Slide 1 :Oxigen: Vital but Dangerous...about oxidative stress in the dog Dominique GRANDJEAN
DVM, PhD, HDR
Colonel, Chief veterinarian, Paris Fire Brigade
Professor, Alfort National Veterinary School
Head of Canine Breeding and Sport Medicine Unit
OXIDATIVE STRESS :OXIDATIVE STRESS State within oxidative reactions overpass the anti-oxidant defenses of the organism Disturbances of cell functions Pathology Acute Chronic
Slide 3 :Acute and temporary oxidative stress Long term disbalance or chronic oxidative stress Base Level Level of Oxidative Stress Acute pathological problems
[Sport Medicine] Chronic diseases Muscle Heart Intestin Cancer Nervous syst Eye Heart Joints
Oxidative stress in racing dogsConsequences and Prevention :Free Radicals Production; consequences
Main related pathological problems
Dogs working in extreme conditions: a model
Nutritional and environmental consequences Oxidative stress in racing dogsConsequences and Prevention
Free Radicals(toxic reactive species of oxigen) :Cellular lesions Lipoperoxidations
Degradation of proteins
Destruction of DNA Free Radicals(toxic reactive species of oxigen) ’O2 O2- H2O2 OH . Antiradicals defense systems Enzymes
Chelators of minerals
Vitamins
Slide 6 :Who are they? Highly unstable toxic compounds,
very short life span,
generated by oxigen Free Radicals
Slide 8 :Examples of free radicals H• Unstable hydrogen CCl•3 Trichloromethyle O• - Superoxide 2 OH• Hydroxyle RS• Thiol
Slide 9 :Normal conditions: Redox reactions in each cell
Induced or increased by environment factors
X rays
Polluants (pesticides,…)
Radiations
Tobacco How are they produced ? Free Radicals
Slide 10 :Free Radicals What are their biological actions? Inactivations of enzymes
Destruction of cell membranes
Destruction of mitochondrias
Hemolysis of red cells
Impairment of protein synthesis
: FREE RADICALS ACTIVATE
INFLAMMATORY CELLS
: CELLULAR AND ORGANS LESIONS INDUCED BY FREE RADICALS
Slide 13 :Free Radicals Chain reaction responsible for the destruction of lipids in membranes, of structure proteins, of nucleic acids
Directly responsible of lesions leading to:
aging
poor performance
cancers
muscle diseases
tendons and joints diseases
heart dysfunctions
neurological dysfunctions What are their consequences?
Slide 14 :Free Radicals Is the organism able to defend itself? Superoxide dismutase Uric acid Betacarotene Polyphenols Catalase Vitamines E and C Glutathion peroxidase Coenzime Q10 Ceruloplasmine Transferrine
Slide 15 :ANTI OXIDANT SYSTEMS
Structure of a normal cell membrane :Structure of a normal cell membrane Fatty acids Glycoproteines
Slide 17 :Structure of a cell membrane destryed by free radicals Malondialdehide
Pentane - Ethane Oxidation of aminoacides Peroxidation Rupture
Slide 19 :NORMAL MUSCLE FIBERS OXIDATIVE STRESS Destruction of muscle fibers Dysfunction of muscle vessels
Slide 20 :Muscle lesions induced by
Cellular Oxidative Stress
Slide 22 :Future stress fracture
Slide 23 :Production of
ROS Oxidative stress Waste
(parts of destoyed compounds) Initiation
Functional abnormalities
Pathological consequences Antioxidants Fixing process
Slide 24 :Production of free radicals
O2- ., H2 O2 , OH. Antioxidants
Vit E, Vit C, Glutathion, SOD... Pathology Health
Oxidative stress in racing dogsConsequences and Prevention :Oxidative stress in racing dogsConsequences and Prevention Free Radicals Production; consequences
Main related pathological problems
Dogs working in extreme conditions: a model
Nutritional and environmental consequences
Slide 26 :Oxidative Stress induced pathologies
Slide 27 :Aging
Neurological degenerations
Diabetis
Arthrosis
Chronic Kidney Failure
Stress Diarrheas
Rhabdomyolysis
Ophtalmological Diseases
…
Slide 28 :EMOTIONAL STRESS AND LIPOPEROXYDATION 150% 200% 1 2 3 Pentanes in outcoming air days [Philipko, 1982] 15 mn 4 5 Emotional Stress
Oxidative stress in racing dogsConsequences and Prevention :Free Radicals Production; consequences
Main related pathological problems
Dogs working in extreme conditions: a model
Nutritional and environmental consequences Oxidative stress in racing dogsConsequences and Prevention
Slide 30 :Stamina in aerobiosis oxidation processes
High level of oxygen consumption free radicals
Source of energy oxidation of lipids
Extreme environments environmental stress
Intense work, motivation psychological stress WHY??? Conditions for an intense cellular oxidative stress
Slide 31 :Intense stamina in cold weather
Intense stamina in altitude HOW?
Slide 32 :Intense
Exercise Aerobic
Power Hypoxia
Ischemia-Reperfusion Inflammation Pathophysiology ?
Slide 33 :Oxydative stress and Stamina in dogs Bardlay, 1991 ROS are a cause of fatigue in racing dogs Jerome, 1994 Ischemia induces production of ROS in the muscle of racing dogs Mo, 1997 Hypoxia generates production of ROS in the muscle Weinstock, 1997 Physical exercise induces the production of inflammatory molecules in the muscle of racing dogs
Slide 34 :• Dogs on a treadmill
• 40 km/d, 5 days/week, 55 weeks
speed : 6-7 km/h, slope : 15 p.100 • Total Glutathion in Muscle and Liver Endurance training improves anti-oxidant reserves of the organism Oxidative Stress and Stamina in Dogs
Slide 35 :Destruction of cell membranes
Lesions Death ± muscle hypoxia
± muscle ischemia-reperfusion High intensity exercise Free Radicals
« Chiens des cimes » : the concept :Increase capacities of « dogs who save lives « Chiens des cimes » : the concept
« Chiens des cimes » : the concept :Increase capacities of « dogs who save lives » Increase working time Prevention of
injuries « Chiens des cimes » : the concept Resistance to stress
« Chiens des cimes » : the concept :Limited practical working time 15 min in 1990
40 min in 2004
? Selection Intense physical work Concentration Motivation Extreme environment Environment Nutrition Ergogenic aids « Chiens des cimes » : the concept
« Chiens des cimes » : the concept :Intense cellular oxidative stress prevention Motivation Intense physical work Concentration Motivation Extreme environment Education Antioxidant substances Dedicated exercises Handler’s stress Outsiders Dual Nutritional Ergogenic « Chiens des cimes » : the concept
« Chiens des cimes » : the concept :Dedicated pathological problems - SDDS
- Rhabdomyolysis
- Water diabetis
- … Nutrition Ergogenic aids Traumatology Propioception Stress pathology « Chiens des cimes » : the concept Intense physical work Extreme environments Handler’s stress Outsiders Concentration Motivation
« Chiens des cimes » : the concept :Why extreme environments ? Real operations can occur everywhere
Very high levels of oxidative stress
Larger biological modifications « Chiens des cimes » : the concept
« Chiens des cimes » : the concept :O2 O2 O2 Stress UV … Polluants Toxic free radicals « Chiens des cimes » : the concept
Slide 43 :«Chiens des cimes - Licancabur» - From sea level 4 500 m in less than 24 hours
- Inca rubbles between 2 500 m and 5 500 m
- No snow
- « Easy » climbing 6000 m Licancabur (Chile)
Slide 44 : Cynotechnic teams . Sapeurs Pompiers de Paris (France)
. Carabinieros de Chile
. UMES
. Royal Canin Scientific teams Logistic team Members of the expedition «Chiens de cimes - Licancabur»
Slide 45 :France: 35/20 Dry food + Vitamin E (500 mg/d) Vitamin C (500 mg/d) Omega 3 (300 mg/d) Chili : 35/20 Dry food Two groups of dogs Scientific protocole «Chiens des cimes - Licancabur»
Slide 46 :Paris (France) Santiago (Chile) Antofagasta 2500 m San Pedro
de Atacama 4500 m Base camp
(Bolivia) 6000 m «Chiens des cimes - Licancabur»
Slide 47 :1- Knowledge about « acute mountain sickness »
2- Efficiency of the nutritional approach
Dry food super premium
Anti oxidant protection
Quality of fatty acids in the food «Chiens de cimes - Licancabur» Conclusions
Licancabur ; Chile ; 6000 meters :Group 2 >> Group 1 Biological and nutritional consequences of worrk at high
altitude in search and rescue dogs :
The scientific expedition Chiens des Cimes-Licancabur
The journal of nutrition, Vol. 128, No 12S, December 1998, 2694S-2697S Plasma vitamin E
Peroxidation Resistance Index
Oxygen transfer to working cells
Clinical problems
- stress diarrhea
- muscle stiffness and rhabdomyolysis
- acute pulmonary oedema Licancabur ; Chile ; 6000 meters
Slide 49 :Chiens des cimes :
Mont Blanc 2004
Slide 50 :Mont-Blanc 2004 Test level 0
Slide 51 :Mont-Blanc 2004
Slide 52 :Mont-Blanc 2004 : Results 1/ Heart function 2/ Cellular oxygenation 3/ Cellular oxidative stress
Slide 53 :Mont-Blanc 2004 : Results Cardiac function OXYGEN DEBT ? PROPENTOFYLLINE PLACEBO
Slide 54 :ECG Sinusal Respiratory Arythmia (SRA) cyclic
Exists in the 2 groups = sporting dogs (trained) Sinusal rythm OK - Dog : Tarun Mont-Blanc 2004 : Results
Slide 55 :ECG Under segment ST :
Base line / segment ST
=> Whitnesses an heart muscle hypoxia
0,2 mV < NORM < + 0,15 mV
Intense altitude work
Treated group : OK P P Q Q S S R R T T Dog : Patcho PQRS complex – Work in altitude – Treated group Base line Mont-Blanc 2004 : Results
Slide 56 :ECG PQRS complex – Work in altitude – Placebo group Dog : Malouk P P Q Q S R R T S T Under segment ST :
Segment ST
=> Whitnesses a heart muscle hypoxia
0,2 mV < NORM < + 0,15 mV
Intense altitude work
Placebo group: severe hypoxia of the heart muscle -0,4 mV Base line Mont-Blanc 2004 : Results
Slide 57 :ECG % of dogs with no myocardial hypoxia in the 2 groups Mont-Blanc 2004 : Results
Slide 58 :1/ Heart function 2/ Cellular oxygenation 3/ Cellular oxidative stress Mont-Blanc 2004 : Results
Slide 59 :Oxygen saturation Evolution of the rate of oxygen saturation of hemoglobin (plaine vs altitude) Mont-Blanc 2004 : Results
Slide 60 :1/ Heart function 2/ Cellular oxygenation 3/ Cellular oxidative stress Mont-Blanc 2004 : Results
Slide 61 :Evolution of plasma vitamin E levels
Sea level vs Altitude Placebo Propentofylline Vitamin E (mol/l)
Slide 62 :Total Plasma Anti-Oxidant)
Sea level vs Altitude Placebo Propentofylline TPAO
(mmol/l)
Slide 63 :ANTIOXIDANT STATUS
OF RACING SLED DOGS
Slide 64 :DOGS : Vitamin E deficiency found in necropsics “... the important thing is all three of the dogs
with muscle inflammation or degeneration had
critically low values of vitamin E...” VITAMIN E : Magic Pill or red herring “... Nutrient may be key to dog health...” FAIRBANKS DAILY NEWS-MINER, SUNDAY, MAY 4, 1997
Slide 65 :Exams find vitamin deficiencies “... three of the five dogs that died running the Iditarod
Trail Sled Dog Race last month suffered from vitamin E
deficiencies...” Ulcers killed Iditarod dogs “... Iditarod needs to pay attention to how much
vitamin E dogs need...” Dead dogs : Veterinary pathologists find deficiencies
in vitamin E “... A buildup of toxic oxidants in the blood, or the
stress placed on the dog’s heart by widespread cellular
and organ failures triggered by the vitamin E storage...” FAIRBANKS DAILY NEWS-MINER, SATURDAY, APRIL 19, 1997
Slide 66 :Vitamin E
Supplement
(mg/kg)
0
5
15
45 Sudden Death
[heart lesion]
+++
+++
++
0 Muscle
Degeneration
+++
++
++
0 Gastric
ulcer
++
++
++
0 [Bengtsson, 1978] Influence of vitamin E supplementatio
on clinical symptoms observed in vitamin E deficient pigs
ALASKA COME BACK RACE 1997 RESEARCH PROTOCOLE UMES-ROYAL CANIN :Jacques PHILIP Helmuth PEER No supplement 800mg Vitamin E / day ALASKA COME BACK RACE 1997 RESEARCH PROTOCOLE UMES-ROYAL CANIN Monique BENE
Slide 68 :40 30 20 10 0 Plasma vit. E (µmol/l) Before [Alaska Come Back 97] After 31,6 41,5 30,9 24,7 Suppl. [1g Vit. E / dog/day] no suppl. EVOLUTION OF PLASMA VITAMIN E IN J. PHILIP’S TEAM
DURING THE ALASKA COME BACK RACE 1997
Slide 69 :40 30 20 10 0 PRI [s] Before [Alaska Come Back 97] After 41,4 42,3 41,0 35,1 Suppl. [1g Vit. E / dog/day] no suppl. EVOLUTION OF THE PEROXIDATION RESISTANCE INDEX
IN J. PHILIP’S TEAM
DURING THE ALASKA COME BACK RACE 1997
Slide 70 :Drop in blood vitamin E in 16 sled dogs racing 57 kms per day for 3 days in a row (Hinchcliff, 1998) 10 (% of contrôl) 20 30 40 50 60 70 80 Control Day 1 Day 3 90 100
Slide 71 :Plasma concentration in isoprostanes in 16 sled dogs racing 57 kms per day
3 days in a row (Hinchcliff, 1998) 100 (% of control) 200 300 400 500 600 700 800 Control Day 1 Day 3
Slide 72 :Vitamin E : 400 to 800 mg / dog / day
correlated to lipidic caloric intake
Alaska Come Back, Wyoming, IRSSDR…
Super Oxide Dismutase : 600 g / dog / day
natural gliadins protected sources
not cheep…
Polyphenols, lignanes, flavonoïds…
actual trials (La Grande Odyssée) Sled dogs in Alaska and Wyoming
Results (IRMSSR) :Results (IRMSSR) Clinical Datas Distance Dogs dropped Definitely Dogs dropped temporarely Group E # 250/dog 0 2** Group P # 120/dog 3* 12*** * - 1 stress fracture
- 2 acute pumonary oedema
** - stress diarrheas
*** - muscle pain, stress diarrheas
Slide 74 :1/ DOES IT INDUCE ANY SIGN OF OXIDATIVE STRESS ? 2/ IS THERE AN INFLUENCE OF THE SIZE OF THE CAGE ? AIR TRANSPORTATION
OF RACING SLED DOGS
Slide 76 :1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8 AIR TRANSPORTATION OF RACING SLED DOGS
UMES-AIR FRANCE RESEARCH PROTOCOLE
Slide 77 :2 weeks supplement
[1g vitamin E/day] DOG
1
2
3
4
5
6
7
8 Kennel
S
S
S
S
L
L
L
L CDG FBK Depart.
30,5
22,5
27,8
31,3
24,6
28,3
31,5
35,2 Arrival
30,5
24,6
27,9
34,4
22,5
27,9
29,2
36,3 Kennel
L
L
L
L
S
S
S
S FBK CDG Depart.
38,6
45,7
40,9
41,5
33,9
41,0
36,8
31,4 Arrival
36,2
40,2
30,1
40,2
33,8
39,9
36,1
30,4 -2,75 -0,72 PLASMA VITAMIN E [µmol/l]
Slide 78 :2 weeks supplement
[1g vitamin E/day] DOG
1
2
3
4
5
6
7
8 Kennel
S
S
S
S
L
L
L
L CDG FBK Depart.
33
35
31
38
30
38
34
38 Arrival
37
33
36
40
31
36
30
34 Kennel
L
L
L
L
S
S
S
S FBK CDG Depart.
41
46
39
43
35
36
37
45 Arrival
32
38
32
36
34
35
41
44 -7,75 -0,75 PEROXIDATION RESISTANCE INDEX (s)
Oxidative stress in racing dogsConsequences and Prevention :Free Radicals Production; consequences
Main related pathological problems
Dogs working in extreme conditions: a model
Nutritional and environmental consequences Oxidative stress in racing dogsConsequences and Prevention
Slide 80 :‘70s ‘80s ‘90s 1997 Rhabdomyolisis on Dunlap’s dogs
Kronfeld moves to high fat diets
Grandjean : High fat diets enhance endurance
Kronfeld : Protein / Energy Ratio
Reynolds : Fat adaptation in racing dogs
Reinhart : Quality of fatty acids
Grandjean : L carnitine, antioxidants
Back to rhabdomyolisis related to high fat diets BIOLOGIC IS IRONIC
Slide 81 :Life in Kennel Musher’s Stress Perception
competition / training Psychological Intensive Training Performance
(races) Repetition
of competitions Physical Nutritional Prevention OXIDATIVE
STRESS Celles and Organs Lesions
Nutritional Anti oxidants :Nutritional Anti oxidants
Nutritional Anti oxidants :Nutritional Anti oxidants TARGETS DNA
Intracell memb
Cell memb hydro
soluble liposoluble both 1 2 3
Slide 84 :ANY QUESTION ? www.dominiquegrandjean.com