8th International Society of
Exercise and Immunology
Symposium
October 25-27th 2007
ISEI 2007
Inflammation in Exercise
Friend or Foe?
SENDAI KOKUSAI HOTEL
Sendai, Japan
http://isei8sendai.jp/
Co-hosted by Japanese Society of Physical Fitness and Sports Medicine (JSPFSM)
Japanese Society of Exercise and Immunology (JSEI)
Dear Colleagues and friends,
On behalf of the local organizing committee of ISEI, we would like to welcome you to
the 8th ISEI conference in Sendai, Japan.
The spirit of modern Japan largely inherits the spirit of Bushido, a code of conduct for
former Samurai, in which the seven virtues of benevolence, rectitude, respect, wisdom, loyalty,
honesty, and courage are respected. In the pursuit of exercise immunology, the spirit of Bushido
will no doubt push us forward for the sake of research directed towards the improvement of health
and the prevention and treatment of disease through physical activity and enhancement of the
athletic endeavor.
The scientific program of the 8th ISEI conference will cover hot topics in exercise
immunology. The title of the first ISEI meeting in Asia will be “Inflammation in Exercise - Friend
or Foe”. Inflammation may largely compromise one’s exercise capacity but may in return support
and trigger adaptation. How should we deal with inflammation? Professor Shizuo Akira, one of
the outstanding scientists in immunological science, will give a keynote lecture to initiate our
inflammatory process through his collection of studies on toll-like receptors. We will eventually
find out where the inflammation will take us to. Finally, the social program of ISEI 2007 will
provide opportunities to discover the culture and traditions of Japan.
We hope that the first ISEI meeting in Asia will offer a productive crucible for established,
new, and potential researchers and practitioners in exercise immunology from multidisciplinary
backgrounds and help them in their endeavors.
Prof. Ryoichi Nagatomi
President of ISEI
Chairman of the local organizing committee
Keynote lecture
Mammalian Innate Immune System and Viral Infection: Shizuo Akira
Symposia
S-I: Trigger of Inflammation: Up and Down of TLRs
1. Regulation of Inflammatory Responses by Nuclear IkB Proteins: Kiyoshi Takeda
2. Role of Toll-Like Receptors in the Pro-Inflammatory Gene Response of Skeletal Muscle during Sepsis:
Basil Petrof
3. Toll-Like Receptors and Exercise: Michael Flynn
S-II: Sick Athletes: Infection or Inflammation?
1. Sick Athletes: Infection or Inflammation?: Stéphane Bermon
2. Infectious Disease Outbreaks in Sports. A Review of the Literature: Sean Turbeville
S-III: Manipulation of inflammation in Exercise Medicine
1. Inflammation, Nutritional Status, Body Composition and Physical Activity in Healthy Adolescents:
Ascensión Marcos
2. Beneficial Versus Harmful Effects of Stress on Immune Function: Firdaus Dhabhar
3. Exercise as a Means of Reducing Inflammation and its Consequences: Jeffrey Woods
S-IV: Inflammation in Skeletal Muscle; Friend or Foe?
1. Role of Inflammation in Muscle Injury: James Tidball
2. Inflammation in Skeletal Muscle- Hormesis? Does Muscle Adaptation Require Inflammation? (No Pain,
No Gain?): Kazunori (Ken) Nosaka
S-V: Players of Innate Immunity: Friend or Foe?
1. Natural Killer Cell Subsets and Exercise: Masatoshi Suzui
2. The Role of Neutrophils in Tissue Injury: Friend or Foe? : Jonathan Peake
S-VI: Cytokines: Friend or Foe?
1. Cytokine Storm: Friend or Foe?: Ganesh Suntharalingam
2. Cytokine Response to Exercise: Katsuhiko Suzuki
Free communication (Slide)
F- 1: Acute stress-induced gut epithelial HSP70 expression requires commensal bacteria
components and intrinsic glucocorticoid: Kaori Matsuo
F- 2: The impact of resistance training and aging on toll-like receptor gene expression in
human skeletal muscle: Jonathan Peake
F- 3: Living Salmonella administration induces an acute reduction in wheel-running activity via TLR5 but not
TLR4 in mice: Takashi Matsumoto
F- 4: Patterns of TLR-expression in leukocyte subpopulations following a marathon run: Elvira Fehrenbach
F- 5: Physical activity and susceptibility to upper respiratory tract infection: Elinor Fondell
F- 6: The relationship between Epstein-Barr virus reactivation and upper respiratory infection
during summer training camp in rugby football: Ryohei Yamauchi
F- 7: Salivary IgA, Upper respiratory tract infections and peripheral blood T cells phenotypic
alterations from elite suimmers during a training season: Ana Maria Teixeira,
F- 8: The effect of exercise on innate mucosal immunity: Nicholas West
F- 9: Effects of exercise, age and gender on SIgA in elderly: Kazuhiro Shimizu
F-10: Cytokine responses to treadmill running in healthy and illness-prone athletes: Maree Gleeson
F-11: Effect of chronic carbohydrate consumption on cytokine responses to cycle ergometry: Amanda Cox
F-12: Effect of dietary antioxidant restriction on oxidative stress and inflammatory responses
to exercise: Robin Callister
F-13: The effects of low-intensity strength training on muscle hypertrophy and markers of
inflammation in very old women: Kishiko Ogawa
F-14: Acute inflammation response varies according to differences in basal C-reactive protein
concentrations: Mary Miles
F-15: Electrical pulse stimulation of C2C12 cultured myotubes induces IL-6 production: Arta Farmawati
F-16: Effects of overexpression of heat shock protein 70 on energy metabolism: Jürgen Steinacker.
F-17: Acute resistance exercise markedly upregulates gene expression of key chemotactic factors in skeletal
muscle: Paul Della Gatta
F-18: Corticosterone Accelerates Atherosclerosis in the Apolipoprotein E Mutant Mouse: Mitsuharu Okutsu
F-19: The effect of exercise intensity on some cytokine responses in women: Liz Gough
F-20: Exercise intensity influences plasma TNF-alpha concentration in response to
lipopolysaccharide in rats: Hiromi Kitamura
F-21: The effect of moderate exercise and ex vivo extracellular acidosis on T lymphocyte and
monocyte activation: João Viana
F-22: The different effects of exhaustive exercise on poly I:C-induced IFN-β and TNF-α
productions in mice: Masataka Uchida
Free communication (Poster)
P- 1: Kinetics of HSP70 expression in the intestine following exhaustive exercise: Ingrid Brenner
P- 2: Effect of exhaustive exercise on TLR4 expression in macrophages in mice: Daisuke Shiva
P- 3: Exhaustive exercise is reduced TNF-α production in response to lipopolysaccharide, but
it does not depend on gene expression in mice: Yohei Tanaka
P- 4: Effects of exercise training on B cells in elderly rats: Keisuke Nokura
P- 5: The effect of Pamper Pole jump on salivary IgA and cortisol responses in adventure
education: Wen-Yu Chan
P- 6: Effect of stimulating saliva flow on the changes in salivary secretion of IgA, lysozyme
and α-amylase with prolonged exhaustive exercise: Judith Allgrove
P- 7: Inquire: Immunocompetence and exercise among people over 65-years-old: Martins Raul
P- 8: Effect of exercise, aging and functional capacity on acute secretory IgA response in
elderly people over 75 years of age: Yuzuru Sakamoto
P- 9: Mucosal immunity after a soccer match: Vahid Sari-Sarraf
P-10: A rat model of saliva secretory immunoglobulin A suppression caused by intense
exercise: Fuminori Kimura
P-11: Supplementations of alpha and gamma-tocopherol have the similar effect on cellular
immune function following moderate exercise training: Mayumi Kaneyasu
P-12: Circulating leukocyte and T-lymphocyte subset response to carbohydrate and protein
feeding after prolonged exercise: Ricardo da Costa
P-13: Why wheat-dependent exercise-induced anaphylaxis is provoked by various wheat processed foods:
Mamoru Tanaka
P-14: Mucosal lesion in small intestine in WDEIA accelerated by gliadin: Hana Kozai
P-15: 10 weeks Endurance Running on Gene Expression of IL-1ß, Mn-SOD and Caspase-3 in
the Heart and Gastrocnemius of Spontaneously Hypertensive Rats: Wanglok Lee
P-16: Eccentric muscle contractions induce greater oxidative stress than concentric
contractions in skeletal muscle: Michihiro Kon
P-17: Physiological, muscle injury-related and leukocyte subset responses to exercise and cold
exposure in short track and inline skaters: Kijin Kim
P-18: Reduced nitric oxide production response to lipopolysaccharide in skeletal muscle is
associated with the surface poor in CD14 expression: Noriaki Kawanishi
P-19: Relationship between leg fatigue and plasma IL-6 concentration on mild intensity of
running exercise: Hiromi Yano
P-20: Biochemical and psychological markers of fatigue and immunodepression in exercise: Richard
Baskerville.
P-21: Non-invasive measures of some cytokines in response to exercise: Lindy Castell, .
P-22: Evaluation of the IMMULITE® Automated Immunoassay System for the detection of IL-6
and IL-10 in Saliva for Sport, Exercise & Health Science Applications: Andrew Misiura
P-23: Exercise training increases adiponectin in elderly males and females: Melissa Markofski
P-24: Low-grade inflammation is not associated with soft-drink consumption in healthy
adolescents: Julia Wärnberg
P-25: Effects of 5-day Kendo practice on lymphocyte level and expression of lymphocyte
apoptosis in young Kendo athletes: Yuko Tanimura
P-26: The effect of prolonged cycling for 15 consecutive days on neutrophil responses in
untrained cyclists: Tzai-Li Li
P-27: Disappearance of an allergenic activity of ovomucoid, a major egg allergen, by formation
of a complex with wheat gluten was assessed by voluntary physical activity in mice: Yasuko Kato
Program
Thursday, October 25
9:00 –
Registration (Sendai Kokusai Hotel) and poster mounting and viewing
9:30 – 10:00
Ryoichi Nagatomi (ISEI President): : Introduction:
10:00 – 11:00
KEYNOTE LECTURE:
Shizuo Akira: Mammalian Innate Immune System and Viral Infection
11:00 – 11:15
Coffee break
11:15 – 12:45
SESSION I: Trigger of Inflammation; Up and Down of TLRs
Kiyoshi Takeda: Regulation of Inflammatory Responses by Nuclear IkB Proteins
Basil Petrof: Toll-like Receptors in skeletal muscle tissue
Michael G. Flynn: Toll-Like Receptors and exercise
12:45 – 14:15
Lunch and Poster (P1-P27)
14:15 – 15:15
Free communication (Slide) (F1-F4)
15:15 – 16:15
SESSION II :
Sick Athletes- Infection or Inflammation?
Stephane Bermon: Sick athletes: Infection or Inflammation ?
Sean Turbeville: Infectious Disease Outbreak in Sports. A Review of the
Literature.
16:15 – 16:30
Coffee break
16:30 – 18:00
Free communication (Slide) (F5-F10)
18:30 –
Welcome Reception
Friday, October 26
8:30 – 10:00
SESSION III: Manipulation of inflammation in exercise medicine
Ascension Marcos: Inflammation, nutritional status, body composition and
physical activity in healthy adolescents
Firdaus Dhabhar: Beneficial versus Harmful Effects of Stress on Immune
Function
Jeffery Woods: Exercise as a Means of Reducing Inflammation and its
Consequences
10:00 – 10:15
Coffee break
10:15 – 11:15
Free communication (Slide) (F11-F14)
11:15 – 12:15
SESSION IV: Inflammation in skeletal muscle; Friend or Foe ?
James Tidball: Myeloid cell function in modulating skeletal muscle response to
injury and disease
Ken Nosaka: Does Muscle Adaptation Require Inflammation? (No Pain, No
Gain?)
12:15 – 13:45
Lunch and Poster (P1-P27)
13:45 – 14:45
Free communication (Slide) (F15-F18)
15:15 –
Social activities: Bus-tour to Matsushima Bay
Saturday, October 27
8:30 –
9:30
SESSION V: Players of Innate Immunity; Friend or Foe?
Masatoshi Suzui: Natural Killer Cell Subsets and Exercise
Jonathan Peake: The Role of Neutrophils in Tissue Injury: Friend or Foe?
9:30 –
9:45
9:45 – 10:45
10:45 – 11:45
Coffee break
Free communication (Slide) (F19-F22)
SESSION VI: Cytokines; Friend or Foe?
Katsuhiko Suzuki: Cytokine Response to Exercise
Ganesh Suntharalingam: Cytokine storm – Friend or Foe?
11:45 – 13:00
Lunch
13:00 - 13:30
Awards
13:30 - 13:45
Conclusion
Ryoichi Nagatomi
13:45 - 14:05
Future directions
Michael Gleeson
14:05 - 14:20
ISEI2009
Hinnak Northoff
14:30
Farewell
KEYNOTE LECTURE
Keynote lecture
Mammalian Innate Immune System and Viral Infection
Shizuo Akira, M.D., Ph.D.
Department of Host Defense, Research Institute for Microbial Diseases, Osaka University,
ERATO, Japan Science and Technology Agency
The innate immune system is an evolutionally conserved host defense mechanism against
pathogens. Innate immune responses are initiated by pattern recognition receptors (PRRs), which
recognize specific structures of microorganisms. Among them, Toll-like receptors (TLRs) are
capable of sensing organisms ranging from bacteria to fungi, protozoa and viruses, and play a major
role in innate immunity. Individual TLRs recognize different microbial components, and give rise
to different patterns in gene expression. The difference in signaling pathways among TLRs is in part
due to selective usage of adaptor molecules. Adaptor MyD88 is essential for inflammatory
responses whereas adaptor TRIF is involved in antiviral response such as induction of type 1
interferon and interferon inducible genes. TLRs recognize pathogens either at the cell surface or in
the lysosome/endosome compartments. Recently, cytoplasmic PRRs have been identified to detect
pathogens that invade cytosols. In the case of RNA virus infection, two helicase domain-containing
proteins, RIG-I and MDA-5 are essential for detection of double-stranded RNA that is generated
during viral replication, and subsequent type I interferon production. RIG-I and MDA-5 recognize
different structure of double-stranded RNA and as a result, different RNA virus species. This
cytoplasmic recognition takes place in a variety of cell types except for plasmacytoid dendritic cells,
where type I interferon production is mainly mediated by TLR system. We recently have obtained
some evidence that double-stranded DNA is also recognized in the cytoplasm by an as yet
unidentified receptor. In this lecture, I focus on the roles of PRRs in viral recognition and their
downstream signaling cascades.
SYMPOSIA
S-I-1
Regulation of Inflammatory Responses by Nuclear IkB Proteins
Kiyoshi Takeda, M.D., Ph.D.
Department of Microbiology and Immunology, Graduate School of Medicine, Osaka University
Toll-like receptor (TLR)-mediated activation of innate immunity, when in excess, induces
development of chronic intestinal inflammation, as demonstrated in mice lacking Stat3 specifically
in innate immune cell populations. Therefore, activity of innate immune cells in the colon of normal
mice is finely regulated to prevent excessive inflammatory responses. Indeed, macrophages residing
in the colonic lamina propria of normal mice show defective TLR-dependent proinflammatory
cytokine production, whereas cells derived from Stat3 mutant or IL-10-deficient mice show robust
cytokine production. An analysis of the mechanisms for the hyporesponsiveness to TLR stimulation
of these cells leads to identification of the nuclear IkB protein IkBNS. IkBNS is constitutively
expressed in colonic lamina proprial macrophages and induced by TLR stimulation in macrophages
of other tissues, and inhibits expression of a subset of NF-kB-dependent genes by terminating
NF-kB activity. IkBNS knockout mice produce higher amounts of IL-6 and IL-12p40, and suffer
from severe intestinal inflammation after dextran sodium sulfate administration, indicating that
IkBNS is responsible for limiting inflammation.
TLR triggers the rapid induction of primary response genes via MyD88-dependent activation of
NF-kB. Then, some of primary response gene products induce secondary response genes, including
IL-6 and IL-12p40. The primary response gene product IkBNS is shown to inhibit induction of
secondary response genes. Analyses of promoters of primary and secondary response genes indicate
that both genes are epigenetically differentially regulated, and chromatin structures of secondary
response gene promoters are remodeled after TLR stimulation. Another primary response gene
product IkBzeta, which is structurally related to IkBNS, mediates induction of the secondary
response genes through histone modification of promoters. Thus, TLR-dependent gene induction is
differentially regulated by nuclear IkB proteins, IkBzeta and IkBNS.
S-I-2
Role of Toll-Like Receptors in the Pro-inflammatory Gene Response of Skeletal
Muscle during Sepsis
Basil J. Petrof, M.D.
Meakins-Christie Laboratories & Respiratory Division, McGill University
Immunologically active molecules such as cytokines and chemokines have been implicated in
skeletal muscle weakness during sepsis as well as recovery from muscle injury. In sepsis,
Toll-Like Receptors (TLRs) act as key sentinel molecules of the innate immune system. Here we
determined skeletal muscle cell responses of two prototypical CC and CXC chemokine genes
(MCP-1 and KC, respectively), to stimulation with specific TLR ligands. In addition, we
examined whether NF-kB and calcineurin signalling are involved in these responses.
Differentiated myotubes and intact whole muscles expressed TLR2, TLR4, TLR5, and TLR9.
Stimulation with ligands for TLR2 (peptidoglycan) or TLR4 (LPS) elicited robust and equivalent
levels of MCP-1 and KC mRNA expression, whereas stimulation of TLR5 (by flagellin) required
IFN-γ priming to induce similar effects. Although both TLR2 and TLR4 ligands activated the
NF-kB pathway, NF-kB reporter activity was approximately 20-fold greater after TLR4 stimulation
as compared to TLR2. Inhibitory effects of NF-kB blockade on TLR-mediated chemokine gene
expression, by either pharmacological (pyrrolidine dithiocarbamate) or molecular (IKKß dominant
negative transfection) methods, were also more pronounced during TLR4 stimulation. In contrast,
inhibitory effects on TLR-mediated chemokine expression of calcineurin blockade (by FK506)
were greater for TLR2 than for TLR4 stimulation. MCP-1 and KC mRNA levels also demonstrated
differential responses to NF-kB and calcineurin blockade during stimulation with specific TLR
ligands. In vivo, multiple pro-inflammatory genes (including MCP-1, KC, TNF-α, IL-1α, IL-1ß,
IL-6) were significantly upregulated in the mouse diaphragm after systemic administration of LPS
or the induction of pneumonia with live Pseudomonas bacteria, and this was associated with
significant diaphragm muscle weakness in both sepsis models. We conclude that skeletal muscle
cells utilize TLRs to differentially regulate signalling pathways controlling the expression of
pro-inflammatory genes under septic conditions.
S-I-3
Toll-Like Receptors and Exercise.
Michael G. Flynn, Ph.D.
Health and Kinesiology, Purdue University
Toll–like receptors (TLR) have a range of NF-kB activating exogenous and endogenous ligands,
which up-regulate innate immunity and inflammation. Our initial interest in TLR emerged from an
attempt to explain an apparent exercise training-induced reduction in monocyte sensitivity to LPS.
We subsequently found that both TLR4 mRNA and TLR4 receptor expression were reduced by
exercise training and were lower in physically active than inactive persons. In general, physically
active subjects have lower TLR4 expression than inactive subjects, but 10 weeks of training
significantly lowers TLR4 expression in previously inactive subjects. We have been focused on
TLR4 as a potential explanation for the anti-inflammatory influence of exercise, but we, and others,
have examined several other potential mechanisms for the inflammation suppressing potential of
exercise. Our emphasis has been the influence of training and/or high physical activity levels on
TLR expression. TLR4 was consistently lower in physically active subjects or after training in
previously sedentary subjects; but TLR2 was not influenced by activity level or training.
Gleeson’s group from Loughborough has focused on the acute, strenuous exercise effects on TLR
expression and has observed a post-exercise reduction in TLR1, TLR2, and TLR4 post- and two
hours post-exercise. We view an exercise training-induced reduction in TLR4 expression as a
positive adaptation and speculate that it provides a possible link between the positive influences of
exercise training on chronic disease. In contrast, Gleeson’s group views the post- acute exercise
TLR down-regulation as a putative explanation for post-exercise immunosuppression. Irrespective
of this apparent difference in interpretation, emerging links between TLR and heart disease,
diabetes, osteoporosis, and other chronic diseases provides a promising topic for future research and
an enticing mechanism for pharmacological or exercise interventions.
S-II-1
Sick Athletes: Infection or Inflammation?
Stéphane Bermon, M.D., Ph.D.
Monaco Institute of Sports Medicine
Upper Respiratory Tract Infection (URTI) is the most common medical condition affecting
athletes in both summer and winter Olympic Games. The causes of these disturbances, also
occurring during training, remain unclear. If virus such as rhinovirus, adenovirus and para-influenza
virus are frequently reported as the source of URTI, some comprehensive laboratory and
epidemiological studies recently reported more than 30% of these illness episodes without
identifiable pathogens. A recent study by Spence et al. (2007) even reported an approximate 60%
ratio in sedentary controls, recreational and elite athletes. This study, that used polymerase chain
reaction typing and a validated respiratory symptom questionnaire, also showed that most of the
unidentified upper respiratory illnesses were shorter in duration and less severe than infectious ones.
This concept of inflammation without infection in athletes is quite new and lead us to consider
other explanatory pathophysiological conditions. Thus, increases in airway neutrophils, eosinophils
and lymphocytes have been described under resting conditions in endurance sports, swimmers and
cross-country skiers. These inflammatory patterns may be due to pollutants or chlorine-related
compounds in swimmers. After intense exercise similar airways cellular profiles have been reported,
with high amount of bronchial epithelial cells. This increase in airway inflammatory cells in athletes
can result from hyperventilation-induced increase in airway osmolarity stimulating bronchial
epithelial cells to release chemotactic factors. Fortunately, in most cases, these inflammatory cells
express rather low level of adhesion molecules, explaining why airway inflammation may appear
blunted in athletes despite numerous inflammatory cellular elements.
The link between exercise-induced airway inflammation and secondary viral infection has been
poorly studied but there are some experimental data supporting this alternative. Adhesion molecules
and ICAM-1 for instance may be over-expressed in post-exercise or chronic asthma situation
theoretically leading to a higher risk of URTI, ICAM-1 being the major rhinovirus receptor.
This lecture also emphasizes the necessity of further basic research to define the variable
etiologies of upper respiratory ilnesses and their relationship with vasomotor disturbance, allergy,
inflammation or asthma. This would help athletes and their entourage for better diagnosis,
treatments, and management strategies of upper respiratory episodes.
S-II-2
Infectious Disease Outbreaks in Sports. A review of the literature
Sean D. Turbeville, Ph.D., Linda D. Cowan, Ph.D., and Ronald A. Greenfeld, M.D.
Department of Biostatistics and Epidemiology, College of Public Health,
University of Oklahoma Health Sciences Center
Recent outbreaks of infectious diseases in athletes in competitive sports have stimulated
considerable interest. The environments in which these athletes compete, practice, receive therapy
for injuries, and travel, both domestically and internationally, provide varied opportunities for the
transmission of infectious organisms. The purpose of the presentation is to report the agents most
commonly reported in the medical literature as responsible for infectious disease outbreaks in
specific sports and their modes of transmission and to guide targeted prevention efforts. A literature
review of English-language articles in medical publications that reported outbreaks of infectious
diseases in competitive athletes was conducted in PubMed MEDLINE from 1966 through May
2005. Outbreaks that were solely food borne were excluded. Fifty-nine reports of infectious disease
outbreaks in competitive sports were identified in the published medical literature. Herpes simplex
virus infections appear to be common among wrestlers and rugby players, with no single strain
responsible for the outbreaks. Methicillin-resistant Staphylococcus aureus was responsible for
several recent outbreaks of soft tissue and skin infections among collegiate and professional athletes.
The most common mode of transmission in outbreaks was direct, person-to-person (primarily
skin-to-skin) contact. Blood-borne exposure was implicated in 2 confirmed outbreaks of hepatitis.
Airborne and vector transmissions were rarely reported. This review provides an overview of
infectious disease outbreaks thought to be either serious enough or unusual enough to report.
Appropriate surveillance of the frequency of infections will allow sports medicine staff to identify
outbreaks quickly and take necessary measures to contain further transmission and prevent future
outbreaks.
S-III-1
Inflammation, Nutritional Status, Body Composition
and Physical Activity in Healthy Adolescents
Ascensión Marcos & Julia Wärnberg
Immunonutrition research group. Department of Metabolism and Nutrition.
Instituto del Frio. Spanish National Research Council (CSIC)
Inflammatory processes are involved in the pathogenesis of the most common chronic
non-communicable diseases and may also play an important initiating role in their development.
Only recently have inflammatory markers been included in epidemiological studies focusing on
nutritional status, body composition and physical activity. We are just starting to understand how
different lifestyles can determine basal levels of inflammatory biomarkers in early ages. This review
aims to summarise what is known about the relationships between lifestyle-related determinants
(focusing on overweight, physical activity and dietary habits) and inflammatory markers in
apparently healthy young populations. Obesity is the most widely studied determinant and several
large-scale studies have now demonstrated that healthy young subjects with more body fat or higher
BMI have moderately higher concentrations of inflammatory markers than their leaner peers,
supporting the idea that obesity should be considered as a state of chronic low-grade inflammation.
Less data is available to allow us to elucidate how physical activity or dietary patterns may have a
direct effect on inflammation in apparently healthy, disease-free young populations.
S-III-2
Beneficial Versus Harmful Effects of Stress on Immune Function
Firdaus S. Dhabhar, Ph.D.
Department of Psychiatry & Behavioral Sciences, Stanford Immunology Program, Comprehensive Cancer Center,
Stanford University
Although stress has a “bad” reputation, it is important to appreciate that a psycho-physiological
stress response is nature's fundamental survival system. Without a fight or flight response, a lion
has no chance of catching a gazelle, just as the gazelle has no chance of escape. Keeping this in
mind, we initially hypothesized that just as the stress response prepares the cardiovascular,
musculoskeletal and neuroendocrine systems for fight or flight, under certain conditions, stress may
also prepare the immune system for challenges (e.g. wounding or infection) that may be imposed by
a stressor (e.g. predator or surgical procedure). This hypothesis was confirmed when studies
showed that short duration stressors induce a large-scale redistribution of leukocytes within the
body and that immune function is significantly enhanced in organs like the skin to which leukocytes
traffic during acute stress. Subsequent studies have identified additional mechanisms involving
immune cell function and cytokine production through which acute stressors enhance immune
function.
In contrast to acute stress, chronic stressors are known to exert detrimental effects on different
dimensions of physiology and health. In keeping with numerous studies, our laboratory has shown
that chronic stressors suppress/dysregulate leukocyte trafficking, surveillance, and function, and
cytokine and chemokine gene expression. Moreover, human subjects who experience chronic
stress show accelerated leukocyte aging and this may be a critical mechanism mediating chronic
stress induced suppression and/or dysregulation of immune function. Recent studies from our
laboratory have also shown that chronic stress increases susceptibility to skin cancer by suppressing
protective immunity on the one hand, and enhancing endogenous immunosuppressive mechanisms
on the other.
In summary, our laboratory investigates molecular, cellular, and physiological mechanisms
mediating the immunoenhancing effects of acute versus the immunosuppressive / dysregulatory
effects of chronic stress. We work with human as well as rodent models of health and disease.
Examination of the bidirectional effects of stress on immune function is important because on the
one hand stress is thought to suppress immunity and increase susceptibility to infections and cancer,
while on the other it is thought to enhance immunopathology and exacerbate autoimmune and
inflammatory disorders.
S-III-3
Exercise as a Means of Reducing Inflammation and its Consequences
Jeffrey A. Woods, Ph.D.
Department of Kinesiology and Community Health, University of Illinois at Urbana-Champaign
Dysregulated immune responses, including exaggerated acute or chronic inflammation,
contribute significantly to disease incidence and severity; especially in populations such as the
elderly, obese, or those with co-morbid conditions (e.g. cancer, autoimmune disease). For over 10
years, my laboratory has been determining the influence of exercise and physical activity as a
means of improving immune responses and reducing acute and chronic inflammation in hopes of
providing a scientifically-based public health message. This lecture will provide a brief overview of
the influence of exercise and physical activity on inflammatory processes (as summarized in Woods
et al. Neurol Clin, 2006). Importantly, I will summarize recent findings from our laboratory
indicating that exercise has the potential to reduce acute and chronic inflammation and disease
morbidity and mortality. We have shown that exercise can reduce inflammation in an animal model
of cancer (Zielinski et al. J Appl Physiol 2004) and influenza virus infection (Lowder et al. Brain
Behav Immun 2005; Lowder et al. Exerc Immunol Rev, 2006) and, in both instances, reduce disease
burden. Moreover, we have recently found that regular exercise reduces inflammatory chemokine
expression in visceral adipose tissue of diet-induced obese mice. This is important because
obesity-associated inflammation is mechanistically linked to co-morbidities such as insulin
resistance and hepatic steatosis. By attending this lecture, you will gain an appreciation for the
emerging role of inflammation in disease progression and the potential for exercise to regulate the
inflammatory process perhaps via increased vagal tone (Vieira et al. J Am Geriat Soc, 2007).
Work supported, in part, by NIH AG-18861
S-IV-1
Myeloid cell function in modulating skeletal muscle response
to injury and disease.
James G. Tidball Ph.D., Bo Deng, Armando Villalta and Michelle Wehling-Henricks.
Departments of Physiological Science, and of Pathology and Laboratory Medicine
David Geffen School of Medicine at UCLA
Los Angeles CA USA
Inflammation is a typical response to modified muscle use, acute injury and disease. Expanding
knowledge of myeloid cell function shows that these cells play complex and sophisticated roles in
modulating the response of muscle to injury and disease. Whether these responses have a net
beneficial or destructive role varies with the numbers of invading cells, the duration of
inflammation, the concentration of free radicals or cytokines they release and their interactions with
the host muscle. The quality of muscle response to injury or disease also varies with the
phenotype of macrophages that invade the muscle; the relative balance between Th1 and Th2
macrophage phenotype playing a central role in determining whether there is further muscle injury
or muscle repair, regeneration and growth. In our investigations, we examine the role of myeloid
cells in muscle injury, repair and regeneration in two models of muscle injury and disease. In the
first, we characterize the inflammatory response in muscle experiencing modified use, and test how
perturbations in the numbers of myeloid cells affects muscle injury or repair. We also test the
effect of modified production of free radicals or cytokines by inflammatory cells on the response of
muscle to modified use. Our data show that the initial inflammatory response to modified use is
dominated by a Th1 response in which high levels of hypochlorous acid exacerbate muscle injury.
This Th1 response is then attenuated by invasion of Th2 macrophages that express CD163, CD206
and leukemia inhibitor factor (LIF). Invasion by Th2 macrophages is required for normal growth
and repair following increased muscle use. However, our second model suggests that a persistent
Th2 response is associated with promoting muscle pathology. Dystrophin-deficient mdx muscles
show chronic muscle inflammation by CD206+ Th2 macrophages and by eosinophils, a myeloid
cell population that is characteristic of the Th2 inflammatory response. Our findings show that the
persistent Th2 inflammatory response is associated with increased muscle pathology and fibrosis,
which can contribute to death in dystrophin-deficiency. (Supported by NIH RO1 AR47855).
S-IV-2
Inflammation in Skeletal Muscle–Hormesis?
Does Muscle Adaptation Require Inflammation? (No Pain, No Gain?)
Kazunori (Ken) Nosaka, Ph.D.
School of Exercise, Biomedical and Health Sciences, Edith Cowan University
Skeletal muscle is injured by various causes such as diseases (e.g. muscular dystrophy),
chemicals (e.g. local anesthetics), ischaemia-reperfusion, excessive heat or cold, and mechanical
force. Degeneration is followed by acute inflammation, which involve vascular and cellular
responses. The vascular responses are initiated by the injured and/or surrounding cells releasing
vasoactive factors (e.g. reactive oxygen species, prostaglandins, cytokines), and inflammatory cells
such as neutrophils and monocytes migrate to the site of injury. It appears that inflammatory
responses differ in the cause or the magnitude of injury. It is reported that neutrophil accumulation
is absent after lengthening contractions of rat EDL muscles; however, massive neutrophil
accumulation occurs prior to macrophage infiltration after injection of bupivacaine. The cardinal
signs of acute inflammation are redness (vasodilation), swelling, heat, pain, and loss of function,
and most of these are evident after performing an accustomed exercise consisting of lengthening
contractions (eccentric exercise). However, it is important to note that the time course of muscle
soreness, swelling, loss of muscle function, and histological changes does not match. Increases in
muscle proteins (e.g. creatine kinase and myoglobin) in the systemic circulation have also been used
as indicators of muscle damage. However, even when a large increase in muscle proteins in the blood
is found after eccentric exercise, little or no changes in cytokines in the blood are evident. This is
different from muscle damage induced by endurance exercise such as marathon or triathlon. It is
known that the magnitude of the symptoms of muscle damage is attenuated when the same or
similar exercise is repeated within several weeks, but no significant difference between bouts for
changes in cytokines in the blood is found. Performing lengthening contractions with light load or
heating the muscle one day prior to eccentric exercise, which do not induce inflammation at least in
the symptom level, provide a protective effect to muscle damage induced by eccentric exercise. In a
training to improve muscle function and hypertrophy muscle, most of the muscle adaptation to the
training occurs when muscles become less susceptible to muscle damage and inflammation. Thus, it
seems unlikely that “pain” or inflammation is hormesis for muscle adaptation.
S-V-1
Natural Killer Cell Subsets and Exercise
Masatoshi Suzui, Ph.D.
Meiji University
Natural killer cells are important components of the innate immune system because of their
cytokine production and cytolytic activity against virus-infected or tumour cells. Since the
mid-90s, researches have been conducted to clarify the functions of membrane receptors and their
signals in NK cells, which regulate the activation or inhibition for recognizing and killing the target
cells. Recent findings suggest that NK cells are involved in the regulation of innate and adaptive
immune systems. NK cells in human can be divided into CD56bright NK cell and CD56dim NK cell.
Majority of NK cells are CD56dim cells and have high lytic activity. In contrast, CD56bright NK
cells have low cytotoxicity but obviously produce cytokines, such as IFN-γ and TNF-α. Thus,
several researchers suspect that CD56bright NK cells works as regulators.
NK cells are also one of the most responsive leukocyte subsets to physiological and
psychological stress. We previously reported decreased total and per cell cytotoxicity of NK cell
at the end of one-month of intensive training, without any changes in the CD56dim NK cell number.
These changes could be partly explained by increased CD56bright NK cell subset. Moreover, these
findings also suggested the possibility that acute exercise might also affect the balance of CD56dim
and CD56bright NK cells. Therefore, we investigated mobilizations of these subsets during
incremental exercise and during and after steady exercise. The four-stages incremental exercise
(50, 80, 110 and 140% ventilatory threshold (VT)-VO2) induced significantly increased proportion
of CD56dim NK cell to the total lymphocyte, but the percentage of CD56bright NK cell remained
unchanged throughout the experiments. Steady exercise (110% VT-VO2) also increased the
proportion of CD56dim NK cells during exercise, and dropped below the resting level after exercise.
On the other hand, the percentage of CD56bright NK cells tended to increase 30 min post-exercise,
but was not statistically significant.
Responses of NK cell to exercise were one of the original sources of “the Open Window Theory”.
This down-regulated condition, however, was quite short-lived in young healthy subjects. The
proportion of CD56bright NK cell seemed to increase when NK cell lytic activity decreased. It
might trigger future recovery and progression.
S-V-2
The Role of Neutrophils in Tissue Injury: Friend or Foe?
Jonathan M. Peake, Ph.D.
School of Human Movement Studies, University of Queensland
Neutrophils comprise the majority (50-60%) of circulating white blood cells. Their half life
within the circulation is 12-20 h. Neutrophils invade damaged or infected tissue within 1 h, and may
remain in the tissue for up to 5 d. These cells play a key role in the phagocytosis of foreign
pathogens and degradation of cellular debris at sites of tissue damage, by releasing serine proteases
(e.g., elastase, myeloperoxidase, cathepsin G) and reactive oxygen species (e.g., superoxide) into
both intra- and extracellular spaces. While neutrophil activation is an integral component of innate
immunity, when released in excess, proteases and reactive oxygen species may damage healthy
bystander tissues. Research has indirectly implicated neutrophils in muscle tissue injury by default
(i.e., presence in damaged muscle) and also through a temporal association between neutrophil
invasion and a decline in muscular strength following lengthening muscle contractions. Using a
more direct approach, other studies have indicated that blocking neutrophil activation using
monoclonal antibodies or gene knockout reduces muscle damage. Furthermore, some studies report
that depletion of circulating neutrophils attenuates muscle damage and proteolytic activity
following single stretch, mixed muscle contractions and ischemia-reperfusion injury. In contrast,
others have indicated no effect of neutrophil depletion on muscle damage after repeated lengthening
contractions. Neutrophils promote renal injury during the early stages of glomerulonephritis, and
lung injury during systemic inflammation without infection. The role of neutrophils in repair of
tissue damage is less certain. Blocking neutrophil activation does not accelerate muscle growth
following muscle unloading/reloading. In contrast, depletion of circulating neutrophils impedes
muscle regeneration and clearance of cellular debris after muscle damage. During sepsis,
neutrophils do not induce lung injury but enhance survival rate by killing bacteria. Therefore,
neutrophils appear to enhance tissue injury under a variety of conditions, whereas evidence that
neutrophils improve muscle repair is currently lacking. The relative role of neutrophils as ‘friend’ or
‘foe’ in tissue injury/repair is likely specific to the type of tissue involved, the nature of stimulus
causing tissue injury, and the number and activation status of invading neutrophils. More work is
needed to establish whether neutrophils play a role in tissue repair.
S-VI-1
Cytokine Storm: Friend or Foe?
Ganesh Suntharalingam, F.R.C.A.
Clinical Director, Critical Care, Northwick Park Hospital
At 9 am on March 13th 2006, six healthy volunteers took part in a first-into-man phase 1 trial of
TGN1412, a novel monoclonal antibody, at a commercial research unit leased and operated by
Parexel International on the site of a UK acute hospital.
The trial drug was a genetically engineered, humanised monoclonal anti-CD28 superagonist
antibody, designed to stimulate and expand type 2 helper T cells independently of T cell receptor
ligation. The drug was designed to provide immunomodulation in T cell mediated chronic
inflammatory conditions, and to expand the collapsed T cell compartment in in haematological
malignancy (B-CLL).
The effect in healthy humans was dramatically different to that anticipated by the drug
development team. All six volunteers developed rapid induction of cytokines accompanied by a
severe systemic inflammatory response and marked organ impairment, resulting in a pattern of
critical illness similar to that seen in severe septic shock. The incident was clinically difficult to
manage given the unknown effect and kinetics of this first-in-man drug. Unusually detailed
real-time cytokine data were available from this incident, due to the high degree of sampling
performed for clinical decision-making at the time.
S-VI-2
Cytokine Response to Exercise
Katsuhiko Suzuki, M.D.
Faculty of Human Sciences, and Consolidated Research Institute for Advanced Science and Medical Care, Waseda
University
It has been documented that strenuous endurance exercise induces leukocytosis (systemic
inflammation), muscle damage and suppression of cellular immunity. To determine the underlying
mechanisms of these phenomena, attention has been focused on cytokines released into the
circulation following exercise; indeed, there has been a tremendous accumulation of new findings.
Many studies have consistently shown that IL-6, IL-1 receptor antagonist (IL-1ra) and IL-10
increased remarkably following endurance exercise longer than 2 h such as marathon and triathlon,
but not so much by short-time intensive exercise and eccentric exercise. These responses are not
dependent on muscle damage (inflammation), but on exercise intensity (stress). Indeed, it has been
demonstrated that IL-6 responses to exercise depend on energy crisis and heat stress, are correlated
with epinephrine responses, but are suppressed by energy supply and cooling. On the other hand,
IL-6 responses might enhance recruitment of energy substrates such as free fatty acids, helping
endurance performance, but IL-6 induces neutrophil mobilization and activation together with
immunosuppressive IL-1ra and IL-10 release. Therefore, IL-6 might be good for athletes in
shortening their race time, but might be bad in terms of susceptibility to infections and systemic
inflammation. It is possible that appropriate countermeasures such as energy and fluid supply and
prevention of body temperature elevation might help to keep endurance performance without
causing harmful side effects on the body.
Suzuki K, et al. Endurance exercise causes interaction among stress hormones, cytokines,
neutrophil dynamics, and muscle damage. J. Appl. Physiol., 87, 1360-1367, 1999.
Suzuki K, et al. Circulating cytokines and hormones with immunosuppressive but
neutrophil-priming potentials rise after endurance exercise in humans. Eur. J. Appl. Physiol., 81,
281-287, 2000.
Suzuki K, et al. Exhaustive exercise and type-1/type-2 cytokine balance in special focus on
interleukin-12 p40/p70. Exerc. Immunol. Rev. 9, 48-57, 2003.
Peake J, Nosaka K, Suzuki K. Characterization of inflammatory responses to eccentric exercise in
humans. Exerc. Immunol. Rev. 11, 64-85, 2005.
Suzuki K, et al. Changes in markers of muscle damage, inflammation and HSP70 after an Ironman
triathlon race. Eur. J. Appl. Physiol. 98, 525-534, 2006.
FREE COMMUNICATION (SLIDE)
F-1
Acute stress-induced gut epithelial HSP70 expression requires commensal bacteria
components and intrinsic glucocorticoid
Kaori Matsuo1, Xiumin Zhang2, Yusuke Ono3 and Ryoichi Nagatomi1
1
Dept. of Medicine and Science in Sports and Exercise, Tohoku University Graduate School of Medicine, Japan
School of Public Health, Jilin University, China
3
Randall Division of Cell and Molecular Biophysics,King's College London, U.K.
2
Background: Induction of Heat Shock Protein 70 (HSP70) is considered to be protective for cells
under stressors such as heat, hypoxia, ultraviolet rays, and oxidative stress or endoplasmic reticulum
stress, serving as molecular chaperones. Physical stressors such as restraint stress also induce
HSP70 in various organs and tissues in vivo. It is not clear how such physical stressors are
translated to cellular stressors. We tried to identify the role of restraint stress-induced glucocorticoid
response on HSP70 expression of gut epithelial cells in which commensal bacteria are known to be
responsible for basal HSP70 expression under conditions without physical stressors. Objective: We
investigated how acute restraint-induced stress responses lead to expression in gut epithelial cells of
HSP70, which is known to suppress gut inflammation and basal expression of which is dependent
on commensal bacteria. The effects of glucocorticoid blockade and elimination of commensal
bacteria on gut epithelial HSP70 expression were examined. We also postulated that tight junction
integrity of gut epithelia may be altered under stress facilitating access by luminal bacterial
components to lamina propria TLR4. Materials and methods: Commensal bacteria of male
C57BL/6 mice were depleted by antibiotic administration. Mice were restrained for 2 h with or
without glucocorticoid antagonist RU486 treatment. Mice were sacrificed and gut HSP70 was
quantified by ELISA and examined by immunohistochemistry. Gut ZO-1 and TLR4 expression
were examined by western blot and immunohistochemistry. Whether LPS administration to
antibiotic-treated mice before restraint could restore gut epithelial HSP70 augmentation was also
examined. Results: Serum corticosterone and gut epithelial HSP70 expression of restrained mice
increased. Antibiotic treatment abrogated augmented HSP70 without affecting its basal expression.
LPS administration partially restored HSP70 augmentation. RU486 treatment abrogated the gut
HSP70 augmentation. TLR4 was expressed in smooth muscle cells at lamina propria. ZO-1
expression was significantly decreased by restraint stress. Conclusions: These results suggest that
both glucocorticoid and commensal bacteria are required for physical stress-induced HSP70
expression in gut epithelia. Decreased gut epithelial ZO-1 expression by restraint stress may
facilitate access of bacterial components to TLR4 in the lamina propria resulting in HSP70
augmentation in epithelial cells.
F-2
The impact of resistance training and aging on toll-like receptor gene expression in human
skeletal muscle.
Jonathan Peake1,
Cameron-Smith2
1
2
Paul
Della
Gatta2,
Michelle
Farnfield2, Andrew
Garnham2,
David
School of Human Movement Studies, University of Queensland, Brisbane, Australia.
School of Exercise and Nutrition Sciences, Deakin University, Melbourne, Australia.
Objective: Toll-like receptors (TLRs) mediate signaling pathways related to the synthesis of
inflammatory cytokines, and are therefore implicated in physical inactivity, inflammation and
disease. We investigated alterations in TLR gene expression in vastus lateralis after a single session
of resistance exercise in young males, and also after 12 weeks of resistance training in young and
old males. Methods: Seven young males (22.1±0.8 yrs) completed three sets of resistance exercise
for the leg muscles. Two sets consisted of 8-12 repetitions at 80% 1-RM, whereas in the final set the
subjects exercised until exhaustion. Muscle biopsies were obtained before exercise, 2, 4 and 24 h
after exercise. Thirteen young males (20.4±2.1 yrs) and 13 old males (67.8±4.3 yrs) completed a
12-week resistance exercise training program. All subjects trained three times per week. Training
involved two sets of each exercise initially at 50% 1-RM, then at 80% 1-RM from 6 weeks onwards.
Muscle biopsies were obtained at rest before and after the training program. Muscle tissue from
both studies was analyzed for the expression of TLR2 and TLR4 genes, in addition to downstream
genes including MyD88, IL-1 receptor associated kinase (IRAK)-1 and interferon-regulatory factor
(IRF)-1. Results: Following acute resistance exercise, TLR2, TLR4 and IRAK-1 mRNA did not
change significantly. MyD88 mRNA was elevated at 2 h (3-fold, P=0.026) and 24 h (4.7-fold,
P=0.039). IRF-5 mRNA was also elevated at 24 h (4-fold, P=0.037). Following 12 weeks of
resistance training, TLR4, MyD88 and IRAK-1 mRNA did not change significantly, and did not
differ between young and old males. TLR2 and IRF-1 mRNA was not detectable. Conclusion:
Acute resistance exercise up-regulated expression of the TLR downstream genes MyD88 and IRF-5.
Follow-up work will determine whether aging skeletal muscle responds in a similar manner. Neither
aging nor chronic resistance training altered the expression of TLR genes in skeletal muscle.
Therefore, TLR genes expressed in skeletal muscle cells may respond less to exercise training than
other cell types, such as circulating monocytes (1), and may not account for the increased
expression of pro-inflammatory cytokines in aging skeletal muscle at rest (2).
1. L.K. Stewart et al. 2005. Brain Behav Immun vol 19: 389-397.
2. B. Przybyla et al. 2006. Exp Gerontol vol 41: 320-327.
F-3
Living Salmonella administration induces an acute reduction in wheel-running activity via
TLR5 but not TLR4 in mice
Takashi Matsumoto1, Daisuke Shiva2, Noriaki Kawanishi3, Yasuko Kato4, Jeffrey A. Woods5,
Hiromi Yano2
1
Microbiology, Department of Infectious Disease, Oita University, Oita
Division of Health Science
3
Department of Health and Sports Science
4
Department of Clinical Nutrition, Kawasaki University of Medical Welfare, Okayama, Japan, and 5Department of
Kinesiology and Community Health, University of Illinois at Urbana-Champaign, IL, USA
2
To clarify a role of toll-like receptor (TLR)s of the sickness behavior induced by Salmonella
infection, male C3H/HeN (wild type) and C3H/HeJ (tlr4 gene mutated) mice were administered
with live (1x106 CFU/kg i.p.) Salmonella enterica serovar Dublin and examined for wheel-running
activity. Wheel-running activity in both C3H/HeJ and C3H/HeN mice was significant lower than
that in uninfected control groups (p<0.01, respectively). Wheel-running activity in C3H/HeJ mice
was not different from uninfected controls when Salmonella was heat-killed prior to administration.
Gentamicin treatment, which removes flagella from living Salmonella, completely inhibited the
reduction of wheel-running activity in C3H/HeJ mice, even though injections of flagella-rich
supernatant of Salmonella or pure flagellin significantly reduced wheel-running activity in these
mice (p<0.01, respectively). Our findings suggest that Salmonella-induced reduction of voluntary
physical activity might be due to flagellin binding to TLR5, but not LPS acting through TLR4 in
mice.
F-4
Patterns of TLR-expression in leukocyte subpopulations following a marathon run
Elvira Fehrenbach1, Perikles Simon2, Petra Buettner3, Karsten Krueger4, Harald Funke3, Hinnak
Northoff1, Andreas M. Niess2 and Frank C. Mooren4
1
Institute of clinical and experimental Transfusion Medicine, University of Tuebingen
Dept. of Sportsmedicine, Internal Medicine, University clinic of Tuebingen
3
University Hospital Jena
4
Dept Sportsmedicine, Institute of Sports Science, University of Giessen
2
Objective: Toll-like receptors (TLR) recognize pathogen-associated molecular patterns and play a
role in protective immunity against infection and inflammation. They activate the innate immune
system and subsequently modulate adaptive immunity. Patterns of TLR expression in different
subpopulations of peripheral blood imply specific roles in each population. A general activation of
the immune system by endurance exercise is well known.
However, it is not quite clear how TLRs are involved in the exercise-related immune response.
Methods: 12 male, healthy subjects performed a marathon. Before, immediately, 3, 24, 48 and 72 h
after the run monocytes (M) and T-lymphocytes (T) were isolated from peripheral blood, separately,
using the Rosette technique with specific antibody cocktails. mRNA of individual subjects was
pooled in three comparable groups. Gene expression was analysed using the microarray U133A
(Affymetrix). Pathway search with the significantly regulated genes (Welch Anova with Benjamini
Hochberg) was performed using KEGG- and GOtree-analysis ("WEB-based GEne SeT AnaLysis
Toolkit", http://bioinfo.vanderbilt.edu/webgestalt/). Selected gene expression changes were
confirmed and quantified by real time PCR. Results: In both, T and M the TLR-pathway was
significantly affected. Genes belonging to TLR-pathway were 6/3.5-fold overrepresented among the
significantly regulated genes in T and M, respectively. Real time PCR of selected genes confirmed
microarray results. It revealed that basal expression of TLR2- and CD14-mRNA was lower in T
than M. However, in T the increase in response to exercise was more pronounced than in M. The
TLR-pathway is directly connected with additional significantly involved pathways. Conclusions:
Up-regulation of TLRs plays an important role in the acute marathon-induced immune response.
Both, innate (M) and adaptive (T) immune cells are involved. The more pronounced stimulation in
T, indicates a particular exercise-induced immunological activation of T-cells. The specific
stimulation of TLR2 implies the involvement of a direct T-helper-1-mediated immune response to
endurance exercise. TLR-increase in T-cells principally allows a direct, fast reaction of the adaptive
immune system by-passing the innate immune response and thereby increasing sensitivity to
endotoxin. This may help contribute to altered susceptibility to infection after prolonged exercise.
F-5
Physical activity and susceptibility to upper respiratory tract infection
Elinor Fondell1,2, Anna L.V. Johansson1, Ylva Trolle Lagerros3, Carl Johan Sundberg4, Mats
Lekander2,5, Olle Bälter6, Kenneth J. Rothman7 and Katarina Bälter1
1
Department of Medical Epidemiology and Biostatistics, Karolinska Institutet, Sweden
Osher Center for Integrative Medicine, Department of Clinical Neuroscience, Stockholm, Sweden 3Department
of Medicine, Clinical Epidemiology Unit, Karolinska Institutet, Sweden
4
Department of Physiology and Pharmacology, Karolinska Institutet, Sweden
5
Department of Clinical Neuroscience, Karolinska University Hospital, Stockholm, Sweden
6
School of Computer Science and Communication, Royal Institute of Technology, Stockholm, Sweden, 7 RTI
Health Solutions, Research Triangle Park, North Carolina, USA, and Department of Epidemiology, Boston
University School of Medicine, Boston, USA
2
Objective: Upper respiratory tract infection, URTI, is by far the most frequent disease in the
industrialized world today, and is the number one reason for work absenteeism. Still, little is known
about strategies to reduce susceptibility. Most studies of this issue have focused on athletes or other
selected groups (Hemilä et al., 2003; Nieman et al., 2000; Novas et al., 2002). We investigated the
potentially beneficial effects of physical activity on the risk of contracting URTI among the general
population. Methods: We studied a population-based cohort recruited via paper mail, using the
Internet as a novel tool for data collection. We followed the cohort of 1509 men and women for
four months, contacting them every three weeks via e-mail. Total physical activity was assessed
through a questionnaire, measuring all activities during 24 hours, giving total MET-hours (multiples
of resting metabolic rate) per day. We also collected data on potential confounding factors,
including age, sex, body mass index, smoking, diet, education, asthma, contact with young children,
use of public transportation, effect of season, among other factors. Results: High levels of physical
activity (53-114 MET-hours per day) were associated with a 27 % reduced risk (IRR 0.73, 95% CI:
0.61-0.88) of contracting URTI compared with low levels of physical activity. Apparent protective
effects of similar magnitude were observed for both non-vigorous and vigorous physical activity.
The effect was present among men and women, and across all ages. The effect of high physical
activity appears stronger among those reporting high levels of chronic stress than among those
reporting lower levels of chronic stress. Conclusion: We found that high physical activity is
associated with a lower risk of upper respiratory tract infection. Highly stressed people might
benefit more from physical activity than those with lower stress levels.
References
Hemilä H. Physical Activity and the Common Cold in Men Administered Vitamin E and
B-carotene. Med Sci. Sports Exerc. 2003;35:1815-1820.
Nieman D.C. et al. Immune function in female elite rowers and non-athletes. Br J. Sports Med.
2000;34:181-187.
Novas A. et al. Total Daily Energy Expenditure and Incidence of Upper Respiratory Tract Infection
Symptoms in Young Females. Int J Sports Med 2002;23:465-470.
F-6
The relationship between Epstein-Barr virus reactivation and upper respiratory infection
during summer training camp in rugby football
Ryohei Yamauchi1, Fuminori Kimura1, Katsuhiko Suzuki2, Takao Akama2, Ichiro Kono1, Takayuki
Akimoto2
1
2
University of Tsukuba
Waseda University,
Objective: It is well known that highly trained athletes suffer from a high incidence of upper
respiratory tract infections (URTI). Thus, it is important for all athletes to manage risk of URTI
through training and competitive periods. In some previous research, salivary secretory
immunoglobulin A (SIgA) has been used for evaluation of the risk of URTI. However, there has
been a moderate relationship between SIgA levels and URTI. Here we established a new index to
monitor the risk of URTI in intensive training period on the field. Methods: We prospectively
investigated the relationship between daily changes in EBV-DNA as well as SIgA level, and
appearance of URTI symptoms in collegiate rugby football players during summer training camp
for a month. Results: The SIgA concentration tended to decrease before the appearance of URTI
symptoms compared to that in the non-infection period. For EBV-DNA, the working curve was
obtained from the result of PCR that used the EBV-DNA inserted vector. It became possible to
quantify the amount of EBV-DNA appearance by being based on working curve that we established.
The detection system of the amount of EBV-DNA appearance was constructed. The appearance of
EBV-DNA was also related to the appearance of URTI symptoms. Conclusion: Our findings
suggest that monitoring of SIgA secretion rate and EBV-DNA may be useful for assessment of risk
status of athletes for URTI.
F-7
Salivary IgA, Upper respiratory tract infections and peripheral blood T cells phenotypic
alterations from elite suimmers during a training
Teixeira, Ana Maria1, Rama, Luís1, Henriques, Ana 2, Azevedo, Sara 2, Rosado, Fátima.2, Paiva,
Artur1 and Alves, Francisco3
1
Center of Biokinetic Studies, Faculty of Sport Sciences and Physical Education, Coimbra University, Portugal.
Histocompatibility Centre of Coimbra, Coimbra, Portugal.
3
Faculty of Human Motricity, Technical University of Lisbon, Portugal
2
Objectives: Changes in lymphocyte subsets after exercise have been well documented, but fewer
studies have looked at the responses of distinct subpopulations that may reflect functional changes
during a training season. The aim of this study was to see how training load might influence the
proportion and phenotypic features of circulating T cells subsets in peripheral blood (PB) of high
competitive levels athletes and how it could affect susceptibility to disease. Methods: To study
the response of T cell subsets, quantification, by flow cytometry, of PB cells from 19 swimmers of
Portuguese elite level (13 men and 6 women, mean age 17+ 1,25 years old), was done in four
different moments of the training season that matched the transitions between preparation cycles.
The absolute counts and percentage of CD4 and CD8 T cells expressing HLA-DR, granzyme B,
CD25, CD119 and CD126; the naïve, central memory, effector memory and effector T cells
compartments by CD45RA and CCR7 expression and a distinct population of CD4+/CD25++T cells
(regulatory T cells) were determined. Salivary IgA concentration and secretion rate were also
determined by ELISA.
The upper respiratory tract infection (URTI) episodes that occurred during the training season were
documented using daily logs. Results: Significant differences were found (particularly between the
first and second moment) in the absolute counts of leucocytes and also in the percentage of CD119
(a decrease) and CD126 (an increase) on CD4 and CD8 T cell subsets. Furthermore, athletes that
suffered more infectious episodes appear to show a lower number of regulatory T cells, an increase
of CD4 T cells expressing CD119 and an increase in granzyme B expression in CD8 T cells,
comparing with those athletes that had no URTIs during the training season. Salivary IgA
concentration and secretion rates were higher in the 3rd and 4th moments that corresponded to a
decrease in training load. Conclusion: This preliminary data seem to indicate that some quantitative
and qualitative alterations observed in T cells subsets may be related with the participation of
athletes on specially intensive and voluminous training programs and their predisposition to be
affected by infections.
F-8
The effect of exercise on innate mucosal immunity
Nicholas P West,1,2 David B Pyne,1,3 Jenelle M Kyd,3 Gillian M C Renshaw,
4
Peter A Fricker,5 Allan W Cripps6
1
Department of Physiology, Australian Institute of Sport
School of Physiotherapy and Exercise Science, Griffith Health, Griffith University
3
School of Medicine, Australian National University
4
School of Health Services, Central Queensland University
5
Executive, Australian Institute of Sport, Canberra, Australia
6
Griffith Health, Griffith University, Australia.
2
Objective: To compare the concentration of salivary lactoferrin and lysozyme in elite rowers and
sedentary individuals (controls) over a training season to determine the chronic effects of exercise
and to examine the time course of changes in salivary lactoferrin and lysozyme in response to a
graded exercise session. Methods: Saliva samples were taken fortnightly over 5 months from a
cohort of elite male and female rowers (n=17, mean age 24.3 ± 4.0 y) and at three time points,
baseline, midpoint and the end of the study, from a sedentary control group (n=18, mean age = 27.2
± 7.1 y) over the same period. In the dose response study salivary lysozyme and lactoferrin
concentrations were measured at rest, at a submaximal workload and after exercise to exhaustion.
Lactoferrin and lysozyme concentrations were determined by ELISA and corrected for changes in
albumin to control for salivary flow rate and total protein concentration differences. Values were
log-transformed prior to analysis. Magnitudes of differences and changes were interpreted as a
standardized (Cohens) effect size (ES). Results: The rowers had approximately 60% lower
concentration of lactoferrin than control subjects at baseline and at the midpoint of the longitudinal
study: baseline (7,913±4948 ng.ml-1 mean±SD, 19,444±22,035 ng.ml-1, P=0.05, ES=0.68,
‘moderate’), midpoint (6,437±5,651 ng.ml-1 mean±SD, 21,585±16,634 ng.ml-1, P=0.001, ES=0.89,
‘moderate’). There was no significant difference between rowers or control subjects in the
concentration of salivary lysozyme over the study. Correcting for albumin had little impact on the
outcomes of the study. In the dose response study there was a significant increase in the
concentration of both lactoferrin (15,322±10,272 ng.ml-1, 30,209±16,914 ng.ml-1, P=0.04) and
lysozyme (13,509±12,629 ng.ml-1, 29,693±13,569 ng.ml-1, P=0.01) from pre-exercise to maximal
exertion only. Conclusions: Lower lactoferrin and lysozyme concentration may be indicative of a
possible impairment of innate protection of the upper respiratory tract during a training season in
elite rowers. The increase in the concentration of lactoferrin and lysozyme following exhaustive
exercise may be a protective response to limit damage to the body from exercise.
F-9
Effects of exercise, age and gender on SIgA in elderly
Kazuhiro Shimizu1, Fuminori Kimura1, Takayuki Akimoto2, Takao Akama3, Takeshi Otsuki4,
Takahiko Nishijima1, Shinya Kuno1, Ichiro Kono1
1
Graduate School of Comprehensive Human Sciences, Tsukuba University, Ibaraki, Japan
Institute for Biomedical Engineering, Consolidated Research Institute for Advanced Science and Medical Care,
Waseda University, Tokyo, Japan
3
Faculty of Sport Sciences, Waseda University, Saitama, Japan
4
Center for Tsukuba Advanced Research Alliance, Tsukuba University, Ibaraki, Japan
2
Objective: The influence of age and gender on salivary secretory immunoglobulin A (SIgA) in
response to moderate exercise training was studied in 158 elderly subjects. Methods: Subjects
were assigned to an exercise training group (EXC: 51 males, 74 females) or a non-exercise
control group (CON: 11 males, 22 females). The subjects in each group were separated into four
age-gender subgroups (60–69-yr-old males, over 70-yr-old males, 60–69-yr-old females, over
70-yr-old females) and compared by age and gender. Subjects in EXC participated in exercise
sessions 5-days a week for 6 months. Saliva samples were collected both before and after the
study period. Results: The SIgA secretion rates were significantly increased after training (p <
0.05) in all the age-gender subgroups of EXC (60–69 males: 41%, over 70 males: 55%, 60–69
females: 40%, over 70 females: 38%); no age- or gender-related differences were observed. On
the other hand, all the age-gender subgroups of CON did not show significant changes in SIgA
secretion rate; also, there were no age- or gender-related differences. Conclusions: Enhancement
of the mucosal immune function following regular moderate exercise training occurs in the
elderly in their 60’s and over 70, and in both males and females.
F-10
Cytokine responses to treadmill running in healthy and illness-prone athletes
Amanda J Cox1,2, David B Pyne1, Philo U Saunders1, Robin Callister2 and Maree Gleeson2
1
2
Department of Physiology, Australian Institute of Sport, Canberra, ACT 2616, AUSTRALIA
School of Biomedical Sciences, Faculty of Health, University of Newcastle, Callaghan, NSW 2308, AUSTRALIA
Objective: Explanations for the increased incidence of upper respiratory symptoms (URS) amongst
some athletes remain unclear. A disruption of inflammatory homeostasis may help to explain
episodes where an infectious aetiology is excluded. The purpose of the current study was to
characterise pro- and anti-inflammatory cytokine responses to treadmill running in illness-prone
athletes and compare them to responses in healthy athletes. Methods: The study involved 10
healthy (≤2 episodes of URS in the past year) and 8 illness-prone (≥4 episodes of URS in the past
year) trained distance runners. Runners had been training for 11.1 ± 6.8 y (mean ± SD) with a
typical training load of 8.8 ± 4.9 h.wk-1 (mean ± SD). Runners completed a standardised bout of 60
min treadmill running at 65% VO2max and blood samples were collected, pre-, post-, 1 h, 10 h and
24 h post-exercise. Plasma concentrations of pro- (IL-6, IL-8) and anti-inflammatory (IL-10,
IL-1ra) cytokines were determined using a Bio-Plex Suspension Array System (Bio-Rad
Laboratories Pty Ltd). Cytokine concentrations and responses to exercise were compared between
healthy and illness-prone groups using paired t–tests (p<0.05). Standardised mean changes and
qualitative descriptors were used to describe the likely range of the true effect. Data are presented as
(% difference; p-value; qualitative descriptor). Results: At rest, the illness-prone runners had
substantially lower IL-8 (-22%; p=0.13; small to large), IL-10 (-29%; p=0.16; trivial to large) and
IL-1ra (-40%; p=0.04; small to large) concentrations compared to the healthy runners. Post-exercise
IL-6 responses were greater in the illness-prone group (45-150%) at all post-exercise time-points. In
contrast, the magnitudes of increase in IL-10 (1-13%) and IL-1ra (-2-20%) concentrations were
similar between the two groups. Conclusions: Cytokine responses to controlled treadmill running
differed between healthy and illness-prone athletes. Illness-prone distance runners showed evidence
suggestive of impaired regulation of inflammatory processes. This alteration in inflammatory
regulation may partly account for the greater frequency of URS experienced by these athletes and
be an explanation for the non-infective aetiology of the symptoms. In contrast, pro- and
anti-inflammatory cytokine responses among healthy athletes appeared to be regulated in a more
proportionate manner.
F-11
Effect of chronic carbohydrate consumption on cytokine responses to cycle ergometry
Amanda J Cox1,3, David B Pyne1, Greg Cox2, Robin Callister3 and Maree Gleeson3
1
Department of Physiology, Australian Institute of Sport, Canberra, ACT 2616, AUSTRALIA
Department of Sports Nutrition, Australian Institute of Sport, Canberra, ACT 2616, AUSTRALIA
3
School of Biomedical Sciences, Faculty of Health, University of Newcastle, Callaghan, NSW 2308, AUSTRALIA
2
Objective: The current study examined the effects of chronic (28-days) carbohydrate (CHO)
supplementation on cytokine responses to cycle ergometry with or without acute CHO beverage
ingestion. The aim was to investigate if acute and chronic CHO supplementation had a synergistic
effect in attenuating cytokine responses to exercise. Methods: The study involved sixteen highly
trained male cyclists/triathletes. One group (n=8) consumed a high-CHO (8.5 g.kg-1 body mass) diet
for 28 days; a second group (n=8) consumed a moderate-CHO diet (5.3 g.kg-1 body mass). Athletes
completed two trials in randomised order, each following a 24 h standardised diet (60% CHO). One
trial involved the consumption of a 10% CHO beverage (15mL.kg-1.hr-1); the other water (WAT).
Both trials involved 100 min steady state cycle ergometry at 70% VO2max followed by a time trial of
~30 min duration. Blood samples were collected pre-, immediately post- and 1 h post-exercise.
Plasma concentrations of pro- (IL-6, IL-8) and anti-inflammatory (IL-10, IL-1ra) cytokine
concentrations were determined using a Bio-Plex Suspension Array System (Bio-Rad Laboratories
Pty Ltd). Cytokine responses were compared between pre-and post-intervention trials for each of
the groups using paired t-tests (p<0.05) and standardised mean changes to describe the likely range
of the effect. Results: Acute CHO beverage ingestion attenuated increases in anti-inflammatory
cytokines at 1 h post-exercise (30-40% reductions, trivial to moderate differences), but had less of
an affect on pro-inflammatory cytokine responses. The 28-day high-CHO diet did not substantially
affect cytokine responses to either WAT or CHO beverage exercise trials. In contrast, following the
28-day moderate-CHO diet, there were substantial reductions in anti-inflammatory cytokine
responses to the WAT trial (30-50% reductions; trivial-moderate differences), as well as reductions
in responses to the CHO beverage trial for both pro-inflammatory (15-45% reductions; trivialmoderate differences) and anti-inflammatory cytokines (20-50% reductions; trivial-moderate
differences). Conclusions: There did not appear to be a synergistic effect between acute and
chronic-CHO consumption in protecting against post-exercise cytokine disruptions. In contrast,
where dietary CHO was lower, acute CHO use attenuated post-exercise disruptions in cytokine
concentrations. This may have implications for athletes undertaking CHO-restricted diets for
participation in sports with defined weight classes.
F-12
Effect of dietary antioxidant restriction on oxidative stress and inflammatory responses to
exercise
Robin Callister, Brendan A Plunkett, Trent A Watson and Manohar L Garg
Nutraceuticals Research Group, School of Biomedical Sciences, University of Newcastle, NSW, Australia
Exercise is known to increase the production of reactive oxygen species (ROS). Dietary
carotenoids have antioxidant properties and may be an important source of exogenous antioxidants
for exercising populations. Objective: The aim of this study was to determine the effect of dietary
antioxidant restriction on a marker of oxidative stress (F2-isoprostane), plasma carotenoid (α- and
β-carotene, lycopene, β-cryptoxanthin, lutein), TNFα and IL-6 concentrations following
submaximal and maximal exercise. Methods: Seventeen endurance-trained males (24 ± 1 years old;
VO2max 58 ± 1 mlO2.kg-1.min-1) performed two separate exercise trials consisting of 30min
treadmill running at 60% of VO2max followed by a progressive exercise test to exhaustion.
Participants followed their habitual (high) antioxidant diet (HAO) prior to the first exercise trial
then followed a reduced antioxidant diet (RAO) for 2 weeks before performing the second exercise
trial. Blood was collected at rest and post-exercise for the analysis of inflammatory markers and
carotenoids. Results: There was no difference due to diet in F2-isoprostane levels prior to exercise,
but levels were 38% higher after submaximal exercise and 45% higher after maximal exercise on
the RAO diet. TNF-alpha concentration was 38-fold higher pre-exercise on the RAO diet but only
14-fold higher post exercise. There was no difference in IL-6 concentrations on the diets
pre-exercise or in response to exercise. Carotenoid concentrations were 35-50% lower on the RAO
compared to HAO diet pre-exercise. Carotenoid concentrations decreased 25-60% following
exercise on the HAO diet, to levels similar to those prior to exercise on the RAO diet, whereas the
decrease in carotenoid concentrations following exercise on the RAO diet was substantially less.
Conclusion: Healthy endurance-trained adults performing short-duration exhaustive exercise may
require higher intakes of carotenoids to combat the oxidative stress and inflammation generated
through exercise. These carotenoids can be provided via a diet containing high-carotenoid foods or
dietary supplements.
F-13
The effects of low-intensity strength training on muscle hypertrophy and markers of
inflammation in very old women.
Kishiko Ogawa1, Jonathan Peake2, Mitsuharu Okutsu1, Katsuhiko Suzuki1
1
Waseda University, Tokorozawa, Japan, 2 University of Queensland, Brisbane, Australia
Background: Aging is associated with low-grade inflammation, which is implicated in a variety of
diseases. The benefits of regular exercise for aging individuals are well established, whereas less is
known about the impact of low-intensity strength exercise on low-grade inflammation in the elderly.
Objectives: We investigated the effects of low-intensity strength training on changes in physical
fitness, muscle hypertrophy, and circulating growth factors and markers of inflammation in very old
women. Methods: Twenty-one very old women, (mean age 85±5 years) participated in 12 weeks of
low-intensity strength exercise. The participants performed the training (vertical traction, leg press,
shoulder press, and chest press) once a week. Physical fitness, muscle thickness, circulating levels
of C-reactive protein (CRP), serum amyloid A (SAA), heat shock protein (HSP)70, tumor necrosis
factor (TNF)-a, interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1, insulin, insulin like
growth factor (IGF)-I, and vascular endothelial growth factor (VEGF) were measured before and
after the exercise intervention. Results: The 12-weeks of low intensity strength training increased
abdominal (P<0.01) and subscapula (P<0.05) muscle thickness, whereas it reduced systolic blood
pressure (P<0.05), and the serum concentration of CRP, SAA (P<0.05), HSP70, IGF-I, and insulin
(all P<0.01). The serum concentrations of MCP-1, TNF-a, IL-6, or VEGF did not change
significantly following the training. The training-induced increase in muscle thickness correlated
with the decrease in the serum concentrations of CRP (r= -0.56) and TNF-a (r= -0.53) (both P<0.05).
Conclusion: Low-intensity exercise training in very old women improved systolic blod pressure,
insulin resistance and muscle thickness, and reduced some markers of low-grade inflammation.
Strength training may assist in maintaining or improving muscle mass in the aging population by
reducing low-grade inflammation.
F-14
Acute inflammation response varies according to differences in basal C-reactive protein
concentrations
Mary P. Miles, Lindsay K. Gordon, Christopher M. Depner, and Jessy R. Kidd
Dept. of Health and Human Development, Montana State University, USA.
While basal levels of C-reactive protein (CRP) associate with risk for several chronic diseases
including cardiovascular diseases and type 2 diabetes, the significance of basal CRP concentrations
in young, apparently healthy individuals is not known. Purpose: To determine whether acute
inflammatory responses differ in young adults with LOW (less than the group median) compared to
HIGH (group median + 1 standard deviation) basal CRP concentrations. Methods: College-aged
men and women (n=44) completed exercise and control conditions in random order and met criteria
for both grouping and a stable basal inflammatory state. The exercise condition consisted of a three
sets of 15 maximal eccentric actions using the elbow flexor muscles of one arm, and blood was
collected pre-, 4, 8, 12, and 24 h post-exercise for inflammatory variables and additionally at 48, 96,
and 120 for muscle markers of muscle damage (maximal isometric force (MIF), muscle soreness,
and circumference of the arm at the mid-biceps level). Blood was collected on a time-matched
schedule without exercise for the control condition. Serum and plasma were analyzed for soluble
tumor necrosis factor receptor 1 (sTNFR1), IL-6, CRP, cortisol, and CK. Additional basal
measurements included soluble intercellular adhesion molecule-1 (sICAM) and IL-10. Results:
Basal CRP concentrations were less than 0.574 mg/L and between 0.574 to 4.430 mg/L in the LOW
and HIGH groups, respectively. Muscle damage markers increased similarly between LOW and
HIGH groups. The HIGH group was similar to the LOW group in basal sTNFR1, IL-6, IL-10, and
cortisol and higher (P<0.05) than the LOW group in sICAM-1 (LOW = 188.0±32.9 verses HIGH =
207.2±25.4 pg/mL, P<0.05) and body mass index (LOW = 22.6±2.4 verses HIGH = 24.9±2.7 kg/m2,
P<0.05). Measured as area under the curve between exercise and control conditions, the acute
inflammatory response was greater (P<0.05) for sTNFR1 and CRP in the HIGH compared to LOW
group. Measured using repeated measures ANOVA for response amplitude relative to baseline,
IL-6 increased (P<0.001) 8 h post-exercise in the LOW but not the HIGH group, and CRP increased
(P<0.001) in the HIGH but not the LOW group. Conclusions: The HIGH CRP group had a lower
amplitude of the IL-6 response to muscle damaging exercise and a subsequent increase in CRP that
did not occur for the LOW group. IL-6 downregulates inflammatory initiating cytokines and
induces synthesis of anti-inflammatory mediators, thus, lack of a robust IL-6 response may allow
low-level inflammation to persist. We conclude that higher levels of CRP in young, apparently
healthy individuals reflects differences in both basal and acute inflammation that should be further
investigated to determine whether these differences influence development of chronic diseases.
Supported by a grant from the American Heart Association Pacific Mountain Affiliate.
F-15
Electrical pulse stimulation of C2C12 cultured myotubes induces IL-6 production
Arta Farmawati1, Taku Nedachi2, Makoto Kanzaki2 and Ryoichi Nagatomi1
1
Department of Medicine & Science in Sports & Exercise, Tohoku University Graduate School of Medicine, Sendai,
Japan
2
TUBERO/Tohoku University Biomedical Engineering Research Organization, Tohoku University, Sendai, Japan
Interleukin-6 is known to be produced and released by contracting muscle. Recently, in vitro
experiment have shown that electrical pulse stimulation (EPS) may lead to contraction of the
muscle cell cultures. This model might be able to open up the physiological and biochemical role of
cellular level in relation to contraction it self in cultured cells. To determine wheather the EPS could
directly induce contraction to produce IL-6 production in in vitro purified C2C12 myotube cultured
cells, we examined the effect of EPS-induced contraction, glucose availability and glycogen content
on IL-6 production in C2C12 myotubes. Our study established that the IL-6 release related to
frequency and duration of stimulation. The level of IL-6 transcript peaked at 24 hours after
stimulation. The differentiated C2C12 myotubes were cultured in medium containing either 5 mM
or 22.5 mM glucose. After stimulation, both IL-6 protein and transcript were induced above the
control level in all conditions, but most remarkably in 5 mM glucose. Again 5 mM glucose
condition was more favourable as compared to higher glucose. The IL-6 protein and transcript
would remarkably increase if glycogen in the lowest level. These results indicate that IL-6
production and IL-6 mRNA expression of the C2C12 myotubes were accelerated by repetitive
muscle contraction by electrical pulse stimulation. Our result strongly support that myocyte is the
main source of IL-6 release, also glucose availability and glycogen content influenced the rate of
IL-6 production in contracting skeletal muscle. The cultured contracting C2C12 myotubes may
serve as a good model for understanding the regulation of myokine IL-6 production.
F-16
Effects of overexpression of heat shock protein 70 on energy metabolism
Jürgen M. Steinacker, Liangli Wang, Larissa Gampert, Olga Prokopchuk, Yuefei Liu
Section of Sports and Rehabilitation Medicine, Department of Internal Medicine II, University of Ulm, Ulm, Germany
Objective: The response of heat shock proteins is stress reaction of muscle cells in intensive
training (Liu et al. JAP 86: 101-104, 1999). Hsp70 induction is related to energy metabolism which
not be distinguished from the chaperoning effects. Possible effects of upregulated Hsp70 on energy
metabolism have not been examined until now. Methods: We established a cell line expressing
different levels of Hsp70. Changes in metabolites and ATP levels were determined thereafter. In
addition, activities of enzymes involved in glycolysis (phosphofructokinase, PFK; lactate
dehydrogenase, LDH), Krebs cycle (citric synthase, CS) and oxidative phosphorylation (NADH
dehydrogenase, CI; ubiquinol:cytochrome-c reductase, CIII) were analyzed. Results: Glucose
consumption and lactate excretion were significantly elevated in cells expressing increased Hsp70.
Simultaneously, the activity of glycolytic enzymes was markedly increased in cells with relatively
high level of Hsp70 (>33.4 ng/321.5 ng total protein), and a good correlation was observed. For
complex I, the activity remained constant except a notable decrease in cells with low level of Hsp70
(LL) (33.4 ng/312.5 ng total protein), however, the activity of CS was significantly reduced except
in cells expressing moderate level of Hsp70 (ML) (89.3 ng/312.5 ng total protein). In addition,
intracellular ATP level was significantly elevated in ML and a clear increase was also observed in
cells with high level of Hsp70 (HL) (163.2 ng/321.5 ng total protein) compared with that in LL.
Conclusion: Upregulated Hsp70 in HeLa cells resulted in an increase in ATP level, which is related
to an enhanced glycolytic capacity and a possible involved mitochondrial biogenesis.
F-17
Acute resistance exercise markedly upregulates gene expression of key chemotactic factors in
skeletal muscle
Paul Della Gatta, Jessica Mathers, Kate Carey and David Cameron-Smith
School of Exercise and Nutrition Sciences, Deakin University, Melbourne, Australia.
Objective: Chemotaxis and activation of leukocytes following muscular injury or eccentric exercise
is critical for skeletal muscle remodeling and satellite cell activation. Our aim was to investigate the
effect of a single bout of resistance exercise on the gene expression of muscle-derived chemotaxis
factors. Methods: Seven young males (22.1±0.26yrs) completed three sets of resistance exercise for
the leg muscles (leg press, leg extension and squat). Two sets consisted of 8-12 repetitions at 80%
1-RM, whereas in the final set the subjects exercised until exhaustion. Muscle biopsies were
obtained before exercise, 2, 4 and 24 h after exercise. Muscle tissue was analyzed for the expression
of Monocyte Chemoattractant Protein-1 (MCP-1;CCL2), Fractalkine (CX3CL1), Urokinase Type
Plasminogen-Activator Receptor (uPAR;CD87), MDC (CCL22) and Vascular Endothelial Growth
Factor (VEGF) [1]. Results: MCP-1 (53 fold), CX3CL1 (1.5 fold), and uPAR (15 fold) were all
elevated significantly 2 hrs after the cessation of exercise. These changes were diminished 4 hrs
post exercise and returned to resting levels by 24 hrs post-exercise. VEGF mRNA expression did
not change significantly. MDC was not detectable at any time point measured. Conclusion: The
protein products of the genes analyzed have previously been identified as the major regulators of
monocyte chemotaxis in cultured myocytes [1]. In the present study, acute resistance exercise
significantly increased the expression of 3 of these identified genes associated with chemotaxis and
leukocyte activation. To our knowledge, this is the first evidence that CX3CL1 and uPAR respond
to muscle contraction. whilst, MCP-1 has previously been reported to be elevated following intense
exercise [2] and is known to exert a significant role in promoting muscle regeneration and function
following injury [3, 4]. Thus a single bout of intense resistance exercise enhances the expression of
several key chemotaxis genes, providing further insight into the mechanisms coordinating the
chemotaxis and activation of leukocytes.
1.
2.
3.
4.
Chazaud, B., et al., Satellite cells attract monocytes and use macrophages as a support to
escape apoptosis and enhance muscle growth. J Cell Biol, 2003. 163(5): p. 1133-43.
Chen, Y.W., et al., Molecular responses of human muscle to eccentric exercise. J Appl
Physiol, 2003. 95(6): p. 2485-94. Epub 2003 Aug 22.
Warren, G.L., et al., Role of CC chemokines in skeletal muscle functional restoration after
injury. Am J Physiol Cell Physiol, 2004. 286(5): p. C1031-6.
Warren, G.L., et al., Chemokine receptor CCR2 involvement in skeletal muscle regeneration.
Faseb J, 2005. 19(3): p. 413-5.
F-18
Corticosterone Accelerates Atherosclerosis in the Apolipoprotein E Mutant Mouse
Mitsuharu Okutsu1, Osamu Terada 2, Katsuhiko Suzuki1,3
1
Institute for Biomedical Engineering, Consolidated Research Institute for Advanced Science and Medical Care,
Waseda University, Saitama, Japan.
2
Graduate School of Human Sciences, Waseda University, Saitama, Japan.
3
Faculty of Human Sciences, Waseda University, Saitama, Japan
Objective: Monocytes in the blood stream eventually differentiate into macrophages, which act as a
primary responder to various pathogens to orchestrate immune responses that finally eliminate the
pathogen. Monocytes, however, are also involved in atherogenesis on the endothelium of arteries,
which leads to cardiovascular events. CC-chemokine receptor 2 (CCR2) is a receptor for
monocyte chemoattractant protein 1 (MCP-1)/CCL2, and it plays a critical role in the early stage of
atherogenesis. Since stress is well known as a risk factor of atherosclerosis, it may influence
CCR2 expression on monocytes. The aim of this study was to examine whether corticosterone
administration influences CCR2 expression on monocytes and the formation of atherosclerotic
lesions. Methods: Male B6.KOR/Stm-Apoe shl mice were obtained at 5 weeks of age. To
determine the contribution of circulating corticosterone in regulation of atherosclerotic lesions, mice
were treated with corticosterone in their drinking water. Corticosterone was first dissolved in 30%
(w/v) 2-hydroxypropyl-b -cyclodextrin (HBC) and then diluted to 25 mg/ml with distilled water.
The final concentration of HBC in the drinking water for all mice was 0.6%. Mice were fed a
high-fat Western-type diet. Drink and food intake were measured 2-3 days/weeks, and body weight
was measured every week. At 18 weeks, mice were sacrificed and peripheral blood, serum and heart
tissue were collected. CCR2 expression on monocytes was measured using flow cytometry.
High-sensitivity lipoprotein profiling was studied with the LipoSEARCH system. Quantification of
atherosclerotic lesions was assessed by Oil Red O staining. Results: Corticosterone administration
increased lesion formation but did not affect monocyte CCR2 expression. Corticosterone
administration also raised the levels of total cholesterol (p<0.05), LDL (p<0.05), VLDL (p<0.05)
and small-dense LDL (p<0.05) in comparison to control mice. Conclusion: These results suggest
that corticosterone regulated serum lipid profile, which may contribute to chronic stress-induced
development of cardiovascular diseases. Corticosterone treatment did not influence monocyte
CCR2 expression. Therefore, factors other than corticosterone may mediate the involvement of
monocytes in atherogenesis.
F-19
The effect of exercise intensity on some cytokine responses in women
Gough, L.,1,2 and Castell, L.1
1
2
CNRG, Nuffield Dept of Anaesthetics, University of Oxford OX2 6HA
School of Sport and Education, Brunel University UB8 3PH
Introduction: The effect of exercise on the cytokine IL-6 is of interest, as it is proposed to mediate
the beneficial effects of regular exercise in terms of systemic, low-grade inflammation. However,
this hypothesis is based on the results of studies which have employed prolonged, strenuous
exercise (Petersen & Pedersen, 2005). In the present study plasma IL-6, IL-1ra and IL-10 were
measured during 30min treadmill running. Female athletes were used because there is a dearth of
studies on them compared with male athletes, mainly due to cyclic variations. Methods: Eleven
endurance-trained, eumenorrhoeic women (mean±SD, age 33±5 yrs, height 164±5cm, weight
60±5kg, VO2max 49±6 ml/kg/min) consented to take part. They completed a 30 min run at lactate
threshold (LT), 90% and 110% LT on separate occasions in the early follicular stage of the
menstrual cycle after an overnight fast. Blood samples were collected at baseline (Pre),
immediately after exercise (Post) and 60 min post (Post-60). Cytokine concentrations were
measured using ELISA kits. Three-way repeated measures ANOVA with pairwise comparisons
indicated changes over time and time x trial interactions. Results: There was a small increase in
plasma IL-6 at Post and Post-60 after the LT and 110%LT trials (p<0.05); but no significant
difference between trials. Plasma IL-1ra was increased after all exercise trials (p<0.05); the
Post-60 increase was greater, but not significant, with increasing exercise intensity (31%, 93% and
155% for the 90%LT, LT and 110%LT trials respectively). Plasma IL-10 did not increase in
response to exercise. Discussion: IL-6 was increased in response to exercise, but the absolute
increase was small (~0.5pg·ml-1).
The increase in IL-1ra, however, appeared to be
intensity-dependent. These data indicate that IL-6 did not mediate the putative anti-inflammatory
effects of regular, moderate exercise. Thus, it is suggested that IL-1ra may have more impact than
IL-6 as an anti-inflammatory cytokine after this type of exercise in women.
Reference
Petersen, AMW & Pedersen, BK (2005).
J. Appl. Physiol. 98:
1154-1162
F-20
Exercise intensity influences
lipopolysaccharide in rats
plasma
TNF-alpha
concentration
in
response
to
Kitamura, H.1, Minato, K.1, Kimura, M.2, Yamauchi, H.3 and Yano, H.4
1
Wayo Women’s University
Kyoritsu University of Pharmacy
3
The Jikei University School of Medicine
4
Kawasaki University of Medical Welfare, Japan
2
Objective: It has been previously shown that moderate exercise can reduce the risk of infectious
disease. In contrast, strenuous exercise has been shown to transiently increase the risk of infection.
Unfortunately, the mechanisms responsible for this observation are still not clear. The purpose of
this study was to determine whether exercise intensity was responsible for reduced tumor necrosis
factor (TNF) -alpha production in response to lipopolysaccharide (LPS) in rats. Methods: Female
10 week-old F344 rats were randomly assigned to one of four groups; R, rest (n=18), L-Ex,
treadmill running at 10 m/min (n=8), M-Ex, at 21 m/min (n=8) and H-Ex, at 26 m/min (n=7) for 30
min at 15 % grade. Immediately after those exercise or resting period, rats were injected with LPS
(1 mg/kg) i.v. Blood samples were obtained from abdominal vein under general anesthesia with
pentobarbital (60 mg/km) immediately after exercise or rest and 1 h after LPS injection. Results:
Plasma TNF-alpha concentrations in both M-Ex and H-Ex groups were significantly lower than that
in R and L-Ex groups, respectively. The exercise intensity did not influence plasma corticosterone
concentration. In contrast, plasma adrenaline concentrations in M-Ex and H-Ex groups were
significantly higher than that in R and L-Ex groups, respectively. In addition, H-Ex group was also
observed high plasma noradrenaline concentration compared with was other three groups.
Conclusion: Middle- and high-, but not low- intensity exercise suppressed TNF-alpha response to
LPS. This immunosuppression might be regulated by changing catecholamine concentration, which
depends on exercise intensity.
F-21
The effect of moderate exercise and ex vivo extracellular acidosis on T lymphocyte and
monocyte activation
João Viana1, Emma Clapp1, Alice C. Smith2 and Nicolette C. Bishop1
1
2
School of Sport and Exercise Sciences, Loughborough University, Loughborough, LE11 3TU, UK
John Walls Renal Unit, Leicester General Hospital, Leicester, LE5 4PW, UK
Objective: Metabolic acidosis, a state in which the blood pH is low, is a common condition among
patients with chronic kidney disease (Kraut and Kurt, 2005). Although these patients have long
been known to suffer from immune dysfunction, few studies have focused on the effect of
extracellular acidosis on immune function (reviewed by Lardner, 2001). Therefore, the purpose of
this study was to investigate the effect of extracelluar acidosis on T lymphocyte and monocyte
activation before and after moderate exercise. Methods: With local ethics committee approval, 8 (4
males and 4 females) recreationally active healthy subjects (mean±SD: age 25±2 years; body mass
65.4±10.0 kg) ran or walked for 30 min on a motorised treadmill at a 1% gradient and at a speed
that elicited a subjective rate of perceived exertion in the range of 12-14 (“somewhat hard”).
Venous blood samples were collected before and after exercise. Peripheral blood mononuclear cells
were cultured in acidotic (pH 7.1) vs. neutral medium (pH 7.4) and lymphocyte phenotypic subsets
(CD4+ and CD8+) activation (CD69 expression) and monocyte (CD14+) activation (HLA-DR and
CD86 co-expression) were determined by flow cytometry following 20 h in vitro stimulation by
staphylococcal enterotoxin B (SEB) and tetanus toxoid and flu vaccines (TT+FLU). Results were
analysed using a 2-factor repeated measures ANOVA (exercise x medium). Fluorescence intensity
for a surface antigen was calculated as geometric mean (GMFI) of all positive-staining cells and
expressed as a ratio to unstimulated cells. Results: HLA-DR and CD86 co-expression on CD14+
monocytes was greater on cells cultured in acidotic vs. neutral medium with both stimuli (main
effect of medium; SEB: 1.47±0.45 vs. 1.27±0.31, P<0.05; TT+FLU: 1.27±0.45 vs. 1.12±0.52,
P<0.01) but there was no significant effect of exercise. There were no significant effects of exercise
or medium on CD69 expression by CD4+ or CD8+ lymphocytes for both stimuli. Conclusion:
These findings suggest that extracellular acidosis up-regulates HLA-DR and CD86 co-expression
by CD14+ moncoytes yet has negligible effect on CD69 expression by CD4+ and CD8+
lymphocytes. In addition, these findings suggest that this particular moderate exercise protocol has
no effect on these activation markers.
References: Kraut, J.A. & Kurtz, I. (2005). American Journal of Kidney Diseases, 45, 978-993.
Lardner, A. (2001). Journal of Leukocyte Biology, 69, 522-530.
Acknowledgments: João Viana is supported by a Portuguese Foundation for Science and
Technology (FCT-MCTES, Portugal) scholarship (SFRH/BD/27838/2006).
F-22
The different effects of exhaustive exercise on poly I:C-induced IFN-β and TNF-α productions
in mice
Masataka Uchidaa, Youhei Tanakaa, Noriaki Kawanishia, Yasuko Katob, Daisuke Shivac, Hiromi
Yanoa,
a
Department of Health and Sports Science, and bDepartment of Clinical Nutrition, and cDivision of Health Science,
Kawasaki University of Medical Welfare, Okayama, Japan
Objective: It is well known that severe exercise induces immune suppression. In fact, we already
reported that lipopolysacchride (LPS)-induced tumor necrosis factor (TNF)-α production as a
bacterial infection model to induce immune response, was inhibited by exhaustive exercise in rats.
However, it remains unclear that immune response to viral infection is also inhibited by the severe
exercise. The polyriboinosinic:polyribocytidylic acid (poly I:C) is a potent inducer of
interferon(IFN)-β, which is an anti virus inflammatory cytokine. The aim of this study was to
determine whether exhaustive exercise inhibits IFN-β and TNF-α productions after Poly I:C
injection in mice. Methods: Male C3H/HeN mice, 9-10 weeks old (n= 27), were divided into two
groups; EX group ran on the treadmill to exhaustion (the means exhaustive running time: 72±5min),
and N-EX group: mice were the rest. Each group was injected poly I:C (5mg/kg) immediately after
exercise or rest. The mice were collected with blood sample from orbital vessel at 0, 1, 3, and 6
hours, to measure IFN-β and TNF-α concentrations in plasma using ELISA kits. Results: TNF-α
concentration in EX group was significantly lower than that in N-EX group at 1 hour after poly I:C
injection (p<0.01). However, IFN-β concentration was not significantly difference between in EX
and N-EX groups, although poly I:C induced an increasing IFN-β in both groups (p<0.01,
respectively). Conclusion: These results suggest that anti-virus response to viral infection might be
maintained despite severe stressful exercise in mice.
FREE COMMUNICATION (POSTER)
P-1
Kinetics of HSP70 expression in the intestine following exhaustive exercise
Ingrid Brenner1,2 and Ashton Gibson2
1
2
Trent/Fleming School of Nursing, Trent University, Peterborough, ON. Canada
Dept. of Biology, Trent University, Peterborough, ON. Canada
It has recently been proposed that heat shock protein (HSP) may be bound to endotoxin and
released from the gut enterocytes into the circulation in response to an acute exercise stress. It is
possible that this complex activates macrophages and may help explain the mechanism by which
HSP70/72 can resolve bacterial infection. Objectives: This study was designed to examine the
kinetics of HSP70/72 expression in the intestine, in plasma and in splenic lymphocytes following an
acute bout of exercise.
A total of 25 young (6 – 9 wk old) female, untrained C57BL/6, mice
participated in the study. Twenty mice performed an exhaustive bout of exercise, whereas 5
served as pre-exercise controls. Methods: Following two brief familiarization sessions, mice
were allocated to one of four groups (immediately following and 1, 2 and 24 hours post-exercise) (n
= 5 per group). Exhaustive exercise was performed for 90 min (30 min each at 22 m/min, 25 m/min
and 28 m/min) on a motorized treadmill inclined at a 9° slope. Mice were euthanized at the time
intervals specified above. The small and large intestine, spleen, and blood samples were removed
for further processing. Intestinal HSP expression was measured using an HSP70 ELISA kit, plasma
HSP was measured by Western Blot and intracellular HSP expression in splenic lymphocytes was
measured by flow cytometry. Intestinal samples were further analyzed by histological staining.
Results: Preliminary results show a biphasic increase in HSP expression in the large intestine
(peaking immediately post and 24 hours after exercise), whereas, in the small intestine HSP
increased and then returned to baseline levels 24 hours following exercise. Intracellular HSP
expression in splenic lymphocytes followed a similar pattern, increasing immediately following the
acute exercise stress. Data for Western Blot and histology are currently being analyzed.
Conclusions: The results of this study are useful in identifying the timing of HSP70/72 release
between different tissue compartments within the body. Further research is required to explain the
biphasic release of HSP from the large intestine and to examine the role of HSP-bound endotoxin in
the activation of macrophages.
P-2
Effect of exhaustive exercise on TLR4 expression in macrophages in mice
Daisuke Shiva1, Noritsugu Tsutsum2, Noriaki Kawanishi2, Yohei Tanaka2, Michel J. Klemenik2,
Takashi Matsumoto3, Hiromi Yano2
1
Division of Health Science
Department of Health and Sports Science, Kawasaki University of Medical Welfare, Okayama 3 Microbiology,
Department of Infectious Disease, Faculty of Medicine, Oita University, Oita, Japan
2
Objective: Previous studies reported that chronic exercise down-regulates Toll-like receptor (TLR)
4 expression in peripheral CD14 positive cells. However, the effect of acute exercise on TLR4
expression remains unclear. TLR4 is down-regulated by receptor agonists, such as
lipopolysaccharide (LPS) and heat-shock proteins (HSPs). Furthermore, several studies suggested
that acute exercise increased plasma LPS and HSPs concentration. Therefore, acute
exercise-induced molecules, including LPS and HSPs, could regulate surface TLR4 expression. The
purpose of this study was to investigate the effect of exhaustive exercise on TLR4 expression of
macrophages using alveolar macrophages and macrophage cell line (RAW264.7 cells). Methods:
C3H/HeN mice were divided into two groups; running exercise group (EX) and resting group
(CON). EX mice ran on a treadmill starting at 9m/min for 3 minutes followed by a 2m/min
increment every 3 minutes until exhaustion (the means of exhaustive running time: 89±5 min).
Immediately after exercise and rest, mice were removed bronchoalveolar lavage fluid (BALF)
including alveolar macrophages, to analysis surface TLR4 expression by flow cytometry. In
addition, blood samples were also collected in both groups, and then RAW264.7 cells were
incubated with the collected serum for twenty-four hours, to evaluate surface TLR4 expression and
TNF-α mRNA expression. Results: Surface expression of TLR4 in alveolar macrophages was no
observed difference between in EX and CON groups. Serum protein concentration in EX mice was
trend toward higher than that in CON group, surface expression of TLR4 in RAW264.7 cells was
not affected by EX serum treatment compared with treatment of CON serum. Furthermore, there
was no significant difference between LPS-induced TNF-∂ mRNA expression in RAW264.7 cells
with EX-serum and CON-serum treatments. Conclusion: These results suggest that acute
exhaustive exercise might not affect on surface TLR4 expression and LPS sensitivity in mouse
macrophages.
P-3
Exhaustive exercise is reduced TNF-α production in response to lipopolysaccharide, but it
does not depend on gene expression in mice
Yohei Tanaka1, Noriaki Kawanishi1 , Yasuko Kato1, Daisuke Shiva1, Hiromi Kitamura2, Hiromi
Yano1
1
2
Kawasaki University of Medical Welfare, Okayama
Wayo Women’s University, Chiba, Japan
Objective: Stressful exercise has been found to immune suppression. In fact, we already reported
that exhaustive-exercised rats showed low level of tumor necrosis factor (TNF)-α in plasma despite
lipopolysaccharide (LPS) injection. However, the mechanism of exercise-induced immune
suppression remains unclear. To clarify the mechanism of exercise-induced reduction of TNF-α
production, we measured TNF-α contents and it’s mRNA expression in several tissues in
exhaustive-exercised mice after LPS stimulation. Our hypothesis was that LPS-induced TNF-α
production in tissues was inhibited by the exhaustive exercise. Methods: Experiment was
conducted on male C3H/HeN mice, 9-10 week-old age (n=42). Exercising mice (EX group) ran on
a treadmill to exhaustion (the mean exhaustive running time: 69±3min). Resting mice (N-EX group)
were sedated for 60 min. The animals were injected LPS (1mg/kg) immediately after exercise
(n=22) or rest. One hour after LPS injection, mouse blood samples were collected, and then liver,
lung and spleen were removed for determination of TNF-α using ELISA kit. Those tissues in some
mice were removed 30 min after the LPS injection for determination of TNF-α mRNA expression
by RT-real time PCR. Results: Plasma TNF-α concentration in EX group was significantly lower
than that in N-EX group one hour after LPS injection (p<0.01). However, TNF-α mRNA expression
response to LPS in liver, lung and spleen was unchanged by the exhaustive exercise. In contrast,
tissue TNF-α contents in liver, lung and spleen of EX group was significantly lower than that in
those of N-EX group (p<0.01, p<0.01 and p<0.05, respectively). Conclusion: There data suggest
that exhaustive-exercise induces reduction of plasma TNF-α concentration despite LPS stimulation,
and then this mechanism might be depend on translation of TNF-α but not the transcription of
tissues in mice.
P-4
Effects of exercise training on B cells in elderly rats
Keisuke Nokura1, Kazuhiro Shimizu2, Fuminori Kimura2, Takayuki Akimoto3, Takao Akama1,
Ichiro Kono2
1
Faculty of Sport Sciences, Waseda University,
Graduate School of Comprehensive Human Sciences University of Tsukuba ,
3
Fuculty of Medicine, The University of Tokyo
2
Objective: To study that the alternation of B cells in elderly rats by exercise training.
Methods: We exercised 12-months-old rats ( male : n=10, female : n=8 ) by a treadmill running
for 30 minutes each day , during 8 weeks. After the training period, we analyzed splenocytes by
flow cytometry. The data were compared with those of non-exercised rats ( male : n= 4, female :
n=4 ). Results: The parts of B cells in splenocytes of the trained rats ( 85±2% ) were significantly
higher than those of non-trained rats ( 62±10% ). Conclusion: In elderly rats, the part of B cells in
splenic cells might be increased by exercise training.
P-5
The effect of Pamper Pole jump on salivary IgA and cortisol responses in adventure education
Wen-Yu Chan and Tzai-Li Li
Department of Sports and Leisure, National Dong-Hwa University, Hualien, Taiwan
Objective: The aim of this study was to determine the effect of anxiety induced by Pamper Pole on
salivary IgA and cortisol responses. Methods: With the approval of the local Ethics Committee, 9
healthy men and 6 women (age 21.8 ± 0.4 years, height 1.66 ± 0.02 m, body mass 63.9 ± 2.3 kg;
means ± SEM ), who were recreationally active and never experienced a high element Pamper Pole
jumping before, participated in this study. Subjects performed Pamper Pole jump in each trial,
separated by at least 6 days. For avoidance of circadian variation, the two experimental trials were
conducted at same time of day. No food and sleep was allowed after 11:30 am. No water was
allowed to consume 30 min before each trial until the trials finished. Subjects were encouraged to
climb to the top of Pamper Pole after wearing safety equipments and then to jump off the pillar.
Saliva samples were collected at 10 min before filling in Beck Anxiety Inventory sheet (Pre-EX),
arrival to the top of Pamper Pole (Pre-Jump), and 30 min (P-30-EX) and 60 min (P-60-EX) after
jumping off. Heart rate was recorded at Pre-EX, just before climbing (Pre-Climb), immediately
arrival to the top of Pamper Pole (Top), Pre-Jump, and immediately after jumping off (Post-Jump)
by radiotelemetry. Results: The main findings of this study were: (1) experiences of Pamper Pole
jump appeared not to influence responses of blood pressure, the scores of Beck Anxiety Inventory,
saliva flow rate, salivary IgA concentration and secretion rate, heart rate, and salivary cortisol to the
pressure of high element in adventure education; (2) the situation of Pamper Pole jump significantly
decreased saliva flow rate but significantly increased heart rate. Conclusion: The findings of this
study suggested that the Pamper Pole jump does not appear to induce substantial anxiety enough to
alter oral immunity and the experience of conducting Pamper Pole may not affect the feeling of
anxiety.
Keywords: Pamper Pole, saliva flow rate, salivary IgA, anxiety, cortisol
P-6
Effect of stimulating saliva flow on the changes in salivary secretion of IgA, lysozyme and
α-amylase with prolonged exhaustive exercise
Judith Allgrove, Marta Oliveira, Alice Gallen and Michael Gleeson
School of Sport and Exercise Sciences, Loughborough University, UK
Objective: A higher rate of upper respiratory tract infection experienced by some athletes may be
due to an exercise-induced decrease in mucosal immune defences. Stimulating saliva flow during
exercise may potentially increase oral immune protection. Therefore, the aim of the present study
was to investigate the salivary secretion rates of immunoglobulin A (s-IgA), lysozyme and
α-amylase in response to strenuous exercise in both stimulated and unstimulated saliva flow
conditions. Methods: Twenty four fit, healthy, men (mean ± SD age: 23 ± 5 years; maximal oxygen
uptake, VO2max: 56.6 ± 9.2 ml/kg/min) cycled for 2.5 hours at 60%VO2max (with regular water
ingestion) and then cycled to exhaustion at 75% VO2max. Timed collections of whole saliva were
made immediately before exercise, mid-exercise, after completion of the 2.5 hour moderate exercise
bout and immediately after the exhaustive exercise bout. After each unstimulated saliva collection a
stimulated saliva flow sample was collected following chewing mint-flavoured gum for one minute.
Saliva was analysed for s-IgA, lysozyme and α-amylase and secretion rates were calculated. Data
were analysed using a 2-way repeated measures ANOVA with post-hoc t-tests. Results: Saliva flow
rate was ~3 times higher when saliva flow was stimulated. Saliva flow rate decreased with exercise
for stimulated saliva flow only (P<0.01). Exercise was associated with increases in lysozyme and
α-amylase concentration and secretion rates in saliva (P<0.01). Stimulating saliva flow caused a
higher secretion rate of lysozyme (>2-fold higher) and α-amylase (>3-fold higher) compared with
unstimulated saliva flow (both P<0.01). S-IgA concentration and secretion rate increased with
exercise but were both lower in stimulated saliva compared with unstimulated (P<0.05). Following
the exhaustive exercise bout, s-IgA secretion rates were 129 ± 92 and 165 ± 97 µg/min in the
stimulated and unstimulated saliva flow conditions, respectively (P<0.05). Conclusion: Stimulating
saliva flow during exercise had positive effects on the quantity of saliva and on the antimicrobial
proteins lysozyme and α-amylase, but resulted in a slightly lower secretion rate for s-IgA. Increases
in lysozyme, α-amylase, and the saliva flow rate suggest that stimulating saliva flow during exercise
by chewing gum may acutely provide increased oral immune protection.
P-7
Inquire: Immunocompetence and exercise among people over 65-years-old
Martins Raul1, Teixeira-Verissimo Manuel2 and Teixeira Ana Maria1
1
2
Department of Biokinetic Studies, Faculty of Sport Sciences and Physical Education, University of Coimbra, Portugal
Faculty of Medicine, University of Coimbra, Portugal
Objective: Scarce papers have been published concerning immunological chronic effects in elderly.
Some cross-sectional studies have attained an improvement in immunity as a consequence of
moderate intensity exercise. However, not all the results have been consistent, particularly when
longitudinal investigations are required. The interest remains about the immunology adaptations to
exercise in elderly.
The purpose of the present research was to analyse the chronic adaptations in plasmatic IgA to a
moderate aerobic exercise program in elderly. Methods: Twenty four women (77.53±7.99 years
old) and seventeen men (75.43±6.64 years old), aged between 65 and 95 years old participated.
Subjects were divided into two groups: aerobic exercise and control. The aerobic program was
constituted by a 3 week sessions, during 16 weeks. Fasting venous blood samples were collected
with a minimum of 48 hours after exercise sessions. Statistical analysis used MANOVA and
Bonferroni post hoc test. Results: The IgA evaluation for the aerobic group was
1.08g.L-1±0.50g.L-1 (initial) 2.29g.L-1±0.93g.L-1 (final). The control group, attained
2.48g.L-1±1.85g.L-1 (initial) and 3.40g.L-1±1.80g.L-1 (final). MANOVA reveals differences
between evaluations, for aerobic group [F(2, 21)=61.60, p=0.00], which reflect an improvement in
immunocompetence in respect to IgA. By the contrary, in the control group the differences were not
significant [F(2, 14)=1.94, p=0.18]. When one looks for the interaction between sex and evaluation,
the results are not significant for both groups: aerobic [F(2, 21)=1.31, p=0.29]; control [F(2,
14)=0.14, p=0.87]. This means that in the control group the inexistence of differences between
evaluations are common to female and male. Concerning aerobic group, the differences between
evaluations are also common to both sexes. Conclusion: Some papers refers an increase (Akimoto
et al., 2003; Gleeson et al., 2000; Karacabey et al., 2005), some refers no changes (Sari-Sarraf et al.,
2007; Whitham et al., 2006), and one refers a decrease in IgA (Tharp &amp;amp; Barnes, 1990).
The intensity associated to each one of this works varies between moderate and high. Our results
seems to be in agreement with a growing body of evidence that supports an immunity improvement
as a consequence of aerobic moderate exercise, even in elderly people.
P-8
Effect of exercise, aging and functional capacity on acute secretory IgA response in elderly
people over 75 years of age
Yuzuru Sakamoto1, Shouzoh Ueki2, Toshiyuki Kasai2, Jinro Takato2, Hideki, Shimanuki3, Haruhiko
Honda2, Tsunehisa Ito4 and Hiroshi Haga5
1
Department of Molecular & Cellular Immunology, Shinshu University Graduate School of Medicine. 2Faculty of
Medical Science and Welfare, Tohoku Bunka Gakuen University.
3
Department of Medicine & Science in Sports & Exercise, Tohoku University Graduate School of Medicine.
4
Junior College Division, Tohoku Seikatsu Bunka College.
5
Department of Gerontology, Obirin University Graduate School of International Studies, Japan.
Objective: Age-associated decline in immune function and regulation, referred to as
immunosenescence, brings about an increased incidence of infectious diseases in the aged. However,
there is little data on the effect of aging and exercise on the mucosal immune function in elderly
people. Moreover, there is no evidence whether the change in functional capacity affects mucosal
immunity in elderly people. The purpose of this study was to examine the effects of exercise, aging,
and functional capacity on the mucosal immune function in elderly people over 75 years of age.
Methods: The subjects were 92 community-dwelling elderly women aged over 75 years who lived
in a rural community, Miyagi prefecture. The subjects periodically performed about 20 min of
low-intensity exercise. Saliva samples were collected before and after exercise, and Saliva flow,
secretory immunoglobulin A (SIgA) concentration and SIgA secretion rate were determined. The
Tokyo Metropolitan Institute of Gerontology Index of Competence was used to measure the
functional capacity. Results: In comparison with before exercise, Saliva flow, secretory
immunoglobulin A (SIgA) concentration and SIgA secretion rate were significantly increased after
exercise in elderly subjects. In addition, when low- and high-value groups of resting SIgA levels
were compared, acute SIgA responses was observed only in the low value group. However, there
was no significant effect of aging and decline in functional capacity on exercise-induced SIgA
response. Conclusion: These results suggest that resting SIgA levels influence the mucosal immune
function response for exercise in elderly people over 75 years of age.
P-9
Mucosal immunity after a soccer match
Vahid Sari-Sarraf 1, Parvaneh Alavi-Namvar 2, Saeed Nikoo-Kheslat 1
1
2
Faculty of Physical Education and Sport Sciences, University of Tabriz, Tabriz/Iran
Dept. of Physical Education and Sport Sciences, Tabriz Azad University, Tabriz/Iran
Objective: Acute exposure to naturalistic stressors reliably produces increases in salivary
Immunoglobulin A (IgA).Soccer players are exposed to a number of physical and psychological
stressors during the competitive season. The aim of this investigation was to scrutinize the salivary
IgA and cortisol alterations in elite soccer players in response to a real soccer-match. Methods:
Twenty-two elite soccer players, head coach and coach of the first division men soccer team were
consenting. After the fist visit to record their body mass, height and percent body fat, 11 players
reported to the pitch; temperature, relative humidity and wind speed were 15 ± 2 oC and 40 ±
10 % RH, 18 km·h to undertaking a match play and 9 players, head coach and coach watched the
match from their bench as control group. Unstimulated saliva samples were collected before and
immediately post-match. Differences were examined using a paired-sampled t-test for dependent
groups. Data are presented as mean values and standard error of the mean (± SEM). Results: The
results showed that salivary osmolality for players and non-players increased from 79.1 ± 32 to
166.5 ± 79 mOsmol.kg-1 (P=0.004) and from 71.3 ± 7 to 95.6 ± 26 mOsmol.kg-1 (P=0.01)
respectively. Salivary IgA concentration for players boosted from 275.9 ± 122 to 786.8 ± 534 mg.l-1
(P=0.014) and for non-players from130.6 ± 74 to 318.3 ± 293 mg.l-1 (P=0.09). Salivary
concentration of cortisol for players displayed an increase significantly (P<0.001). This increased
pattern for non-players was not displayed significant (P>0.05). Salivary IgA to osmolality ratio and
protein in neither soccer players or non-players was significantly increased (P>0.05). Conclusion:
It is concluded that soccer-match, appears to stimulate the athletes’ mucosal immunity because of
higher concentration of salivary IgA at least by immediately post- match. These data indicate also
that the match results in significant elevations of salivary cortisol.
P-10
A rat model of saliva secretory immunoglobulin A suppression caused by intense exercise
Fuminori Kimura1, Katsuji Aizawa1, Kai Tanabe1, Kazuhiro Shimizu1, Michihiro Kon1, Hoseong Lee2,
Takayuki Akimoto3, Takao Akama4 and Ichiro Kono1
1
Graduate School of Comprehensive Human Sciences, Tsukuba University, 1-1-1 Tennodai, Tsukuba, 305-8577, Japan
Department of Physical Education, Dankook University, San 29 Anseo-dong, Cheonan-si, Choongnam, 330-714,
Korea
3
Division of Biomedical Materials and Systems, Center for Disease Biology and Integrative Medicine Faculty of
Medicine, Tokyo University, 7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan
4
Faculty of Sport Sciences, Waseda University, 2-579-15 Mikajima, Tokorozawa, 359-1192, Japan
2
Objective: To develop a valid model of immunosuppression induced by intense exercise in rats.
Methods: Rats were divided into three groups. In the Rest group (N = 5), saliva was collected from
resting rats on 4 consecutive days. In the Ex group (N = 11), rats ran on a treadmill to exhaustion at
15~30 m.min-1 (exercise time: 60.0 ± 3.7 min), and their saliva was collected before and after
exercise; the salivary glands were removed after exercise. In the Con (control) group (N = 11),
saliva collection and gland removal were also performed, but the rats did not exercise. Secretory
immunoglobulin A (SIgA) concentrations in saliva and polymeric immunoglobulin receptor (pIgR)
mRNA expression in the glands were measured. Results: There was no significant change in SIgA
concentration in the Rest group over 4 days. In the Ex group, SIgA concentration decreased
significantly after exercise compared with before (p < 0.05), whereas there was no significant
change in the Con group. Expression of pIgR mRNA was significantly lower in the Ex group
post-exercise than in the Con group (p < 0.05). Conclusion: Our procedure for saliva collection
appeared suitable, and the exercise-induced SIgA suppression was probably caused by a decline in
pIgR mRNA expression. We propose to use this reproducible and reliable rat model of
exercise-induced SIgA suppression in future studies.
P-11
Supplementations of alpha and gamma-tocopherol have the similar effect on cellular immune
function following moderate exercise training.
Mayumi Kaneyasu1, Saiko Ikeda2, Tomono Uchida2, Mio Ichikawa3, Hiroyuki Yoshimura3 and
Satoru Moriguchi1
1
Department of Human Nutrition, Faculty of Nursing and Human Nutrition, Yamaguchi Prefectural University
Department of Nutritional Sciences , Nagoya University of Arts and Science
3
Eisai Food & Chemical Co., Ltd.
2
Objective: We have previously found that alpha- tocopherol (Toc) has an ability to restore the
decrease of cellular immune function following a bout of exercise. From this result, alpha-Toc may
have a beneficial effect on cellular immune function following exercise training. As there are 4
types of tocopherols (alpha, beta, gamma and delta) in nature, we have investigated the effect of
alpha or gamma-Toc supplementation on cellular immune function of healthy young women
following moderate exercise training. Methods: Double-blind study was performed on subjects
(n=19, aged 18-21 yr) divided into three groups. Seven subjects were orally given 100 mg of
alpha-Toc once a day for 3 weeks, seven subjects took 100 mg of gamma-Toc, and five subjects
took placebo for the same period. Then, all subjects started exercise training (walking for 30 min at
6 km/h, 5 d/wk) for 4 weeks. Blood samples were collected three times during experimental period
(0, 3 and 7 weeks). The study was approved by the Research Ethics Committee of Yamaguchi
Prefectural University, Yamaguchi, Japan. Results: Following 3-week supplementation of
alpha-Toc, plasma alpha-Toc concentration of alpha- Toc supplementation group has significantly
increased by 1.6 times compared to Pre. On the other hand, plasma gamma-Toc concentration
significantly decreased. And plasma gamma-Toc concentration following 3-week supplementation
of gamma-Toc has significantly increased by 4.8 times compared to Pre, but plasma alpha-Toc
concentration significantly decreased. Although exercise training had no effect on the plasma
alpha-Toc concentration in both supplemental groups, plasma gamma-Toc concentration in
gamma-Toc supplemented group significantly decreased following 4-week exercise training. NK
activity of peripheral blood lymphocyte (PBL) decreased in both supplemental groups after exercise
training. And Con A or PHA-induced lymphocyte proliferation of both supplemental groups
increased after 3-week supplementation, and decreased after 4-week exercise training. Conclusion:
It has been suggested that if moderate exercise training preferentially spends gamma-Toc rather
than alpha-Toc in the high plasma concentration of alpha and gamma-Toc. However, alpha and
gamma-Toc supplementations have the similar effect on cellular immune function following
moderate exercise training.
P-12
Circulating leukocyte and T-lymphocyte subset response to carbohydrate and protein feeding
after prolonged exercise
Ricardo da Costa1, Anthony Blanchfield1, Eifion Williams1, Jennifer Heaney1, Lucy Bywater1,
Samuel Oliver1, Stewart Laing1, James L.J. Bilzon2 and Neil Walsh1
1
2
University of Wales, Bangor, UK
Headquarters Army Training and Recruiting Agency, Upavon, UK
Objective: To determine the effects of immediate and delayed carbohydrate (CHO) and protein
(PRO) feeding on circulating leukocyte and T-lymphocyte subset responses after prolonged
exercise. Methods: Seven trained male runners completed three feeding interventions in
randomised order after 2 h running at 75% VO2max. On the control trial (CON), participants
received water equal to 12 ml.kg-1 body mass immediately post-exercise and 1 h post-exercise. On
the immediate feeding trial (IF), participants received a CHO-PRO solution equal to 1.2 g CHO·kg-1
and 0.4 g PRO·kg-1 immediately post-exercise and water 1 h post-exercise. On the delayed feeding
trial (DF), participants received water immediately post-exercise and CHO-PRO solution 1 h
post-exercise. Venous blood samples were collected pre-exercise, post-exercise, and at 20 min
intervals until 2 h post-exercise. Total and differential leukocyte counts and T-lymphocyte subsets
were measured and corrected for changes in plasma volume. Results: The leukocytosis (main effect
of time (meot): P<0.01) following exercise (mean ± SEM immediately post-exercise: 10.0 ±
1.1x109.L-1) remained elevated to a similar extent throughout the feeding period (20-120 min post
samples) on CON (11.2 ± 1.1 x109.L-1), IF (11.5 ± 1.3 x109.L-1) and DF (11.3 ± 0.8 x109.L-1). CD3+
and CD8+ subsets decreased during the 2 h recovery period (meot: P<0.01). No interaction or main
effects were observed for circulating CD4+. Conclusion: Ingestion of a CHO-PRO solution equal to
1.2g CHO•kg-1 and 0.4g PRO•kg-1 either immediately after or 1 h after prolonged strenuous
exercise does not influence short-term recovery of circulating leukocyte or T-lymphocyte subsets.
P-13
Why wheat-dependent exercise-induced anaphylaxis is provoked by various wheat processed
foods
Tanaka, M.1, Yano, H.2 and Kato, Y. 1
1
2
Department of Clinical Nutrition
Department of Health and Sports Science, Kawasaki University of Medical Welfare, Okayama, Japan
Objective: Wheat-processed food is divided into two patterns depending on the cooking processing
form. One is dough forming such as pasta and bread, and another is a form-suppressed
dough-formation, such as crepes and pancakes (batter). Large particle gluten is formed with gliadin
and glutenin during the formation of the dough. On the other hand, the batter controlled the
formation of the gluten as much as possible. That is, it is thought the possibility that the allergenic
activity of the gliadin, a major allergen, decreases according to the formation of insoluble
heat-coagulated gluten. We have already reported wheat flour-dependent exercise
induced-anaphylaxis, but the effect of wheat-processed food on the anaphylaxis remains unclear. In
this study, the difference of the allergenic activity of two types of wheat-processed foods was
examined using mice, and the reasons were discussed. Methods: Heated-dough and
unheated-batter were made from wheat flour. The sensitized B10.A mice were injected
intraperitoneally with gliadin in an aluminium hydroxide with an interval of 7 days between five
injections. The specific anti-gliadin IgE in mouse serum was evaluated by ELISA. The sensitized
mice were orally administrated heated-dough or unheated-batter containing 10 mg gliadin, then ran
on a treadmill for 30 min. After the exercise, these mice were placed individually in a cage with a
rotation wheel, and were monitored for voluntary physical activity for 15 hr. A voluntary
momentum assessed the allergic symptoms. Ethanol extract was obtained from various
wheat-processed foods with or without dough-formation and examined by SDS-PAGE and
immunoblotting. Results: The voluntary physical activity of gliadin-sensitized mice ingested with
heated dough or unheated batter decreased more than the control. However, no significant
difference was observed between the activity in heated dough and unheated batter groups.
Comparing ethanol soluble proteins (total gliadin) in wheat-processed foods, gliadin was detected
more in batter-form foods than in dough-form. However, omega-gliadin was detected from
dough-form foods just the same as from batter-formed ones. Conclusion: It was suggested that the
presence of free omega gliadin without dough-formation in various processed wheat-foods might
cause the development of wheat dependent exercise-induced anaphylaxis by eating various
wheat-processed foods.
P-14
Mucosal lesion in small intestine in WDEIA accelerated by gliadin
Hana Kozai1, Hiromi Yano2, Tsukasa Matsuda3 and Yasuko Kato4
1
Department of Comprehensive Rehabilitation, Osaka Prefecture University
Department of Health and Sports Sciences, Kawasaki University of Medical Welfare
3
Graduate school of Applied Molecular Bioagricultural Sciences, Nagoya University
4
Graduate school of Medical Professions, Kawasaki University of Medical Welfare
2
Objective: We reported that the major allergens in wheat-dependent exercise-induced anaphylaxis
(WDEIA) were gliadin and glutenin in B10.A mice. The mucosal lesion in small intestine was
observed in gliadin and glutenin treated mice. In this study, we investigated the histological damage
in small intestine using mice in WDEIA. Moreover, the mechanism of mucosal lesion was
examined by measurement of the inflammatory cytokine concentrations in intestine cannel and
portal blood. The permeability of the Caco-2 monolayer as a model of intestinal epithelium was
measured after addition of wheat proteins by transepithelial electrical resistance (TER). Methods &
results: B10.A mice were sensitized with each fractionated wheat protein, salt-soluble protein,
gliadin and glutenin. Sensitized mice were ingested each fractionated wheat protein following
30min-treadmill running. In sensitized mice after gliadin ingestion and exercise, all part of small
intestine was injured with bleeding. The upper part of small intestine in unsensitized mice after
gliadin ingestion and exercise was also injured with bleeding. The mucosal lesion area in small
intestine of both sensitized- (42%) and unsensitized- (32%) gliadin groups were significantly higher
than the other groups (p<0.01). Moreover, the mucosal lesion in unsensitized mice ingested gliadin
without exercise was also observed. It was considered that gliadin ingestion might be influence the
mucosal lesion. The TER of Caco-2 layer treated with gliadin and gliadin-digest decreased to 3 and
7%, whereas it decreased to 82 and 45% by treatment with salt-soluble protein and the digest,
respectively. Caco-2 permeability was significantly accelerated treatment with gliadin and the
digest. Inflammatory cytokine levels in portal blood and small intestine canal were measured in
sensitized mice after allergen ingestion and exercise. The concentrations of TNF-alpha and
IFN-gamma in portal blood and small intestine canal in Gliadin group were significantly higher
than the others, respectively (p<0.01). Conclusion: Our study revealed that the small intestine of
gliadin-ingestion mice in WDEIA was injured with bleeding as similar histological mucosal lesion
in small intestine reported in celiac disease by model mice. It might be caused that the increase of
inflammatory cytokine, and the permeability of mucosa epithelial cell in small intestine accelerated
by gliadin ingestion.
P-15
10 weeks Endurance Running on Gene Expression of IL-1ß, Mn-SOD and Caspase-3 in the
Heart and Gastrocnemius of Spontaneously Hypertensive Rats
Wanglok Lee, Huigeun Park, Junhyun Jeong, Yongsoo Jeon, Soonmi Kwon, Youngran Lee,
Jungmin Whang, Aram Yoon, Jaewoo Yoon, Jongkui Jun
Department of Community & Recreation, College of Science, Chungnam National University, Daejeon, Korea
Objective: The present research was to investigate the effect of a long-term endurance exercise
training on the expression of IL-1ß, Mn-SOD and Caspase 3 in the heart and gastrocnemius of
Spontaneously Hypertensive Rats (SHR).
Methods: Twenty Wistar-Kyoto (WKY) rats and twenty SHRs (age, 5 weeks) were used and
randomly divided into 4 groups; WKY Control (WC, n=10), WKY exercise (WE, n=10), SHR
control (SC, n=10) or SHR exercise (SE, n=10). Endurance exercise was performed by a
treadmill-running program (15m/min, 0% grade, 40mins/day, 5 days/week, 10 weeks). Total RNA
was extracted from heart and gastrocnemius tissues using TRIzol method. After cDNA was
synthesized, Gene expression of IL-1ß, Mn-SOD and Caspase-3 were measured by Real-Time PCR
after 10 weeks, while systolic blood pressure (BP) and heart rate (HR) were monitored by tail-cuff
pressure measurement for the whole experimental period. Repeated measures ANOVA and
One-way ANOVA were used and p value under 0.05 was considered as statistical significance.
Results: 10 weeks running significantly inhibited the increment of systolic blood pressure in SE.
IL-1ß gene expression of WC (0.98 ± 0.17) and WE (0.15 ± 0.01) was significantly higher than that
of SC (0.04 ± 0.008) and SE(0.04 ± 0.007) in heart. Mn-SOD gene expression of WE (26.79 ± 2.09)
was significantly increased. Caspase-3 gene expression of WE (5.90 ± 0.73) was significantly
higher than that of the others. Further, Caspase-3 of WC (3.18 ± 0.51) was significantly increased
comparing to that of SC (0.84 ± 0.15) and SE (1.12 ± 0.16). However, IL-1ß, Mn-SOD and
Caspase-3 gene expression of gastrocnemius were not significant in the all groups.
Conclusion: 10 weeks endurance running had positive effect on the systolic pressure, but not on the
gene expression of heart in SHRs. IL-1ß, Mn-SOD and Caspase-3 gene expression of heart were
might impaired by chronic high blood pressure. However, IL-1ß, Mn-SOD and Caspase-3 gene
expression of gastrocnemius might not be affected by chronic high blood pressure.
P-16
Eccentric muscle contractions induce greater oxidative stress than concentric contractions in
skeletal muscle
Michihiro Kon1, Kai Tanabe1, Hoseong Lee2, Fuminori Kimura1, Takayuki Akimoto3 and Ichiro
Kono1
1
Graduate School of Comprehensive Human Sciences, Doctoral program of Sports Medicine, University of Tsukuba,
Japan
2
Taekwondo Major School of Sports Science, University of Dankook, Korea
3
Center for Disease Biology and Integrative Medicine, Fuculty of Medicine, The University of Tokyo, Japan
The purpose of this study was to examine oxidative stress in skeletal muscle after eccentric and
concentric muscle contractions. Eight week-old Institute of Cancer Research (ICR) mice (n=90)
were divided into three groups: eccentric muscle contraction group (ECC, n=42), concentric muscle
contraction group (CON, n=42), and control group (pre, n=6). The tibialis anterior muscle was
stimulated via the peroneal nerve to contract either eccentrically or concentrically. The tibialis
anterior muscle was isolated before and 0, 6, 12, 18, 24, 72, and 168 hours after muscle contraction.
Immediately after muscle contractions, thiobarbituric acid reactive substances (TBARS) in skeletal
muscle significantly increased (p< 0.05) in both ECC and CON conditions. However, in ECC group
alone, the TBARS level peaked at 12 and 72 hours after the contractions. There was greater
migration of mononuclear cells in ECC than CON muscle. In addition, there were a correlation
between TBARS in skeletal muscle and migration of mononuclear cells in ECC muscle (r= 0.773,
p<0.01), but not in CON (r=0.324, p=0.12). The increased mononuclear cells may reflect
inflammatory cells. These data suggest that eccentric muscle contraction induces greater oxidative
stress in skeletal muscle, which may in turn be due to enhanced generation of reactive oxygen
species by migrating inflammatory cells.
P-17
Physiological, muscle injury-related and leukocyte subset responses to exercise and cold
exposure in short track and inline skaters
Kijin Kim1, Katsuhiko Suzuki2, 3, Jonathan Peake 4, Nayoung Ahn1, Junghee Hong5, Hyunjeong
Kim 5, Kishiko Ogawa2, Changbae Hong1, Sanghyun Kim1, Inseon Lee5, Yoonjung Shin1 and
Kyungmin Gong1
1
Department of Physical Education, College of Physical Education, Keimyung University, Daegu, Korea
Faculty of Human Sciences, Waseda University, Tokorozawa, Japan
3
Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, Tokyo, Japan
4
School of Human Movement Studies, University of Queensland, Brisbane, Australia
5
The Center for Traditional Microorganism Resources, Keimyung University, Daegu, Korea
2
Objective: We investigated physiological responses, possible development of muscle injuries and
changes in circulating immune cells following exercise in cold and warm conditions. Methods:
Participants were short track skaters (n=9) who were acclimatized to cold conditions, and inline
skaters (n=10) who were not acclimatized. All skaters were young, and skating at a recreational
level three days per week for at least one year. Using a cross-over design, study variables were
measured during 60 min of submaximal cycling (65%VO2max) in cold (ambient temperature:
5±1ºC, relative humidity: 41±9%) and warm conditions (ambient temperature: 21±1ºC, relative
humidity: 35±5%). Heart rate, blood lactate and tympanic temperature were measured at rest,
during exercise and recovery. Plasma cortisol, calprotectin, lipid profiles and circulating blood cell
numbers were measured before and after 60 min of cold or warm conditions, and during recovery
from exercise. To estimate possible muscle injury in the cold condition, creatine kinase (CK),
lactate dehydrogenase (LDH) and myoglobin (Mb) were also measured. Results: Heart rate was
lower in both groups during exercise in cold versus warm conditions (P<0.05). The inline skater
group was significantly (P<0.05) higher CK values in the cold than warm condition, while the short
track skater group showed no significant difference between cold and warm conditions. Level of
Mb in inline skater group significantly elevated during recovery phase in the cold compared with in
the warm condition. The increase in total leukocytes during recovery was primarily due to an
increase in neutrophils in both groups. The short track skater group increased neutrophils after
exercise in cold exposure, whereas the non-acclimatized group increased lymphocyte and cortisol
after exercise in cold exposure. Lymphocyte subsets significantly changed in both groups over time
during recovery as compared to rest. Conclusion: Acclimatization to exercise in the cold attenuates
exercise-induced injury in cold conditions. The greater lymphocyte and cortisol responses in the
non-acclimatized group after exercise in cold conditions may be associated with acute phase
responses of the immune system due to lack of adaptation to cold conditions, while acclimatization
increases transiently an immunosurveillance through neutrophil recruitment after exercise
independent of muscle injury in cold conditions.
Key words: cold, warm, CK, myoglobin, neutrophil, lymphocyte
P-18
Reduced nitric oxide production response to lipopolysaccharide in skeletal muscle is
associated with the surface poor in CD14 expression
Noriaki Kawanishi1, Daisuke Shiva2 and Hiromi Yano 1
1
2
Department in Health and Sports Science, Kawasaki University of Medical Welfare, Okayama, Japan
Division of Health Science, Kawasaki University of Medical Welfare, Okayama, Japan
Objective: Infection decreases behavior performance. In fact, lipopolysaccharide (LPS), which is
outer membrane of gram-negative bacteria, induces skeletal muscle contractile dysfunction. LPS
binds CD14/toll like receptor (TLR) 4, and then actives NF-kB and IRF-3, which induce nitric
oxide (NO) and pro-inflammatory cytokines productions. However, LPS-induced NO production in
skeletal muscle was largely unexplained. It is known that NO increases risk of bacterial infection
although NO can protect invasion of neutrophil and macrophage. The purpose of this study was to
clarify the effect of LPS on NO production in skeletal muscle cells. Materials and methods:
C2C12 cells, mouse skeletal muscle cell line, and RAW 264.7 cells, mouse macrophages cell line,
were used to assess LPS-induced NO, IFN-r and MCP-1 productions. Furthermore, we evaluated
inducible NO synthese (iNOS) mRNA expression using RT-PCR, and cell surface expression of
CD14 and TLR4 using flowcytometry. Results: In C2C12 cells, LPS stimulation neither induced
NO nor IFN-β, which was one of NO inducers. Recombinant IFN-β stimulation enhanced
LPS-induced NO production. In contrast, MCP-1 production was increased by LPS stimulation, but
combined treatment of LPS and NO doner, NONOate, significantly inhibited its production in
C2C12 cells. Interestingly, we found that surface expression of CD14, which regulates IFN-r
production, in C2C12 cells was greatly lower than that in RAW 264.7 cells, although TLR4
expression in C2C12 cells was similar to that in RAW 264.7 cells. Conclusion: These data suggest
that reduced NO production response to LPS might depend on low expression of CD14 on skeletal
muscle, and that reduced NO production might enhance LPS-induced MCP-1 production. Together,
those function of skeletal muscle might be decreased risk of bacterial infection by promoting
inflammation reaction.
P-19
Relationship between leg fatigue and plasma IL-6 concentration on mild intensity of running
exercise
Hiromi Yano1, Syogo Watanabe2, Kodai Harada1, Daisuke Shiva1, Noriaki Kawanishi1, Makoto
Okada2, Hiroshi Kato3, Tomomi Kitawaki2, Takao Nakamura2, Satoshi Ikeda2, Hisao Oka2
1
Kawasaki University of Medical Welfare
Okayama University Medical School
3
Kibi International University, Okayama, Japan
2
Objective: It is well known that severe prolonged exercise increases plasma interleukin (IL)-6,
which may be induced from skeletal muscle, mainly. Furthermore, it was reported that anti-IL-6
receptor antibodies reduced the symptoms of chronic fatigue in healthy humans. However, the
relationship between leg fatigue and changes in plasma IL-6 concentration remains unclear at mild
intensity of exercise. The purpose of this study was to clarify whether plasma IL-6 depend on leg
fatigue during and after mild intensity of exercise. Methods: Voluntary healthy male subjects
(n=10) performed a running exercise at 143 m/min for 90 min on a treadmill. Borg CR10 scale, on
which leg fatigue depends, was measured at 0, 60 and 90 min during exercise, and then 0.5, 1, 3, 6,
24 and 48 hours after the exercise. The collected blood samples at each measuring time of Borg
CR10 scale were examined concentration of plasma IL-6 and serum CK and LDH activities.
Results: In this study, the exercise could induce muscle damage, because we observed that serum
activity of CK, which leaked from skeletal muscles, significantly increased after exercise. In
addition, Borg CR10 scale was significantly increased during and immediately after the exercise
compared with the value of pre-exercise. Plasma IL-6 concentration was significantly increased
during and until 3hour after the exercise. Interestingly, the significant correlation between Borg
CR 10 scale and plasma IL-6 concentration was observed (r = 0.618, p<0.01). Conclusion: These
results suggest that exercise-induced IL-6 in plasma might be involved in leg fatigue on mild
intensity of running exercise.
P-20
Biochemical and psychological markers of fatigue and immunodepression in exercise
Brown, D.,1 Simpson, J.,1 Gough, L., 1,3 Baskerville, R.,2 and Castell L.M.1
1
2
CNRG, University of Oxford OX2 6HE, School of Sport and Education, Brunel University, UB8 3PH
27 Beaumont Street, Oxford OX1 2NR, School of Sport and Education, Brunel University, UB8 3PH
Introduction: This 8-week pilot study investigated potential biochemical and psychological
markers of fatigue and immunodepression in rowing and treadmill running. Methods: Study 1:
Open weight (OW, n=14) and light weight (LW, n=7) female rowers consented to participate.
Questionnaires were completed daily on incidence of upper respiratory tract infections (URTI),
gastrointestinal (GI) disturbances, subjective mood states (POMS). Rate of perceived exertion
(RPE) was measured in OW during a 1hr/wk ergometer session; resting/fasting blood samples were
taken in Wks 1, 4, 8. Plasma C-reactive protein (CRP) was measured on an auto-analyser; plasma
glutamine (p[Gln]) and glutamate concentrations (p[Glu]) were analysed with separate enzymatic
assays. Study 2: Male runners consented to 40 min treadmill running at 5% below lactate
threshold (LT). Blood samples were taken Pre, Post, 30min Post and 1hr Post exercise. p[Gln],
p[Glu], p[Alanine] were measured.
RPE was measured before and after exercise. Results: Study
1: Compliance was poor and complete questionnaire data was only obtained from 5 OW and 5
LW; complete blood sample data was obtained from 12 OW. No significant changes were seen
overall in mood state or incidence/duration of illness in either OW or LW, but LW had significantly
higher motivation, depression and fatigue ratings cf. OW (p<0.01). p[Glu] significantly increased
from wk 1-8 (p<0.001, n=12); the p[Gln]/Glu] ratio significantly decreased concomitantly (p<0.01,
n=12) with an increase in RPE. This correlated positively with URTI (p<0.05, n=5) and all profile
of mood state (POMS) parameters (p<0.01, n=5). The incidence and severity of gastrointestinal
disturbances correlated strongly with percentage body fat in 5 OW (r=0.976, p=<0.01).
It is
speculated this might be due to high caloric intake. CRP did not change throughout the study.
Study 2:
p[Gln/Glu] decreased significantly at 30 min post exercise (50%; p<0.01); this did not
correlate with increased RPE during exercise. p[Ala] (18%, p<0.05) increased post-exercise,
correlating with RPE.
Five athletes with high RPE (fatigue) had significantly higher p[Ala]
(p<0.05) compared with those who had low RPE (n=6). The p[Gln/Ala] ratio was significantly
decreased at 30 min post exercise (p<0.05) but did not correlate with RPE. Conclusions:
p[Gln/Glu] and RPE show promise as potential markers of fatigue and/or immunodepression and
mood state disturbance in female rowers and male runners.
In addition, p[Ala] may be a marker of fatigue.
P-21
Non-invasive measures of some cytokines in response to exercise
Castell, L.,1,2 Gough, L. 1 and Godfrey, R.2
1
2
CNRG, Nuffield Dept of Anaesthetics, University of Oxford OX2 6HA
School of Sport and Education, Brunel University UB8 3PH
Introduction: Venous blood sampling is invasive and not always suitable for data collection from
athletes. Saliva and urine are simply collected non-invasively, not requiring a phlebotomist.
Markers relevant to physiological monitoring of athletes are present in saliva and urine, including
cytokines. This study investigated whether the salivary concentrations of growth hormone (GH)
and IL-6, and the urinary concentrations of GH, IL-6, IL-1ra and IL-10 reflected circulating
concentrations before and after an acute bout of exercise. Methods: Eleven healthy males (mean
±SD, age 26 ±5 yrs, height 179 ±7cm, weight 77 ±9kg, VO2 peak 57 ±6 ml/kg/min) consented to a 40
min run at 5% below lactate threshold after overnight fasting. Blood and timed, unstimulated
saliva samples were collected at baseline (Pre), immediately after exercise (Post), 30 min post
(Post-30) and 60 min post (Post-60). Midstream urine was collected Pre, Post and Post-60. Water
intake was controlled. Cytokine concentrations were measured using ELISA kits. Repeated
measures ANOVA with pairwise comparisons indicated changes with time; correlations indicated
relations between variables. Results: Circulating concentrations of IL-6, IL-1ra and IL-10 were
increased by the exercise (p<0.001). There was no effect of exercise on salivary and urinary IL-6,
or IL-1ra. Serum GH was increased at Post (p<0.001); salivary GH was increased at Post, with a
further increase at Post-30 (p<0.001); urinary GH was increased at Post (p<0.01) with a further
increase at Post-60 (p<0.05). Salivary and urinary GH correlated with serum GH (rs=0.412 and
0.434 respectively). Discussion: In contrast to clinical studies, non-invasive measures of IL-6,
IL-1ra and IL-10 do not appear to be a suitable alternative to venous blood sampling after exercise
of this type. In studies on intensive care patients there was a marked correlation between plasma
and urinary IL-6 (Adam et al., 2006).
In the present study, salivary and urinary GH
concentrations were responsive to exercise and correlated with serum GH.
However, serum GH
peaked immediately post-exercise, whereas salivary GH peaked at 30 min post-exercise and urinary
GH was increased further at one hr post-exercise.
The functional significance of the
exercise-induced increase in salivary and urinary GH is unclear and may be related to the clearance
of GH from the circulation.
Reference
Adam Z, Wardman S, Garrard C, Tomlin G, Castell L.M. (2006).
Intensive Care & Emergency Med., Brussels, March, 2006
26th Int. Meeting
P-22
Evaluation of the IMMULITE® Automated Immunoassay System for the detection of IL-6
and IL-10 in Saliva for Sport, Exercise & Health Science Applications
Andrew Misiura
University of Gloucestershire, Department of Health & Physical Activity, Gloucester, UK.
Objective: To evaluate the use of the Immulite® automated, chemiluminescent enzyme
immunometric assay system for the determination of saliva IL-6 and IL-10 levels. Methods: 20
healthy University Sport, Exercise & Health Science Students (mean age 20) volunteered to
exercise on treadmill using an intermittent protocol: 5 minutes warm up at 50% maximum heart
rate (MHR), followed by 10x30s bouts of inclined (-30%) treadmill running at 95% MHR and then
a 10 minute cool down of slow jogging in an air conditioned laboratory (18’C). Samples of non
artificially stimulated saliva were provided at rest, after the intense exercise period and then again at
20, 40 and 60 minutes post exercise by dribbling into a 2ml microcentrifuge tube (Eppendorf®).
Samples were loaded onto the IMMULITE® immediately following collection and cytokine was
measured after 60 minutes incubation with the antibody coated bead and agitation at 37’C (IL-10)
or 90 minutes (IL-6). The assay utilizes monoclonal anti-IL-6 and IL-10 antibodies on the solid
phase and an enzyme-labeled monoclonal detection anti-body that are coincubated with saliva.
Results: Neither IL6 nor IL10 were detectable at rest (sample 1,< 5pg/ml), IL6 peaks at the end of
the exercise at a mean value of 28.8 pg/ml (p<0.05) (sample 2) and then declines post exercise
(samples 3(p<0.05) and 4(p<0.05)). IL10 peaked 20 minutes post exercise with a mean value of
7.6mg/ml (sample 3(p<0.05) and then declines. Conclusion: IL6 and IL10 are detectable down to
7.6pg/ml following intermittent maximal exercise using the Immulite automated immunoasay
system with a reported detection limit of 5pg/ml using unstimulated and unprocessed 1.5ml saliva
samples. It appears that both the magnitude and the timing of the IL6 and IL10 responses differ,
although as anticipated, the pro-inflammatory IL6 increase is associated with an anti-inflammatory
IL10 rise, albeit delayed. Saliva is a more convenient medium and changes in salivary interleukins
have been detected where there were no detectable changes in blood.
P-23
Exercise training increases adiponectin in elderly males and females
Melissa M. Markofski1, Michael G. Flynn1, Kyle L. Timmerman2, Paul M. Coen1, Brandt Pence1
1
2
Wastl Human Performance Laboratory, Health & Kinesiology Department, Purdue University, West Lafayette, IN
Sealy Center on Aging, University of Texas Medical Branch, Galveston, TX USA
Objective: The purpose of this study was to examine the effect of a 12-week exercise training
program on adiponectin, sTNFαRII, and unstimulated TNFαproduction in whole blood cultures.
Methods: Fifteen physically inactive males and females between the ages of 65-80 were recruited
for the intervention portion of the study. Fifteen age-matched physically active subjects served as a
comparison group and participated in two rounds of testing separated by 12 weeks. The 12-week
intervention (3d/wk) consisted of 20 minutes of treadmill walking at 70%-80% of heart rate reserve
and eight resistance exercises at 80% of 1-RM. Blood was collected at baseline and at least 72 hours
following the last training session. Heparinized whole blood was diluted (1:10) with RPMI and
incubated (37ºC, 5% CO2) for 24 hours. The supernatant was centrifuged (10 minutes 800g),
filtered, and stored for later TNFα analysis. Serum was collected and stored (-80ºC) for subsequent
analysis of adiponectin and sTNFαRII using ELISA. Results: Twelve weeks of exercise training
led to a significant (p<0.05) increase in muscular strength (leg press 1RM 153.2±46.2 kg pre vs.
177.5±44.1 post) and adiponectin concentration (13.9 ±9.2µg/mL pre vs. 21.6±17.3 post).
Adiponectin was correlated with indices of body morphology at baseline, body weight (r =-0.45;
p=0.02) percent body fat (r =0.51, p=0.01), and at the end of the study period, percent fat (r =0.60,
p=0.002). Adiponectin was negatively correlated with markers of strength (leg press 1-RM) at
baseline (r =-0.64, p=0.0006) and at the end of the 12 week study period (r =-0.53, p=0.01), a
finding that was somewhat unexpected. After training, sTNFαRII, a marker of TNFα activity, was
negatively correlated with VO2max (r =-0.38, p=0.04). Unstimulated TNFα and weight (r =0.44,
p=0.02) were associated, but surprisingly adiponectin and sTNFαRII were also positively correlated
(r =0.45, p=0.01). Conclusion: Exercise training increased adiponectin concentration in elderly
males and females without a significant change in body composition. Adiponectin was, as expected,
correlated to indices of fat mass, but also related to measures of strength.
P-24
Low-grade inflammation is not associated with soft-drink consumption in healthy adolescents
Julia Wärnberg1, Javier Romeo1, Imma Palma2, Ligia E. Diaz1, Sonia Gomez-Martinez1, Maria
Isabel Mesana3, Francisco B. Ortega4, Carlos Redondo5, Ascensión Marcos1 and the AVENA study
group.
1
Immunonutrition research group. Dept. Metabolism and Nutrition. Instituto del Frio. Spanish National Research
Council (CSIC). Madrid. 2 Centro de Enseñanza Superior de Nutrición y Dietética (CESNID), Barcelona. 3 E. U.
University of Zaragoza, Zaragoza. 4 University of Granada, Granada 5University of Cantabria, Santander. Spain
Objectives
Inflammation plays a significant role in the pathogenesis of cardiovascular disease, and low-grade
chronic inflammation has been detected already in adolescents. There are ongoing debates of the
implication soft-drink consumption may have on obesity and obesity related diseases. The aim of
this report was to examine how soft drink consumption, obesity and low grade inflammation are
interrelated in young healthy adolescents.
Methods
The Spanish study AVENA is a cross sectional multi-centre study, with adolescents aged 13-18 y
recruited from schools. Body fat percentage was estimated through skinfold thicknesses and a 24h
recall established water and soft drink consumption of the previous day. To study inflammation,
serum C-reactive protein (CRP) was analysed in a fasting blood sample. Subjects were divided in
non risk <1mg/L or risk ≥1mg/L groups, according to AHAs and CDCs adult cut-off points.
Results
291 adolescents (54% males) had CRP values and data on soft drink consumption and were
included in this report. 32% of the subjects had CRP concentrations ≥1mg/L which puts them in
risk for future cardiovascular disease. The CRP risk group drank less soft drinks compared to the
non risk group (mean values were 74mL vs 107mL in females, and 114mL vs 166mL in males,
respectively without reaching significantly difference: P>0.05), while the risk group had
significantly higher body fat percentage.
Conclusion
The interrelationships between determinants and consequences of overweight in adolescence are
complex and single foods such as soft drinks derived from a 24h recall cannot be associated in
cross-sectional studies of apparently healthy subjects. The mechanisms should be studied and
elucidated in complex obesogenic environments, and in longitudinal settings.
Study supported by the Spanish Ministry of Health, FIS nº 00/0015, and student grants from the
Consejo Superior de Deportes (05/UPB32/0, 109/UPB31/03 and 13/UPB20/04) the Spanish
Ministry of Education (AP2003-2128;AP-2004-2745), and grants from Coca-Cola España, Panrico,
Madaus and Procter and Gamble
P-25
Effects of 5-day Kendo practice on lymphocyte level and expression of lymphocyte apoptosis
in young Kendo athletes
Yuko Tanimura1, Michihiro Kon1, Kazuhiro Shimizu1, Fuminori Kimura1, Ichiro Kono1 and
Ryuichi Ajisaka1.
1
Graduate School of Comprehensive Human Sciences, University of Tsukuba, Ibaraki, Japan
Introduction: It is controversial whether lymphocyte apoptosis contributes to post-exercise
lymphocytopenia. No research has demonstrated that athletes had lymphocytopenia with
lymphocyte apoptosis during repetitive prolonged exercise. Objective: The purpose of the present
study is to examine the effects of 5-day Kendo practice on lymphocyte level and expression of
lymphocyte apoptosis in young Kendo athletes. Methods: The subjects were eight healthy,
well-trained males (age: 19.7±1.0 years, athletic career: 12.4±1.9 years) belonging to a college
Kendo club. All subjects underwent the Kendo practice of about 5 hours (AM; 140 minutes, PM;
170 minutes) a day for 6 days and took the same meal during the training camp. Blood samples
were collected at two weeks before (PRE), the first day (Day 1), third day (Day 3), fifth day (Day 5),
and 1 week after the training (POST) to determine lymphocyte level and CD95 expression on
lymphocyte (CD4+, CD8+) by flow cytometry. Sequential changes were assessed by repeated
measures analysis of variance (ANOVA) with Bonferroni/Dunn post-hoc test. Statistical
significance was accepted at p<0.05 in ANOVA, and at p<0.005 in post-hoc test. Results:
Lymphocyte counts tended to decrease at Day 1 (1650±420 cell/µl, p=0.045 ) and significantly
decreased at Day 3 (1494±506 cell/µl, p<0.005) compared with PRE (1919±583 cell/µl)and
gradually recovered thereafter (Day 5 ;1729±465 cell/µl, POST; 1749±366 cell/µl). The
percentage of CD95 positive lymphocytes significantly increased (p < 0.05) at Day 1 (57.9±11.2 %)
and Day 3 (76.5±6.1 %) compared with PRE (41.8±6.3 %). The percentage of CD4+ CD95 + cells
significantly increased (p < 0.05) at Day 1 (24.9±6.3 %) and Day 3 (34.0±3.6 %) compared with
PRE (17.3±4.6 %). The percentage of CD8+ CD95 + cells did not significantly change during the
training. Conclusion: It was suggested that 5-day Kendo practice induced lymphocytopenia with
apoptosis, especially that of CD4+ CD95 + cells in young Kendo athletes.
P-26
The effect of prolonged cycling for 15 consecutive days on neutrophil responses in untrained
cyclists
Tzai-Li Li1, Yen-Chen Chiu2, Wei Hung2, Chi-Wei Chiang3, Ching-Lin Wu4, and Cheng-Kang
Chang4
1
Department of Sports and Leisure, National Dong-Hwa University, Hualien, Taiwan
Department of Exercise Health Science, National Taiwan College of Physical Education
3
Sport Science Research Center, National Taiwan College of Physical Education
4
Department of Athletics, National Taiwan College of Physical Education
2
Objective: The aim of this study was to determine the effect of consecutive bouts of prolonged
exercise on neutrophil responses. Methods: With the approval of the local Ethics Committee, 20
healthy men and 7 women (age 20.30 ± 0.37 years, body mass 62.07 ± 1.65 kg; body fat percentage
17.47 ± 1.28%; VO2max 40.60 ± 1.65 mL.kg-1.min-1; means ± SEM ) voluntarily participated in
this study. Subjects cycled de Taiwan for 15 consecutive days. Venous blood samples were
collected at 06:30 before exercise on day 1 (0 km), day 5 (282 km), day 10 (704 km), and day 15
(1099 km). Haematological analysis was performed using an automated cell counter. Phorbol
myristate acetate (PMA) induced oxidative burst activity was measured using a chemiluminescence
(CL) assay (Kinght Scientific Limited, Plymouth) and bacteria-stimulated neutrophil degranulation
(elastase release) was determined as described by Li and Gleeson (2005). Results were analyzed
using a repeated one way ANOVA with post hoc Bonferroni adjusted multiple comparisons.
Results: The main findings of this study were: (1) there were no significant alterations in
circulating neutrophil counts, but significantly decreased leukocyte and lymphocyte numbers (both
P<0.01) during the period of de Taiwan; (2) neutrophil oxidative burst activity was increased at day
5, day 10, and day 15 and neutrophil degranulation was increased at day 5 and day 10 on per cell
basis (all P<0.05) compared with day 1. Conclusion: The findings of this study suggested that the
consecutive bouts of prolonged exercise may enhance neutrophil functions at resting status.
Keywords: neutrophil, consecutive bouts of prolonged exercise
Reference: Li, T. L., & Gleeson, M. (2005). Eur. J. Appl. Physiol., 95, 391-399.
Acknowledgement: This study was supported by Taiwan National Science Council
(NSC-95-2413-H-028-002)
P-27
Disappearance of an allergenic activity of ovomucoid, a major egg allergen, by formation of a
complex with wheat gluten was assessed by voluntary physical activity in mice
Yasuko Kato and Kana Joo
Graduate School of Medical Professions, Kawasaki University of Medical Welfare, Kurashiki, Okayama 701-193 Japan
Objective: Ovomucoid (OM) is a major egg allergen and the allergic activity is not disappeared by
long-time heating, because OM is soluble after the heating. We reported that the complex of OM
and wheat gluten lose the solubility based on the disulfide bond between them. It is therefore
necessary to clarify that an allergic symptom after absorbed eating the egg does not develop. In this
study, we assessed the allergenic activity of the complexes after oral administration in mice with
voluntary physical activity.
Methods: OM-specific IgE values in OM-sensitized mouse sera were measured by ELISA. A
complex of OM and gluten was prepared as follows; a mixture of egg and wheat flour in water was
kneaded for 30 min, benched for 1 hr and boiled in salt solution for 15 min. Three groups of the
sensitized mice (n=21) were orally administrated OM, OM-gluten mixture or heated OM-gluten
complex (Dosage of OM were assumed to be the same; 10mg/mouse). The mice administrated
samples were housed individually in cages with a rotation wheel, and the revolutions were counted
for 15 hr.
Results: The systematic anaphylaxis of the sensitized mouse administrated samples was assessed
by voluntary physical activity, comparing with rectal temperature and anaphylaxis score. The every
hourly revolution-counts of the sensitized mice orally ingested with OM and OM-gluten mixture,
significantly decreased and the lower count were continued for 12 and 9 hr, respectively. Whereas
the activity of OM-gluten complex group did not decreased.
Comparing 15-hr count among
them, the mean counts of OM and mixture of OM-gluten groups decreased to 19% and 26%,
respectively, however the count of the OM-gluten complex group indicated a slight decrease. These
results agreed well with the assessment by rectal temperature and anaphylaxis score. The voluntary
physical activity would be a novel indicator for the analyses of the active systemic anaphylaxis and,
the allergenic activity of OM was disappeared by the formation of insoluble OM-gluten complex
based on the S-S bond.
Conclusion: These results suggested the possibility that allergenic activity of egg in wheat
processed foods such as bread and pasta is disappeared by assessment of the voluntary activity.
ISEI2007 Local Organizing Committee
Chairman
Ryoichi Nagatomi, (Tohoku Univ., President of ISEI)
Vice Chairman
Masatoshi Suzui, (Meiji Univ.)
Hiromi Yano, (Kawasaki Univ. Medical Welfare)
Katsuhiko Suzuki, (Waseda Univ.)
Mitshuharu Okutsu, (Waseda Univ.)
Yuzuru Sakamoto, (Shinshu Univ.)
Takayuki Akimoto, (Univ. of Tokyo)
Hiromi Kitamura, (Wayo Women’s Univ.)
Kishiko Ogawa, (Waseda Univ.)
Satoru Moriguchi, (Yamaguchi Pref. Univ.)
Scientific Committee
Michael Gleeson (Loughborough Univ., Vice President of ISEI)
Ryoichi Nagatomi, (Tohoku Univ., President of ISEI)
Acknowledgement of Sponsorship
The ISEI2007 Local Organizing Committee gratefully acknowledges the financial support and/or sponsorship that
has been provided by the following institutions:
•
Asian Office of Aerospace Research & Development/Air Force Office of Scientific Research
•
Otsuka Pharmaceutical Co., Ltd
•
POLA Pharma Inc.
•
SYSMEX CORPORATION
•
Gonryo Medical Foundation