WO2012135227A2

WO2012135227A2 - Methods and systems for assessing exposure to heavy metals

Info

Publication number: WO2012135227A2
Application number: PCT/US2012/030752
Authority: WO
Inventors: Eddie Reed
Original assignee: Eddie Reed
Priority date: 2011-03-29
Filing date: 2012-03-27
Publication date: 2012-10-04
Also published as: WO2012135227A3

Abstract

Embodiments of the present invention relate to methods and systems for assessing exposure of a subject to an environmental source of heavy metals. Some embodiments include determining the level of at least one metal in a subject. Some embodiments include systems for assessing the exposure of a plurality of subjects to an environmental source of heavy metals.

Description

METHODS AND SYSTEMS FOR ASSESSING EXPOSURE TO HEAVY METALS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 61/516070, entitled "DETECTION METHOD FOR CRUDE OIL EXPOSURE USING HEAVY METALS", filed March 29, 201 1, the disclosure of which is incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0002] Embodiments of the present invention relate to methods and systems for assessing exposure of a subject to an environmental source of heavy metals. Some embodiments include determining the level of at least one heavy metal in a subject. Some embodiments include systems for assessing the exposure of a plurality of subjects to an environmental source of heavy metals.

BACKGROUND OF THE INVENTION

[0003] The BP-related Gulf oil disaster of 2010, resulted in an estimated 4.4 million barrels of crude oil being spilled into the Gulf of Mexico (1). The long term health effects of that event have yet to unfold. Previous studies of the health effects of major oil spills have focused on social disruptions to the respective community, and to the mental health of individuals. Studies of the biology of health effects from major oil spills have been somewhat limited (2,3,4,5). The Gulf Oil Spill of 2010 has potential implications for the public health of the affected community; as well as implications for the medical health of specific individuals who may be directly exposed to crude oil. Accordingly, there is a need to assess exposure of an individual to environmental sources of heavy metals, such as crude oil.

SUMMARY OF THE INVENTION

[0004] Some embodiments of the methods and systems provided herein include a method for assessing the exposure of a subject to an environmental source of heavy metals comprising measuring the level of at least one heavy metal in a biological sample obtained from said subject.

[0005] Some embodiments also include comparing the level of the at least one heavy metal in the biological sample to a control level of said at least one heavy metal, wherein said control level of said at least one heavy metal is selected from the group consisting of the level of said at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals, the level of said at least one heavy metal in a sample which has a known exposure to an environmental source of heavy metals, a predetermined reference level of said at least one heavy metal which is known to be indicative of exposure to an environmental source of heavy metals and a predetermined reference level of said at least one heavy metal which is known to be indicative of a lack of exposure to an environmental source of heavy metals.

[0006] In some embodiments, an increase in the level of said at least one heavy metal in the biological sample compared to the level of said at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals, or compared to a predetermined reference level of said at least one heavy metal which is known to be indicative of a lack of exposure to an environmental source of heavy metals is indicative of the exposure of the subject to an environmental source of heavy metals.

[0007] In some embodiments, the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium.

[0008] In some embodiments, the method comprises measuring the levels of at least two heavy metals in the biological sample. In some embodiments, the method comprises measuring the levels of at least three heavy metals in the biological sample. In some embodiments, the method comprises measuring the levels of at least four heavy metals in the biological sample. In some embodiments, the method comprises measuring the levels of at least five heavy metals in the biological sample.

[0009] In some embodiments, measuring the level of at least one heavy metal comprises performing a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively- coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

[0010] In some embodiments, the environmental source of heavy metals comprises an oil spill.

[0011] In some embodiments, the presence, absence or level of the at least one heavy metal in the biological sample is indicative of the environmental source of heavy metals.

[0012] Some embodiments also include measuring the level of at least one heavy metal in a plurality of biological samples, wherein each biological sample is obtained from a different subject.

[0013] In some embodiments, the biological sample comprises nucleic acid.

[0014] In some embodiments, the biological sample comprises protein. [0015] In some embodiments, the level of said at least one heavy metal in said biological sample is measured ex vivo.

[0016] In some embodiments, the subject is mammalian.

[0017] In some embodiments, the subject is human.

[0018] In some embodiments, the subject resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals.

[0019] Some embodiments also include providing a treatment for exposure to heavy metal, or recommending a preventative or diagnostic regimen if the level of said at least one heavy metal in said biological sample from said subject indicates that said subject has been exposed to an environmental source of heavy metals.

[0020] Some embodiments of the methods and systems provided herein include a method for assessing the likelihood of a subject developing a disorder associated with exposure to an environmental source of heavy metals comprising measuring the level of at least one heavy metal in a biological sample obtained from said subject.

[0021] Some embodiments also include comparing the level of the at least one heavy metal in the biological sample to a control level of said at least one heavy metal, wherein said control level of said at least one heavy metal is selected from the group consisting of the level of said at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals, the level of said at least one heavy metal in a sample which has a known exposure to an environmental source of heavy metals, a predetermined reference level of said at least one heavy metal which is known to be indicative of exposure to an environmental source of heavy metals and a predetermined reference level of said at least one heavy metal which is known to be indicative of a lack of exposure to an environmental source of heavy metals.

[0022] In some embodiments, the disorder associated with exposure to an environmental source of heavy metals is selected from the group consisting of kidney damage, nerve damage, pulmonary toxicity, cardiac toxicity, skin rash, skin lesion, an immunological disorder, and cancer.

[0023] In some embodiments, the presence, absence, or level of the at least one heavy metal in the biological sample is indicative of the likelihood of the subject developing a disorder associated with exposure to an environmental source of heavy metals.

[0024] In some embodiments, the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium. [0025] In some embodiments, the method comprises measuring the levels of at least two heavy metals in the biological sample. In some embodiments, the method comprises measuring the levels of at least three heavy metals in the biological sample. In some embodiments, the method comprises measuring the levels of at least four heavy metals in the biological sample. In some embodiments, the method comprises measuring the levels of at least five heavy metals in the biological sample.

[0026] In some embodiments, measuring the level of at least one heavy metal comprises performing a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively- coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

[0027] In some embodiments, the environmental source of heavy metals comprises an oil spill.

[0028] Some embodiments also include measuring the level of at least one heavy metal in a plurality of biological samples, wherein each biological sample is obtained from a plurality of subjects.

[0029] In some embodiments, the biological sample comprises nucleic acid.

[0030] In some embodiments, the biological sample comprises protein.

[0031] In some embodiments, the level of said at least one heavy metal in said biological sample is measured ex vivo.

[0032] In some embodiments, the subject is mammalian.

[0033] In some embodiments, the subject is human.

[0034] In some embodiments, the subject resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals.

[0035] Some embodiments also include providing a treatment for exposure to heavy metal, or recommending a preventative or diagnostic regimen if the level of said at least one heavy metal in said biological sample from said subject indicates that said subject has been exposed to an environmental source of heavy metals.

[0036] Some embodiments of the methods and systems provided herein include a system for assessing exposure of a plurality of subjects to an environmental source of heavy metals comprising: (a) at least one data acquisition unit comprising hardware accessories and software that facilitates interface with a subscriber and the entry of subject demographic data, and test equipment configured to measure test data comprising the level of at least one heavy metal in a biological sample obtained from a subject, said test data in combination with said subject demographic data comprising heavy metal data; and (b) at least one data processing unit communicatively coupled to the at least one data acquisition unit, said at least one data processing unit configured to systematically receive cumulative heavy metal data from the plurality of subjects, analyze cumulative heavy metal data to identify correlations between subject demographic data and test data, and identify a subject exposed to an environmental source of heavy metals.

[0037] In some embodiments, the at least one data processing unit is configured to compare the cumulative heavy metal data with a reference model.

[0038] In some embodiments, the at least one data processing unit is configured to modify a reference model.

[0039] Some embodiments also include a plurality of data acquisition units distributed over a geographical area.

[0040] Some embodiments also include a plurality of data processing unit distributed over a geographical area.

[0041] In some embodiments, the at least one data processing unit is communicatively coupled to the at least one data acquisition unit over a network.

[0042] In some embodiments, subject demographic data is selected from the group consisting of personal identification data, physical characteristics data, health profile data, family health history data, drug and vitamin/mineral supplement data, health baseline data, diet and nutritional data, environmental exposure data, and behavioral data.

[0043] In some embodiments, the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium.

[0044] In some embodiments, the test equipment is configured to measure the levels of at least two heavy metals in the biological sample. In some embodiments, the test equipment is configured to measure the levels of at least three heavy metals in the biological sample. In some embodiments, the test equipment is configured to measure the levels of at least four heavy metals in the biological sample. In some embodiments, the test equipment is configured to measure the levels of at least five heavy metals in the biological sample.

[0045] In some embodiments, the test equipment is configured to perform a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively-coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

[0046] In some embodiments, the environmental source of heavy metals comprises an oil spill. [0047] In some embodiments, the test data comprises the level of heavy metal- nucleic acid in said biological sample.

[0048] In some embodiments, the test data comprises the level of heavy metal- protein in said biological sample.

[0049] In some embodiments, the system is configured to receive test data comprising the level of said at least one heavy metal in said biological sample which has been measured ex vivo.

[0050] In some embodiments, the test data has been obtained from a mammalian subject.

[0051] In some embodiments, the test data has been obtained from a human subject.

[0052] In some embodiments, the test data has been obtained from at least one subject who resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals.

[0053] Some embodiments of the methods and systems provided herein include a method for assessing exposure of a population of subjects to an environmental source of heavy metals comprising (a) obtaining heavy metal data from at least one data acquisition unit comprising hardware accessories and software that facilitates interface with a subscriber and the entry of subject demographic data, and test equipment configured to measure test data comprising the level of at least one heavy metal in a biological sample obtained from a subject, said test data in combination with said subject demographic data comprising said heavy metal data; and (b) transmitting the heavy metal data to at least one data processing unit from the at least one data acquisition unit, the at least one data processing unit configured to: systematically receive cumulative heavy metal data from a plurality of subjects, analyze cumulative heavy metal data to identify correlations between subject demographic data and test data, and identify a subject exposed to an environmental source of heavy metals.

[0054] In some embodiments, the at least one data processing unit is configured to compare the cumulative heavy metal data with a reference model.

[0055] In some embodiments, the at least one data processing unit is configured to modify a reference model.

[0056] Some embodiments also include a plurality of data acquisition units distributed over a geographical area.

[0057] Some embodiments also include a plurality of data processing unit distributed over a geographical area. [0058] In some embodiments, the at least one data processing unit is communicatively coupled to the at least one data acquisition unit over a network.

[0059] In some embodiments, subject demographic data is selected from the group consisting of personal identification data, physical characteristics data, health profile data, family health history data, drug and vitamin/mineral supplement data, health baseline data, diet and nutritional data, environmental exposure data, and behavioral data.

[0060] In some embodiments, the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium.

[0061] In some embodiments, the test equipment is configured to measure the levels of at least two heavy metals in the biological sample. In some embodiments, the test equipment is configured to measure the levels of at least three heavy metals in the biological sample. In some embodiments, the test equipment is configured to measure the levels of at least four heavy metals in the biological sample. In some embodiments, the test equipment is configured to measure the levels of at least five heavy metals in the biological sample.

[0062] In some embodiments, the test equipment is configured to perform a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively-coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

[0063] In some embodiments, the environmental source of heavy metals comprises an oil spill.

[0064] In some embodiments, the test data comprises the level of heavy metal- nucleic acid in said biological sample.

[0065] In some embodiments, the test data comprises the level of heavy metal- protein in said biological sample.

[0066] In some embodiments, the system is configured to receive test data comprising the level of said at least one heavy metal in said biological sample which has been measured ex vivo.

[0067] In some embodiments, the test data has been obtained from a mammalian subject.

[0068] In some embodiments, the test data has been obtained from a human subject.

[0069] In some embodiments, the test data has been obtained from at least one subject who resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals. DETAILED DESCRIPTION

[0070] Crude oil has many chemical components including heavy metals, such as, lead, nickel, vanadium, cadmium, and copper, single chain hydrocarbons, and a range of polycyclic aromatic hydrocarbons (6-12). Studies from a number of geographic locations indicate that after a crude oil spill, environmental contaminants contain high levels of PAHs and heavy metals (6-12).

[0071] Environmental contamination with heavy metals has been studied following a number crude oil spills of at sites including Nigeria and India. After an oil spill, heavy metals including lead, nickel, copper, vanadium, cadmium, are detected at high levels on the surface, in sub-surface soils, and in plants (13-18). In the Niger River Delta, heavy metals were detected in surface, in sub-surface soils, and in plants in the following ranges: lead: 0.32 to 0.80 mg/kg; nickel: 0.53 to 18.0 mg/kg; copper: 0.15 to 0.30 mg/kg; vanadium: up to 0.20 mg/kg; and cadmium: up to 0.20 mg/kg. Measurements in plants also showed an uptake of these metals to levels higher than the surrounding soil. In a similar study following crude oil spill off the coast of Spain, heavy metals were found in water and sediment that were not visibly soiled by oil (17). Metals that were measured included: cadmium, zinc, copper, lead, arsenic, and others. High levels of heavy metals such as lead, chromium, cadmium have been linked to medical disorders that are manifested by kidney damage, nerve damage, pulmonary toxicity, cardiac toxicity, skin rashes and lesions, immunologic disorders, teratogenesis, and cancer (20).

[0072] Polycyclic aromatic hydrocarbons may exist as two-ring (naphthalenes), three-ring (phenanthrene), four-ring (pyrene), or higher-complexity hydrocarbons. Reports in the literature suggest that the molecular weight of the polycyclic aromatic hydrocarbon, may influence the tendency to evaporate, to be soluble, and to be absorbed into the food chain. In a study that followed a major oil spill along the coastline of India, more than 39 different polycyclic aromatic hydrocarbons were measured by GC-mass spectrometry in open sea samples (6). These polycyclic aromatic hydrocarbons included two ring polycyclic aromatic hydrocarbons (naphthalenes), three ring polycyclic aromatic hydrocarbons (phenanthrenes), four ring polycyclic aromatic hydrocarbons (pyrenes), and other very high molecular weight hydrocarbons (6). In the India study, the low saline conditions in Coastal waters appeared to result in increased uptake of polycyclic aromatic hydrocarbons into the plant and animal flora of the area. This effect was enhanced even further when oil dispersants were used.

[0073] Applicant has discovered at least nine heavy metals including cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium, present in samples taken from oil- contaminated shorelines. [0074] It is possible that any individual might be exposed to significant levels of one or more heavy metals, as a matter of random activity. If it is assumed that random exposure to any heavy metal, such as cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium, is independent of random exposure to any other heavy metal such as cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium, the mathematical probability of random exposure for any single person, to two, or three, or more of these nine heavy metals can be determined.

[0075] For example, if it is assumed that the chance of random exposure to any one heavy metal is independent of the chance of random exposure to any other heavy metal, and the chance of random exposure is the same for each heavy metal, and if the chance of random exposure to any one heavy metal is 1 in 100, then the chance of a random exposure to any two heavy metals is, 2: ==> 0.001997079. The chance of random exposure to any three of the heavy metals is, 3: ==> 3.362086e-05. The chance of random exposure for any four of the heavy metals is, 4: ==> 3.396047e-07. If the chance of random exposure to any one heavy metal is 1 in 500, then, the chance of a random exposure to any two heavy metals is 2: ==> 0.00008316. The chance of exposure to any three heavy metals is 3: ==> 2.777667e-07. The chance for random exposure for any four heavy metals is 4: ==> 5.566467e-10. If the chance of random exposure to any one is 1 in 1000; then, the chance of a random exposure to any two heavy metals is 2: ==> 0.0000209. The chance of random exposure to any three heavy metals is 3: ==> 3.48602 le-08 (or 0.0000000349). The chance of random exposure to any four heavy metals is 4: ==> 3.4895 lOe-11. The p- values associated with exposure to more than one heavy metal are summarized in Table 1.

TABLE 1

[0076] If an individual has evidence of exposure to any two heavy metals listed, such as cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, or chromium, the exposure is unlikely to be due to random chance. Moreover, if an individual has evidence of exposure to three or more, of any of the nine heavy metals listed, it is even more unlikely that the exposure is due to random chance, and strong evidence of exposure to a crude oil spill. Exposure to a crude oil spill may be direct (skin contact or direct ingestion), or indirect (through the food chain, etc). Therefore, heavy metal measurements could be coupled with social history, history of potential exposures, clinical parameters, and other assessments.

Metal-protein levels and metal-DNA levels can be measured in human tissues

[0077] Measurements of a foreign metal, such as platinum, cadmium, and gallium in biological samples such as human tissues and tissue culture are well known in the art (22,24,28- 31 ,38). Platinum-DNA adducts have also been measured in a wide range of tissues, including white blood cells, bone marrow, liver, kidney, cancer tissues, etc, in cord blood (39) and intraocular tumors of childhood (40). Platinum levels have been measured in whole blood, in protein-free supernatant from blood, and in cellular DNA from white blood cells. The measurement of platinum in whole blood (or any whole tissue) is easier, because the platinum levels are higher, and less tissue is needed for the analysis. The lower limits of detection for platinum, for cadmium, and for gallium are all similar.

[0078] Using AAS to measure platinum-DNA damage, different levels of clinical activity in human subjects can be distinguished. For example, platinum-DNA damage was measured by AAS, in a cohort of 49 persons with 24 different tumor types (26). The data showed that metal-DNA damage level, in white cell DNA, was related to the human clinical outcome, regardless of tumor type. Patients with complete tumor response had higher levels of metal-DNA damage than patients with partial tumor response; who, in turn, had higher metal- DNA damage levels than patients with no tumor response to therapy. These data suggest that it is not only possible to answer exposure questions in a "yes-no" fashion. Thus, in some embodiments, the levels of heavy metal, metal-protein , and/or the levels of metal-DNA damage is determined, for one or more of the nine heavy metals described above in order to identify or diagnose meaningful biological endpoints in human individuals exposed to crude oil.

[0079] In some embodiments, metal-protein damage and/or metal-DNA damage is measured in humans exposed to crude oil, in order to identify or diagnose clinically important medical problems that such individuals might experience. As an example, a person with measurable levels of one or more heavy metals covalently bound to blood proteins, may also have an elevated serum creatinine indicating kidney damage; or clinical evidence of nerve damage; or other medical effects.

[0080] One characteristic of heavy metals, when found in human tissues, is persistence. Measurable levels of platinum compounds can persist in human and animal tissues for months to years (38,41). This is also true for metals such as lead, mercury, or platinum bound to biological molecules in humans or other mammals (20). This suggests the possibility that humans who may not have had an acute exposure to crude oil, may have an ultimate exposure to crude oil, indirectly, through food chain exposure, or other possible sources.

Environmental sources of heavy metals

[0081] Some embodiments of the methods and systems provided herein include assessing a subject's exposure to an environmental source of heavy metals. There are a variety of environmental sources that a subject may be exposed to. In general, an environmental source of heavy metals includes materials with a high concentration of heavy metals. In some embodiments, an environmental source of heavy metals can include crude oil, the products of crude oil, products of industrial methods to purify metals such as smelting of copper, products of the industrial preparation of nuclear fuels. In some embodiments, a subject's exposure to an environmental source of heavy metals is direct. For example, a subject may come in contact with an environmental source of heavy metals. In some embodiments, a subject's exposure to an environmental source of heavy metals is indirect. For example, a subject may consume material that has contacted an environmental source of heavy metals.

Biological samples

[0082] Some embodiments of the methods and systems provided herein include biological samples. Biological samples include any biological material that may be used to measure the presence, absence or level of a heavy metal in an organism. One skilled in the art would know methods for selecting a particular biological sample and how to collect the sample. Examples of sources of biological samples include animals, plants, and microbes. In some embodiments, sources of biological samples include biopsy or other in vivo or ex vivo analysis of prostate, breast, skin, muscle, facia, brain, endometrium, lung, head and neck, pancreas, small intestine, blood, liver, testes, ovaries, colon, skin, stomach, esophagus, spleen, lymph node, bone marrow, kidney, placenta, or fetus. In some embodiments, a biological sample includes a fluid sample, such as peripheral blood, lymph fluid, ascites, serous fluid, pleural effusion, sputum, cerebrospinal fluid, amniotic fluid, lacrimal fluid, stool, or urine. Samples include single cells, whole organs or any fraction of a whole organ, in any condition including in vitro, ex vivo, in vivo, post-mortem, fresh, fixed, or frozen. In some embodiments, a biological sample includes nucleic acid derived from a subject. Methods to obtain nucleic acids from cells or tissues are well known in the art. Methods for measuring heavy metals

[0083] Some embodiments of the methods and systems provided herein include measuring the presence, absence or level of one or more heavy metals in a biological sample. Examples of heavy metals include arsenic, cadmium, cobalt, chromium, copper, mercury, manganese, nickel, lead, tin, and thallium. In preferred embodiments, heavy metals include cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium. Methods to measure heavy metals in biological samples are well known in the art and include atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively-coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

[0084] In techniques that include measuring atomic absorption, the "ground state" atom absorbs light energy of a specific wavelength as it enters the "excited state." As the number of atoms in the light path increases, the amount of light absorbed also increases. By measuring the amount of light absorbed, a quantitative determination of the amount of analyte can be made. The use of special light sources and careful selection of wavelengths allow the specific determination of individual elements. The basic components of an atomic absorption instrument include: a light source that emits the spectrum of the element of interest; an absorption cell in which atoms of the sample are produced (e.g., flame, graphite furnace, MHS cell, FIAS cell, FIMS cell); a monochromator for light dispersion; a detector, which measures the light intensity and amplifies the signal; and a display that shows the reading after it has been processed by the instrument electronics. In addition, there are two basic types of atomic absorption instruments: single- beam and double-beam. The main sources used for atomic absorption are the hollow cathode lamp (HCL) and the electrodeless discharge lamp (EDL). Quantitative measurements in atomic absorption are based on Beer's Law, which states that concentration is proportional to absorbance (C = kA). It is well known that for most elements, particularly at high concentrations, the relationship between concentration and absorbance deviates from Beer's Law and is not linear. Modern atomic absorption instruments have the ability to calibrate and compute concentrations using absorbance data from linear and nonlinear curves (See e.g., "Analytical Methods for Atomic Absorption Spectroscopy" Perkin Elmer Atomic Absorption Manual Part No. 0303-0152, Release D, September 1996, incorporated by reference herein in its entirety).

[0085] In techniques that include measuring atomic emission, the sample is subjected to a high-energy thermal environment in order to produce excited-state atoms. This environment can be provided by a flame or a plasma. However, since the excited state is unstable, the atoms spontaneously return to the "ground state" and emit light. The emission spectrum of an element consists of a collection of emission wavelengths called emission lines because of the discrete nature of the emitted wavelengths. The intensity at an emission line will increase as the number of excited atoms of the element increases. In particular, ICP-MS employs an inductively coupled argon plasma as an ionization source and a mass spectrometer to separate and measure analyte ions formed in the ICP-MS source. See e.g., U.S. Patent No.s 6,265,717, 6639665, incorporated by reference herein in their entireties. Normally, the sample is taken into solution and pumped into a nebulizer, which generates a sample aerosol. The sample aerosol passes into the ICP-MS, where it is desolvated, atomized and ionized. The resulting sample ions are then transferred from the plasma at atmospheric pressure, to the mass spectrometer that is situated inside a vacuum chamber, via a differentially pumped interface. The ions pass through two orifices in the interface, known as sampling and skimmer cones, and are focused into a quadrupole mass analyzer. The analyzer separates the ions based on their mass/charge ratio prior to measurement by an electron multiplier detection system. Each elemental isotope appears at a different mass with a peak intensity directly proportional to the initial concentration of that isotope in the sample; thus elemental concentrations in the sample can be measured. Inductively coupled plasma atomic emission spectroscopy (ICP-AES), also referred to as inductively coupled plasma optical emission spectrometry (ICP-OES) is an another example of a technique that may be used to measure heavy metals. Stefansson A, et al, (2007). "New methods for the direct determination of dissolved inorganic, organic and total carbon in natural waters by Reagent-Free Ion Chromatography and inductively coupled plasma atomic emission spectrometry". Anal. Chim. Acta 582 (1): 69-74, incorporated by reference herein in its entirely.

[0086] Some embodiments of the methods and systems provided herein include methods for assessing the exposure of a subject to an environmental source of heavy metals comprising measuring the level of at least one heavy metal in a biological sample obtained from the subject. It will be appreciated that the terminology "measuring the level of at least one heavy metal" includes measuring the level of one heavy metal or a plurality of heavy metals. Some embodiments include measuring in a biological sample obtained from a subject the level of at least one heavy metal, at least two heavy metals, at least three heavy metals, at least four heavy metals, at least five heavy metals, at least one six metals, at least seven heavy metals, at least eight heavy metals, at least nine heavy metals, at least ten heavy metals, or more.

[0087] Some embodiments also include comparing the level of at least one heavy metal in a biological sample with the level of at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals, the level of at least one heavy metal in a sample which has a known exposure to an environmental source of heavy metals or to a predetermined reference level which is known to be indicative of exposure to an environmental source of heavy metals or to be indicative of a lack of exposure to an environmental source of heavy metals.

[0088] In some embodiments, an increase in the level of at least one heavy metal or a plurality of heavy metals in a biological sample compared to the level of at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals or to a predetermined reference level known to be indicative of a lack of exposure to an environmental source of heavy metals is indicative of the exposure of the subject to an environmental source of heavy metals. In some embodiments, the increase in the level of at least one heavy metal in a biological sample compared to the level of at least one heavy metalin a sample which has not been exposed to an environmental source of heavy metals or to a predetermined reference level known to be indicative of a lack of exposure to an environmental source of heavy metals can be at least about 2-fold greater, at least about 5-fold greater, at least about 10-fold greater, at least about 10-fold greater, at least about 20-fold greater, at least about 30-fold greater, at least about 40-fold greater, at least about 50-fold greater, at least about 60-fold greater, at least about 70-fold greater, at least about 80-fold greater, at least about 90-fold greater, at least about 100-fold greater, at least about 1000-fold greater, at least about 10,000-fold greater, or more. In some embodiments, the level of at least one heavy metal in the biological sample compared to the level of at least one heavy metal in a sample which has a known exposure to an environmental source of heavy metals or to a predetermined reference level known to be indicative of exposure or a lack of exposure to an environmental source of heavy metals is indicative of the exposure of the subject to an environmental source of heavy metals.

[0089] In some embodiments, the presence, absence or level of a particular heavy metal or combination of heavy metals is indicative of the environmental source of heavy metals. For example, in some embodiments, the presence, absence or level of a particular heavy metal or combination of heavy metals can identify the source of an oil spill. It is envisaged that different oil spills comprise different levels and combinations of heavy metals. Such differences can be used to identify the source of an oil spill and the environmental source of at least one heavy metal.

[0090] In some embodiments a subject comprises a biological material which can be used as the source of a biological sample. Examples of subjects include mammals. In some embodiments, a subject includes a human. In some embodiments, a subject resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals. In some embodiments, a subject is located, resides, works or has been at a location no further than about 1 mile, no further than about 2 miles, no further than about 5 miles, no further than about 10 miles, no further than about 15 miles, no further than about 20 miles, no further than about 30 miles, no further than about 50 miles, no further than about 60 miles, no further than about 70 miles, no further than about 80 miles, no further than about 90 miles, no further than about 100 miles, no further than about 150 miles, no further than about 200 miles, from an environmental source of heavy metals. In some such embodiments, an environmental source of heavy metals includes a coastline contacted with an oil spill. In some embodiments, a subject is located, resides works or has been at a location greater than about 1 mile, greater than about 2 miles, greater than about 5 miles, greater than about 10 miles, greater than about 15 miles, greater than about 20 miles, greater than about 30 miles, greater than about 50 miles, greater than about 60 miles, greater than about 70 miles, greater than about 80 miles, greater than about 90 miles, greater than about 100 miles, greater than about 150 miles, greater than about 200 miles, or more, from an environmental source of heavy metals. In some such embodiments, an environmental source of heavy metals includes a coastline contacted with an oil spill.

[0091] Some embodiments include measuring the level of at least one heavy metal in a plurality of biological samples, wherein each biological sample in the plurality of biological samples is obtained from a different subject. In some such embodiments, a plurality of subjects includes at least 2 subjects, at least 5 subjects, at least 10 subjects, at least 50 subjects, at least 100 subjects, at least 200 subjects, at least 500 subjects, at least 1000 subjects, at least 5000 subjects, at least 10000 subjects, or more.

Methods for assessing risk

[0092] Some embodiments of the methods and systems provided herein include methods for assessing the likelihood of a subject developing a disorder associated with exposure to an environmental source of heavy metals. Disorders associated with exposure to an environmental source of heavy metals include disorders in which a subject is exposed to high levels of at least one heavy metal. Examples of such disorders include kidney damage, nerve damage, pulmonary toxicity, cardiac toxicity, skin rash, skin lesion, an immunological disorder, and cancer.

[0093] In some embodiments, the level of at least one heavy metal in a biological sample obtained from said subject is measured, and compared to the level of at least one heavy metal in a sample with known potential of developing a disorder associated with exposure to an environmental source of heavy metals or to a predetermined reference level known to be indicative of a risk of developing or not developing a disorder associated with exposure to an environmental source of heavy metals. In some embodiments, the level of at least one heavy metal in a biological sample obtained from said subject is compared to the level of at least one heavy metal known to be sufficient to increase the likelihood for a subject develop a disorder associated with exposure to an environmental source of heavy metals. In some embodiments, the likelihood for a subject develop a disorder associated with exposure to an environmental source of heavy metals is increased at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%..

Systems for assessing exposure to a source of heavy metals

[0094] Some embodiments of the methods and systems provided herein include systems for assessing exposure of a population of subjects to an environmental source of heavy metals. Some such systems include at least one data acquisition unit and at least one data processing unit.

[0095] In some embodiments, a data acquisition unit can include hardware accessories and software that facilitate interface with a subscriber and the entry of subject demographic data. Examples of hardware and software that facilitate the entry of subject demographic data into a system are well known in the art. In some embodiments, hardware can include a computer, a central processing unit, a display, memory, and an input device. In some embodiments, a subscriber can include any individual, for example, a subject, a physician, a technician, a researcher, or a scientist. In some embodiments, subject demographic data includes subject information that is known to be or is likely to be correlative to a subject's exposure to an environmental source of heavy metals. Exemplary data categories include personal identification data, physical characteristics data, health profile data, family health history data, drug and vitamin/mineral supplement data, health baseline data, diet and nutritional data, environmental exposure data, and behavioral data.

[0096] The personal identification data comprises essential personal information that facilitates the identification of a subject and the establishment of core demographic characteristics. For example, the personal identification data includes name, address, residence history, age, gender, race, ethnicity, education, sexual preference, martial status, living arrangements, marital history, children, occupation, work history, home and work environments, travel history, military service history, genealogy, relationships, recreational activities and the like. Sensitive information, such as, name and street address, maybe segregated and coded for privacy and security.

[0097] The physical characteristics data includes height, weight, body fat ratio, body symmetry and dimensions, skin shade and texture, eye color, hair growth/color and texture, strength symmetry, endurance, coordination, posture, gait, nail growth and features, feet size, physical peculiarities, physical deformities, growths, blemishes, teeth and gums, flexibility, and the like. These characteristics are useful in establishing a subject's physical uniqueness as well as membership in population groups that share certain characteristics.

[0098] The health profile data comprises a broad array of information pertaining to a subject's psychological and physiological characteristics and condition, medical history data, hazardous and toxic material exposure data, allergies data, disabilities, reproductive history, depression data, family (genealogy) health history data stress level data, mental condition data, current health conditions data, physical sensitivities and pain data, activity level and physical fitness data, illnesses history data, history of injuries data, chronic conditions data, visual acuity data, night vision data, hearing acuity data, reaction time data and the like. The health profile data also incorporates the results of quantitative tests including blood chemistry tests, breath analysis (i.e. laser absorption spectroscopy), medical imaging (i.e. x-rays, magnetic resonance imaging, lithotripsy, computed tomography, fluorescence spectroscopy, ultrasounds, thermographs, and others), photographic imaging, and other psychological, physical, and other physiological tests.

[0099] The family health history addresses the health conditions and unique characteristics of a subject's living and deceased blood relatives. The family health history data is comprised of personal description data, physical description data, physical characteristics, demographic data, occupational data, disabilities, behaviors, health and medical histories, and the like. The family history data collection includes names, birthdates, place of birth, number of children (including genders and birthdates), places of residency, health histories, ages at death, height, weight, physical and health peculiarities, chronic conditions, sensitivities/allergies, disease history, cause of death, health conditions at time of death, history of injuries, deformities, visual acuity, hearing acuity, mental condition and acuity, disabilities, occupations/professions, medication history (including diagnosis, treatments, test results, evaluations, and the like), reproductive histories, alcohol and drug usage, blood types, and other psychological, physical, physiological and behavioral details that would be useful in the identification of genetic characteristics and predispositions. [0100] The drug and vitamin/mineral supplement data comprises a detailed history of prescription and non-prescription drugs, vitamin supplements, herbs, and mineral supplement usages. Included in the history may be the item description, dosage, frequency taken, date started, reason for taking, date stopped, reason for stopping, and observed effects, side effects, reactions, and the like.

[0101] The health baseline data may, in part, be derived from information compiled in previous databases including, the personal identification data, physical characteristics data and health profile data. Certain key psychological characteristics (i.e. depression, confusion, neurosis and other like mental conditions or mental cognitive peculiarities), physical characteristics (i.e. height, weight, body fat ratio, posture, flexibility, mobility, hair growth, hair color, skin color/tone, eye color and the like), physiological characteristics (i.e. visual performance, hearing performance, blood pressure, heart rate, repertory rate, heart rhythm, blood chemistry, and other major organ system performance characteristics), and medical conditions that are recorded over an extended period of time.

[0102] The diet and nutritional data records a subject's dietary and nutritional intake and eating practices over time. For example, data may be obtained by periodically requesting information on what, how much, and when the subscriber ate and/or drank.

[0103] The environmental exposure data comprises environmental characteristics that describe both natural environmental considerations, such as, natural occurrences such as outside air temperature, humidity, sunlight, naturally occurring toxic/hazardous emissions, terrain, rain, water temperature, and others; manmade or man influenced environments considerations, such as, air conditioning, heating, ergonomics, lighting, pollution and contamination, traffic, and the like; and hazardous environments, such as, intentional and unintentional manmade or man caused environmental considerations such as exposure to dangerous situations and dangerous substances such as nuclear materials, toxic or hazardous biological substances, and toxic or hazardous chemicals, and the like. In some embodiments, environmental exposure data includes a subject's exposure to an environmental source of heavy metals.

[0104] The behavior data documents a variety of behaviors that are known to affect wellness and longevity. To simplify behavior assessment, and, as used herein, "behaviors^' are strictly defined as the actions taken by a person to relax, deal with stress, and occupy free time. The behaviors, amount of time spent in these behaviors, and the degree or intensity in which the subscriber participates in a behavior may be registered. The behaviors may be divided into three general categories. The first category includes behaviors that involve taking a substance (alcohol, tobacco, drags, food, coffee, and the like). The second category includes behaviors that require doing something (jogging, watching sports, gambling, watching TV, playing golf, conversations, sewing, and the like). The third category addresses coping impulses which include impulsive reactions to anger, affection, fear, confusion, and embarrassment. The data acquired may be subjected to a pattern-analysis to identify repetitive patterns and tendencies.

[0105] In some embodiments, a data acquisition unit includes test equipment. In some embodiments, the test equipment is communicatively coupled to the hardware and software that facilitate the entry of subject demographic data. For example, the test equipment is communicatively coupled to the hardware and software that facilitate the entry of subject demographic data directly or over a network. In some embodiments, the test equipment is configured to measure test data comprising the level of at least one heavy metal in a biological sample obtained from a subject. Examples of test equipment include any equipment that can be used to measure the presence, absence or level of at least one heavy metal in a biological sample. In some embodiments, test equipment includes a spectrometer that is capable of performing atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP- MS), or inductively-coupled plasma atomic emission mass spectrometry (ICP-AES). In some embodiments, test equipment can measure X-ray fluorescence. Examples of test equipment include PinAAcle™ 900 AA Spectrometer, Optima™ 8x00 ICP-OES Spectrometer, and NexION® 300 ICP-MS Spectrometer (Perkin Elmer). In some embodiments, the test data in combination with said subject demographic data comprises heavy metal data.

[0106] In some embodiments, systems for assessing exposure of a population of subjects to an environmental source of heavy metals also include at least one data processing unit. In some embodiments, the data processing unit is communicatively coupled to the at least one data acquisition unit. For example, the data processing unit is communicatively coupled to the data acquisition unit directly or over a network. In some embodiments, the data processing unit can include a computer, a central processing unit, a display, memory, and an input device. In some embodiments, heavy metal data is obtained from at least one data acquisition unit, or a plurality of data acquisition units. The plurality of data acquisition units can be dispersed over a geographical area. Some embodiments include a plurality of data processing units. In some such embodiments, the plurality of data processing units can be communicatively coupled to one another directly or over a network. In some embodiments, a plurality of data processing units is dispersed over a geographical area. [0107] In some embodiments, a data processing unit is configured to systematically collect cumulative heavy metal data from a plurality of subjects. In some embodiments, the heavy metal data can be stored and maintained by the data processing unit.

[0108] In some embodiments, a data processing unit is configured to analyze cumulative heavy metal data. In some embodiments, the data analysis function organizes and digests data and searches for correlations between the data elements themselves and between the data elements and reference materials (for example, medical encyclopedias, studies, prescription drug reference materials, standard heavy metal levels in and the like). The data elements are evaluated individually and as a set or group. As a group, certain relationships may become evident and their cumulative effect may suggest a condition or abnormality that may not otherwise be recognized. Upon the discovery of a possible abnormality, the symptoms may be further assessed to identify probable causes, such as exposure to an environmental source of heavy metals, and possible interventions. Also, the data analysis process may be programmed to detect patterns or trends that provide early indications of a condition that may require attention, such as a disorder associated with a subject's exposure to an environmental source of heavy metals. When compiled and evaluated collectively, these data elements contribute to establishment of a health baseline against which further changes may be measured and the effectiveness or response to prescribed medical therapies may be evaluated. The data elements are subjected to five analytical processes: comparative analysis; patterns and trends analysis; reference data correlation; relative condition; abnormality detection.

[0109] In some embodiments, analysis of cumulative heavy metal data can include a comparative analysis process in which a subject's heavy metal data may be compared against a reference model. In some embodiments, a reference model can include a model in which one or more data characteristics are known to be sufficient for a subject to develop a disorder associated with exposure to an environmental source of heavy metals. In some embodiments, a reference model can include a model in which a subject is unlikely to develop a disorder associated with exposure to an environmental source of heavy metals. In some embodiments, the reference model may be modified in view of cumulative heavy metal data.

[0110] In some embodiments, analysis of cumulative heavy metal data can include analysis of patterns and trends. In such embodiments, data may be mathematically assessed in order to calculate patterns and determine whether there is evidence of a trend. Patterns include measurements, values or events that fluctuate over time, in a predictable manner, within an established range. In some cases, patterns may be linked to regularly or irregularly occurring causes, such as, at least one subject having a disorder associated with exposure to an environmental source of heavy metals and the subject's habitual contact with a shoreline contacted with an oil spill, or the subject's diet contaminated with an environmental source of heavy metals. Trends include directional deviations from an established pattern. Exemplary categories of information, addressed by the patterns and trends analysis process, includes fluctuations in psychological, physical, physiological, and behavioral characteristic as well as the frequency and severity of specific illnesses and the frequency and severity of specific injuries. Other factors may also be incorporated into the patterns and trends analysis. These include recent diet, recent travel history, seasonal events, season, recent weather, changes in environment, marital relationships, vacations, recreational activities, and the like.

[0111] In some embodiments, analysis of cumulative heavy metal data can include comparison of reference data to determine meaningful relationships or correlations between a subject's unique health characteristics and authoritative sources that include demographic data, census data, health statistics, nutritional data, environmental pollutants and contaminants affects data, nuclear contamination affects data, hazardous/toxic biological substance exposure affects data, hazardous/toxic chemical exposure affects data, drug usage affects and toxicity data, and other health-related source materials. These correlations enhance the probability of linking the subscriber's abnormal psychological, physical, physiological, and behavioral characteristics to certain known conditions and ultimately to their causes and to recognized health behavior modifications and treatments.

[0112] The relative condition process continuously compares a subject's most recent psychological, physical, physiological, and behavioral characteristics against the reference model and similar population groups in order to gauge a subject's relative condition. A variation of this process includes the comparison of the subject to a population group that share certain key characteristics that may include the same gender, race, ethnicity, geographical region, and the like.

[0113] In some embodiments, analysis of cumulative heavy metal data can include abnormality detection analysis. In some embodiments, searches are performed for correlation between data elements that may suggest evidence of a condition or abnormality. The abnormality detection process may be designed to detect early evidence of an abnormality or unusual change in a subject's condition, such as a disorder associated with exposure to an environmental source of heavy metals. In one embodiment, the abnormality detection comprises the following activities: abnormality screening, pattern deviation; behavioral changes; and adverse reaction detection. In abnormality screening, a subject's psychological, physical, and physiological characteristics may be continuously screened in order to identify those that fall outside of the reference model or norms. In pattern deviation, current health characteristics may be screened against previous established ranges of fluctuation or patterns in order to identify deviations. Deviations can include of those most recent health characteristics that fall outside of the normal range of fluctuation and therefore suggest a change in a subject's condition. In behavioral changes, a subject's most recent behaviors and relationships are compared against previous behaviors and relationships in order to identify abnormal behavior, relative to a reference model or norms of a similar population group, and unusual changes or precipitating events that could affect or be affected by a subject's health condition. In adverse reaction detection, a subject's initial participation in a prescription drug program triggers an assessment of their health history. This assessment identifies evidence of any previous adverse reactions or side effects to the particular drug being used or similar drug. Also, subject that use prescription drugs have their psychological, physical, physiological, and behavioral characteristics closely monitored to detect early evidence of known adverse reactions and side effects. In some embodiments, analysis of cumulative heavy metal data includes analysis is predictive of a subject developing a disorder associated with exposure to an environmental source of heavy metals.

[0114] Some embodiments of the methods and systems provided herein include methods for assessing exposure of a population of subjects to an environmental source of heavy metals. Some such methods include use of the systems provided herein. In some embodiments a method for assessing exposure of a population of subjects to an environmental source of heavy metals includes obtaining test data test equipment configured to measure test data comprising the level of at least one heavy metal in a biological sample obtained from a subject. Examples of test equipment include any equipment that can be used to measure the presence, absence or level of at least one heavy metal in a biological sample. In some embodiments, test equipment includes a spectrometer that is capable of performing atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), or inductively-coupled plasma atomic emission mass spectrometry (ICP-AES).

[0115] In some embodiments a method for assessing exposure of a population of subjects to an environmental source of heavy metals includes obtaining heavy metal data from at least one data acquisition unit comprising hardware accessories and software that facilitates interface with a subscriber and the entry of subject demographic data, and test equipment configured to measure test data comprising the level of at least one heavy metal in a biological sample obtained from a subject, said test data in combination with said subject demographic data comprising said heavy metal data. [0116] Some embodiments also include transmitting the heavy metal data to at least one data processing unit from the at least one data acquisition unit. In some embodiments, the heavy metal data can be transmitted to the at least one data processing unit directly, or over a network. In some embodiments, a plurality of data acquisition units is distributed over a geographical area. In some embodiments, each data processing unit of a plurality of data processing units is communicatively coupled to one another directly or over a network. In some embodiments, a plurality of data processing units is distributed over a geographical area.

[0117] In some embodiments, the at least one data processing unit is configured to systematically collect cumulative heavy metal data from a plurality of subjects. In some embodiments, the at least one data processing unit is configured to analyze cumulative heavy metal data. In some embodiments, the at least one data processing unit is configured to identify correlations between subject demographic data and test data. In some embodiments, the at least one data processing unit is configured to identify a subject exposed to an environmental source of heavy metals. In some embodiments, the at least one data processing unit is configured to identify a subject with a disorder associated with exposure to an environmental source of heavy metals, or a subject likely to develop a disorder associated with exposure to an environmental source or heavy metals. In some embodiments, the at least one data processing unit is configured to compare the cumulative heavy metal data with a reference model. In some embodiments, the at least one data processing unit is configured to modify a reference model.

EXAMPLES

Example 1— Heavy metal content of environmental samples

[0118] The heavy metal content of environmental samples was measured. Samples were taken from three sites in Louisiana state: Lake Raccourci, Lake Pointe Aux Chien, and Lake Devil Bayou. The lakes lie within a 10-mile radius of one another and in close proximity to the Gulf of Mexico. At the time samples were taken, in December 2010, from Lake Raccourci and Lake Pointe Aux Chien, both lakes were visibly contaminated with crude oil. There was no visible contamination with crude oil when samples were taken from Lake Devil Bayou on October 16, 2010. Samples included: top soil, subsurface soil (3 inches deep), marsh soil, coffee grounds -like soil, surface water, subsurface water, oysters, shrimp, and snails. Other test samples included a light sweet crude oil from the Discover Enterprise site in the Gulf of Mexico ("BP-mc252" Sample #01 1) collected on May 20, 2010, at 1 1 :15 AM. This sample was stored at -20 °C protected from light. Another sample included 10w40 commercial motor oil. [0119] Each sample was assayed by inductively coupled plasma - mass spectrometry, using methods well known in the art. See e.g., Mendiguchia C, et al. Screening of dissolved heavy metals (Cu, Zn, Mn, Al, Cd, Ni, Pb) in seawater by a liquid-membrane-ICP-MS approach. Anal Bioanal Chem. 2008; 391 :773-8; Milne A, et al., Determination of Mn, Fe, Co, Ni, Cu, Zn, Cd and Pb in seawater using high resolution magnetic sector inductively coupled mass spectrometry (HR-ICP-MS). Anal Chim Acta. 2010; 665:200-7; Hussain T, et al, Monitoring and assessment of toxic metals in Gulf War oil spill contaminated soil using laser-induced breakdown spectroscopy. Environ Monit Assess. 2008; 136:391-9). The mean heavy metal concentrations were calculated for samples from Lake Raccourci and Lake Pointe Aux Chien, for samples from Lake Devil Bayou, motor oil, and light sweet crude oil. The results and the National Institute of Standards and Technology (NIST) published acceptable levels of heavy metal contamination for consumable oysters are shown in Table 2.

TABLE 2

[0120] In samples from oil-contaminated shorelines (Lake Raccourci and Lake Pointe Aux Chien), Fe, Mn, Cr, Pb, As, Zn, and Cu were detected at levels many times greater than any measurable levels of those heavy metals in samples from an uncontaminated shoreline.

[0121] In shrimp from the oil-contaminated sites (Lake Raccourci and Lake Pointe Aux Chien), high levels of arsenic, zinc, and copper were detected; cadmium and lead were not detected in measurable amounts. In soil from the uncontaminated site (Lake Devil Bayou), arsenic at 1.41 ppm was detected; cadmium, lead, zinc, or copper were not detected in measurable amounts. In motor oil, arsenic and zinc were detected; cadmium, lead, and copper were not detected in measurable amounts. In light sweet crude oil, arsenic was detected; cadmium, lead, zinc, or copper were not detected in measurable amounts. Motor oil and the light sweet crude oil, were negative for nickel, iron, manganese, and chromium. Soil from Lake Devil Bayou was positive for high levels of iron; but negative for nickel, manganese, and chromium. Soils from visibly contaminated lakes were positive for high levels of iron, manganese, and chromium; but negative for nickel, with the exception of marsh soil. Oyster, shrimp, and snail were positive for very high levels of iron, but negative for nickel, manganese, and chromium; with the exception that snail measured positive for manganese.

[0122] It is striking that the sample of light sweet crude oil from the Discover Enterprise site, was positive for measureable levels of arsenic; but there were no measureable levels for the other eight heavy metals investigated. Light sweet crude oil contains a very complex mixture of volatile substances including, polycyclic aromatic hydrocarbons and other volatile hydrocarbons. As oil transversed the Gulf of Mexico, there was a loss of a substantial portion of the original mixture, as a function of aerosolized dissipation, water-mediated dissipation, microbial breakdown, and possibly other processes. These various processes of dissipation may have resulted in concentrating the heavy crude oil residual material that reached the shoreline. This could result in substantially elevated levels of heavy metals observed at the visibly contaminated sites, and would be consistent with what has been observed in other oil- contaminated shorelines. The finding of high levels of heavy metals in environmental samples, such as samples collected from sites at visibly-contaminated lakes on the Gulf of Mexico, is consistent with what has been reported after other oil-contaminated shorelines. The dissipation of a large portion of the Gulf Oil Spill of 2010, paradoxically resulted in concentrating heavy metals, and potentially other non-volatile toxins, so that they collected in sites such as the oil- contaminated shorelines studied and described herein. Example 2— Clinical analysis of a population's exposure to heavy metals

Methods

[0123] A cohort of 500 individuals is identified in particular geographic area known to have been exposed to environmental oil. Each individual is tested each year to determine the level of at least one heavy metal in a biological sample, for a total of four consecutive years; year 1, year 2, year 3, and year 4. Year five of the study is used for follow-up blood testing and data analyses. For each person that tests positive for elevated heavy metal levels or heavy metal- protein damage, that person is re-assessed for possible heavy metal-DNA damage from that specific heavy metal(s).

[0124] Individuals who test positive for elevated heavy metal levels, heavy metal- protein damage, and/or for heavy metal-DNA damage, has their records reviewed for the possible presence of a medical condition that might be related to that specific heavy metal. Given the limited reports of heavy metal toxicity in persons that have been exposed to high levels of crude oil, a possible population rate of testing positive for one or more heavy metals, at 15-20% is anticipated.

[0125] If the rate of percent positivity in the study population is sufficiently great (as discussed in statistical considerations herein), one blood specimen from each person in the study, each year, for the first four years of the five year duration of the study is collected. Changes in heavy metal levels or heavy metal-protein levels over time assessed, and changes in metal-DNA levels over time are assessed. These temporal changes in individuals are assessed, : a) by community; b) by ethnic group; c) by gender; and, d) by other potentially important socioeconomic and demographic markers.

[0126] Baseline comparisons of data are made comparing different groups to one another. The possible relationships between heavy metal levels, heavy metal-protein levels, and heavy metal-DNA levels in individuals are assessed, with clinical and social parameters. Heavy metal levels, heavy metal-protein levels and heavy metal-DNA levels for groups as a whole areassessed. Such groups will include: the different ethnic groups; differences in socioeconomic status; and other demographic parameters.

[0127] Each year, for five years, measure for heavy metal levels, heavy metal-protein damage or heavy metal-DNA levels for the same 500 persons is measured. This will allow for the assessment of the potential/theoretical "flow" of exposures, through the population. For example a person may be negative for evidence of exposure on year 1 , but as a consequence of "flow" of crude oil through the environment, that person may prove to be positive on year 2 and/or year 3, and/or year 4, and/or year 5. That "flow" might be a result of food chain effects, recreational activities in contaminated areas, etc.

[0128] Control samples include stored blood samples from 100 control individuals known to be free of exposure to environmental oil. For example, the control individuals may be individuals who donated prior to the beginning of the Gulf Oil Spill of 2010.

[0129] Metal-protein analysis is performed as follows: 6-8 ml heparinized blood is collected from a subject and separated into three specimens: buffy coat (nucleated white blood cells), plasma, and red cells. Buffy coat and red cell fractions are stored at -70 °C, with 10% DMSO. Plasma is wet-ashed for assay by atomic absorbance spectrometry (AAS) with Zeeman background correction. The use of wet-ashing for measuring heavy metals in human cells, fluids, and tissues has been described (1,2,3,4). An aliquot of plasma is placed in a polyurethane tube, and sequentially: a) mixed 1 : 1 with concentrated nitric acid; b) submerged in a water bath at 90 °C degrees for 5 minutes; c) cooled to room temperature; d) this is mixed 1 : 1 with hydrogen peroxide; and this mixture is ready for assay by AAS. The Perkin Elmer Aanalyst 600 Atomic Absorbance Spectrometer (model year 2008) has specific settings for the measurement of specific metals. The basic AAS parameters for the measurement of the nine heavy metals in crude oil have been determined (34). Example parameters are shown in Table 3.

TABLE 3

[0130] Similar parameters have been utilized for metals such as platinum, gallium, and cadmium for measurement in a human biological matrix (22,27-30,31). Cadmium-protein damage and cadmium-DNA damage can be measured following low micromolar exposures to cadmium (28,29).

[0131] Metal-DNA analysis is performed as follows: blood samples from a subject having a positive measurement for metal-protein damage are obtained for metal-DNA damage analysis. DNA is obtained from the buffy coat of a 30-35 ml blood specimen by cesium chloride density gradient centrifugation (5,6). Isolated DNA is 99.4% pure and any metal present is covalently bound to the DNA. The DNA is dialyzed, quantitated at 260 nm, and sonicated. The DNA solution is assayed by AAS. For example, for platinum, cadmium, and gallium, levels of metal as low as 50-60 pg per sample can be detected. By adjusting the amount of DNA per assay, as few as 3 lesions per million DNA bases for platinum, cadmium, and gallium can be detected with the assay.

[0132] Statistics: If fewer than 5 individuals test positive for two or more heavy metals, and 500 persons are tested, there is 90% confidence and with >84% power, that the true percent positivity rate is less than 2%. If the true positive rate of testing is less than 2%, these studies are not conducted beyond year 1.

[0133] If 37 or more persons test positive for two or more heavy metals, there is 90% confidence and >84% power, that the true positivity rate is at least 5%, or 1 in 20. A rate of 1 in 20 persons testing positive, warrants further detailed study of those specific individuals; and warrants longer follow-up of the entire population. If this occurs, each of these 500 individuals each year are tested for five consecutive years of the study. If a person tests positive in year 1, that person should be tested in greater detail, to determine the time course of positive heavy metal testing, as well as determining whether meaningful biological sequelae may occur. It may be possible that a person who tested negative in year 1 , may test positive in subsequent years, from exposures that may be direct or indirect.

[0134] If the rate of true positivity is between 2% and 5%, sub-groups would be analyzed within the data to determine whether there were one or more sub-groups that may have significantly higher numbers of positive individuals.

[0135] Individuals are interviewed or complete a questionnaire during the study to determine medical history, any changes in medical history, ethnicity, socioeconomic background, any events or activities that may have resulted in exposure to the results of the oil spill.

Results

[0136] Samples of heparinized blood are obtained from 500 adult persons. Each blood sample is assayed by atomic absorbance spectrometry (AAS) for metal-protein damage for heavy metals including: cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium. Measurable metal-protein damage from any two of the above heavy metals indicates a likelihood of exposure by a subject to an oil spill. Measurable metal -protein damage from any three of the above heavy metals indicates a strong likelihood of exposure by a subject to an oil spill. In a population including residents of an area known to have been exposed to an oil spill, 15-20 % of the population has an indication of at least a likelihood of exposure by a subject to an oil spill.

[0137] In subjects who test positive for metal-protein damage, metal-DNA damage is assayed by atomic absorbance spectrometry (AAS) for heavy metals including: cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium. Approximately > 60% subjects who have measurable metal-protein levels in blood, also have measurable levels of metal-DNA in one or more tissues, such as blood.

[0138] The source of crude oil exposure is identified through interviews with individuals and other investigative techniques.

[0139] Data gathered during the study through interviews with subjects and measuring heavy metal levels in biological samples can be used to determine: (a) the percentage of the population having evidence for the presence of one or more heavy metals; (b) the percentage of the population having evidence for probable crude oil exposure; (c) the percentage of the population having evidence for more than probable crude oil exposure; and (d) the relationship between clinical/work/community experiences and the presence of metal-DNA damage, the levels of metal-DNA damage, metal-protein assessment, and the levels of metal- protein exposure.

[0140] In addition, data gathered during the study through interviews with subjects and measuring heavy metal levels in biological samples can be used to determine: (A) Whether exposure to heavy metals, as assessed by metal -protein levels and/or metal-DNA damage, varies by ethnicity, and whether any ethnicity-related exposure correlate with a clinical history of (i) direct exposure to the BP oil spill, (ii) occupational exposure unrelated to the BP oil spill, (iii) exposure through the food chain, (iv) recreational exposure, and (v) DNA repair. (B) Whether there is any correlation between a biological/molecular marker and clinical self-report change in health status for subjects who report the most dramatic changes in health status, and for any correlation (i) whether such correlations are also related to ethnicity, socioeconomic status, and/or education, and (ii) whether such correlations fit within known molecular mechanisms. Example 3— Assessing a subject's risk of developing a disorder associated with exposure to an environmental source of heavy metals

[0141] Test data comprising the levels of at least two heavy metals is obtained from 500 test subjects residing within 20 miles of a geographical area contacted with material deposited during the BP oil spill. The test data comprises the heavy metal concentration in plasma samples obtained from the test subject, and heavy metal concentrations in DNA purified from samples obtained from the test subjects. Demographic data is obtained from the 500 test subjects. Test data in combination with demographic data comprises heavy metal data. Heavy metal data is obtained from each test subject each year, for five years.

[0142] Cumulative heavy metal data shows subjects with certain demographic data develop disorders associated with exposure to materials deposited by the BP oil spill. Cumulative heavy metal data shows subjects with certain demographic data do not develop disorders associated with exposure to materials deposited by the BP oil spill within the period of the study. Analysis of heavy metal data provides a reference model of a subject with a likelihood of developing a disorder associated with exposure to materials deposited by the BP oil spill. Analysis of heavy metal data shows patterns and trends indicative of a subject having or with a likelihood of developing a disorder associated with exposure to materials deposited by the BP oil spill.

[0143] Each of the following references is incorporated herein by reference in its entirety.

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24. Parker, R.J., Gill, I., Tarone, R., Vionnet, J., Grunberg, S., Muggia, F., and Reed, E. Platinum-DNA damage in leukocyte DNA of patients receiving carboplatin and cisplatin chemotherapy, measured by atomic absorption spectrometry. Carcinogenesis, 12: 1253-1258, 1991.

25. Perera, F.P., Tang, D., Reed, E., Parker, R.J., Warburton, D., ONeill, Albertini, R., Bigbee, W., Santella, R., Tsai, W.-Y., Simon-Cereijido, G., Randall, C, Bosl, G., and Motzer, R. Multiple biologic markers in testicular cancer patients treated with platinum-based chemotherapy. Cancer Res, 52:3558-3565, 1992.

26. Reed, E., Parker, R.J., Gill, I., Bicher, A., Dabholkar, M., Vionnet., J.A., Bostick-Bruton, F., Tarone, R., and Muggia, F.M. Platinum-DNA adduct in leukocyte DNA of a cohort of 49 patients with 24 different types of malignancy. Cancer Res, 53:3694-3699, 1993.

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31. Senderowicz A, Reid R, Headlee D, Abernathy T, Horti J, Lush R M, Reed E, Figg W D, and Sausville E A. A phase II trial of gallium nitrate in patients with androgen-independent metastatic prostate cancer. Urologia Internationalis, 63 : 120- 125, 1999.

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39. Koc, O., McFee, M., Reed, E., and Gerson, S.L. Detection of platinum-DNA adducts in cord blood lymphocytes following in utero platinum exposure. Eur J Cancer, 30A:716-717, 1994. 40. Murphree, A.L., Sato, J.K., Malogolowkin, M., Fisher, A., Parker, R., Reed, E., Deegan III, W.F., Gomer, C.J., Villablanca, J. Chemotherapy plus local therapy in the treatment of intraocular retinoblastoma. Arch Ophthalmol, 1 14:1348-1356, 1996.

41. Poirier, M.C., Reed, E., Litterst, C.L., Katz, D., and Gupta-Burt, S. Persistence of platinum- ammine-DNA adducts in gonads and kidneys of rats and multiple tissues from cancer patients. Cancer Res 52:149-153, 1992.

[0144] The term "comprising" as used herein is synonymous with "including," "containing," or "characterized by," and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps.

[0145] All numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification are to be understood as being modified in all instances by the term "about." Accordingly, unless indicated to the contrary, the numerical parameters set forth herein are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of any claims in any application claiming priority to the present application, each numerical parameter should be construed in light of the number of significant digits and ordinary rounding approaches.

[0146] The above description discloses several methods and materials of the present invention. This invention is susceptible to modifications in the methods and materials, as well as alterations in the fabrication methods and equipment. Such modifications will become apparent to those skilled in the art from a consideration of this disclosure or practice of the invention disclosed herein. Consequently, it is not intended that this invention be limited to the specific embodiments disclosed herein, but that it cover all modifications and alternatives coming within the true scope and spirit of the invention.

[0147] All references cited herein, including but not limited to published and unpublished applications, patents, and literature references, are incorporated herein by reference in their entirety and are hereby made a part of this specification. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and/or take precedence over any such contradictory material.

Claims

WHAT IS CLAIMED IS:

1. A method for assessing the exposure of a subject to an environmental source of heavy metals comprising measuring the level of at least one heavy metal in a biological sample obtained from said subject.

2. The method of claim 1, further comprising comparing the level of the at least one heavy metal in the biological sample to a control level of said at least one heavy metal, wherein said control level of said at least one heavy metal is selected from the group consisting of the level of said at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals, the level of said at least one heavy metal in a sample which has a known exposure to an environmental source of heavy metals, a predetermined reference level of said at least one heavy metal which is known to be indicative of exposure to an environmental source of heavy metals and a predetermined reference level of said at least one heavy metal which is known to be indicative of a lack of exposure to an environmental source of heavy metals.

3. The method of claim 2, wherein an increase in the level of said at least one heavy metal in the biological sample compared to the level of said at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals, or compared to a predetermined reference level of said at least one heavy metal which is known to be indicative of a lack of exposure to an environmental source of heavy metals is indicative of the exposure of the subject to an environmental source of heavy metals.

4. The method of any one of claims 1-3, wherein the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium.

5. The method of any one of claims 1-4, wherein said method comprises measuring the levels of at least two heavy metals in the biological sample.

6. The method of any one of claims 1-5, wherein said method comprises measuring the levels of at least three heavy metals in the biological sample.

7. The method of any one of claims 1-6, wherein said method comprises measuring the levels of at least four heavy metals in the biological sample.

8. The method of any one of claims 1-7, wherein said method comprises measuring the levels of at least five heavy metals in the biological sample.

9. The method of any one of claims 1-8, wherein measuring the level of at least one heavy metal comprises performing a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively-coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

10. The method of any one of claims 1-9, wherein the environmental source of heavy metals comprises an oil spill.

1 1. The method of any one of claims 1-10, wherein the presence, absence or level of the at least one heavy metal in the biological sample is indicative of the environmental source of heavy metals.

12. The method of any one of claims 1-11, further comprising measuring the level of at least one heavy metal in a plurality of biological samples, wherein each biological sample is obtained from a different subject.

13. The method of any one of claims 1-12, wherein the biological sample comprises nucleic acid.

14. The method of any one of claims 1-13, wherein the biological sample comprises protein.

15. The method of any one of claims 1-14, wherein the level of said at least one heavy metal in said biological sample is measured ex vivo.

16. The method of any one of claims 1-15, wherein the subject is mammalian.

17. The method of any one of claims 1-16, wherein the subject is human.

18. The method of any one of claims 1-17, wherein the subject resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals.

19. The method of any one of claims 1-18, further comprising providing a treatment for exposure to heavy metal, or recommending a preventative or diagnostic regimen if the level of said at least one heavy metal in said biological sample from said subject indicates that said subject has been exposed to an environmental source of heavy metals.

20. A method for assessing the likelihood of a subject developing a disorder associated with exposure to an environmental source of heavy metals comprising measuring the level of at least one heavy metal in a biological sample obtained from said subject.

21. The method of claim 20, further comprising comparing the level of the at least one heavy metal in the biological sample to a control level of said at least one heavy metal, wherein said control level of said at least one heavy metal is selected from the group consisting of the level of said at least one heavy metal in a sample which has not been exposed to an environmental source of heavy metals, the level of said at least one heavy metal in a sample which has a known exposure to an environmental source of heavy metals, a predetermined reference level of said at least one heavy metal which is known to be indicative of exposure to an environmental source of heavy metals and a predetermined reference level of said at least one heavy metal which is known to be indicative of a lack of exposure to an environmental source of heavy metals.

22. The method of any one of claims 20-21, wherein the disorder associated with exposure to an environmental source of heavy metals is selected from the group consisting of kidney damage, nerve damage, pulmonary toxicity, cardiac toxicity, skin rash, skin lesion, an immunological disorder, and cancer.

23. The method of any one of claims 20-22, wherein the presence, absence, or level of the at least one heavy metal in the biological sample is indicative of the likelihood of the subject developing a disorder associated with exposure to an environmental source of heavy metals.

24. The method of any one of claims 20-23, wherein the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium.

25. The method of any one of claims 20-24, wherein said method comprises measuring the levels of at least two heavy metals in the biological sample.

26. The method of any one of claims 20-25, wherein said method comprises measuring the levels of at least three heavy metals in the biological sample.

27. The method of any one of claims 20-26, wherein said method comprises measuring the levels of at least four heavy metals in the biological sample.

28. The method of any one of claims 20-27, wherein said method comprises measuring the levels of at least five heavy metals in the biological sample.

29. The method of any one of claims 20-28, wherein measuring the level of at least one heavy metal comprises performing a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively-coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

30. The method of any one of claims 20-29, wherein the environmental source of heavy metals comprises an oil spill.

31. The method of any one of claims 20-30, further comprising measuring the level of at least one heavy metal in a plurality of biological samples, wherein each biological sample is obtained from a plurality of subjects.

32. The method of any one of claims 20-31, wherein the biological sample comprises nucleic acid.

33. The method of any one of claims 20-32, wherein the biological sample comprises protein.

34. The method of any one of claims 20-33, wherein the level of said at least one heavy metal in said biological sample is measured ex vivo.

35. The method of any one of claims 20-34, wherein the subject is mammalian.

36. The method of any one of claims 20-35, wherein the subject is human.

37. The method of any one of claims 20-36, wherein the subject resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals.

38. The method of any one of claims 20-37, further comprising providing a treatment for exposure to heavy metal, or recommending a preventative or diagnostic regimen if the level of said at least one heavy metal in said biological sample from said subject indicates that said subject has been exposed to an environmental source of heavy metals.

39. A system for assessing exposure of a plurality of subjects to an environmental source of heavy metals comprising:

(a) at least one data acquisition unit comprising:

hardware accessories and software that facilitates interface with a subscriber and the entry of subject demographic data, and

test equipment configured to measure test data comprising the level of at least one heavy metal in a biological sample obtained from a subject, said test data in combination with said subject demographic data comprising heavy metal data; and

(b) at least one data processing unit communicatively coupled to the at least one data acquisition unit, said at least one data processing unit configured to:

systematically receive cumulative heavy metal data from the plurality of subjects,

analyze cumulative heavy metal data to identify correlations between subject demographic data and test data, and

identify a subject exposed to an environmental source of heavy metals.

40. The system of claim 39, wherein the at least one data processing unit is configured to compare the cumulative heavy metal data with a reference model.

41. The system of any one of claims 39-40, wherein the at least one data processing unit is configured to modify a reference model.

42. The system of any one of claims 39-41 further comprising a plurality of data acquisition units distributed over a geographical area.

43. The system of any one of claims 39-42 further comprising a plurality of data processing unit distributed over a geographical area.

44. The system of any one of claims 39-43, wherein the at least one data processing unit is communicatively coupled to the at least one data acquisition unit over a network.

45. The system of any one of claims 39-44, wherein subject demographic data is selected from the group consisting of personal identification data, physical characteristics data, health profile data, family health history data, drug and vitamin/mineral supplement data, health baseline data, diet and nutritional data, environmental exposure data, and behavioral data.

46. The system of any one of claims 39-45, wherein the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium.

47. The system of any one of claims 39-46, wherein the test equipment is configured to measure the levels of at least two heavy metals in the biological sample.

48. The system of any one of claims 39-47, wherein the test equipment is configured to measure the levels of at least three heavy metals in the biological sample.

49. The system of any one of claims 39-48, wherein the test equipment is configured to measure the levels of at least four heavy metals in the biological sample.

50. The system of any one of claims 39-49, wherein the test equipment is configured to measure the levels of at least five heavy metals in the biological sample.

51. The system of any one of claims 39-50, wherein the test equipment is configured to perform a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively-coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

52. The system of any one of claims 39-51, wherein the environmental source of heavy metals comprises an oil spill.

53. The system of any one of claims 39-52, wherein the test data comprises the level of heavy metal-nucleic acid in said biological sample.

54. The system of any one of claims 39-53, wherein the test data comprises the level of heavy metal-protein in said biological sample.

55. The system of any one of claims 39-54, wherein the system is configured to receive test data comprising the level of said at least one heavy metal in said biological sample which has been measured ex vivo.

56. The system of any one of claims 39-55, wherein the test data has been obtained from a mammalian subject.

57. The system of any one of claims 39-56, wherein the test data has been obtained from a human subject.

58. The system of any one of claims 39-57, wherein the test data has been obtained from at least one subject who resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals.

59. A method for assessing exposure of a population of subjects to an environmental source of heavy metals comprising:

(a) obtaining heavy metal data from at least one data acquisition unit comprising: hardware accessories and software that facilitates interface with a subscriber and the entry of subject demographic data, and

test equipment configured to measure test data comprising the level of at least one heavy metal in a biological sample obtained from a subject, said test data in combination with said subject demographic data comprising said heavy metal data; and

(b) transmitting the heavy metal data to at least one data processing unit from the at least one data acquisition unit, the at least one data processing unit configured to:

systematically receive cumulative heavy metal data from a plurality of subjects,

identify a subject exposed to an environmental source of heavy metals.

60. The method of claim 59, wherein the at least one data processing unit is configured to compare the cumulative heavy metal data with a reference model.

61. The method of any one of claims 59-60, wherein the at least one data processing unit is configured to modify a reference model.

62. The method of any one of claims 59-61 further comprising a plurality of data acquisition units distributed over a geographical area.

63. The method of any one of claims 59-62 further comprising a plurality of data processing unit distributed over a geographical area.

64. The method of any one of claims 59-63, wherein the at least one data processing unit is communicatively coupled to the at least one data acquisition unit over a network.

65. The method of any one of claims 59-64, wherein subject demographic data is selected from the group consisting of personal identification data, physical characteristics data, health profile data, family health history data, drug and vitamin/mineral supplement data, health baseline data, diet and nutritional data, environmental exposure data, and behavioral data.

66. The method of any one of claims 59-65, wherein the heavy metal is selected from the group consisting of cadmium, lead, arsenic, zinc, copper, nickel, iron, manganese, and chromium.

67. The method of any one of claims 59-66, wherein the test equipment is configured to measure the levels of at least two heavy metals in the biological sample.

68. The method of any one of claims 59-67, wherein the test equipment is configured to measure the levels of at least three heavy metals in the biological sample.

69. The method of any one of claims 59-68, wherein the test equipment is configured to measure the levels of at least four heavy metals in the biological sample.

70. The method of any one of claims 59-69, wherein the test equipment is configured to measure the levels of at least five heavy metals in the biological sample.

71. The method of any one of claims 59-70, wherein the test equipment is configured to perform a technique selected from the group consisting of atomic absorbance spectroscopy (AAS), inductively-coupled plasma mass spectrometry (ICP-MS), inductively-coupled plasma atomic emission mass spectrometry (ICP-AES), and X-ray fluorescence.

72. The method of any one of claims 59-71 , wherein the environmental source of heavy metals comprises an oil spill.

73. The method of any one of claims 59-72, wherein the test data comprises the level of heavy metal-nucleic acid in said biological sample.

74. The method of any one of claims 59-73, wherein the test data comprises the level of heavy metal-protein in said biological sample.

75. The method of any one of claims 59-74, wherein the system is configured to receive test data comprising the level of said at least one heavy metal in said biological sample which has been measured ex vivo.

76. The method of any one of claims 59-75, wherein the test data has been obtained from a mammalian subject.

77. The method of any one of claims 59-76, wherein the test data has been obtained from a human subject.

78. The method of any one of claims 59-77, wherein the test data has been obtained from at least one subject who resides at, is located at, works at or has been at a location or within close proximity to the location of an environmental source of heavy metals.