Contact-Thermography and Live Blood Analysis and Clot Retraction Test Techniques – response to queries Prepared by: Luis Vitetta1 Marilyn Johnson 2 Fernando Cortizo 3 Avni Sali4
Graduate School of Integrative Medicine, Swinburne University 1 Director of Research 2 Research Assistant 3 Senior Lecturer and Research Supervisor 4 Professor and Head of School
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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Contact-Thermography and Live Blood Analysis Techniques – response to queries
Graduate School of Integrative Medicine Swinburne University 9 Frederick Street Hawthorn Vic 3122 Telephone: + 61 3 9214 5296 Facsimile: + 61 3 9214 8009 Email: [email protected] Website: www.swin.edu.au/gsim
© 2003
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Overall Summary:
We have been asked to comment in the form of an official scientific report on
three separate screening techniques that are part of a current proceeding by the
ACCC against Australian Biologics – Testing Services Pty Ltd.
The screening techniques in question include:
1. contact thermography
2. live blood analysis
3. clot retraction test
For this review of the scientific evidence we have utilised the rating system that is
recommended by the Quality of Care and Health Outcomes Committee and has been
adapted from the system developed by the US Preventive Services Task Force,
which has also been adopted by the National Health and Medical Research Council
of Australia. The rating system consists of the following:
• Level I Evidence is obtained from a systematic review of all relevant
randomised controlled trials – meta analyses.
• Level II Evidence is obtained from at least one properly designed
randomised controlled clinical trial.
• Level III Evidence is obtained from well designed controlled trials without
randomisation or from well designed cohort or case control analytic design
studies, preferably from more than one centre or research group or from
multiple time series with or without an intervention.
• Level IV Evidence represents the opinions of respected authorities based
on clinical experience, descriptive studies or reports of expert committees.
As far as contact thermography is concerned there is ample level II, III and IV
evidence to show that this is a most important modality that can be considered to
be most useful in determining peripheral blood flow in numerous disease states.
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We refer you to our previously prepared document.1 Further, the evidence to date
demonstrates that contact thermography has applications which have been
successfully applied in the diagnosis and treatment of various disease states that
include breast cancer, cardiovascular disease, diabetes, and general blood flow
problems. Contact thermography has general adjuvant diagnostic properties that
need to be fully exploited by the medical profession. Moreover, contact
thermography has been advocated as a rapid, non-invasive diagnostic modality in
evaluating patients at risk for breast cancer.91 In that study which was conducted
approximately twenty years ago it was reported that thermograms should be
reported as either being normal or abnormal, criteria that would allow the modality
to show a high degree of reliability.
We did not identify any research evidence at levels I to III associated with the
Live Blood Analysis Test on searching the peer reviewed medical literature.
However, this should not constitute a lack of validity or scientific principle
associated with this technique. Our report comprises level IV evidence and is part
of an expert opinion in the area of the Live Blood Analysis Test. Alterations in
erythrocyte (red blood cells) and leukocyte (white blood cells) morphology can be
easily detected and the results have a high level of reproducibility when expert
technicians are used to employ the technique.
Along similar lines of scientific reasoning, recently, the erythrocyte agglutination
test has been successfully employed to demonstrate combined leukocyte and
erythrocyte aggregation in the peripheral venous blood.2-6 Further, also recently it
was concluded that the association between increased erythrocyte
adhesiveness/aggregation, higher concentrations of acute phase proteins, and
increased atherosclerotic risk factors points to a possible clinical applicability of
the erythrocyte adhesiveness/aggregation test to reveal the presence of both low-
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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grade subclinical smoldering inflammation and morbid biology in individuals with risk
factors for atherosclerosis.7
The Live Blood Analysis Test requires further scientific validation however, from
our own research on the technique we have shown that there is a high degree of
correlation between screening results obtained by The Live Blood Analysis Test and
Standard Diagnostic Medical (haematological) Tests in the area of erythrocyte and
leukocyte morphology [Refer Appendix I). Further, the detection of specific
precipitates in peripheral blood identified with the Live Blood Analysis Test has
shown strong correlations with gastrointestinal dysfunction and immune function
[Graduate School of Integrative Medicine, Swinburne University Database].
In our experience The Live Blood Analysis Test that employs the dark field
illumination microscopy technique is a most useful and accurate method as a first
line screening for digestive system and immune function dysfunction. Hence the
employment of dark field illumination microscopy provides the observer the ability
to distinguish all the major subtypes of white blood cells. This leads to a complete
differential screening – analysis of the leukocytes and thus is gained a significant
insight into the immunologic function of the patient. The Live Blood Analysis Test
that employs dark field illumination microscopy method allows the screening of
blood cells in a single session and clinical setting.
Further, for the benefit of patients The Live Blood Analysis Test requires the
employment of scientific technicians with a detailed knowledge of pathological
haematology and who communicate in the first instance with qualified medical
practitioners for the proper management of their patients. As our knowledge is
enhanced from further research and development in this area of medical research,
the general practice clinician who represents the first line contact with the
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community is hence provided with a powerful, sensitive, and comprehensive
screening health method that in the first instance can be used as a screening tool
for promoting lifestyle changes in their patients with the ultimate goal being one of
disease prevention and health promotion.
Responses to Queries:
1. thermography does not provide assessment of the degree of illness
(slight to serious) (ACCC interpretation “thermography does not
provide an assessment of the degree of all illness)
We have reviewed this screening technique in a previous report that
has been delivered to Australian Biologics.
In Summary: from our interpretation of the evidence of this
technique we believe contact thermography to be a most useful
adjuvant screening procedure for a number of different medical
problems.
At no time has there been an interpretation that this screening
technique would provide an assessment of the degree of all illness.
Moreover there is no current medical test/procedure that was or is
currently available, that provides an assessment or measure of the
degree of ALL illness.
Excerpt from our review:1 In terms of breast pathology we conclude in
our report that from the evidence that is currently available that contact
thermography may constitute a low cost adjuvant screening intervention that
could be a yearly evaluation as part of a routine physical assessment for
women who find mammography a painful and difficult procedure. Therefore
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as soon as a suspicious (positive) breast thermal examination is performed,
the appropriate follow-up diagnostic and clinical testing can be ordered. This
would include mammography and other imaging tests, clinical laboratory
procedures, nutritional and lifestyle evaluation and training in breast self
examination. Thermography is a simple, non-invasive, highly accurate,
inexpensive form of screening imaging technique as well as being a ‘breast
friendly’ procedure.
Moreover, as thermography is capable of measuring changes in temperature
that can reflect blood supply and inflammation which are general
physiological changes, it is then possible to envisage its adaptation to screen
other physiological changes that are associated with blood flow. An
additional search of the current medical literature showed that contact
thermography has been applied to other medical conditions. Namely,
cardiovascular disease, comorbidities associated with diabetes and as an
adjuvant method to better define cancer diagnoses.
We have identified numerous studies from the medical literature that hence
emphasize the importance of this technique in medical practice.1-175
In addition to breast screening studies we have identified other peer
reviewed studies that have employed contact thermography in other areas
of medicine and these include:
cardiovascular disease and circulation/blood flow
• Coloured contact thermography has been employed as an additional
method for the diagnosis of diseases of the abdominal aorta and its
major branches.171
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• Thermography has been used to study myocardial temperature during
infusion of cold cardioplegic solutions. Slow cooling was recorded
distal to coronary stenosis or occlusions, thereby indicating
insufficient protection of the myocarium in these areas169, 171-173
• In a recent study of thermograms in army personnel regional
hothermia was investigated with thermograms.160 Further, in another
recent study of thermograms in army personnel by the same group of
researchers investigated thermograms during and after basic military
training evaluating the temperature changes in thermograms whilst
differentiating between those with normal foot parameters, those
with musculoskeletal stress and others with foot stress. The
thermograph showed that normal foot cooling differed between the
three groups. In conclusion thermography did not reveal an exact
diagnosis however it was of greatest benefit monitoring severity and
healing of the foot.160
• Liquid Crystal contact Thermography has been shown to be very
reliable for as a method of investigating skin blood flow and for a
screening method in the diagnosis of deep venous thrombosis.59,164
• The use of thermography has been used to evaluate the peripheral
circulatory function in the diagnosis of diabetic complications.163
Thermography has been employed to observe wound temperature and
hence monitor healing in surgical patients. 162, 163
Contact thermography in conjunction with Doppler sonograph resulted
in the highest diagnostic accuracy when investigating retrograde
blood flow in the internal spermatic vein.152
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Diabetic neuropathy
• Contact thermography has been successfully employed to investigate
and identify diabetic patients who are at increased risk for
neuropathic foot ulceration.16
2. thermography can only assess some aspects of peripheral blood flow
and it does provide an assessment of the circulation, disturbances of
cerebral and/or peripheral blood circulation and vascular occlusion.
(ACCC interpretation – “thermography can only assess some aspects of
peripheral blood flow and it does not provide an assessment of the
circulation of the blood as a whole”
There is substantial scientific evidence that thermography can assess
various aspects of peripheral blood flow.1, 8-165 That is, that
microcirculatory disturbances can be clinically assessed by thermography.
Further, the patho-physiological analysis of the peripheral circulation using
thermography has been investigated as an example of functional body
imaging. Moreover, since vascular damage is the main complication of
diseases such as diabetes, in studies with lower limb amputations for
diabetic foot have shown that the amputation level of the limb was
determined by skin thermography. Thermography was useful in avoiding
deep and systemic circulatory problems which are commonly associated with
diabetic limb amputations. Hence, any interpretation that thermography is
only useful for some aspects of peripheral blood flow and that it does not
provide an assessment of the circulation of the blood as a whole is without
merit as there are numerous peer reviewed studies indicating its successful
employment.
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3. thermography does not provide assessment of the strain a person is
able to bear before he be subject to an operation or vaccination
Thermography provides a useful assessment of peripheral blood flow
abnormalities. Therefore when combined with medical expertise these
results may have a useful adjuvant and indirect application in the assessment
of the surgical patient.
4. thermography does not provide an assessment of the success of
therapies ACCC interpretation – “thermography does not provide an
assessment of the success of all therapies”
As an example of thermography providing success of a therapy can be seen
in a recent study121 that assessed temporary stellate ganglion block
thermographically and its effectiveness during cardio-surgical procedures.
The assumption behind this method was that the increase in the
temperature of the upper extremity on the side of the blockade was due to
the broadening of the arterial bed. It was shown that thermography was a
useful method for the assessment of stellate blockade effectiveness.
Effective blockade resulted in increased blood flow in the radial artery.
Further, low temperature is an important factor in protecting the
myocardium during an operation on the heart. This can be difficult to
accomplish if the cold cardioplegic solution is hindered by occlusions or
stenosis of the coronary arteries. Thermography has been used to study
myocardial temperature during infusion of cold cardioplegic solution. Slow
cooling was recorded distal to coronary stenosis or occlusions, thereby
indicating insufficient protection of the myocarium in these areas.162-165
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5. thermography is not suitable for diagnostic purposes in the cardiac
field
Thermography has been employed extensively in the cardiac field. Namely,
thermography has been used to study myocardial temperature during
infusion of cold cardioplegic solutions. Slow cooling was recorded distal to
coronary stenosis or occlusions, thereby indicating insufficient protection of
the myocarium in these areas.162-165 Further, in order to understand, treat,
and prevent acute coronary syndromes it has been reported that there is a
need to improve the ability to identify the rupture-prone, vulnerable
atherosclerotic coronary plaque. In a recent review it was noted that
thermography used in clinical trials provided useful results.176 Moreover, it
has also been recently reported that an atherosclerotic plaque is considered
vulnerable when it is at higher risk of inducing acute cardiac events. The
early detection and follow-up of the vulnerable plaque are crucial to prevent
these events from happening. Arterial wall thermography, which traces the
heat signature of the activated macrophages, is a new and promising method
in this direction.177
6. thermography is not suitable for analytical purposes in the cardiac
field
Thermography is a very suitable analytical technique in the cardiac field.
The identification of vulnerable plaque is one of the primary goals in
cardiology during the last years. Several techniques have been developed
for the anatomic and functional assessment of atherosclerotic plaques.
Thermography is a new method for the evaluation of the inflammatory
process locally within the atherosclerotic plaque. Several animal and clinical
studies demonstrated the value of thermography not only for the detection
of inflamed atherosclerotic plaques, but its use in new fields like in the
evaluation of inflammation in the coronary vascular bed and the
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cardiovascular system. A recent article has reviewed the developments and
the clinical implications of thermography.178
7. thermography does not represent true preventative medicine (ACCC
interpretation of this “thermography does not prevent people from
becoming ill”)
To claim outright that contact thermography has no preventative medicine
benefits is both unsubstantiated and erroneous given the current peer
reviewed literature that is available. To claim that it is a technique that
prevents people from becoming ill is further erroneously misrepresented.
Contact thermography is a very useful adjuvant technique that can assist
the clinician to provide guidance to the patient with breast cancer or a
suspicious breast lump.1 Further, it has been shown that contact
thermography has other useful applications in medicine as has been pointed
previously in points 4 and 5.
8. thermography does not provide an indication of unrecognised disease,
hidden cause and dangerous sequelae (complications).
From the available evidence it has already been shown that contact
thermography has been successfully used to monitor newly developing breast
disease.1 These results support contact thermography as being useful in
detecting previously unrecognised disease. Therefore contact thermography
does provide some evidence for previously undiagnosed disease with serious
sequalae (eg. Breast cancer).
9. thermography does not provide a valuable aid in monitoring the
restoration of health.
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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This statement is inaccurate. Refer further to points 1, 2, 3, 4, and 8
previously.
10. CRT does not indicate the degree of oxidation in the body
11. CRT cannot determine whether a patient’s treatment regime is
effective within a short period of time by way of a simple finger prick
test (ACCC interpretation of this – “CRT is not capable of accurately
measuring the effectiveness of a treatment regime”)
12. CRT does not indicate
(i) the degree of free radicals
(ii) the function of the heart
(iii) the function of the ovary
(iv) the function of the prostate
(v) the function of any other human organ
(vi) physical stress
(vii) psychological stress
(viii) allergies
(ix) arthritis
(x) vitamin levels
(xi) mineral levels
(xii) heavy metal level
With respect to items 10, 11 and 12 we note that the clot retraction test
(CRT) or Bolen’s Test has been extensively investigated in Europe.179-195 That
the CRT cannot accurately measure the effectiveness of a treatment regime
is inaccurate. Our research indicates otherwise.
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Reactive Oxygen Toxic Species are by-products of virtually every
dysfunction or challenge to cells. Most of these reactive oxygen species are
normally produced as a consequence of metabolic function however abnormal
species are known to be produced. Their causes may be either exogenous or
endogenous. The term free radical has been generally used to encompass
these species. Reactive Oxygen Toxic Species are molecular substances
that have modifications of the oxygen molecule, are highly reactive to
biochemical structures and include all free radicals. The most important
Reactive Oxygen Toxic Species which is not a free radical is hydrogen
peroxide.
Oxidation of biomolecules as a result of Reactive Oxygen Toxic Species can
play a role in susceptibility to a number of diseases. Reactive Oxygen Toxic
Species may result in a metabolic challenge to every cell in the body. It is
the interactions of these toxins with body cells which produce the systemic
effects of metabolic dysfunctions and challenges. Moreover, in affecting
the body’s basic cellular structures and biochemical pathways they react
with blood constituents to form various metabolic by-products which can be
detected. These morphological changes in the blood as well as the
constituents of metabolic by-products and the specific Reactive Oxygen
Toxic Species vary as a function of the metabolic dysfunction or challenge,
strength of the immune system and the level of Reactive Oxygen Toxic
Species being generated.
Oxidative stress is caused by the presence of free radical or radical-
generating agents in concentrations that overwhelm natural radical-blocking
or -scavenging mechanisms. Sources of oxidative stress include exogenous
factors, such as cigarette smoke, and endogenous factors, such as the
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oxidative burst from activated macrophages. Antioxidant mechanisms
include antioxidant enzymes and plasma antioxidants, many of which are
determined by dietary antioxidant intake. Oxidative stress, in turn, can
cause oxidative damage to DNA, proteins, and lipids, and many clinical
conditions are associated with increased indices of oxidant stress; this
suggests that overwhelming the antioxidant defence system initiates and
propagates processes involved in the pathogenesis of many diseases.196-198
Free radicals are in fact potent deleterious agents causing cell death or
other forms of irreversible damage, eg, free radicals appear to modify <
~10,000 DNA base pairs every day.197
From a research perspective the Graduate School of Integrative Medicine,
Swinburne University is focusing on fully investigating this technique with
the Live Blood Analysis Test to further set guidelines for its implementation.
We have currently data available for approximately 2500 patients of which
160 patients have had these tests correlated with standard blood tests
In summary we have observed through the CRT test, physiological and
biochemical changes that were consistent with medical investigations and
reports provided by physicians that were consistent with the degree of
free radicals associated with a normal patient profile, the function of
the heart, the function of the ovary, the function of the prostate, the
function of any other human organ, physical stress, psychological
stress, allergies, arthritis, vitamin C levels, mineral levels, and heavy
metal levels. (Refer Appendix I - Plates 1 to 11 for examples of these).
Further, we have identified and correlated the following results obtained
from CRTs.
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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1. Reactive Oxygen Toxic Species – these are white masses
seen in drops and if relevant organs are affected these are
noted and a detailed report provided to medical practitioner.
2. Heinz bodies – small black dots seen in Reactive Oxygen
Toxic Species masses – possibly due to lysing red blood cells.
3. Adrenal stress – small translucent masses spread throughout
the drop of blood.
4. Reactive Oxygen Toxic Species Linkage – indicative of
inflammation process.
5. Allergic reactions – indicated by spreading of small masses in
the centre of the drop of blood.
6. Advanced cancer profiles.
Currently this technique is being employed to follow up patients diagnosed by
Medical Practitioners with conditions such as chronic fatigue, hormonal
dysfunction, psychological distress, physical stress, cancer and a
micronutrient inbalance. We have employed qualified technicians with
expertise in haematology to conduct and report on the Live Blood Analysis
Test and the CRT.
13. LBA does not provide valuable information about
(i) the human digestive system
(ii) the human immune system
14. LBA does not show
(i) the oxygen carrying capacity of red cells
(ii) the efficiency of protein metabolism
(iii) the efficiency of fat metabolism
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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(iv) liver function
(v) gall bladder function
(vi) the degree of bowel toxicity
With respect to items 13 and 14, The Live Blood Analysis Test encompasses
innovative scientific research in areas of science and medicine not previously
part of conventional medicine. We have not identified any peer reviewed
research articles at the level I to III evidence that were associated with
the Live Blood Analysis Test. However, this should not constitute a lack of
validity or scientific principle associated with this technique. Our own
experience of several years investigating the Live Blood Analysis Test
constitutes the best evidence that is deemed level IV evidence and this will
serve as a template for further discussions herein.
Orthodox medicine has always attempted to treat the affected part of the
body only, in the belief that by doing so illness would be eliminated. Medical
statistics are based solely on conventional medical treatments giving rise to
a significant decrease in acute infectious diseases whilst chronic diseases
such as cancer, cardiovascular disease and viral infections have increased
significantly over the past 50 years.203 This trend has fostered the
development of new immunology and endocrinology based research ideas that
have given rise to the disciplines of psychoneuroimmunology and
psychoneuroendocrinology. As a result, immune system functions, hormonal
function and their relationship to the nervous system have been related to
the effect of the body as a ‘whole’. Hence, adverse environmental sequelae
(eg. heavy metal toxicity), high energy nutrition with poor food choices,
psychological, life and traumatic stresses have been shown to have negative
impacts on overall health. Within this framework and scientific paradigm,
The Live Blood Analysis Test can primarily aid the physician screen, then
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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diagnose and then monitor progress of their patients’ chronic conditions as
they recommend and follow up lifestyle changes that are made to their
patients.
The Live Blood Test that adopts the technique of dark field microscopy can
successfully detect various alterations of erythrocyte or leukocyte
morphology. Further, it can also be used to identify compon(ts(te-5.ek)2.8(ocyte 0.073 -2.0011 TD0.00171 Tc0.someir )45(a( t)d cyc6(mi-6.)-6.5e2.8(o.6(26]TJ10.6(26]T189.96 572.7.4399 88.22039Tm-0.0007.43TD09(ne )]TJ10.6779 0 202.4Tj67.867.42 9022035 TD0.00145 Tc0.0(m)9(mlrtizolowinso4(nh)6.6( dry)-6.5(( l)5.rs.6(ct2(p)1.6(ot)1.2(ion6(cts(suc6(ct2(p)1.6inso4(mntif7(mn4 t)u-7.274.8n)-1.4(t-6.uc6(ct(( l)5eir8i-6uuc6(ct2.8(6.6( a p8n)-1athe0.3ie.8n)-1t whiy)-6.5h ca)-4.0 70.07te )]TJ0 -2.0973 TD0.0018 Tc-0.04.3(ya( l8(thge)-6.4126(is (-)0.nr8i)-6ben4 t)d )5.6(o )54((f)--6.)1(c)-ow)1(c)-en4 t)d )56( u )56(p( lt)d )5.6wf)--6i4126)-6..2( h)1(s)5.6(s.4126(isen4 t)r).6754.3(3( .6(f)2(nr8i)-v).6780.nr8i)-6..2( i4126)-onr8i)-ar8i1((f)--6 s (-)0ar8i1(en4 t)04.3(ya.4126(io.)1(c)-og).67-6i47(3(3( ar8i1((f)--6 .4126(ien4 t)s6..2( s)1(c3g)4he0. )15.55(n )]TJ-0.073 -2Tm-0.0007 TD0 Tc( )Tj0 -2.093 TD0.00131 Tc0.D4.34Gradu8)r)9u8)5( (da)-1(c)-en4 26.)-68(e8n)-12(p14(s)mnso4()-1(c)-cr)9u8)os).675.6)9u)4.3( c9(p(72.2(a4 t)d p14(s)o).2745.6( w)-68(en4 26t2(o)-5(p)1.6a)-1(c)-lmnso4(5(ons)5.8(9 )5.6-can al4e Sch14(s)oo).274e8n)-12(pcan h) t)6rad3)7.1-6bl4e Sen4 26.nr72.2 us).675.ed(3( en4 26x)mi-6t2(o)-en4 26ns).675.if)--6v).671(ech14(s)y a)mi4(nrmog)e8nan
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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and activity is also well observed. Blood platelets, fibrin crystals (appearing
as thin slightly refractile needles or spicules), and chylous material are all
observed and clearly discerned.
At this point I would like to extend this discussion to include our research
experience of The Live Blood Analysis Test.
What can The Live Blood Analysis Test do?
1. micronutrient deficiencies
The relative sizes of erythrocytes can reveal important factors in
cytogenesis that can affect all body cells. Vitamins such as folic acid and B12
are essential for the synthesis of nucleic acids. A deficiency in both of
these vitamins can lead to asynchronous development of all body cells as the
development of the nucleus will be delayed while the maturation of the
cytoplasm continues relatively unaffected. This pathogenic process may
progress as new generations of abnormally large cells are formed. The
relatively uniform size of red cells provides an unambiguous and ready
contrast from the generation of cells affected by such a state of deficiency
presenting with larger sized cells. These large red cells are termed
macrocytes and display diameters exceeding normal values and in the range
of 8.5 – 9.0 µm.200, 205, 207 This erythrocyte abnormality is the principle
characteristic of frank folate and or vitamin B12 deficiency in pernicious
anemia where the macrocytes take on an oval conformation.187 The relatively
short life of red blood cells provides for this characteristic before many
other tissues are significantly affected. If the deficiency state is not
corrected an overt disease termed megaloblastic anemia will ensue. Similarly
neutrophilic white blood cells and platelets will present with an abnormally
large morphology that can further assist the screening of the haematologic
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condition. This can be simply discerned under dark field illumination
microscopy. In a study reported in the Journal of Clinical Pathology199 it was
reported that significant nuclear hypersegmentation of the neutrophil
provides a reliable and sensitive parameter by which to screen for folic acid
deficiencies. The normal neutrophil may have as many as four nuclear
segments. These nuclear segments are sharply contrasted from the highly
refractive granules of the neutrophil cytoplasm. Hence hypersegmentation
is defined by the presence of five or more nuclear segments within the
neutrophil. Thus the discernment of a significant increase in number of
nuclear segments in the hypersegmented neutrophil provides a screening
parameter of folic acid deficiency second only to a folic acid serum assay or
red blood cell folate assay in sensitivity.
Further, the dark field illumination microscopy method used in The Live
Blood Cell Analysis technique also provides a ready made discernment of
abnormally small red blood cells by a physical contrast with normal sized
cells. Microcytes are red cells that are usually less than 6µm in diameter
and are liberated as such into the peripheral blood or may be the result of
cellular fragmentation.200, 205 Microcytes are the classical hallmark of iron
deficiency anemia.205 Microcytosis results from the incorporation of
insufficient levels of haemoglobin into maturing erythrocytes and can also
result from aluminium or lead excess body burden200, 202 where these two
potentially toxic metals have been reported to interrupt porphyrin synthesis.
2. Unbalanced and excessive oxidation of the internal environment of
the body is manifest by features that affect red blood cells. The lipid
component of the cellular membranes is readily degraded by excessive
oxidation. The oxidised lipids which comprise the majority of the structural
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property of the cellular membranes lose much of their structural integrity
and strength. The red cell may then assume a change in conformational
shape that is most probably due to the inability to counter osmotic pressure
changes caused by the cytoplasm as a response to the oxidative degradation
of the lipids within the cell membranes. Conditions of excessive oxidation
may overwhelm the red cell mechanism of antioxidation that protects the
cell and its haemoglobin. This denaturation commonly results in the
formation of precipitate aggregates of the protein globin. These bodies of
aggregated globin are easily discerned within the red cell and are termed
Heinz bodies. These haematological features can be distinguished from
many ‘non-live’ and stained specimens. Further, making use of the dark field
illumination technique of Live Blood Analysis of a blood sample it is possible
to observe the aggregation of red blood cells.
Moreover one also is able to observe the aggregation of thrombocytes
(platelets) and thus enable the discernment of an important indicator of
potential thrombosis. Thrombocyte aggregation and ‘stickiness’ is one of the
hallmarks of possible impending cardiovascular disease that may lead to
stroke and heart attack. Dark field illumination microscopy of a live blood
sample that is unstained or altered by anticoagulants may demonstrate the
potentially thrombotic activation of thrombocytes with the observation of
aggregates. With proper technique and knowledge in the area of
haematological pathology this analysis enables a powerful indicator of risk
for an ischemic crisis not reliably available from other routine means of
study. If significant aggregation of thrombocytes is indicated effective
preventive measures by a medical physician can be employed (eg. prescribing
of aspirin). This form of analysis has important sequelae in potentially
screening at risk populations for such indicators of risk.
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3. Digestive System and Liver Problems
Dietary fats and oils are absorbed by an active formation of micelles in the
small intestine. At this stage these micelles are termed chylomicrons with
an approximate size of 1µm. The chylous material enters the systemic
circulation from the lymphatic system at the left subclavian vein and can be
demonstrated in the peripheral blood after approximately 15 to 20 minutes
of ingestion. In individuals with malabsorption the appearance of the lipid
micelles in the blood may be delayed or diminished in proportion to the lipid
content amount of a meal. The chylomicrons are normally transformed into
smaller micelles with high efficiency by the healthy liver and are then
termed chylous material. In instances of significant liver dysfunction the
efficiency of this process may be impaired and chylomicrons may be
detected in the peripheral blood in greater proportions than in trace
amounts. Usually these liver modified micelles will range in size from
between 0.1µm to 0.4µm and can be readily visualised and identified by dark
field microscopy. Under dark field illumination microscopy they will appear
as small highly refractile particles. Moreover, the relative size of the
chylous particle indicates its constituent lipids. Hence, the smaller particles
will typically be of the higher density lipoproteins while the larger micelle
particles will be composed largely of the lower density lipoproteins.
Conventional medical research has shown that increased proportions of low
density lipoproteins have been associated with increased risk for
atherosclerosis and cardiovascular disease. Conversely, high levels of high
density lipoproteins have been associated with a reduction in risk.
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The chylous material is normally removed from the circulation by cellular
absorption after approximately four hours. In individuals with a lipid
intolerance may demonstrate significant amounts of chylous material in the
peripheral blood after six or more hours of fasting. The relative ease of
discerning the blood chylous material under dark field microscopy
illumination enables a screening analysis of lipid absorption and tolerance.
Such results are then referred back to medical practitioners for follow up
and or further quantitative analytical testing.
4. Immune function
Conditions of inflammation or a significant immune response typically
increase the concentration of the blood protein by the introduction of acute
phase globulins such as histamine or immunoglobulins. Increasing the blood
concentration of these proteins may produce a cohesive linkage of individual
red cells into ordered chains termed rouleaux (vide infra Appendix I –
Patient A Plate A) or into tightly aggregated masses in approximate
proportion to blood concentrations of these globulins. The cohesive cross –
linkage of the red cells can significantly impede the microcirculatory
perfusion of the tissues’ capillaries. A profound degree of rouleaux is a
common feature in the clinical condition termed shock that may result from
extensive physical trauma.
Rouleaux may be evaluated from preparations of peripheral blood by
observation with dark field microscopy of stacked chains of red cells by the
percentage of involvement and the degree of complexity. This type of
screening may be of clinical importance in the evaluation of soft tissue
trauma or to reveal unknown indications of inflammation that may be present
in the instance of a malignancy. The quantitative degree of rouleaux is
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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indicated by the sedimentation rate procedure of common haematological
practice. Hence while the sedimentation rate is the quantitative procedure
of choice for analysis, it is a less direct method of analysis. Dark field
illumination microscopy provides a means of discerning a haematological
condition that is a first line screening procedure.
Leukocytes or white blood cells are nucleated cells normally present in the
circulating peripheral blood. The principle function of these cells is
immunologic. This principle function normally involves body defence against
principally infectious agents that comprise bacteria and viruses or any other
foreign agent. White blood cells may be classified according to their
morphological appearances or their immunological functions. At the most
basic level of classification these cells can be classified as either
granulocytes or agranulocytes. Hence, granulocytes represent a distinct
classification of white blood cells that are involved with immunological
surveillance that is carried out by a phagocytosis mechanism that is
augmented by the antigen – immunoglobulin and complement systems. These
cells contain high concentrations of granules containing proteases and
peroxides that are used to destroy phagocytised target antigens (vide infra
Appendix I – Patient D Plate A).
These granules are readily distinguishable by dark field microscopy by the
high cytoplasmic refractivity. Three classes of granulocytes are defined
from their relative staining affinities. The neutrophil may be characterised
by the regular, fine, and white granules densely occupying the cytoplasm of a
blood cell that is typically polynuclear. Neutrophils are normally the most
abundant subtype of granulocytes comprising ~40 – 65% of the total white
blood cell count. Neutrophils are mobilised within a few hours of exposure
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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to an immune challenge, principally to an elevated white blood cell count early
during an immune response and challenge by an antigen(s). The neutrophil
provides an important function by their potent phagocytosis, albeit
unselective without the specific directing influence of the more specific
immunoglobulins. The nuclei of the neutrophils are poorly refractive by dark
field illumination microscopy and sharply contrasted from the white
cytoplasmic granules. This high contrast provides a simple method of
counting the nuclear segments and enables an index of the relative maturity
of individual neutrophils.
The neutrophil is capable of multidirectional motility (vide infra Appendix I –
Patient E - Plates A to E) that enables an active surveillance and a migration
from the systemic circulation into interstitial spaces and the mucosa. A
diminished capacity for this active immune surveillance can be indicated by
the lack of apparent motility of the neutrophils after several minutes in the
Live Blood Analysis Test specimen.
We have examined extensively the motility of neutrophils and have observed
that neutrophil motility that is not recovered within a few minutes following
preparation of the blood sample for Live Blood Analysis constitutes a sample
from a patient that may have a diminished capacity for this active immune
surveillance response. Further, we have observed a diminished capacity for
neutrophil motility in patients diagnosed by a physician to have depression
(unpublished data and communication).
The eosinophil is a classification of the granulocyte based upon an affinity
for the acidic stain eosin. Eosinophils are the 4th most abundant subtype of
white blood cell, normally constituting 0 – 4% of the total white blood cell
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differential count. These cells have been demonstrated to have a response
to foreign protein substances such as allergic antigens, protozoan or fungal
infections and certain chemical exposures. Certain malignancies will
demonstrate an increase in the ratio of eosinophils. The physical features
of an eosinophil’s cytoplasmic granules enable selective identification of this
white blood cell subtype. Hence, the eosinophil’s cytoplasmic granules are
very refractive and appear more brilliant pink - yellow tinged in colour and
are approximately twice the size of the cytoplasmic granules of the
neutrophils. Active eosinophils may be indicated by a hypodense character
that may be viewed under dark field illumination microscopy by a more
diffuse distribution of the bright granules in the eosinophil cytoplasm.
Basophils are normally those white blood cells with the lowest presence in
the blood. This subgroup of leukocytes comprises less then 1% of the total
white cell differential count in the peripheral blood. These cells are distinct
but similar to the tissue mast cells. They contain large granules that are
reported to contain histamine that may be released as the cells degranulate
typically as part of immediate hypersentitive reaction. Under dark field
illumination microscopy the basophil can be clearly distinguished from other
granulocytes by the comparatively large size of their cytoplasmic granules,
typically appearing hollow and yellow in colour.
We have correlated an increase in the number of eosinophils and basophils
observed with the Live Blood Analysis Test in patients previously diagnosed
by a physician to be allergic to an environmental stimulus.
Dark field illumination microscopy also allows the distinction of lymphocytes
to be observed. Lymphocytes are a classification of the agranulocytes that
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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normally represent the second most abundant subtype of white blood cell
observed. Lymphocytes normally range from ~18 – 45% of the total
differential white blood cell count. The lymphocytes may be differentiated
by dark field microscopy by their single nucleus and notable thick nuclear
membrane and diffusely distributed off – white cytoplasmic granules.
Moreover, natural killer cells can be distinguished from other lymphocytes
by a distinctive cytoplasmic marker, that is a small brightly refractive
cytoplasmic granule/particle structure (vide infra Appendix I – Dark Field
Microscopy Blood Cell Images).
The monocytes are a further classification of the agranulocytes that
normally represent the third most abundant form of white blood cells. The
monocytes normally range from 4 – 8% of the total differential white blood
cell count. The principal activity of the monocyte is reported to be a
selective phagocytosis of micro-organisms or cellular debris than that of the
neutrophil. Blood monocytes are the principle source of leukocyte interferon
and lysozyme. It has also been reported that these cells are transitory cells
progressing to full development as a macrophage and hence incorporation
into the reticuloendothelial system. The monocyte can be distinguished by
dark field microscopy from the single nucleus presenting a roughened tree
bark like texture with a thick and well defined nuclear membrane and a
diffuse distribution of cytoplasmic granules seen as blue-grey in colour.
Hence the employment of dark field illumination microscopy provides the
observer the ability to distinguish all the major subtypes of white blood
cells. This leads to a complete differential screening – analysis of the
leukocytes and thus is gained a significant insight into the immunologic
function of the patient. The Live Blood Analysis Test that employs dark
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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field illumination microscopy method allows the screening of blood cells in a
single session and clinical setting. As our knowledge is enhanced from
further research and development in this area of medical research, the
general practice clinician who represents the first line contact with the
community is hence provided with a powerful, sensitive, and comprehensive
screening health method that in the first instance can be used as a
screening tool for promoting lifestyle changes in their patients with the
ultimate goal being one of disease prevention and health promotion.
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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169. Salmi, A. M.; Tukiainen, E., and Asko-Seljavaara, S. Thermographic mapping of perforators and skin blood flow in the free transverse rectus abdominis musculocutaneous flap. Ann Plast Surg. 1995; 35(2):159-64.
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Appendix I Live Blood Analysis Images from the Graduate School of Integrative Medicine, Swinburne Universtity. Sample images of live blood samples before and after interventions by General Practitioners practicing nutritional medicine are herewith presented. Part of a Database of 2500 patients with correlations so far conducted on 160 patients. Patient A / Plate A – before intervention (dark field microscopy)
Patient A / Plate B – after intervention ~ 6 months (dark field microscopy)
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Patient B / Plate A – before intervention (direct light microscopy)
Patient B / Plate B – after intervention (direct light microscopy)
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Patient C / Plate A – Patient seeks Medical advice from a General Practitioner because of chronic diarrhoea. GP suspects and then diagnosis gastrointestinal dysfunction associated with an irritable bowel. Live Blood Analysis Test confirms the presence of large bowel toxicity crystals (yellow arrows). Further, the patient also exhibits poor nutritional absorption represented by erythrocytes appearing as ‘Ghost’ like cells (green arrows). Compare with ‘healthy’ erythrocytes in a patient with no bowel toxicity [Plate D]. Note granulocyte (red arrow).
Patient D / Plate A – no bowel toxicity
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Patient E / Plates A – E: Time photography plates of neutrophil motility (red arrow - site of specific point of reference). A. (0 secs – time reference)
B. (+ 15 secs)
C. (+ 20 secs)
D. (+ 22 secs)
E. (+ 25 secs)
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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Dark Field Microscopy Images:
A. Neutrophil (magnification factor X 600)
B. Eosinophil (magnification factor X 600)
C. NK cell (Natural Killer) lymphocyte [red arrows denote
distinctive refractive cytoplasmic granule/particle markers] (magnification factor X 600)
D. Non NK cell lymphocytes [note absence of refractive
granule/particle] – Blood monocytes (red arrow) (magnification factor X 600)
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Clot Retraction Test (Bolen’s test) Plate 1 – A Normal Patient Profile
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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Clot Retraction Test (Bolen’s test) Plate 2 - Patient with “line fractures or spokes” (arrows) observed on clot retraction test – patient at risk for osteoporosis with hypercalcemia confirmed by a standard blood test showing an elevated blood calcium level.
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Plate 3 – CRT of blood sample of a patient with small to medium Reactive Oxygen Toxic Species masses (arrows) – in this patient indicative of adrenal stress.
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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Plate 4 – CRT of blood sample of a patient with Reactive Oxygen Toxic Species linkage (arrows) – when the masses tend to become linked is often an indication of inflammation – for this patient with an inflammation - query arthritis and report to medical practitioner.
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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Plate 5 – CRT of blood sample of a patient with Reactive Oxygen Toxic Species (arrows) correlated with hormonal dysfunction - report to medical practitioner for further follow up.
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Plate 6 – CRT of blood sample of a patient with an allergy profile.
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Plate 7 – CRT of blood sample of a patient with an inflammatory profile.
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Plate 8 – CRT of blood sample of a patient with a primary bile duct cancer. Refer Arrows – areas of extensive free radical activity.
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Plate 9 – CRT of blood sample of a patient with suspected heart disease. Refer Arrows – areas of extensive free radical activity.
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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Plate 10 – CRT of blood samples of patients with different vitamin C consumptions. Refer Arrows – red blood cell with and without dispersion.
a) – normal\elevated vitamin C consumption
b) – low vitamin C consumption
Contact-Thermography and Live Blood Analysis Techniques – response to queries Vitetta L, Johnson M, Cortizo F, Sali A. Graduate School of Integrative Medicine
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Appendix II
Current Data from the Graduate School of Integrative Medicine (N = 160 patients)
Live Blood Analysis Test comparisons with Standard Haematological* Tests
and General Practitioner Diagnoses
Condition
LBA Test
(Observed)
Standard Haematological
Tests r
Microcytosis √ 0.90* Macrocytosis √ 0.90* Immune Function
- multi-lobed granulocytes - neutrophil motility - eosinophilia (allergies)
√ √ √
0.85* 0.70* 0.80
Digestive System - platelet adhesion - liver function - erythrocyte aggregation - rouleaux formation - severe erythrocyte aggregation - essential fatty acids deficiency
√ √ √ √ √ √ √
0.80* 0.80 0.85 0.85* 0.65 0.70
Fungal infections √ 0.70