Compositions, Apparatus and Methods for Monitoring Biomarkers

Abstract

A single device to collect, transfer, and measure salivary nitric oxide analyte and metabolite, nitrite, a biomarker for nitric oxide, as well as, a method to assess the effects of diet and exercise on changing an individual's nitric oxide status and health.

What is claimed is 
     
         1 . A device for detecting the presence of a bioniarker in a bodily fluid wherein the device comprises a single unit for collecting the bodily fluid, analyzing the contents of the bodily fluid, and correlating the analysis with physiological status. 
     
     
         2 . The device of  claim 1 , wherein the bodily fluid comprises saliva, sputum, tears, sweat, mucus, serum, semen, mine, or blood. 
     
     
         3 . The device of  claim 1 , wherein the contents of the bodily fluid comprise nitric oxide analytes and metabolic products, including nitrite, nitrate, other reactive nitrogen intermediates, leukotriene, B4, aberrant gene methylation (p16 and/or O 6 -methylguanine-DNA methyltransferase promoters), mir-21 expression, EGF, lysozyme, lactoferin, cytokines, INFganima, TNFalpha, eotaxin, adhesion molecules (ICAM-1, selectin etc.), cytokines, neuropeptides, substance P, calcitonin-gene-related peptide, vasoactive intestinal peptide, mucin, cytokines, growth factors, tumor markers, growth factors, proteins, immunoglobulins, enzymes, enzyme inhibitors, antigens, proteolytic enzymes, citric acid, acid phosphatase, lipids, prostate specific antigen, metabolites, sugars, urinary carcinogen metabolite, trans, trans-muconic acid (tt-MA) or S phenylmercapturic acid (metabolites of benzene), 1- and 2-naphthol, hydroxyphenanthrenes or phenanthrene dihydrodiols, 1-hydroxypyrene (1-HOP), metabolites of benzo[a]pyrene, aromatic amines or heterocyclic aromatic amines, N-nitrosoproline, 4-(methylnitrosamino)-1-(3-pyridyl)-1-hutanol and its glucuronides (NNAL and NNAL-Glue), 8-oxodeoxyguanosine, thioethers, mercapturic acids, alkyladenines, nitric oxide, heavy metals, hormones, cortisol, dehydroxyepiandrosterone, toxins, toxin metabolites, cotinine, enzymes, lysozyme, α-amylase, immunoglobulins, RNA, or DNA. 
     
     
         4 . The device of  claim 1 , wherein the bodily fluid comprises saliva and the biomarker comprises saliva-derived nitric oxide analytes or saliva-derived nitric oxide metabolites. 
     
     
         5 . The device of  claim 1 , wherein the single unit comprises an elongated strip, wherein the strip optionally contains a scored mark at the midpoint of the strip and wherein the strip contains a first absorbent pad at one end and a second absorbent pad at the opposite end. 
     
     
         6 . The device of  claim 5 , wherein the first absorbent pad comprises a fluid collection pad, and wherein the second absorbent pad comprises a test pad. 
     
     
         7 . The device of  claim 6 , wherein the fluid collection pad comprises a wicking pad, membrane, paper, resin, sponge, immunoabsorbent pad, ionic, or other suitable platform that absorbs saliva analytes to be transferred to the test reagent pad. 
     
     
         8 . The device of  claim 6 , wherein the test pad enables dry reagent detection chemistry comprising components modified from the Ciri ess diazotization reaction, comprising mixtures of N-naphthylenediamine-dihydrochloride and sulphanilamide or N-(naphthyl)ethylenediammonium-dihydrochloride and sulphanilatnide or para-arsanilic acid N-ethylenediamine tetrahydroquinoline or para-arsanilic acid N-(1-naphthyl)ethylenediamine dihydroehoride. 
     
     
         9 . The device of  claim 4 , wherein saliva-derived nitric oxide analytes comprise nitrite anion, nitrogen oxide anions, nitrogen dioxide, or nitrate. 
     
     
         10 . The device of  claim 6 , wherein the bodily fluid comprises saliva, wherein the biomarker comprises nitric oxide analyte, nitrite, wherein the fluid collection pad comprises a wicking pad, and wherein the test pad enables dry reagent detection chemistry comprising components modified from the Griess diazotization reaction. 
     
     
         11 . A method for detecting the presence of a biomarker in a bodily fluid wherein the device comprises a single unit for collecting the bodily fluid, analyzing the contents of the bodily fluid, and correlating the analysis with physiological status. 
     
     
         12 . The method of  claim 11 , wherein the bodily fluid comprises saliva, sputum, tears, sweat, mucus, serum, semen, urine, or blood. 
     
     
         13 . The method of  claim 11 , wherein the contents of the bodily fluid comprise nitric oxide analytes and metabolic products, including nitrite, nitrate, other reactive nitrogen intermediates, leukotriene, 134, aberrant gene methylation (p16 and/or O 6 -methylguanine-DNA inethyitransferase promoters), mir-21 expression, EGF, lysozyme, lactoferin, cytokines, INFgamma, TNFalpha, cotaxin, adhesion molecules (ICAM-1, selectin etc.), cytokines, neuropeptides, substance P, calcitonin-gene-related peptide, vasoactive intestinal peptide, mucin, cytokines, growth factors, tumor markers, growth factors, proteins, immunoglobulins, enzymes, enzyme inhibitors, antigens, proteolytic enzymes, citric acid, acid phosphatase, lipids, prostate specific antigen, metabolites, sugars, urinary carcinogen metabolite, trans, trans-muconic acid (tt-MA) or S phenylmercapturic acid (metabolites of benzene), 1-or 2-naphthol, hydroxyphenanthrenes or phenanthrene dihydrodiols, 1-hydroxypyrene (1-HOP), metabolites of benzo[a]pyrene, aromatic amines or heterocyclic aromatic amines, N-nitrosopro ne, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronides (NNAL and NNAL-Gluc), 8-oxodeoxyguanosine, thioethers, mercapturic acids, alkyladenines, nitric oxide, heavy metals, hormones, cortisol, dehydroxycpiandrosterone, toxins, toxin metabolites, cotinine, enzymes, lysozyme, a-amylase, immunoglobulins, RNA, or DNA. 
     
     
         14 . The method of  claim 13 , wherein the bodily fluid comprises saliva and the biomarker comprises nitric oxide, saliva-derived nitric oxide analytes, nitrite anion, nitrogen oxide anions, nitrogen dioxide, nitrate, nitric oxide, or saliva-derived nitric oxide metabolites. 
     
     
         15 . The method of  claim 11 , wherein the single unit comprises an elongated strip, wherein the strip contains a scored mark at the midpoint of the strip and wherein the strip contains a first absorbent pad at one end and a second absorbent pad at the opposite end. 
     
     
         16 . method of  claim 15 , wherein the first absorbent pad comprises a fluid collection pad, and wherein the second absorbent pad comprises a test pad. 
     
     
         17 . The method of  claim 16 , wherein the fluid collection pad comprises a wicking pad, membrane, paper, resin, sponge, immunoabsorbent pad, ionic, or other suitable platform that absorbs saliva analytes to be transferred to the test reagent pad. 
     
     
         18 . The method of  claim 16 , wherein the test pad enables dry reagent detection chemistry comprising components modified from the Griess diazotization reaction, comprising mixture of N-naphthylenediamine-dihydrochloride and sulphanilamide or N-(naphthyl)ethylenediammonium-dihydrochloride and sulphanilamide or para-arsanilic acid N-ethylenediamine tetrahydroquinoline or para-arsanilic acid N-(1-naphthy)ethylenediamine dihydrochoride. 
     
     
         19 . method of  claim 16 , wherein the bodily fluid comprises saliva, wherein the biomarker comprises nitric oxide analyte, nitrite, wherein the fluid collection pad comprises a wicking pad, and wherein the test pad enables dry reagent detection chemistry comprising components modified from the Griess diazotization reaction 
     
     
         20 . The method of  claim 19 , wherein the fluid collection pad is inserted into the mouth for a first predetermined period of time to absorb saliva, wherein the device is then folded so that the fluid collection pad and test pad make contact for a second predetermined period of time, wherein the contact between the fluid collection pad and test pad results in a color change on the test pad following a third predetermined period. 
     
     
         21 . The method of  claim 20 , wherein the color change is compared to a color scale and wherever the color falls within the scale, a representative physiological nitric oxide content is evaluated. 
     
     
         22 . The method of  claim 21 , wherein the representative physiological nitric oxide content is utilized to enable the assessment of vascular wellness and fitness, 
     
     
         23 . The method of  claim 22 , wherein vascular wellness and fitness comprises the ability to maintain a normal blood pressure range, lower blood pressure, extend time-to-exhaustion, reduce the need for oxygen consumption, improve cellular respiration, or mitochrondial efficiency. 
     
     
         24 . The method of  claim 22 , wherein the representative physiological nitric oxide content is utilized to enable modification of nitric oxide levels comprising daily physical exercise, anaerobic training, aerobic training, consuming a diet consisting of leafy green vegetables and other nitrate-rich vegetables, fruit, whole foods supplement intended to increase nitric oxide levels, consuming a Mediterranean. diet, or consuming a DASH diet, 
     
     
         25 . The method of  claim 20 , wherein the first and second predetermined period of time are about 3-5 seconds; and wherein the third predetermined period of time is about 45-60 seconds. 
     
     
         26 . The method of  claim 20 , wherein the test pad provides a concentration range of the nitric oxide analyte, nitrite, ranging from 12 to>400 umol/L with visibly distinct colorimetric ranges of 0 to 25, 25 to 100, 100 to 200, 200 to 350, or greater than 400 umol/L. 
     
     
         27 . The method of  claim 20 , wherein the test pad comprises more than one zone or dry chemistry enabling pact that can detect more than one biomarker for a single collection, wherein the multiple zones or test pad assesses for difference ranges in sensitivity of the metabolite or different biomarkers for health status.