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Dark Chocolate Intake Acutely Enhances Neutrophil Count in Peripheral Venous Blood

Authors:

Abstract

Beside the well-established impact on decreasing the risk of cardiovascular diseases (1), recent attention has been paid to the relationship between cocoa-containing foods and the immune system (2), showing that dark chocolate consumption enhances the systemic defense against bacterial (3) and viral (4) infections. Hence, the current study aimed at investigating the acute effect of dark chocolate intake on peripheral blood leukocytes.
Letter to The Editor| Iran J Pathol. 2017; 12 (3): 311-312
Vol.12 No.3 Summer 2017 IRANIAN JOURNAL OF PATHOLOGY
Iranian Journal of Pathology | ISSN: 2345-3656
Dark Chocolate Intake Acutely Enhances Neutrophil Count in
Peripheral Venous Blood
Martina Montagnana1, Elisa Danese1, Gabriel Lima-Oliveira1, Gian Luca Salvagno1,
Giuseppe Lippi1*
1. Section of Clinical Biochemistry, University of Verona, Verona, Italy
KEYWORDS
ABSTRACT
Chocolate
Cocoa
Leukocytes
Neutrophils
Infection
Beside the well-established impact on decreasing the risk of cardiovascular
diseases (1), recent attention has been paid to the relationship between cocoa-
containing foods and the immune system (2), showing that dark chocolate
consumption enhances the systemic defense against bacterial (3) and viral (4)
infections. Hence, the current study aimed at investigating the acute effect of dark
chocolate intake on peripheral blood leukocytes.
Key Words: Chocolate; Cocoa; Leukocytes; Neutrophils; Infection
Article Info
Received 14 Oct 2016;
Accepted 28 Dec 2016;
Published Online 01 Jul 2017;
Corresponding Information: Prof. Giuseppe Lippi, Section of Clinical Biochemistry,University Hospital of Verona,Piazzale LA Scuro,,37100
Verona,ITALY,E-mail: giuseppe.lippi@univr.it; ulippi@tin.it
Copyright © 2017, IRANIAN JOURNAL OF PATHOLOGY. This is an open-access article distributed under the terms of the Creative Commons Attribution-noncommercial
4.0 International License which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
Dear Editor
All participants provided an informed consent
for participating in this study, which was carried out
in accordance with the Declaration of Helsinki, and
approved by the local ethics committee. The study
population consisted of 18 consecutive healthy
male volunteers (37±10 years), recruited from the
laboratory staff, in alphabetical order. Exclusion
criteria were history of hematological disorders,
abnormalities of routine blood tests and signs or
symptoms of disease, including weakness, cough,
and fever. Each volunteer ingested 50 g of 90%
cocoa chocolate (Noir Prodigieux, Lindt,
Kilchberg, Switzerland) within 3 to 5 minutes. The
nutrition information of the 50 g chocolate was as
follows: 1242 kJ (i.e., 296 kcal), 27.5 g of lipids (15
g of saturated fat acids), 7 g of carbohydrates (3.5 g
of sugars), and 5 g of proteins. Blood was drawn,
immediately before chocolate intake and 4 hours
after consumption, in K2EDTA evacuated blood
tubes (Vacutest Kima, Padova, Italy). All subjects
accomplished 8 hours of overnight fasting before
chocolate ingestion, and were then refrained from
eating, drinking, and smoking for the next 4 hours
after chocolate intake. White Blood Cell (WBC)
count and differential count was performed using
Siemens ADVIA 2120 (Siemens Healthcare
Diagnostics, Tarrytown NY, USA). Differences of
laboratory data before and after chocolate intake
were evaluated with paired Wilcoxon’s signed-rank
test and Bland-Altman plots (Analyze-it Software
Ltd, Leeds, UK).
The results of this study are shown in Table 1.
A significant increase of both WBC and
neutrophils counts was observed 4 hours after dark
chocolate intake, whereas the other parameters of
WBC remained unmodified. The Bland-Altman
plot analysis revealed a mean increase of 17% (95%
CI, 12% to 23%) for WBC and 14% (95% CI, 6%
to 22%) for neutrophils, respectively.
312 Dark Chocolate Intake
Vol.12 No.3 Summer 2017 IRANIAN JOURNAL OF PATHOLOGY
Table 1. White Blood Cell Count and Differential Count Four Hours after Dark Chocolate Ingestion in Eighteen
Ostensibly Healthy Subjects
Parameter
Baseline
Mean±SD
Mean±SD
p
White blood cells (x109/L)
5.9±0.9
7.0±0.9
<0.001
Neutrophils (x109/L)
3.4±0.8
3.9±0.7
0.001
Lymphocytes (x109/L)
2.1±0.7
2.3±0.5
0.056
Monocytes (x109/L)
0.4±0.2
0.4±0.1
0.379
Eosinophils (x109/L)
0.2±0.2
0.2±0.2
0.146
Basophils (x109/L)
0.1±0.1
0.1±0.1
0.086
LUC (x109/L)
0.1±0.1
0.1±0.1
0.234
SD, standard deviation; LUC, Large and Unstained Cells
In conclusion, the acute effect of dark chocolate
intake should be regarded as a potential source of
biological (preanalytical) variability (5).
Furthermore, a recent study showed that acute
cocoa consumption decreases the expression of
adhesion molecules (6), thus leading to reduced
migration of neutrophils in peripheral tissues
(including atherosclerotic plaques), which would
hence contribute to reduce local inflammation,
whereas their increase in the circulation may
promote more efficient response against pathogens.
Acknowledgements
There was no financial support for this study.
Conflict of interest
The authors declare that there was no conflict of
interest.
References
1. Lippi G, Franchini M, Montagnana M,
Favaloro EJ, Guidi GC, Targher G. Dark chocolate:
consumption for pleasure or therapy? J Thromb
Thrombolysis 2009; 28:482-8.
2. Pérez-Cano FJ, Massot-Cladera M, Franch
A, Castellote C, Castell M. The effects of cocoa on the
immune system. Front Pharmacol 2013;4:71.
3. Niehues M, Stark T, Keller D, Hofmann T,
Hensel A. Antiadhesion as a functional concept for
prevention of pathogens: N-Phenylpropenoyl-L-amino
acid amides as inhibitors of the Helicobacter pylori
BabA outer membrane protein. Mol Nutr Food Res
2011;55:1104-17.
4. Kamei M, Nishimura H, Takahashi T,
Takahashi N, Inokuchi K, Mato T, Takahashi K. Anti-
influenza virus effects of cocoa. J Sci Food Agric
2016;96:1150-8.
5. Simundic AM, Cornes M, Grankvist K,
Lippi G, Nybo M. Standardization of collection
requirements for fasting samples: for the Working
Group on Preanalytical Phase (WG-PA) of the
European Federation of Clinical Chemistry and
Laboratory Medicine (EFLM). Clin Chim Acta
2014;432:33-7.
6. Ellinger S, Stehle P. Impact of Cocoa
Consumption on Inflammation Processes-A Critical
Review of Randomized Controlled Trials. Nutrients
2016;8(6). pii: E321.
How to Cite This Article:
Montagnana M, Danese E, Lima-Oliveira G, Salvagno GL, Lippi G. Dark Chocolate Intake Acutely
Enhances Neutrophil Count in Peripheral Venous Blood. Iran J Pathol. 2017;12(3):311-312.
... Indeed, animals fed the C10 diet had even higher leukocyte counts than those fed REF diet, which was due to a higher granulocyte proportion. These results agree with the results obtained in a clinical study about acute exercise [58] and could be due to a lower granulocyte migration to tissues due to an effect of cocoa flavanols on adhesion molecules as reported [59]. ...
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Dark chocolate: consumption for pleasure or therapy? The effects of cocoa on the immune system Antiadhesion as a functional concept for prevention of pathogens: N-Phenylpropenoyl-L-amino acid amides as inhibitors of the Helicobacter pylori BabA outer membrane protein
  • G Lippi
  • M Franchini
  • M Montagnana
  • Ej Favaloro
  • Gc Guidi
  • G Targher
  • Fj Pérez-Cano
  • M Massot-Cladera
  • A Franch
  • C Castellote
  • M Castell
  • T Stark
  • D Keller
  • T Hofmann
  • A Hensel
Lippi G, Franchini M, Montagnana M, Favaloro EJ, Guidi GC, Targher G. Dark chocolate: consumption for pleasure or therapy? J Thromb Thrombolysis 2009; 28:482-8. 2. Pérez-Cano FJ, Massot-Cladera M, Franch A, Castellote C, Castell M. The effects of cocoa on the immune system. Front Pharmacol 2013;4:71. 3. Niehues M, Stark T, Keller D, Hofmann T, Hensel A. Antiadhesion as a functional concept for prevention of pathogens: N-Phenylpropenoyl-L-amino acid amides as inhibitors of the Helicobacter pylori BabA outer membrane protein. Mol Nutr Food Res 2011;55:1104-17.
Antiinfluenza virus effects of cocoa Standardization of collection requirements for fasting samples: for the Working Group on Preanalytical Phase (WG-PA) of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)
  • M Kamei
  • H Nishimura
  • T Takahashi
  • N Takahashi
  • K Inokuchi
  • T Mato
  • K M Takahashi
  • K Grankvist
  • G Lippi
  • M Nybo
Kamei M, Nishimura H, Takahashi T, Takahashi N, Inokuchi K, Mato T, Takahashi K. Antiinfluenza virus effects of cocoa. J Sci Food Agric 2016;96:1150-8. 5. Simundic AM, Cornes M, Grankvist K, Lippi G, Nybo M. Standardization of collection requirements for fasting samples: for the Working Group on Preanalytical Phase (WG-PA) of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM). Clin Chim Acta 2014;432:33-7. 6. Ellinger S, Stehle P. Impact of Cocoa Consumption on Inflammation Processes-A Critical Review of Randomized Controlled Trials. Nutrients 2016;8(6). pii: E321.
Dark chocolate: consumption for pleasure or therapy?
  • G Lippi
  • M Franchini
  • M Montagnana
  • E J Favaloro
  • G C Guidi
  • G Targher
  • F J Pérez-Cano
  • M Massot-Cladera
  • A Franch
  • C Castellote
  • M Castell
Lippi G, Franchini M, Montagnana M, Favaloro EJ, Guidi GC, Targher G. Dark chocolate: consumption for pleasure or therapy? J Thromb Thrombolysis 2009; 28:482-8. 2. Pérez-Cano FJ, Massot-Cladera M, Franch A, Castellote C, Castell M. The effects of cocoa on the immune system. Front Pharmacol 2013;4:71.
Standardization of collection requirements for fasting samples: for the Working Group on Preanalytical Phase (WG-PA) of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)
  • M Kamei
  • H Nishimura
  • T Takahashi
  • N Takahashi
  • K Inokuchi
  • T Mato
  • K Takahashi
  • M Cornes
  • K Grankvist
  • G Lippi
  • M Nybo
Kamei M, Nishimura H, Takahashi T, Takahashi N, Inokuchi K, Mato T, Takahashi K. Antiinfluenza virus effects of cocoa. J Sci Food Agric 2016;96:1150-8. 5. Simundic AM, Cornes M, Grankvist K, Lippi G, Nybo M. Standardization of collection requirements for fasting samples: for the Working Group on Preanalytical Phase (WG-PA) of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM). Clin Chim Acta 2014;432:33-7.