U.S. patent application number 10/463465 was filed with the patent office on 2003-10-23 for screening test and procedure using apocrine sweat.
Invention is credited to Berlin, Stuart M..
Application Number | 20030199743 10/463465 |
Document ID | / |
Family ID | 26940684 |
Filed Date | 2003-10-23 |
United States Patent
Application |
20030199743 |
Kind Code |
A1 |
Berlin, Stuart M. |
October 23, 2003 |
Screening test and procedure using apocrine sweat
Abstract
Marker compounds indicative of variations in physiological
conditions, genetic defects or disease states have been found to be
present in apocrine sweat and, in many conditions, these markers
are present in apocrine sweat in elevated levels when compared to
blood. These markers are collected and/or detected in apocrine
sweat and used as an indicator that a disease state or particular
physiological condition exists. A skin patch, wipe or other
collection means is used to collect apocrine sweat. The apocrine
sweat is then used to indicate the existence of a disease state or
changing physiological condition by the presence of specific marker
compounds. Apocrine sweat can also be collected from individuals
known to have a specific disease or physiological condition and, by
comparison with apocrine sweat from normal individuals, used to
identify new markers indicative of the specific disease state or
physiological condition.
Inventors: |
Berlin, Stuart M.; (Thousand
Oaks, CA) |
Correspondence
Address: |
Michael J. Ram
KOPPEL, JACOBS, PATRICK & HEYBL
Suite 107
555 St. Charles Drive
Thousand Oaks
CA
91360
US
|
Family ID: |
26940684 |
Appl. No.: |
10/463465 |
Filed: |
June 16, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10463465 |
Jun 16, 2003 |
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09994535 |
Nov 26, 2001 |
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6585646 |
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60250206 |
Nov 29, 2000 |
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Current U.S.
Class: |
600/346 |
Current CPC
Class: |
G01N 33/57434 20130101;
G01N 33/528 20130101 |
Class at
Publication: |
600/346 |
International
Class: |
A61B 005/05 |
Claims
I claim:
1. A diagnostic device for collecting and indicating the presence
or elevated state of a marker compound in a mammalian body, the
marker compound indicative of a particular disease state or
physiological condition and being present in apocrine sweat in
elevated concentrations when compared to eccrine sweat, comprising
a patch for placement on the skin of a living mammal, the patch
comprising a. an absorbent means for collecting sweat gland output,
b. one or more detector means which interacts with the elevated
concentrations of the marker compound present in the collected
sweat, and c. an indicator evidencing the interaction of the marker
in such elevated concentrations with the detector means.
2. The diagnostic device of claim 1 further including means for
removably attaching the patch to the skin for a period sufficient
to collect a predetermined quantity of sweat gland output and the
marker compound contained therein.
3. The diagnostic device of claim 1 wherein the marker compound is
present in concentrations above normal, said elevated levels
indicative of particular disease states or physiological
condition.
4. The diagnostic device of claim 1 wherein the marker compound is
present only when a particular disease states or physiological
condition is present.
5. The diagnostic device of claim 1 wherein the detector means
provides a color change indicative of the presence of at least a
specified quantity of the marker compound.
6. A method of diagnosing a disease state or physiological
condition in a living mammal comprising: a. collecting liquid or
vapor components of apocrine gland secretions in a collection means
positioned in the underarm, groin or mammary area of the mammal
and, after a sufficient collection period, b. removing the
collection means and analyzing the liquid or vapor components
collected for the presence of a marker compound or compounds in the
apocrine gland secretions, said marker compound or compounds being
indicative of the existence of the diseased state or physiological
condition and not being present in the absence of said disease
state or physiological condition.
7. The method of claim 6 wherein the presence of the marker is
determined by visible indicators, colorimetric indicators, chemical
or monoclonal antibody detection techniques, electronic sensors,
clinical laboratory analysis or instrumental techniques.
8. A method of diagnosing a disease state or physiological
condition in a living mammal comprising: a. collecting liquid or
vapor components of apocrine gland secretions in a collection means
positioned in the underarm, groin or mammary area of the mammal
and, after a sufficient collection period, b. removing the
collection means and analyzing the liquid or vapor components
collected for the presence of a marker compound or compounds in the
apocrine gland secretions, said marker compound or compounds being
indicative of the existence of the diseased state or physiological
condition by being present in quantities elevated over levels
existing in apocrine sweat in the absence of said disease state or
physiological condition, the presence of the marker compound in
apocrine sweat also being proportionally greater than in eccrine
sweat simultaneously collected when compared to apocrine sweat and
eccrine sweat under normal physiological conditions or in the
absence of the disease.
9. The method of claim 8 wherein the physiological condition is
ovulation or premenstrual syndrome in a female.
10. A method of determining histocompatability between two
individuals comprising: a. collecting from each individual liquid
or vapor components of apocrine gland secretions in a collection
means positioned in the underarm, groin or mammary area of the
mammal and, after a sufficient collection period, b. removing the
collection means and analyzing the liquid or vapor components
collected for the presence of a marker compound or compounds in the
apocrine gland secretions of each individual said marker compound
or compounds being indicative of the existence of
histocompatability between said individuals.
11. A method of identifying markers or antibodies against said
markers indicative of a specific disease state or physiological
condition in an individual comprising: a. collecting a first sample
of liquid or vapor components of apocrine gland secretions in a
collection means positioned in the underarm, groin or mammary area
of the individual known to have the specific disease state or
physiological condition, b. collecting a second sample of liquid or
vapor components of apocrine gland secretions in a collection means
positioned in the underarm, groin or mammary area of an individual
known to not have the disease state or physiological condition, and
c. comparing the first sample with the second sample collected to
determine compounds in the first sample not existing, or existing
at lesser concentrations, in the second sample, said compounds
being marker compounds or antibodies against said marker compounds
indicative of the existence of the specific diseased state or
physiological condition.
12. The method of claim 11 wherein the presence of the marker
compound is determined and is identified by calorimetric,
electronic, clinical laboratory analysis or instrumental
techniques.
Description
[0001] This is a Continuation-in-Part of U.S. application Ser. No.
09/994, 535, filed Nov. 26, 2001, now U.S. Pat. No. ______ issued
______, which claims benefit of Provisional Application 60/250, 206
filed Nov. 29, 2000.
[0002] The present invention relates primarily to a test for
diagnosing the existence of certain disease states or physiological
conditions in mammals based on the collection and analysis of
apocrine sweat.
BACKGROUND OF THE INVENTION
[0003] It is known that certain disease states or variations from
normal physiological conditions can be detected or monitored by
analyzing body fluids for variations in constituents of those body
fluids. Blood is a highly complex suspension with many elements,
which can be measured and analyzed to measure normal homeostasis
and variations from normality. Blood is frequently analyzed for
variations of various constituents (cholesterol, glucose, etc.), as
well as its cellular constituents, as indicators of possible
abnormal health conditions. In a like manner, a urine analysis may
be used to detect various chemical imbalances, pregnancy or the
presence of various legal or illegal drugs by testing for
metabolites of the target drug. In addition, there are numerous
examples of calorimetric test strips, typically used on urine, for
indicating various concentrations of normal fluid constituents or
environmental contaminants such as various metals, glucose, protein
and ketones. The strips provide a visible indication of a threshold
level of normally present reference analyte such as IgG, albumin or
the like.
[0004] Diagnostic studies of chemical imbalances in other body
fluids, such as sweat, can also be indicative of different disease
states. For example, an increase in the level of chloride ion in
sweat is indicative of cystic fibrosis. U.S. Pat. No. 3,552,929
describes a band-aid type test patch for collecting perspiration.
The test patch is applied to an accessible skin surface for
collecting eccrine, not apocrine sweat. It is then removed and
subjected to a series of analytical tests to determine the
concentration of chloride ions as an indicator of cystic fibrosis.
U.S. Pat. No. 4,329,999 to Phillips is a further example of a sweat
collection device for the detection of drug or alcohol use. The
patch was applied to shaved skin surfaces (ankle or calf) which do
not have apocrine sweat glands and remained in place for ten days.
U.S. Pat. No. 4,706,676 discloses a third example of a dermal patch
for detecting exposure to various different environmental
chemicals. No attempt was made to collect apocrine sweat.
[0005] U.S. Pat. No. 5,676,144 to Schoendorfer and related patents
(U.S. Pat. Nos. 5,638,815; 5,465,713; 5,438,984; 5,441,048;
5,203,327 and 4.957,108) describe skin patches which contain
various immobilized specific binding agents for certain reference
analytes as well as compounds to be detected. Schoendorfer
recognizes that there are both eccrine and apocrine sweat glands,
that perspiration can be either sensible or insensible and that
various locations on the body sweat at different rates. However, he
fails to recognize the unique diagnostic function of apocrine sweat
glands and proceeds to collect eccrine sweat from the chest because
large quantities of sweat can be collected. After a sufficient
volume of sweat is collected the patches are then removed and
subjected to various different laboratory procedures to detect the
presence or increased levels of various substances such as creatine
kinase MB (CK-MB). CK-MB is released from cardiac muscle during
myocardial infarction and other cardiac stress and therefore is an
indicator of cardiac injury. As indicated in U.S. Pat. No.
5,203,327, many chemical species detectable in blood or urine, such
as sodium, calcium, chloride and potassium, are also detectable in
lower concentrations in sweat. Table 1 of the '327 patent also
lists numerous other normally occurring components detectable in
various concentrations in sweat. Affinity chemistry can be
developed and incorporated in skin patches for monitoring the
concentration of these various chemicals. In addition, various
metabolites of drugs of abuse such as THC, morphine, cocaine or
methadone can be detected in sweat. However, with the exception of
colorimetric indication of a normally present reference compound,
such as IgG or albumin, which is used to provide an indication that
a minimum volume of sweat has entered the patch, the compounds of
interest are merely collected by the patch (U.S. Pat. No.
5,203,327, col. 16, lines 4-30). Detection and analysis of the
target component is then performed in a laboratory setting.
[0006] The FDA has approved sweat collection patches for use in
drug abuse screening programs to test for amphetamines, cocaine,
marijuana, PCP, heroin methamphetamines, their metabolites and
other opiates. Manufacturers of these products include the Sudormed
Sweat Specimen Container, made by Sudormed, of Santa Anna, Calif.,
the EIA Micro-Plate Assay, made by SolarCare Technologies
Corporation, Bethlehem, Pa. and the PharmaChek.TM. Sweat Patch from
Pharmachem Laboratories Inc. of Menlo Park, Calif. These patches
are waterproof, adhesive pads about the size of a playing card
which is worn on the back, upper arm or lower chest. As such, it
collects eccrine sweat. The patch has a tamper-proof feature so it
can be applied only once and not removed and reapplied later. It is
not designed for use by individuals; instead it is intended for use
by trained drug abuse testing professionals in clinical and
rehabilitation centers. After removal, the test patch is sent to a
test laboratory where the presence and quantity of the target
compound is detected by a special laboratory assay. It must be worn
for 5 to 14 days to collect an adequate quantity of chemicals
indicative of drugs of abuse used during that period. Sweat testing
for drugs of abuse provides a convenient and considerably less
invasive method for monitoring drug exposure than blood or urine.
Typically, ELISA immunoassay is used to detect opiates collected on
the patch from eccrine sweat and GC/MS is used to determine
specific opiates. For comparison urine specimens are subjected to
qualitative analysis by EMIT. However, the percentage of false
negative results with the patches indicates that weekly sweat
testing may be less sensitive than thrice weekly urine testing in
detecting opiate use. (Huestis M. A., Cone, E. J., Wong, C. J.,
Umbricht, A., and Preston, K. L. Journal of Analytical Toxicology,
24, pp. 509-521, 2000). This is most likely because those skilled
in the art selected an inappropriate source of sweat (eccrine
sweat), not recognizing the unique diagnostic capacity of apocrine
sweat. Further, because of the extended wear requirement (up to 14
days), the ability to rapidly test for the presence of drugs using
presently available sweat collection patches and procedures is not
possible.
[0007] Besides the use of skin patches to collect sweat, there are
also several skin patch designs for use to deliver chemicals to the
skin surface, such as anti-infectives or allergens, for treating
various maladies or for analyzing sensitivity to various
allergens.
[0008] It is also known to collect an analyte on a test strip and
then perform various tests on the analyte in a laboratory
environment. U.S. Pat. No. 5,459,080 sets forth numerous prior
techniques for analysis, both quantitatively and qualitatively, of
the presence of various compounds found in body fluids. In
particular, the use of specific binding reagents is discussed. Such
analytes include, but are not limited to, antibodies, antigens,
drugs and hormones. One technique for analyzing these materials,
referred to as immunoassay procedures, uses antibodies and
antigens, i.e. immunoreactants, which are capable of binding with
each other, creating a highly specific reaction product indicative
of the presence of the antigen. To use these techniques the analyte
must first be collected and then transported to a laboratory where
the analytical procedure can be performed. Typically, the analyte
on the test strip is then contacted with an indicator reagent which
is then subjected to a detection means. The requirement for such a
procedure results in a major deficiency for diagnostic test strips
as it is often desirable to be able to read the test strip on the
patient or immediately upon removal of the test strip without
performing a further analytical procedure.
[0009] Other patents directed to test strip detection systems
include U.S. Pat. No. 5,900,379 to Noda et al, U.S. Pat. No.
5,965,458 to Kouvonen et al and U.S. Pat. No. 5,998,221 to Milleck
et al.
[0010] In a related approach, to test for pregnancy, monoclonal
antibodies are used to bind to specific chemicals, such as human
chorionic gonadotropin. A detector agent is then added to the urine
/antibody combination to produce a visible color change.
[0011] It is also known that certain cancers generate marker
compounds, or levels of such compounds elevated above normal, which
can be detected in blood and quantified. Elevated levels of
prostate specific antigen (PSA) may be an indication of the
presence of prostate cancer. Likewise elevated levels of prostate
acid phosphatase are found in some patients with prostate cancer.
The protein NPM66 is a marker for breast cancer. CA 125 is produced
by a variety of cells; elevated levels are found in many women with
ovarian cancer. While human chorionic gonotropin (HCG) is normally
produced by the placenta during pregnancy, it can also indicate a
rare form of uterine cancer. Porter-Jordan et al (Hematol. Oncol.
Clin. North Amer. May 8, 1993, 28-44) suggests that tumerigenesis
and tumor growth can be assessed by measuring cell proliferation
markers (Ki67, cyclin D1 and proliferating epidermal nuclear
antigen (PCNA)). Elevated levels of urokinase plasminogen activator
(upa) has been reported in patients with breast carcinomas. CA 15-3
is shed by breast tumor cells into the blood stream. While the
clinical significance is not clear, the FDA has approved test kits
to measure CA 15-3-like antigen for the early detection of
recurrent disease in breast cancer patients. CA 27-29 is also
frequently present in breast cancer patients, as well as in the
blood of patients with colon, stomach, kidney, lung, ovary,
pancreas, uterus or liver cancer. CA 19-9 has been identified in
patients with colorectal, pancreatic, stomach and bile duct cancer.
Measurement of neuron-specific enolase (NSE) has been tracked as a
marker for neuroblastoma, small cell lung cancer, Wilms' tumor, and
cancers of the thyroid, kidney, testicle and pancreas. Her2/neu
(also known as v-erb2) is amplified in 20-30% of breast cancer
patients. It is also known that a certain percentage of cancers,
particularly breast cancer, are hereditary. These patients may
exhibit genetic markers. Mutations of BRCA1 or BRCA2 are found in
80% of these patients and a high percentage of women with mutated
BRCA1 or BRCA 2 show a greatly increased risk of developing breast
or ovarian cancer. Again, these markers are detected and quantified
by blood tests conducted in a laboratory setting. It must be noted
that some of these markers are normally present in small amounts in
healthy patents and increased levels in cancer patients while
others seem to be present only when cancer is present.
[0012] U.S. Pat. No. 5,798,266 describes the analysis of mammary
fluids for breast disease markers such as described in
Porter-Jordan, but particularly addresses carcinoembryonic antigen
(CEA), HMFG, MCA, vasopressin or cathepsin D as markers. Also,
protein Ig, suggested as a breast cancer marker, has been detected
in tears.
SUMMARY OF INVENTION
[0013] Many variations in physiological conditions, genetic defects
or disease states result in the release or increase of specific
biological and/or chemical moieties, i.e., marker compounds, into
body fluids, particularly blood and urine. I have now documented
several of these markers are present in apocrine sweat and are
present in apocrine sweat in elevated levels when compared to
blood. These markers can be collected and/or detected and used as
an indicator that a disease state or particular physiological
condition exists. In a first embodiment, the invention contemplates
a skin patch to collect sweat, the patch including monoclonal
antibodies or other chemical compounds that can indicate the
presence of specific markers. This may include indicators which
render visible, the existence of a reaction between the monoclonal
antibody or chemical compound and the marker compound to indicate
the existence of the condition being tested for. In a second
embodiment a wipe to collect sweat may be used instead of a patch.
A third embodiment uses an electronic sensor to collect and sample
vaporized apocrine sweat or aroma emitted from the apocrine sweat
gland. Various different disease states can be detected including,
but not limited to heart disease, cancer, autoimmune disease,
infectious disease, renal disease, drug intoxication and other
systemic illnesses. In particular, occult cancers of the breast,
colon, stomach, pancreas, lung, uterus, prostate, bile duct and
ovary and neuroblastoma as well as various forms of leukemia can be
detected by this technique.
DESCRIPTION OF INVENTION
[0014] It is generally believed that human sweat has a chemical
composition which reflects chemical changes in blood. In other
words, when a chemical compound not normally present in blood, or a
compound with cyclical presence, appears in the blood stream of an
individual, it is believed that that compound, or a corresponding
compound, analog, metabolite or reaction product, referred to as a
by-product, of that compound which can be correlated with a
systemic disease state or a change in physiological function, such
as ovulation, can usually be identified in sweat of that
individual. However, it has now been discovered that this is not
generally so.
[0015] The human body has two different kind of sweat glands which,
while having some similar functions, also now appear to have
certain functions which are completely different. The eccrine sweat
glands which are present over much of the body (arms, legs, feet,
hands, abdomen, chest, back, forehead, etc.) excrete large
quantities sweat and have a primary function of maintaining proper
body temperature as a result of the cooling effect which
accompanies evaporation of perspiration. On the other hand the
apocrine sweat glands, which are located under the arms (the
axilla) and in the pubic and mammary areas have a unique and
unexpected function and morphology which is not demonstrated by the
eccrine sweat glands. The morphology of the apocrine gland is very
important. Every apocrine gland opens at the base of a hair cell.
This means that the contents of the apocrine gland are deposited on
a hair shaft and that the hair shaft acts as a saturated wick. The
warm enclosed space of the under-arm provides a microenvironment
which causes vaporization of the apocrine gland secretions. This
vapor persists in the armpit which acts as a persistent smell
chamber.
[0016] The apocrine gland also behaves differently from the eccrine
sweat glands in how it responds to certain body conditions. The
response to elevated prostate specific antigen (PSA) levels is an
example. As set forth in Example 1 below, when the serum PSA level
is normal, for example less than 1.0 ng/ml, the apocrine PSA level
is essentially insignificant. However, when the serum prostate
specific antigen level is abnormally elevated the PSA in apocrine
sweat increases to a significantly greater extent and to a
significant therapeutically abnormal range. An active transport ATP
dependent mechanism takes place to concentrate PSA in the apocrine
secretion. As a result of the morphology of the apocrine gland, the
apocrine gland functions to saturate the hair follicle cells with
PSA transmitting a message detectable by smell as a result of the
PSA levels being elevated to an unhealthy level in the armpit smell
chamber. With the discovery of this phenomena, readily readable
tests providing an early warning of this serious medical problem
can be developed.
[0017] I have now found that apocrine sweat glands contain
chemicals in concentrated quantities which are also uniquely
related to disease or physiological state. Such function or
capacity is not demonstrated by the eccrine sweat glands. In other
words, apocrine sweat emit odors or aromas and contain chemicals
which can be directly correlated with disease state or the state of
certain body functions which have, in the past only been determined
by blood or urine tests or other more complicated diagnostic
procedures. In comparison, eccrine sweat does not contain these
chemical compounds or, if present, they exist in much lower
concentrations. It is believed that this function is part of the
normal evolutionary development of humans and provides to a second
individual the capacity to sense if the first individual has a
disease or body malfunction. The utilization of the function of the
apocrine sweat gland as an olfactory signaling system to
communicate the health or physiological status of an individual has
profound implications for the understanding of evolution in humans,
and presumably other animals as well. It is unlikely that the only
evolutionary pressure which shaped this system is mate selection.
Once the range of physiological conditions that the apocrine gland
is sensitive to is identified it may become clearer. However, the
demonstration of an olfactory system to signal health status means
there is an evolutionary advantage in identifying who is ill.
Caring for the sick is a human trait,that confers significant
evolutionary advantage. It is entirely possible that humans in
pre-historic times, who did not wash and hide their smells behind
deodorants may have actually been better than modem man in
diagnosing each other's illness.
[0018] These by-products can also be detected by instrumental means
or other indicators early in the course of the disease to allow
diagnosis while the disease is readily treatable. Apocrine sweat
has been discovered to be the body fluid that has evolved for the
purpose of communicating the health status of an individual. Also,
female apocrine sweat aromas emitted during ovulation may provide a
stimulation toward intercourse and procreation; male apocrine sweat
aroma is sensed by a female in the selection of a suitable mate for
breeding, specifically in terms of the immune system of the
offspring.
[0019] Several biomarkers have been identified for breast cancer.
These can be utilized in an apocrine sweat patch test to signal
occult breast cancer. It is believed that analysis of apocrine
sweat will lead to the discovery of entirely new and as yet unknown
biomarkers for various cancers. A patch can also be produced which
can indicate the presence of more than one marker for more than one
disease. Once a biomarker is identified it can be used in
immunohistopathological evaluation of malignant tumors to determine
its location in the tumor cell.
[0020] Once a biomarker is identified an apocrine sweat diagnostic
modality, using the teachings herein, can be established. This will
result in a diagnostic screening test using the patch and then a
secondary confirmatory test can be performed to locate the tumor in
the body.
[0021] The availability of an apocrine test patch incorporating
markers identified as indicative of breast cancer can supplement or
be used as a prescreen for mammogram. It is expected that there
will be a number of individuals considered by mammography to be
normal who will actually have occult breast cancer. These
instances, when detected by an apocrine skin patch, should not be
considered as false positives for breast cancer but instead an
early warning of the future appearance of the disease. Mammography
is limited in its ability to detect very small or dispersed tumors.
However, tumors of any size generate biological markers, which will
be present in the blood and concentrated in the apocrine sweat. The
presence of the marker is indicative of an abnormal physiological
state whether or not such condition can be determined by other
presently available less sensitive diagnostic modalities. Breast
cancer now occurs in one out of nine women and is generally
considered to have been present for five or six years before
presentation (detection by present detection modalities). It is
generally accepted that a mammogram has limited sensitivity and
that there are individuals who have occult breast cancer that the
mammogram will not detect.
EXAMPLE 1
[0022] The superior and unexpected diagnostic utility of apocrine
sweat collection and analysis in cancer detection is demonstrated
by the analysis for PSA in apocrine sweat when compared to eccrine
sweat or blood. Approximately 0.5 cc of eccrine sweat, apocrine
sweat, and blood of two individuals was tested for PSA. One
individual had normal PSA blood levels while the other had
significantly elevated blood PSA. Values for PSA in blood, eccrine
sweat and apocrine sweat are as follows:
1 PSA level (ng/ml) eccrine sweat apocrine sweat blood RK 0.34 10.3
3.91 SB 0.08 0.16 0.92
[0023] RK has an abnormally high blood PSA level. This was
reflected in a significantly greater quantity of PSA in apocrine
sweat and a 29.4 to 1 ratio of PSA in apocrine compared to eccrine
sweat. Still further, PSA is elevated in apocrine sweat when
compared to blood with a 2.6 magnification of PSA in apocrine
sweat. This means that there is an active process taking place in
the apocrine gland to increase the concentration in the sweat
produced by the apocrine gland. In the normal individual there is
more PSA in apocrine than eccrine sweat but the ratio is not as
pronounced, and there is no concentration over that in blood. It
can also be seen that the amount of PSA in eccrine sweat is
significantly lower than in blood (a dilutive effect) and the
increase in the amount of PSA in eccrine sweat remains in
proportion to the increase in blood. However, the increase in PSA
in blood is reflected by a significantly greater, increase in
apocrine sweat (a magnifying effect).
[0024] Another diagnostic patch for use in indicating a disease
presence contemplated by the invention is for the detection of
abnormal levels of creatine phosphokinase (CPK). CPK is an enzyme
that is present in all human muscle tissue. However, there are
subtypes of this enzyme of which CPK-MB is the subtype that is
present in cardiac muscle. When there is damage to cardiac muscle
the cells leak CPK-MB. This is used to evaluate individuals who
present with chest pain. Serial blood samples are typically drawn
to see if CPK-MB (and also other enzymes) become elevated in serum
over a period of time. While this test has significant diagnostic
value, it has the disadvantage that the results take an
unacceptable time to generate, which is critical when an individual
is having a heart attack. CPK-MB is very similar to PSA as a
biomarker in that it is associated with a specific kind of cell
(cardiac muscle cell compared to prostate cell). It is believed
that it can be readily and rapidly detected and measured in
apocrine sweat as the CPK-MB should also be elevated in apocrine
sweat when CPK-MB is elevated in serum. This is supported by
anecdotal stores of dogs able to determine that his master is ill
and thus saving his master's life. In reported incidents, tests
confirmed that the man had coronary artery disease and was able to
be treated in a timely manner. However, it was the keen sense of
smell of the dog that detected that something was wrong. Based on
applicant's testing it is believed that the dog was able to detect
CPK-MB in his master's apocrine sweat by his sense of smell.
[0025] Since CPK-MB is a known chemical entity, the invention
contemplates an apocrine sweat collection patch incorporating a
reactant chemical or biological material sensitive and specific to
concentrations of CPK-MB in excess of predetermined threshold
levels, preferably containing a color generating compound which can
be correlated with CPK-MB presence. This device would have
considerable value as a rapid, early screening tool for patients
complaining of chest pain. Since these patients are often
diaphoretic (sweating profusely) such a device could be used by
emergency medical pers An added value of a CPK-MB apocrine patch
test would be in diagnosing onnel even before the patient reaches
the emergency room of a hospital to warn medical staff of
likelihood and severity of a heart attack. This test patch also
offers tremendous advantage for home use and could result in much
earlier diagnosis than currently available.
[0026] Unfortunately for mortality from coronary artery disease,
most individuals are asymptomatic until their cardiac disease
presents as death. An added value of a CPK-MB apocrine patch test
would be in diagnosing asymptomatic individuals. Individuals who
secreted CPK-MB during exercise using the patch could be identified
as having occult coronary disease. Ultimately this would be most
useful for individuals at home participating in weekend exercise
activity who could then place an apocrine CPK-MB patch on their
armpits while exercising. A positive result during exercise would
then diagnose a cardiac condition while the individual can still do
something about it.
[0027] It must be recognized that all chemicals found in the body
may not be markers of a disease. One example is the various
chemicals generated in an individual suffering from diabetes.
Example 2 reports an analysis of various compounds found in
elevated levels in a diabetic individual at a point where the blood
sugar levels are under control.
EXAMPLE 2
[0028]
2 Blood Serum Eccrine Apocrine Normal Sodium (mEq/l) 138 (N) 119
(VL) 127 (L) 135-149 Potassium (mEq/l) 4.2 (N) 65.0 (VH) 69.2 (VH)
3.5-5.2 Chloride (mEq/l) 99 (N) 80 (L) 96 (N) 98-108 CO.sub.2
(mEq/l) 25 (N) 2 (VL) 21 (N) 22-32 Glucose (mg/dl) 121 (H) 52 (L)
22 (VL) 61-114 BUN (mg/dl) 60 (H) 437 (VH) 401 (VH) 6-25 Creatinine
2.4 (H) 2.2 (H) 2.2 (H) 0.6-1.5 (VL = Very low; L = Low; N =
Normal; H = High; VH = Very High)
[0029] Standard analysis indicates that both eccrine sweat,
apocrine sweat and blood have about the same values for the various
analytes with the exception of BUN that the body must excrete, and
glucose which the body must conserve. However, apocrine BUN levels
can be readily used to signal an abnormally high concentration.
Apparently a marker for diabetes was not identified or, because
glucose was near normal, no marker was magnified in apocrine sweat.
The conclusion that can be reached from this data is that a proper
marker has not been identified. It can not be concluded that
apocrine sweat is not a suitable test solution for diabetes. The
results may be significantly different if glucose is significantly
elevated.
[0030] This effect is not limited to cancer marker detection or
detection of markers generated by other disease states. An apocrine
response is seen when an individual consumes a drug not normally
present in the body.
EXAMPLE 3
[0031] Samples of apocrine and eccrine sweat were collected prior
to oral consumption of a THC containing product. No THC was
detected in either sample. The test individual then orally consumed
(smoked) a quantity of a THC containing product. Small quantities
(0.5 cc) of apocrine and eccrine sweat was separately collected
over a period of 30 minutes under ambient conditions to induce
sweating. When the collected samples were tested using standard
laboratory tests for THC metabolites no detectable levels were
found in either sample. However, samples tested for THC (rather
than metabolites) using standard laboratory tests showed the
following significant differences between eccrine and apocrine
sweat, compared with THC concentrations in blood and urine.
3 Blood eccrine apocrine urine Delta 9 THC (nanograms/ml) 21 4 107
undetect- able
[0032] The quantity of THC in apocrine sweat was significantly
greater than in eccrine sweat (26.75 times) and blood (5.1
times).
[0033] The unique diagnostic capability of apocrine sweat has been
demonstrated with at least three distinctly different biomarkers
capable of indicating an abnormal physiological condition. PSA,
THC, and urea nitrogen, have been found to be subject to active ATP
transport with concentration in apocrine sweat. These markers,
disease related biochemicals or indicators of compromised normal
physiological condition are indicative of three different,
unrelated physiologic situations, namely prostate cancer or benign
prostatic hypertrophy, chronic renal failure, marijuana and opiate
intoxication. These are merely representative of the disease state
or physiological condition which can be signaled by functioning of
the apocrine gland through the release of detectable marker
constituents present in the smell chamber. Collection, analyzing or
signaling abnormal constituents of apocrine gland output by
collecting aroma samples (vaporized apocrine gland secretions),
clinical laboratory analysis of dissolved or vaporous components of
apocrine sweat, or the in situ signaling of presence of selected
target components by, for example, antibody detection techniques
and/or calorimetric indicators now becomes possible in a highly
reproducible and specific manner. As such the false positive and
false negative results obtained by the general and indiscriminate
collection of sweat, resulting from the failure of prior techniques
to recognize the difference between eccrine and apocrine sweat, can
be eliminated. Collection of only apocrine sweat now renders
useful, for use in many clinical situations and drug testing, what
was previously an unreliable technique.
[0034] The techniques and inventive procedures reported herein are
based upon the testing of liquid apocrine sweat as a:highly
indicative media when compared to liquid eccrine sweat and blood or
urine. An the apocrine sweat can be collected on a moisture
absorbing patch adhered to the collection site to receive apocrine
sweat followed by a detection or visible indication. Alternatively,
topically pad or wipe can be used to collect, in a rapid manner,
the sweat that is on the surface of the axillary (under arm) or
groin hairs (i.e., collect apocrine sweat). Chemical or monoclonal
antibody detection systems can then be used to identify specific
chemicals or biomarkers in apocrine sweat associated with specific
disease states or body functions.
[0035] However, another methodology is the use of electronic aroma
or smell detector systems, referred to as an electronic nose, to
take advantage of the discovery of specific function of the
apocrine gland. Examining the vaporized apocrine gland secretions
using such devices provides a new diagnostic modality which is
rapid, sensitive and non-invasive. It also has the advantage of
evaluating the product of the apocrine gland in the form (vapor)
intended.
[0036] Research reported by others has documented the excretory
function of eccrine sweat. Urea nitrogen and creatinine have now
been documented to have substantially the same values in both
apocrine and eccrine sweat in chronic renal failure. Both urea
nitrogen and creatinine are toxins and there is an obvious
advantage to eliminating them from the body by rapid perspiration.
However, the volume of apocrine sweat is minimal compared to
eccrine sweat and cannot remove a significant amount of any
chemical from the body. It is therefore interesting to note that
the body does not respond to delta nine tetrahydrocannibinol as a
toxin. It remains to be seen whether all foreign chemicals which
are administered to the body are treated as toxins by the eccrine
sweat glands, or whether their presence will just be signaled
solely or primarily by the apocrine glands. Active transport
concentration has only been demonstrated to be shared with both
apocrine and eccrine glands with urea nitrogen. Creatinine is also
interesting in that the values of both eccrine and apocrine gland
secretions in Example 2 above were just below that of serum values,
2.2 compared to 2.4. This is clearly different from the situation
of beta HCG (see below) or PSA in the normal range where the values
of both eccrine and apocrine gland secretions were minuscule
compared to serum values. Creatinine excretion in sweat
demonstrates that there is a situation where a chemical may be
excreted in eccrine sweat or signaled in apocrine sweat with
concentrations that are less than but approaching serum levels.
This is different from the biomarkers described above which are
concentrated by active transport but which still may have
significant clinical application for diagnostic purposes.
[0037] It has been documented that ovulating women were able to
identify the histocompatability type of men by using the smell
deposited upon t-shirts that were saturated with their sweat. Based
on the laboratory data regarding the ability of women to detect
histocompatability of males from apocrine sweat, it is believed
that comparative analysis of apocrine sweat of multiple
individuals, either electronically or chemically, can be used to
identify an individual's histocompatability type. Where this type
of testing is used in transplant medicine for screening organ
donors for a possible histocompatible match, the ability to match
donors using apocrine sweat analysis can be significant.
[0038] Also, based on the data listed herein for other compounds
concentrated in apocrine sweat it is expected that the apocrine
gland secretion of a pregnant woman will not reflect an abnormal
condition, pregnancy not being a condition requiring a biological
alert. This is confirmed by analysis that shows beta HCG blood
levels in a pregnant women to be 1944 .mu.m/ml but the levels in
eccrine and apocrine sweat to be 2 and 6 .mu.m/ml respectively.
However, ovulating women should have elevated levels of a marker,
such as luteinizing hormone which triggers ovulation, or other
compounds, such as estrogen known to be elevated in blood to
evidence a state of fertility suitable for conception, this being a
physiological state that a female body is desirous of transmitting
to a male. On the other hand, significantly low levels of estrogen
and/or progesterone prior to menstruation, often suggested as
triggering physiological and psychological disturbances, referred
to as premenstrual syndrome (PMS), could also be signaled by
absence of these markers in apocrine sweat.
[0039] Specific Applications for the test modality described herein
include testing of apocrine sweat for cancer markers such as
elevated PSA serum levels or other cancer markers. This may be both
a diagnostic tool for detecting the presence of prostate cancer and
an early warning of a potential disease state. Based on data
collected it is believed that the apocrine concentration of PSA, or
other cancer markers, in normal individuals will reflect (be
proportional to) the concentrations in blood serum. However, at a
threshold level, not yet determined, which the body recognizes as
abnormal, the ratio of PSA in apocrine sweat, when compared to
blood or eccrine sweat will increase significantly. This can be
readily detected with a simple patch or wipe collection device or
electronic nose or other electronic sensor as described herein.
[0040] The invention also contemplates the utilization of an
apocrine sweat collection patch which will include a reactive
marker, for example sensitive to PSA levels, other cancer markers,
or other disease markers in excess of a threshold level, which is
the point where the apocrine gland starts to concentrate PSA.
[0041] The invention herein contemplates applicability to many
other known, or presently unknown, biomarkers which can be applied
to an apocrine sweat collection patch. It is expected that analysis
of apocrine sweat will identify presently unknown biomarkers for
many cancers or other illnesses. These biomarkers will have
diagnostic value both in the testing of apocrine sweat as well as
expanding the utility of analysis of other body fluids such as
blood and urine. For example, the patch could be used as a
diagnostic test for HIV, the virus that causes AIDS. Because AIDS
is a readily treatable disease, particularly if detected early,
that early identification is very important. A very effective blood
test already exists which looks for the presence of antibodies
against HIV. This antibody, if it is in the blood should also be in
apocrine sweat in elevated. levels. A rapid patch test, using the
technology that is used in blood testing, is therefore
contemplated. The advantages of a rapid apocrine sweat test
compared to blood are many. Many individuals who would not readily
submit to a blood test would accept such a non-invasive test. Also,
since the result would be rapid there would be no delay, as is
common with blood test, to obtain an indication of a potential
disease state. The test patch could be available as an
over-the-counter product from a pharmacy for self administration in
the privacy of a users home. It could also be used privately by an
individual to test their sexual partners prior to becoming
intimate. It is also likely that an antibody present in the blood
in response to a bacterial or viral vector, such as anthrax, could
be readily and rapidly detected in apocrine sweat. This could
provide a modality to differentiate between viral and bacterial
illness and therefore make the clinical decision to use antibiotics
more scientifically based.
[0042] In several disease modalities contemplated by the invention,
it is possible that this test can be further accelerated by
applying the patch and then stimulating the patient to sweat by a
standard exercise protocol or by placing the patient in a room with
an elevated temperature, or even a sauna.
[0043] Screening studies described above indicate that the
technique and devices described herein have broad utility and are
not limited to the specific examples set forth. It is anticipate
that there are many biomarkers or antibodies against biomarkers
discovered in apocrine sweat that are as yet unidentified but which
are present in apocrine sweat in various abnormal physiological
conditions including but not limited to Alzheimer's, stroke,
Parkinson's, AIDS, Cardiac ischemia, malaria, various cancers other
than prostate or breast cancer, or which reflect changing body
functions such as ovulation, PMS, pregnancy or fetal stress during
pregnancy. These markers are susceptible to identification or
indication by apocrine sweat detection and analysis. By comparing
the apocrine sweat of an individual known to have a specific
disease to that of a normal individual, differences can be
identified and markers indicative of the disease can be isolated.
Similar comparisons can be conducted for different physiological
states. These bio-markers can then be used in diagnostic procedures
as described above to identify the diseased or changed
physiological condition. Also, once the markers are identified,
vaccines, therapy or other treatments can be developed or the
markers can be used to monitor the effect of known or
investigational treatment modalities. Additionally, once markers
are identified in apocrine sweat known techniques can be used to
identify cells within the body with those biomarkers to provide
cell specific treatment.
[0044] A skin patch incorporating features of the invention would
comprise a sensor portion which includes one or more marker
sensitive chemicals or antibodies (a detector chemical) and a
visible agent incorporated therewith which will indicate that the
detector chemical has reacted with or become bound to the
by-product, referred to as an indicator patch or wipe. The patch
could also include means to attach it to the skin surface such as
an adhesive strip. Any of numerous prior known skin patch
constructions can be used. The invention is not directed to the
specific design or use of a skin patch to collect body chemicals.
Instead it is directed to a method of detecting cancer, other
disease states, or physiological conditions by placing a skin
patch, using a wipe, or applying an electronic sensor which detects
and identifies vaporizing apocrine sweat so as to collect emissions
(liquid or vapors) from the apocrine sweat gland. The wipe or patch
can have a detector specific to one or more compounds present in
apocrine sweat which are only present, or are present in
significant elevated levels, when that condition being diagnosed
for exists. The patch, wipe or detector may be combined with an
agent which provides a visible indication, such as a color change,
when the compound exists in collectable and measurable quantities
in apocrine sweat. It is contemplated that the visible indicia of
the presence of a target compound can be ascertained while the
patch is still attached to the skin (i.e., by viewing the outer
surface of the patch) so that a time dependent response can be
monitored or, alternatively, immediately upon removing the patch
and observing the skin contacting surface of the patch. Still
further, the invention contemplates that the agent which provides a
visible indication of marker presence can be applied to the patch
or wipe immediately prior to collecting the sweat or immediate
after the sweat is collected.
[0045] The skin patch would be applied to a portion of the body
where apocrine sweat is generated, such as the underarm or groin,
so as to require the shortest period of time to collect an adequate
sample of sweat. Topical or systemic compounds which stimulate the
generation of perspiration can also be used to increase the
generation of sweat. Alternatively or in addition to these
compounds, exercise sufficient to increase sweat may also be useful
to increase sweat production and reduce the time to collect a
sufficient quantity of sweat. Because the technique and device
described herein is non-invasive it may have particular utility in
acute or urgent care facilities or in instances where blood samples
are difficult to obtain or diagnose, such as in pediatrics or rural
and third world settings. While the technique and products set
forth herein are primarily intended for use in humans, there is no
reasons why the same product and technique can not be adapted for
use on other mammals which produce sweat or other skin exudates,
the techniques also be applicable in a veterinary setting.
[0046] While the principal embodiments are directed to substances
generated by the apocrine gland which are analyzed or detected
contemporaneously with the collection of such materials, the
invention does not exclude the collection for remote primary or
secondary analysis. By secondary analysis, applicant contemplates
the generation of a signal, upon collection of the apocrine gland
sweat, that a target substance has been detected followed by a
laboratory analysis of the collected substance to specify and/or
quantify the collected substance. Further, while apocrine sweat has
been identified as a unique source of particular markers, it is
also known that eccrine sweat may have the same markers but in
lower concentrations. The indicator patch or wipe therefore can
also be utilized in the same manner on eccrine sweat, even though
such use may be less efficient or effective.
[0047] It is evident from the foregoing that there are many
additional embodiments of the present invention which, while not
expressly described herein, are within the scope of this invention
and may suggest themselves to one of ordinary skill in the art. For
example,
[0048] It is therefore intended that the invention be limited
solely by the appended claims.
* * * * *