Mitochondrial Function: Is it the Key to Preventing and Reversing...
52
Decker Weiss, NMD FASA Mitochondrial Function: Is it the Key to Preventing and Reversing Neurological and Cardiovascular Disorders?
Transcript of Mitochondrial Function: Is it the Key to Preventing and Reversing...
Decker Weiss NMD FASA
Mitochondrial
Function Is it
the Key to
Preventing and
Reversing
Neurological and
Cardiovascular
Disorders
The Start ldquoEndosymbiont Theoryrdquo
ldquoTwo cells began to live together
exchanging some sort of substrate or
metabolite [product of metabolism like
ATP] The association became mandatory
so that now the host cell cannot live separatelyrdquo
1967 paper ldquoOn the Origins of Mitosing Cellsrdquo published in the Journal of Theoretical Biology
scientist Lynn Margulisas viewed httpswwwlivesciencecom50679-
mitochondriahtml
Sept1 2017
Mitochondria have their own DNA
As organelles they are capable of translating messages
encoded in their genes to proteins without using any of
the resources of the eukaryotic cell
httpswwwkhanacademyorgsciencebiologystructure-of-a-celltour-of-organellesvmitochondria-video
Mitochondria are responsible for creating more than 90 of the energy needed
by the body to sustain life and support organ function
Mitochondrial DNA
Mitochondrial disorders Challenges in diagnosis amp treatment
Nahid Akhtar Khan1 Periyasamy Govindaraj1 Angamuthu Kannan Meena2 Kumarasamy Thangaraj1
1 CSIR-Centre for Cellular amp Molecular Biology Nizams Institute of Medical Sciences Hyderabad India2 Department of Neurology Nizams Institute of Medical Sciences Hyderabad India Indian Journal of Research2015 Vol 141 Issue 1 Page 13-26
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
bull Superoxide dismutase 1 (SOD1)
bull Superoxide dismutase 2 (SOD2)
bull Superoxide dismutase 3 (SOD3)
bull Glutathione peroxidase 1
(GPx1)
bull Catalase (CAT)
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
The Start ldquoEndosymbiont Theoryrdquo
ldquoTwo cells began to live together
exchanging some sort of substrate or
metabolite [product of metabolism like
ATP] The association became mandatory
so that now the host cell cannot live separatelyrdquo
1967 paper ldquoOn the Origins of Mitosing Cellsrdquo published in the Journal of Theoretical Biology
scientist Lynn Margulisas viewed httpswwwlivesciencecom50679-
mitochondriahtml
Sept1 2017
Mitochondria have their own DNA
As organelles they are capable of translating messages
encoded in their genes to proteins without using any of
the resources of the eukaryotic cell
httpswwwkhanacademyorgsciencebiologystructure-of-a-celltour-of-organellesvmitochondria-video
Mitochondria are responsible for creating more than 90 of the energy needed
by the body to sustain life and support organ function
Mitochondrial DNA
Mitochondrial disorders Challenges in diagnosis amp treatment
Nahid Akhtar Khan1 Periyasamy Govindaraj1 Angamuthu Kannan Meena2 Kumarasamy Thangaraj1
1 CSIR-Centre for Cellular amp Molecular Biology Nizams Institute of Medical Sciences Hyderabad India2 Department of Neurology Nizams Institute of Medical Sciences Hyderabad India Indian Journal of Research2015 Vol 141 Issue 1 Page 13-26
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
bull Superoxide dismutase 1 (SOD1)
bull Superoxide dismutase 2 (SOD2)
bull Superoxide dismutase 3 (SOD3)
bull Glutathione peroxidase 1
(GPx1)
bull Catalase (CAT)
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
httpswwwkhanacademyorgsciencebiologystructure-of-a-celltour-of-organellesvmitochondria-video
Mitochondria are responsible for creating more than 90 of the energy needed
by the body to sustain life and support organ function
Mitochondrial DNA
Mitochondrial disorders Challenges in diagnosis amp treatment
Nahid Akhtar Khan1 Periyasamy Govindaraj1 Angamuthu Kannan Meena2 Kumarasamy Thangaraj1
1 CSIR-Centre for Cellular amp Molecular Biology Nizams Institute of Medical Sciences Hyderabad India2 Department of Neurology Nizams Institute of Medical Sciences Hyderabad India Indian Journal of Research2015 Vol 141 Issue 1 Page 13-26
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
bull Superoxide dismutase 1 (SOD1)
bull Superoxide dismutase 2 (SOD2)
bull Superoxide dismutase 3 (SOD3)
bull Glutathione peroxidase 1
(GPx1)
bull Catalase (CAT)
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondria are responsible for creating more than 90 of the energy needed
by the body to sustain life and support organ function
Mitochondrial DNA
Mitochondrial disorders Challenges in diagnosis amp treatment
Nahid Akhtar Khan1 Periyasamy Govindaraj1 Angamuthu Kannan Meena2 Kumarasamy Thangaraj1
1 CSIR-Centre for Cellular amp Molecular Biology Nizams Institute of Medical Sciences Hyderabad India2 Department of Neurology Nizams Institute of Medical Sciences Hyderabad India Indian Journal of Research2015 Vol 141 Issue 1 Page 13-26
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
bull Superoxide dismutase 1 (SOD1)
bull Superoxide dismutase 2 (SOD2)
bull Superoxide dismutase 3 (SOD3)
bull Glutathione peroxidase 1
(GPx1)
bull Catalase (CAT)
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial DNA
Mitochondrial disorders Challenges in diagnosis amp treatment
Nahid Akhtar Khan1 Periyasamy Govindaraj1 Angamuthu Kannan Meena2 Kumarasamy Thangaraj1
1 CSIR-Centre for Cellular amp Molecular Biology Nizams Institute of Medical Sciences Hyderabad India2 Department of Neurology Nizams Institute of Medical Sciences Hyderabad India Indian Journal of Research2015 Vol 141 Issue 1 Page 13-26
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
bull Superoxide dismutase 1 (SOD1)
bull Superoxide dismutase 2 (SOD2)
bull Superoxide dismutase 3 (SOD3)
bull Glutathione peroxidase 1
(GPx1)
bull Catalase (CAT)
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial disorders Challenges in diagnosis amp treatment
Nahid Akhtar Khan1 Periyasamy Govindaraj1 Angamuthu Kannan Meena2 Kumarasamy Thangaraj1
1 CSIR-Centre for Cellular amp Molecular Biology Nizams Institute of Medical Sciences Hyderabad India2 Department of Neurology Nizams Institute of Medical Sciences Hyderabad India Indian Journal of Research2015 Vol 141 Issue 1 Page 13-26
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
bull Superoxide dismutase 1 (SOD1)
bull Superoxide dismutase 2 (SOD2)
bull Superoxide dismutase 3 (SOD3)
bull Glutathione peroxidase 1
(GPx1)
bull Catalase (CAT)
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
Genetic Expression on Mitochondrial
Function
(ROS Degradation)
bull Superoxide dismutase 1 (SOD1)
bull Superoxide dismutase 2 (SOD2)
bull Superoxide dismutase 3 (SOD3)
bull Glutathione peroxidase 1
(GPx1)
bull Catalase (CAT)
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Superoxide Dismutase 1
SOD1 (CuZnSOD)(Encoded Chrom 21)
Found in cytoplasmic and mitochondrial intermembrane
space(inserted)
Converts superoxide radicals (O2-) to hydrogen peroxide
(H2O2)
Needs copper and zinc as cofactors
Apoptosis Oxidative Stress ALS Myocardial Infarction
Macular Degeneration a marker for heavy Metal toxicity
O2- H2O2SOD1SOD1
Cu Zn
1Murray CJ Lopez AD (May 1997) Alternative projections of mortality and disability by cause 1990-2020 Global Burden of Disease Study Lancet 349 (9064) 1498ndash504 PMID 9167458 doi101016S0140-6736(96)07492-2
2Jump up ^ Braunwald E Kloner RA (November 1985) Myocardial reperfusion a double-edged sword The Journal of Clinical Investigation 76 (5) 1713ndash9 PMC 424191 PMID 4056048 doi101172JCI112160
3uller FL Lustgarten MS Jang Y Richardson A Van Remmen H (August 2007) Trends in oxidative aging theories Free Radical Biology amp Medicine 43 (4) 477ndash503 PMID 17640558 doi101016jfreeradbiomed200703034
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Superoxide Dismutase 2
SOD2 (MnSOD) (Chrom 6)
Found in mitochondria
Converts superoxide radicals (O2-) to
hydrogen peroxide (H2O2)
Needs manganese as a cofactor
Heart Disease Myocardial Infarction
Tumor metastasis neurodegenerative
diseases O2
- H2O2SOD2SOD2
Mn
Pias EK Ekshyyan OY Rhoads CA Fuseler J Harrison L Aw TY (Apr 2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced
apoptosis in PC-12 cells The Journal of Biological Chemistry 278 (15) 13294ndash301
Perry JJ Hearn AS Cabelli DE Nick HS Tainer JA Silverman DN (Apr 2009) Contribution of human manganese superoxide dismutase tyrosine 34 to structure and
catalysis Biochemistry 48 (15) 3417ndash24
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Superoxide Dismutase 3
SOD3
Found extracellularly
Converts superoxide radicals (O2-) to hydrogen
peroxide (H2O2)
Needs copper and zinc as cofactors
Chronic skin disorders Diabetic Neuropathy Folate
Metab Ischemic Heart Disease
O2- H2O2
SOD3SOD3
Cu Zn
Extracellular superoxide dismutase for the treatment of inflammatory skin diseasesSunghwan Kim amp Tae-Yoon KimExpert Review of Dermatology Vol 8 Iss 62013
Juul K Tybjaerg-Hansen A Marklund S Heegaard N H H Steffensen R Sillesen H Jensen G Nordestgaard B G Genetically reduced antioxidative protection and increased
ischemic heart disease risk the Copenhagen City Heart Study Circulation 109 59-65 2004
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Glutathione Peroxidase 1
GPx1(Chrom 3)
Found in the cytoplasm of nearly all mammalian tissues
Reduce hydrogen peroxide (H2O2) to water (H2O) and oxygen (O2)
Uses glutathione an antioxidant as a cofactor
Increased breast cancer risk esophageal cancer Diabetes (2) teleomere expression
H2O2 H2O O2GPx1GPx1
Glutathione
Entrez Gene GPX1 glutathione peroxidase 1 Vats P Sagar N Singh TP Banerjee M (Jan 2015) Association of Superoxide dismutases (SOD1 and SOD2) and Glutathione peroxidase
1 (GPx1) gene polymorphisms with type 2 diabetes mellitus Free Radical Research 49 (1) 17ndash24 Szebeni A Szebeni K DiPeri T Chandley MJ Crawford JD Stockmeier CA
Ordway GA (Oct 2014) Shortened telomere length in white matter oligodendrocytes in major depression potential role of oxidative stress The International Journal of
Neuropsychopharmacology 17 (10) 1579ndash89
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Catalase
CAT(Chrom 11) Found in the cytoplasm of cells
Needs iron and selenium as cofactors
Reduce hydrogen peroxide (H2O2) to water
(H2O)
High turnover coverts millions of molecules of
H2O2 to H2O and oxygen (O2) each second
Hypertension Type 2 Diabetes AcatalasemiahttpsghrnlmnihgovgeneCATconditions
H2O2 H2O O2CATCAT
Fe Se
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial Function Assessment
Mitochondrial Superoxide
(O2-)
Plasma Peroxide Assay
(PPA)
Mitochondrial Membrane
Potential (MMP) Assay
with Oxidative Challenge
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial Superoxide (O2-)
Superoxide is considered to be a major factor
in oxidant toxicity and mitochondrial MnSOD
enzymes constitute an essential defense
against superoxide
High levels indicate the proanti-oxidant
system is out of balance and oxidative stress
is present
Mitochondria are the major source of
superoxide
Superoxide is the origin of reactive oxygen
(ROS)and nitrogen species (RNS) and as
such causes various redox related diseases
and aging
J Clin Biochem Nutr 2015 Jan 56(1) 1ndash7 Published online 2014 Dec 23 doi 103164jcbn14-42 PMCID PMC4306659 A mitochondrial superoxide theory for oxidative stress
diseases and aging Hiroko P Indo123
Hsiu-Chuan Yen4
Ikuo Nakanishi5
Ken-ichiro Matsumoto5Masato Tamura
6Yumiko Nagano
6Hirofumi Matsui
6Oleg Gusev
789Richard
Cornette9
Takashi Okuda9
Yukiko Minamiyama10
Hiroshi Ichikawa11
Shigeaki Suenaga1
Misato Oki2
Tsuyoshi Sato1
Toshihiko Ozawa12
Daret K St Clair3
and Hideyuki J
Majima12dagger
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
SuperOxide (O2-) facts
57 of the tested population has
superoxide levels in the
elevatedhigh range leading to
high risk for certain clinical
conditions
Individuals who are homozygous
recessive(CC) for SOD2 may have
impaired SOD2 function resulting in
higher levels of the free radical
superoxide and require antioxidant
support
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Common Scale Measurement
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Plasma
Peroxidase
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Plasma Peroxide Assay
Low Levels may indicate
oxidative stress and possible
need for antioxidants
High levels may indicate
immune challenges
Can correlate with uric acid
Potential hypertension risk
Related to lipid peroxides
Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension
Lacy Fred1 OConnor Daniel T2 Schmid-Schoumlnbein Geert W13
Journal of Hypertension March 1998 - Volume 16 - Issue 3 - p 291ndash303
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Plasma Peroxidase
37 of the population studied had low
peroxidase activity indicating an impaired
ability to defend against free radicals
34 of the studies population had
elevatedhigh activity potentially indicating
the presence of oxidative stress
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial Membrane Potential
Biological Variability
Subject Day 1 Day 2 Day 3 Mean SD CV
1 91 91 91 91 0 0
2 88 91 88 89 2 2
3 90 93 90 91 1 2
4 93 93 92 93 1 1
5 90 91 92 91 1 2
There is very little variability
between health subjects indicating
a good marker for clinical utility
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial Membrane PotentialNovel Biomarker of Environmental Oxidative
Stress
N o n S m o k e r s (n = 1 3 ) S m o k e r s (n = 8 )
7 5
8 0
8 5
9 0
9 5
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
()
bull Assessed baseline potential between non-smokers and current smokers
bull Significantly different of p lt 0005
Low baseline mitochondrial
membrane potential results in a
decreased capacity to defend
against excessive ROS and may lead
to Oxidative Stress
Similar results to published findingsMuriel Vayssier-Taussat Environmental Health Perspectives
2002
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial Membrane Potential
Novel Biomarker of CVD-Derived Oxidative
Stressbull Males and Females age
45-65
bull All diagnosed with Cardiovascular disease
bull Non-Diabetic and within the past 90-days bull NO Chemotherapeutics
NO antioxidants NO Tanning or Excessive Sun
bull Non-Smokers and NO Excessive use of Alcohol
bull Fatigue was a major complaint
7 5
8 0
8 5
9 0
9 5
B a se l in e
Mit
oc
ho
nd
ria
l M
em
br
an
e P
ote
nti
al
() H e a lth y (5 5 )
C a rd ia c (1 3 )
Maack and Bohm 2011 Madamanchi et al 2005
Patients with CVD
have low membrane
potential The
integrity of their
mitochondria has
been compromised by
oxidative stress
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Mitochondrial Membrane Potential
Assessment and Treatment Monitoring
bull The influence of HIV infection and antiretroviral therapy on the mitochondrial membrane potential
bull Increase of Membrane Potential post Anti-Retroviral Therapy is significant at p lt 002
Schmid Antivir Ther 2007
Mitochondrial membrane
potential can be improved
and can be used to monitor
treatment effectiveness
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Simplified Scale
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Simplified Scale
Assesses the mitochondrias ability to respond to stress This is accomplished
with low and high oxidative challenges using hydrogen peroxide A decreasing
ability to respond indicates an increasing susceptibility to additional stressors
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Clinical Applications
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Oxidative Stress Free
Radical Damage
Heart Hypertension
Ischemia Heart Disease
Skin Skin aging
Psoriasis Melanoma
Kidney Kidney
Disease Nephritis
Joints Arthritis
Rheumatoid Osteo
Lung Asthma Allergies Cancer
Brain AD PD ADHD ASD
Cancer Migraine
Immune Lupus MS AI
Cancer
Multi-Organ Diabetes
Ageing CFS ME
Eyes Macular Degeneration
Cataracts
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Important
STOP ALL CURRENT
THERAPIES THAT DO
NOT PROVIDE SAFETY
STOP IMMEDIATE
DEGENERATION
PROTECT SIGNIFICANT
FUNCTION OR THE
PATIENT ABSOLUTELY
NEEDS
Wait a week
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Lyme Disease Case 44 year old female
Dx Lyme with Babesia Bartonella
Fatigue
Depression
Hormones unable to balance
Low functioning even in high
points on cycling infection
Antibiotic Use
Joint pain
Neuropathy
Insomnia
Palpitations
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
InfectionInfection InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
Neurotransmitter Dysfunction
bullNeurotransmitter Depletion
bullSex Hormone Dysfunction
bullSymptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
PREGNENOLONE
ProgesteroneDehydroepiandrosterone
(DHEA)
Cortisol
Testosterone
Estradiol
Estrone
Estriol
Mitochondrio
n
CHOLESTER
OLP450SC
C
P450SCC P450 Side-Chain Cleavage
Enzyme
bull located on the inner mitochondrial
membrane
Velarde Longevity amp Healthspan
2014
HORMONES
mROS
Mitochondrion membrane potential
Maintain Mitochondrial Function
Mitochondria Regulate
Steroid Hormone Biosynthesis
ROS (H2O2)
Peroxidase
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Tryptopha
nMitochondrio
n
Free
radicals
ROSRNS
Oxidativ
e Stress
Oxidation
DNA
Protein
Lipid Tissue
Necrosis
Inflammatio
n
Immune
Response
Pro-
Inflammatory
Cytokines
BH4
co-factor for
TH and TRPH
Dopamin
e
Serotoni
n
Tyrosin
e
REGULATE
HORMONE
BIOSYNTHESIS
Kynurenin
e
IDO
Kynureni
c AcidQuinolonic
Acid
PREGNENOLO
E
Progestero
neDHEA)
Cortisol
Testostero
ne
Estradiol
Estrone
Estriol
Cholesterol
P450SC
C
MITOCHONDRIONREGULATE
NEUROTRANSMITTE
RS
HORMONES
NEUROTRANSMITTE
RS
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Lyme Disease
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
SOD1 GG (Increased activity)
SOD1 AA (Ancestral levels)
SOD2 TT (Decreased activity)
SOD3 CC (Ancestral levels)
CAT CT (Decreased levels)
GPX1 CT (Ancestral levels)
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
90 Day Plan
Microbiome Reset Nutritional Program
BotanicalSilver Protocol for infection reduction
Ubiquinol 200 am and 200 at noon
Resveratrol dinner and bedtime
Combination fat soluble antioxidants dinner and bedtime
Vitamin C 200 dinner and 200 at bedtime
No glutathione
Appropriate bio-identical
hormones
Methylation Support (active Brsquos
and Betaine)
Lymphatic work starting with
Infrared Sauna moving to
mechanical massagehellipslowly
Catecholamine support AM and
noon
SerotoninGABAMelatonin at
bedtime
Stretching program
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
90 Days Later
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
90 Days Later
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
90 day Symptom Update
Depression lifted
Palpitations minimal
Joint pain improved 50
Fatigue a ldquolittlerdquo better
Sleep ldquoimprovedrdquo 4-6 hours
per night compared to 2-
continuing to improve
Sex drive returned ndash
partner needs Viagra
Optimistic
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
61 year Old Male Parkinsonrsquos X 5 Years- Quick Case
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Carbidopa levadopa25250 5 times daily
Notice that there is more DOPAC than dopamine
Inflammation
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Loss of mitochondrial function equals high oxidation and inflammation
Typical findings
in ALS and
Parkinsonrsquos
patients
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
58 Year Old with Coronary Artery
DiseaseNegative for traditional cardiovascular markers
Stent X 2
Borderline EF for CHF
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Loss of mitochondrial function equals high oxidation and inflammation
Patient has high oxLDL of 110 (0-60)
Decreased functioning SOD2 and GPX1
Chronic inflammation represented by low norepi low epi and low serotonin
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Heart disease is a disease of oxidation and
inflammation now we can diagnose treat
and reverse it
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Central Dogma
Infection or Microbiome
Issue
Infection or Microbiome
Issue
InflammationInflammationFree Radicals
SkyrocketFree Radicals
SkyrocketMitochondrial
StressMitochondrial
Stress
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
Neurotransmitter Dysfunction
bull Neurotransmitter Depletion
bull Sex Hormone Dysfunction
bull Symptoms
