. 2. Papandreou MA,
Dimakopoulou A, Linardaki ZI, Cordopatis P, KlimisZacas D,
Margarity M, Lamari FN. Effect of a polyphenol-rich wild
blueberry extract on cognitive performance of mice, brain
antioxidant
markersandacetylcholinesteraseactivity.BehavBrainRes.2009;19
8:352–8. 3. DeFuria J, Bennett G, Strissel KJ, Perfield JW II,

Milbury PE, Greenberg AS, Obin MS. Dietary blueberry
attenuates whole-body insulin resistance in high fat-fed mice by
reducing adipocyte death and its inflammatory sequelae. J Nutr.
2009;139:1510–6. 4. Martineau LC, Couture A, Spoor D,
Benhaddou-Andaloussi A, Harris C, Meddah B, Leduc C, Burt
A, Vuong T, et al. Anti-diabetic properties of the Canadian
lowbush blueberry Vaccinium angustifolium Ait.
Phytomedicine. 2006;13:612–23.
5. Vuong T, Martineau LC, Ramassamy C, Matar C, Haddad PS.
Fermented Canadian lowbush blueberry juice stimulates glucose
uptake and AMP-activated protein kinase in insulin-sensitive
cultured muscle cells and adipocytes. Can J Physiol Pharmacol.
2007;85: 956–65. 6. Youdim KA, Shukitt-Hale B, MacKinnon S,
Kalt W, Joseph JA. Polyphenolics enhance red blood cell
resistance to oxidative stress: in vitro and in vivo. Biochim
Biophys Acta. 2000;1523:117–22. 7. Hosseinian FS, Beta T.
Saskatoon and wild blueberries have higher anthocyanin
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2007;55:10832–8. 8. Faria A, Oliveira J, Neves P, Gameiro P,
Santos-Buelga C, de Freitas V, Mateus N. Antioxidant
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J Agric Food Chem. 2005;53:6896–902. 9. Blanck HM,
Gillespie C, Kimmons JE, Seymour JD, Serdula MK. Trends in
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A, Howard LR. Whole berries versus berry anthocyanins:
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The requirements are:
BIO 119 – Biology for Health Professionals
Journal Assignment
The assignment is worth
60 points
(39 drafts/21 final)
This is an INDIVIDUAL assignment!
The objective of this assignment is to provide you with
experience in evaluating scientific research papers. The
abilities to understand and critically analyze the results of a
scientific journal article are critical in the healthcare field. For
this assignment, you will evaluate how the scientific method has
been applied in a peer-reviewed journal article.
You will complete this assignment in
four sections (39 points)
and then submit a
final revised paper (21 points)
. Due dates for all components are listed on your syllabus – do
not forget!! Every section is graded and details are explained
below and in the rubric.
For this assignment, you are required to provide a summary of
the scientific method as it was presented by the journal authors.
Refer to the following information when completing your paper.
Format:
The paper shall be about 2-4 pages in length, double-spaced,
and 12-font type.
Include your name at the top right hand corner of the first page.

Title: The title should be centered at the top of the first page of
your paper, single-spaced, in 12-font type. Choose a title that
concisely explains the topic of your paper.
A title page is not required.
Grammar and spelling
:
The paper shall be written with proper grammar and free of
spelling errors. Use spell check, and carefully read your paper
prior to turning it in. ½ point will be deducted for each spelling
or grammatical error.
Evaluation of Scientific Method and Research:
Identify and describe the following components of the scientific
method in the body of your paper:
Section 1:
Definitions and Observations (8 points)
Definition of Terms and Concepts:
Define any important terms and concepts so that the reader
understands the topic of the journal article. You will likely
need to reference additional sources of information. Make sure
you properly cite the source(s) of your information.
Observations and Previous Studies:
Describe the observations that led the authors to conduct this
study. This information can usually be found in the background
information and discussion sections of the article. Describe at
least one previous study that led the authors to investigate the
current topic.
Question
: In your own words, state the question that the authors are
trying to answer in their experiment. The question should be
specific enough that it leads to the framing of a good
hypothesis.

This should be written in the form of a question.
Section 2
:
Hypothesis and Experimental Design (10 points)
Hypothesis
: State the hypothesis provided by the authors of the article.
Make sure that you identify the statement as the hypothesis
(i.e., “The hypothesis is…”). The hypothesis should be written
in your own words. It should be specific and written in the
proper format.
Experiment
: Describe the experiment as it is outlined in the journal
article. The following components must be included:
Independent variable:
State the independent variable.
Dependent variable:
State the dependent variable.
Controlled (constant) variables:
List the controlled variables used by the authors during the
experiment.
Control and experimental groups:
Identify the control and experimental groups.
Description of experiment
and data collection
:
Describe the steps of the experiment. Include a description of
how data and results were collected. You will need to cite the
journal article as the source for the details.
Section 3:
Summary and Interpretation of
Results
(12 points)

This section requires that you read the article, summarize the
results with specific details from graphics in the paper, and
explain the results of the study. This section will be 1 – 3
paragraphs long, depending on the paper. This section must
contain the following:
Summary of results:
Describe the results of the authors’ experiment for each
dependent variable. Be specific and include examples of the
qualitative and/or quantitative results. Compare and contrast
the results of the control and experimental groups.
Reference to graphic:
Choose one
results
table, graph, or diagram from the paper and
refer to it during your summary of results
. The graphic must describe results, NOT the experimental
design nor the demographics of the subjects.
Explain what information is being presented in the graphic.
This information is often found in the title of the graphic and/or
the caption.
Describe the results presented in the graphic. Give SPECIFIC
examples of quantitative and/or qualitative data presented in the
graphic. Cite your journal article as the source of this data.
Explain what the results presented in the graphic mean in detail.
Explanation of results:
Explain the results as if you were explaining them to a patient
or family member. Interpret and summarize them in lay person’s
language. What is the significance and health/medical
application of this study? Write this as if you are actually
explaining to the patient.
Analysis of
hypothesis
:
Discuss whether the hypothesis was supported or refuted.
Why or why not?

Section 4:
Discussion (9 points)
Discuss the validity and future implications of the research.
You should discuss
at least two
of the following points. You must cite outside sources to
support your statements.
Was the experiment designed and conducted according to proper
scientific method? Give specific examples of how it did or did
not follow the scientific method.
Are the authors’ results supported by or in opposition to
research conducted by other scientists?
What additional research or experimentation is needed? What
would you recommend to further investigate this topic?
What are the implications of this research (i.e., what is its use
in the “real world?”)? How does this research add to the basic
field of study or to the disease process specifically?
Additional Resources
:
Additional resources are required in this paper. They may be
helpful in understanding the observations (background) and
analyzing the results of the experiments. An additional source
is required for both of your discussion points. Additional
resources must be cited in the paper and reference page.
Citations within paper (
in-text or
parenthetical citations):
You must provide proper citation of all references (including
your article) throughout the paper. In-text citations tell the
reader where paraphrased or quoted information came from.
Paraphrasing is putting the ideas from a resource in your own
words. When paraphrasing, it is not sufficient to change only

one word of a sentence! See me if you are unclear on how to
paraphrase. If you copy word-for-word from a resource, you
must put the quoted (word-for-word) material in quotation
marks. The majority of the paper should be in your own words
or paraphrasing, not direct quotations from resources.
Follow APA style for in-text citations. Refer to the APA
Citation Guide and the following HCC webpage for more
information on how to cite sources within the body of your
paper:
http://www.harford.edu/library/citation_resources/style_guides/
apaintext.pdf
Failure to appropriately cite resources within the body of your
paper or to include quotation marks around any word-for-word
quotations will be considered
plagiarism
. Plagiarism may result in loss of points and possibly a zero for
the assignment.
http://www.harford.edu/library/tutorials/captivate_tutorials/apa.
htm
provides a tutorial on how to create a reference page
and
how to cite sources within your paper.
You should complete this tutorial as part of your preparations
for this assignment
. Additional APA citation information can be found at
http://www.harford.edu/library/citation_resources/
and at
http://www.apastyle.org/learn/tutorials/basics-tutorial.aspx
Reference
Page
:
This reference page will include the citation for the source of

the article and for
all
sources consulted. Follow APA style for the citations. Use the
APA Citation Guide on Blackboard to complete your reference
page and in-text citatio
Name:
Draft Section 1: Definitions of Terms and Concepts,
Observations, Previous Studies, Question (8 points):
3 Points
2 Points
1 Points
0 Points
Points Earned
Definition of Terms and Concepts
The paper defines terms and concepts necessary for
understanding the experiment.
The paper defines few key terms and concepts.
The paper defines few key terms and concepts and/or the
information is inaccurate.
The paper fails to define key terms and concepts.
Observations and Previous Studies
The paper describes the observations and previous studies that
led the authors to conduct this study.
The paper does not completely describe both the observations
and previous studies.
The description of the observation and previous studies is
incomplete or inaccurate.
The paper fails to describe any observations or previous studies.
Question

Not applicable.
Not applicable.
The paper states the question that the authors are trying to
answer.
The paper fails to state the question that the authors are trying
to answer.
In-text Citations
Not applicable.
Not applicable.
All information is appropriately cited in APA format.
In-text citations are absent or cited incorrectly.
Draft Section 2: Hypothesis and Experiment (10 points)
Name: ____________________________
3 Points
2 Points
1 Points
0 Points
Points Earned
Hypothesis
Not applicable.
Not applicable.
The hypothesis is stated in the correct format.
The hypothesis is missing or is not stated in the correct format.

Independent Variable
Not applicable.
Not applicable.
The independent variable is described.
The independent variable is missing or is incorrect.
Dependent Variable
Not applicable.
Not applicable.
The dependent variable is stated.
The dependent variable is missing or is incorrect.
Controlled (Constant) Variables
Not applicable.
Not applicable.
Controlled variables are stated.
Controlled variables are missing, incorrect, or incomplete.
Control and Experimental Groups
Not applicable.
Not applicable.
The paper identifies the control and experimental groups.
The paper does not identify the control and experimental
groups.
Description of Experiment
The paper thoroughly describes the steps of the experiment.
It describes how the control and experimental groups were
treated.
The description of the steps of the experiment and how the
control and experimental groups were treated is incomplete.
The paper fails to describe either the steps of the experiment
OR how control and experimental groups were treated.
Description of experiment and how control and experimental
groups were treated is missing.

Experiment:
Data Collection
Not applicable.
Not applicable.
The paper describes how data was collected.
The paper does not describe how data was collected.
In-text Citations
Not applicable.
Not applicable.
In-text citations are absent.
Draft Section 3: Interpretation and Summary of Results (12
points)
Name:
_____________________________________________________
____________________________
3 Points
2 Points
1 Points
0 Points
Points Earned
Summary of Results
Results are completely described for each dependent variable.

Differences between the control and experimental groups are
described.
Description of results is mostly complete. Some differences
between the control and experimental groups are described.
Description of results for each dependent variable and between
the control and experimental groups is incomplete or inaccurate.
Description of results is missing.
Graphic Explanation
As part of the summary, the paper refers to a graphic and: 1)
describes what type of information is presented in the graphic;
2) provides and explains example data; 3) explains the overall
meaning of information in the graphic.
The paper fails to thoroughly address one of the required points.
The paper fails to thoroughly address two of the required points.
The paper fails to describe a graphic.
Explanation of Results
The results are explained in a manner that could be easily
understood by a patient. Description demonstrates an excellent
understanding of the results and their application in health care.
The results are explained in a manner that could be understood
by a patient. Description demonstrates an adequate
The results are not explained in a manner that could be easily
understood by a patient. Description demonstrates poor or
inaccurate knowledge of the results and/or does not address
their application in health care.
The explanation is missing.
Analysis of Hypothesis
The paper states if the hypothesis is supported or refuted by the
data and why.
The paper states whether the hypothesis is supported or refuted
by the data but does not explain why.

The paper does not analyze if the hypothesis is supported or
refuted.
In-text Citations
Not applicable.
Not applicable.
Draft Section 4: Discussion and References (9 points) Name:
_________________________________
3 Points
2 Points
1 Points
0 Points
Points Earned
Discussion:
The paper includes a discussion of the validity and future
implications of the research.
Two
of the following points are thoroughly and accurately
discussed:
1) Was the experiment designed and conducted according to
proper scientific method?
2)

3)
What additional research or experimentation is needed?
4)
What are the implications of the research (i.e., what is its use in
the “real world”)?
N/A
The discussion of the first point demonstrates careful
consideration and thoughtful analysis of the topic with use of
additional resources as necessary.
The discussion of the first point demonstrates an average
analysis of the topic with use of additional resources as
necessary.
Appears discussion was written with little thought or purpose.
Minimal effort put into addressing required components.
Discussion of first point is inaccurate or missing.
The discussion of the second point demonstrates careful
The discussion of the second point demonstrates an average
analysis of the topic with use of minimal additional resources as
necessary.
Discussion of second point is inaccurate or missing.
Not applicable.
Not applicable.
At least one additional resource is used for the paper.

No additional resources were used in the paper.
In-text Citations
Not applicable.
Not applicable.
All information is appropriately cited.
In-text citations are absent or incorrectly cited.
Reference Page
Not applicable.
Not applicable.
All sources are cited in the reference page using the correct
APA format.
The reference page is absent or incorrectly cited.
Final Paper: Submit revisions from all drafts (21 points)
Name: ____________________________
2 Points
1.5 Points
1 Points
0.5 Points
0 Points
Points Earned
Format
Not applicable.

Not applicable.
Not applicable.
The paper is formatted according to instructions.
The paper is not formatted according to instructions.
Grammar and Spelling
(½ point deduction for each error)
The work is written in proper English and uses proper
grammatical structure and correct spelling. Work effectively
communicates ideas. There are no spelling or grammatical
errors.
The work is written in mostly proper English. Some
grammatical and/or spelling errors are present. Work
effectively communicates ideas. (One error.)
communicates ideas. (2 errors.)
effectively communicates ideas. (3 errors.)
The work is written in poor English and poor grammatical
structure. Spelling errors may be present. Work does not
effectively communicate ideas. (4 or more errors.)
Definitions of Terms and Concepts
Not applicable.
Not applicable.

Question
Not applicable.
Not applicable.
Not applicable.
answer.
to answer.
Hypothesis
Not applicable.
Not applicable.
Not applicable.
The hypothesis is stated in correct format.
The hypothesis is missing or is not stated in correct format.
Not applicable.
Not applicable.
Not applicable.
Dependent Variable
Not applicable.
Not applicable.
Not applicable.

Not applicable.
Not applicable.
Not applicable.
Not applicable.
Not applicable.
Not applicable.
groups.
Not applicable.
The paper thoroughly describes the steps of the experiment. It
describes how the control and experimental groups were treated.
2 Points
1.5 Points
1 Points
0.5 Points
0 Points
Points Earned
Experiment: Data Collection
Not applicable.

Not applicable.
Not applicable.
Summary of Results
Not applicable.
described.
Graphic Explanation
Not applicable.
Result Explanation
Not applicable.

Not applicable.
Not applicable.
data and why.
refuted.
2 Points
1.5 Points
1 Points
0.5 Points
0 Points
Points Earned
Discussion:
implications of the research. Two of the following points are
discussed:

2) Are the authors’ results supported by or in opposition to
3) What additional research or experimentation is needed?
4) What are the implications of the research (i.e., what is its
use in the “real world”)?
Point 1
necessary.
Point 2
necessary.
Not applicable.
Not applicable.
Not applicable.

No additional resources are used.
In-Text Citations
Not applicable.
Not applicable.
Not applicable.
All sources are cited in the text using the correct APA format.
In-text citations are absent or are formatted incorrectly.
Reference Page
Not applicable.
Not applicable.
APA format.
Some sources are missing from the reference page or the
incorrect format is used for citations.
Reference page is absent.
BIO 119 – Biology for Health Professionals
Journal Assignment
The assignment is worth
60 points
(39 drafts/21 final)
This is an INDIVIDUAL assignment!
The objective of this assignment is to provide you with
experience in evaluating scientific research papers. The
abilities to understand and critically analyze the results of a
scientific journal article are critical in the healthcare field. For

this assignment, you will evaluate how the scientific method has
been applied in a peer-reviewed journal article.
You will complete this assignment in
four sections (39 points)
and then submit a
final revised paper (21 points)
. Due dates for all components are listed on your syllabus – do
not forget!! Every section is graded and details are explained
below and in the rubric.
For this assignment, you are required to provide a summary of
the scientific method as it was presented by the journal authors.
Refer to the following information when completing your paper.
Format:
The paper shall be about 2-4 pages in length, double-spaced,
and 12-font type.
Include your name at the top right hand corner of the first page.
Title: The title should be centered at the top of the first page of
your paper, single-spaced, in 12-font type. Choose a title that
concisely explains the topic of your paper.
A title page is not required.

Grammar and spelling
:
The paper shall be written with proper grammar and free of
spelling errors. Use spell check, and carefully read your paper
prior to turning it in. ½ point will be deducted for each spelling
or grammatical error.
Evaluation of Scientific Method and Research:
Identify and describe the following components of the scientific
method in the body of your paper:
Section 1:
Definitions and Observations (8 points)
Definition of Terms and Concepts:
Define any important terms and concepts so that the reader
understands the topic of the journal article. You will likely
need to reference additional sources of information. Make sure
you properly cite the source(s) of your information.
Observations and Previous Studies:
Describe the observations that led the authors to conduct this
study. This information can usually be found in the background
information and discussion sections of the article. Describe at
least one previous study that led the authors to investigate the
current topic.
Question

: In your own words, state the question that the authors are
trying to answer in their experiment. The question should be
specific enough that it leads to the framing of a good
hypothesis.
This should be written in the form of a question.
Section 2
:
Hypothesis and Experimental Design (10 points)
Hypothesis
: State the hypothesis provided by the authors of the article.
Make sure that you identify the statement as the hypothesis
(i.e., “The hypothesis is…”). The hypothesis should be written
in your own words. It should be specific and written in the
proper format.
Experiment
: Describe the experiment as it is outlined in the journal
article. The following components must be included:
Independent variable:
State the independent variable.
Dependent variable:
State the dependent variable.
Controlled (constant) variables:
List the controlled variables used by the authors during the
experiment.
Control and experimental groups:
Identify the control and experimental groups.

Description of experiment
and data collection
:
Describe the steps of the experiment. Include a description of
how data and results were collected. You will need to cite the
journal article as the source for the details.
Section 3:
Summary and Interpretation of
Results
(12 points)
This section requires that you read the article, summarize the
results with specific details from graphics in the paper, and
explain the results of the study. This section will be 1 – 3
paragraphs long, depending on the paper. This section must
contain the following:
Summary of results:
Describe the results of the authors’ experiment for each
dependent variable. Be specific and include examples of the
qualitative and/or quantitative results. Compare and contrast
the results of the control and experimental groups.
Reference to graphic:
Choose one
results
table, graph, or diagram from the paper and
refer to it during your summary of results
. The graphic must describe results, NOT the experimental
design nor the demographics of the subjects.
Explain what information is being presented in the graphic.
This information is often found in the title of the graphic and/or

the caption.
Describe the results presented in the graphic. Give SPECIFIC
examples of quantitative and/or qualitative data presented in the
graphic. Cite your journal article as the source of this data.
Explain what the results presented in the graphic mean in detail.
Explanation of results:
Explain the results as if you were explaining them to a patient
or family member. Interpret and summarize them in lay person’s
language. What is the significance and health/medical
application of this study? Write this as if you are actually
explaining to the patient.
Analysis of
hypothesis
:
Discuss whether the hypothesis was supported or refuted.
Why or why not?
Section 4:
Discussion (9 points)
Discuss the validity and future implications of the research.
You should discuss
at least two
of the following points. You must cite outside sources to
support your statements.
Was the experiment designed and conducted according to proper
scientific method? Give specific examples of how it did or did
not follow the scientific method.

What additional research or experimentation is needed? What
would you recommend to further investigate this topic?
What are the implications of this research (i.e., what is its use
in the “real world?”)? How does this research add to the basic
field of study or to the disease process specifically?
:
Additional resources are required in this paper. They may be
helpful in understanding the observations (background) and
analyzing the results of the experiments. An additional source
is required for both of your discussion points. Additional
resources must be cited in the paper and reference page.
Citations within paper (
in-text or
parenthetical citations):
You must provide proper citation of all references (including
your article) throughout the paper. In-text citations tell the
reader where paraphrased or quoted information came from.
Paraphrasing is putting the ideas from a resource in your own
words. When paraphrasing, it is not sufficient to change only

one word of a sentence! See me if you are unclear on how to
paraphrase. If you copy word-for-word from a resource, you
must put the quoted (word-for-word) material in quotation
marks. The majority of the paper should be in your own words
or paraphrasing, not direct quotations from resources.
Follow APA style for in-text citations. Refer to the APA
Citation Guide and the following HCC webpage for more
information on how to cite sources within the body of your
paper:
http://www.harford.edu/library/citation_resources/style_guides/
apaintext.pdf
Failure to appropriately cite resources within the body of your
paper or to include quotation marks around any word-for-word
quotations will be considered
plagiarism
. Plagiarism may result in loss of points and possibly a zero for
the assignment.
http://www.harford.edu/library/tutorials/captivate_tutorials/apa.
htm
provides a tutorial on how to create a reference page
and
how to cite sources within your paper.
You should complete this tutorial as part of your preparations
for this assignment
. Additional APA citation information can be found at
http://www.harford.edu/library/citation_resources/
and at

http://www.apastyle.org/learn/tutorials/basics-tutorial.aspx
Reference
Page
:
This reference page will include the citation for the source of
the article and for
all
sources consulted. Follow APA style for the citations. Use the
APA Citation Guide on Blackboard to complete your reference
page and in-text citatio
Name:
Draft Section 1: Definitions of Terms and Concepts,
Observations, Previous Studies, Question (8 points):
3 Points
2 Points
1 Points
0 Points
Points Earned
Definition of Terms and Concepts

Question
Not applicable.
Not applicable.
answer.
to answer.
In-text Citations
Not applicable.
Not applicable.
In-text citations are absent or cited incorrectly.

Draft Section 2: Hypothesis and Experiment (10 points)
Name: ____________________________
3 Points
2 Points
1 Points
0 Points
Points Earned
Hypothesis
Not applicable.
Not applicable.
The hypothesis is stated in the correct format.
The hypothesis is missing or is not stated in the correct format.
Not applicable.
Not applicable.
Dependent Variable
Not applicable.
Not applicable.

Not applicable.
Not applicable.
Not applicable.
Not applicable.
groups.
The paper thoroughly describes the steps of the experiment.
It describes how the control and experimental groups were
treated.
Experiment:
Data Collection
Not applicable.
Not applicable.
In-text Citations

Not applicable.
Not applicable.
Draft Section 3: Interpretation and Summary of Results (12
points)
Name:
_____________________________________________________
____________________________
3 Points
2 Points
1 Points
0 Points
Points Earned
Summary of Results

described.
Graphic Explanation
Explanation of Results
data and why.

refuted.
In-text Citations
Not applicable.
Not applicable.
Draft Section 4: Discussion and References (9 points) Name:
_________________________________
3 Points
2 Points
1 Points
0 Points
Points Earned
Discussion:
implications of the research.

Two
of the following points are thoroughly and accurately
discussed:
2)
3)
What additional research or experimentation is needed?
4)
What are the implications of the research (i.e., what is its use in
the “real world”)?
N/A
necessary.

necessary.
Not applicable.
Not applicable.
No additional resources were used in the paper.
In-text Citations
Not applicable.
Not applicable.
All information is appropriately cited.
In-text citations are absent or incorrectly cited.
Reference Page
Not applicable.
Not applicable.
APA format.
The reference page is absent or incorrectly cited.

Final Paper: Submit revisions from all drafts (21 points)
Name: ____________________________
2 Points
1.5 Points
1 Points
0.5 Points
0 Points
Points Earned
Format
Not applicable.
Not applicable.
Not applicable.
The paper is formatted according to instructions.
The paper is not formatted according to instructions.
Grammar and Spelling
(½ point deduction for each error)
communicates ideas. There are no spelling or grammatical
errors.
effectively communicates ideas. (One error.)

communicates ideas. (2 errors.)
effectively communicates ideas. (3 errors.)
The work is written in poor English and poor grammatical
structure. Spelling errors may be present. Work does not
effectively communicate ideas. (4 or more errors.)
Definitions of Terms and Concepts
Not applicable.
Not applicable.
Question
Not applicable.
Not applicable.
Not applicable.
answer.
to answer.

Hypothesis
Not applicable.
Not applicable.
Not applicable.
The hypothesis is stated in correct format.
The hypothesis is missing or is not stated in correct format.
Not applicable.
Not applicable.
Not applicable.
Dependent Variable
Not applicable.
Not applicable.
Not applicable.
Not applicable.
Not applicable.
Not applicable.
Not applicable.
Not applicable.
Not applicable.
groups.

Not applicable.
The paper thoroughly describes the steps of the experiment. It
describes how the control and experimental groups were treated.
2 Points
1.5 Points
1 Points
0.5 Points
0 Points
Points Earned
Experiment: Data Collection
Not applicable.
Not applicable.
Not applicable.
Summary of Results
Not applicable.
described.

Graphic Explanation
Not applicable.
Result Explanation
Not applicable.
Not applicable.
Not applicable.
data and why.

refuted.
2 Points
1.5 Points
1 Points
0.5 Points
0 Points
Points Earned
Discussion:
implications of the research. Two of the following points are
discussed:
2) Are the authors’ results supported by or in opposition to
3) What additional research or experimentation is needed?
4) What are the implications of the research (i.e., what is its
use in the “real world”)?
Point 1

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