Abstract

A cost-effective optical cancer screening and monitoring technique was demonstrated in a pilot study of canine serum samples and was patented for commercialization. Compared to conventional blood chemistry analysis methods, more accurate estimations of the concentrations of albumin, globulins, and hemoglobin in serum were obtained by fitting the near UV absorbance and photoluminescence spectra of diluted serum as a linear combination of component reference spectra. Tracking these serum proteins over the course of treatment helped to monitor patient immune response to carcinoma and therapy. For cancer screening, 70% of dogs with clinical presentation of cancer displayed suppressed serum hemoglobin levels (below 20mg/dL) in combination with atypical serum protein compositions, that is, albumin levels outside of a safe range (from 4 to 8g/dL) and globulin levels above or below a more normal range (from 1.7 to 3.7g/dL). Of the dogs that met these criteria, only 20% were given a false positive label by this cancer screening test.

© 2007 Optical Society of America

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References

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    [CrossRef] [PubMed]

2005 (2)

A. Statnikov, C. Aliferis, I. Tsamardinos, D. Hardin, and S. Levy, "A comprehensive evaluation of multicategory classification methods for microarray gene expression cancer diagnosis," Bioinformatics 21, 631-643 (2005).
[CrossRef]

B. Leca-Bouvier and L. J. Blum, "Biosensors for protein detection: a review," Anal. Lett. 38, 1491-1517 (2005).
[CrossRef]

2004 (1)

E. P. Diamandis, "How are we going to discover new cancer biomarkers? A proteomic approach for bladder cancer," Clin. Chem. 50, 793-795 (2004).
[CrossRef] [PubMed]

2003 (2)

R. Aebersold and M. Mann, "Mass spectroscopy-based proteomics," Nature (London) 422, 198-207 (2003).
[CrossRef] [PubMed]

L. Pena, M. D. Perez-Alenza, A. Rodriguez-Bertos, and A. Nieto, "Canine inflammatory mammary carcinoma: histopathology, immunohistochemistry and clinical implications of 21 cases," Breast Cancer Res. Treat. 78, 141-148 (2003).
[CrossRef] [PubMed]

2002 (2)

J. Li, Z. Zhang, J. Rosenzweig, Y. Y. Wang, and D. W. Chan, "Proteomics and bioinformatics approaches for identification of serum biomarkers to detect breast cancer," Clin. Chem. 48, 1296-1304 (2002).
[PubMed]

K. P. H. Pritzker, "Cancer biomarkers: Easier said than done," Clin. Chem. 48, 1147-1150 (2002).
[PubMed]

1999 (1)

C. V. Sapan, R. L. Lundblad, and N. C. Price, "Colorimetric protein assay techniques," Biotechnol. Appl. Biochem. 29, 99-108 (1999).
[PubMed]

1997 (1)

P. Jichlinski, M. Forrer, J. Mizeret, T. Glanzmann, D. Braichotte, G. Wagnières, G. Zimmer, L. Gulillou, F. Schmidlin, P. Graber, H. van den Bergh, and H. J. Leisinger, "Clinical evaluation of a method for detecting superficial transitional cell carcinoma of the bladder by light-induced fluorescence of protoporphyrin IX following topical application of 5-aminolevulinic acid," Lasers Surg. Med. 20, 402-408 (1997).
[CrossRef] [PubMed]

1995 (1)

M. Weiss, C. L. Loprinzi, E. T. Creagan, R. J. Dalton, P. Novotny, and J. R. O'Fallon, "Utility of follow-up tests for detecting recurrent disease in patients with malignant melanomas," J. Am. Med. Assoc. 274, 1703-1705 (1995).
[CrossRef]

1993 (3)

T. S. Mang, C. McGinnis, C. Liebow, U. O. Nseyo, D. H. Cream, and T. J. Dougherty, "Fluorescence detection of tumors," Cancer 71, 269-276 (1993).
[CrossRef] [PubMed]

M. Kondo, N. Hirota, T. Takaoka, and M. Kajiwara, "Heme biosynthetic enzyme activities and porphyrin accumulation in normal liver and hepatoma cell lines of rat," Cell. Biol. Toxins 9, 95-105 (1993).
[CrossRef]

R. Beri and R. Chandra, "Chemistry and biology of heme: effect of metal salts, organometals, and metalloproteins on heme synthesis and catabolism, with special reference to clinical implications and interactions with cytochrome P-450," Drug Metab. Rev. 25, 49-152 (1993).
[CrossRef] [PubMed]

1990 (1)

M. R. Hubmann, M. J. P. Leiner, and R. J. Schaur, "Ultraviolet fluorescence of human sera: I. Sources of characteristic differences in ultraviolet fluorescence spectra from sera of normal and cancer-bearing patients," Clin. Chem. 36, 1880-1883 (1990).
[PubMed]

1987 (1)

D. M. Harris and J. Werkhaven, "Endogenous porphyrin fluorescence in tumors," Lasers Surg. Med. 7, 467-472 (1987).
[CrossRef] [PubMed]

1973 (1)

B. R. Munson and R. J. Fiel, "A review: biochemical alterations associated with mouse spleen cells infected with Friend virus," J. Med. 4, 354-370 (1973).
[PubMed]

Anal. Lett. (1)

B. Leca-Bouvier and L. J. Blum, "Biosensors for protein detection: a review," Anal. Lett. 38, 1491-1517 (2005).
[CrossRef]

Bioinformatics (1)

A. Statnikov, C. Aliferis, I. Tsamardinos, D. Hardin, and S. Levy, "A comprehensive evaluation of multicategory classification methods for microarray gene expression cancer diagnosis," Bioinformatics 21, 631-643 (2005).
[CrossRef]

Biotechnol. Appl. Biochem. (1)

C. V. Sapan, R. L. Lundblad, and N. C. Price, "Colorimetric protein assay techniques," Biotechnol. Appl. Biochem. 29, 99-108 (1999).
[PubMed]

Breast Cancer Res. Treat. (1)

L. Pena, M. D. Perez-Alenza, A. Rodriguez-Bertos, and A. Nieto, "Canine inflammatory mammary carcinoma: histopathology, immunohistochemistry and clinical implications of 21 cases," Breast Cancer Res. Treat. 78, 141-148 (2003).
[CrossRef] [PubMed]

Cancer (1)

T. S. Mang, C. McGinnis, C. Liebow, U. O. Nseyo, D. H. Cream, and T. J. Dougherty, "Fluorescence detection of tumors," Cancer 71, 269-276 (1993).
[CrossRef] [PubMed]

Cell. Biol. Toxins (1)

M. Kondo, N. Hirota, T. Takaoka, and M. Kajiwara, "Heme biosynthetic enzyme activities and porphyrin accumulation in normal liver and hepatoma cell lines of rat," Cell. Biol. Toxins 9, 95-105 (1993).
[CrossRef]

Clin. Chem. (4)

M. R. Hubmann, M. J. P. Leiner, and R. J. Schaur, "Ultraviolet fluorescence of human sera: I. Sources of characteristic differences in ultraviolet fluorescence spectra from sera of normal and cancer-bearing patients," Clin. Chem. 36, 1880-1883 (1990).
[PubMed]

K. P. H. Pritzker, "Cancer biomarkers: Easier said than done," Clin. Chem. 48, 1147-1150 (2002).
[PubMed]

E. P. Diamandis, "How are we going to discover new cancer biomarkers? A proteomic approach for bladder cancer," Clin. Chem. 50, 793-795 (2004).
[CrossRef] [PubMed]

J. Li, Z. Zhang, J. Rosenzweig, Y. Y. Wang, and D. W. Chan, "Proteomics and bioinformatics approaches for identification of serum biomarkers to detect breast cancer," Clin. Chem. 48, 1296-1304 (2002).
[PubMed]

Drug Metab. Rev. (1)

R. Beri and R. Chandra, "Chemistry and biology of heme: effect of metal salts, organometals, and metalloproteins on heme synthesis and catabolism, with special reference to clinical implications and interactions with cytochrome P-450," Drug Metab. Rev. 25, 49-152 (1993).
[CrossRef] [PubMed]

J. Am. Med. Assoc. (1)

M. Weiss, C. L. Loprinzi, E. T. Creagan, R. J. Dalton, P. Novotny, and J. R. O'Fallon, "Utility of follow-up tests for detecting recurrent disease in patients with malignant melanomas," J. Am. Med. Assoc. 274, 1703-1705 (1995).
[CrossRef]

J. Med. (1)

B. R. Munson and R. J. Fiel, "A review: biochemical alterations associated with mouse spleen cells infected with Friend virus," J. Med. 4, 354-370 (1973).
[PubMed]

Lasers Surg. Med. (2)

D. M. Harris and J. Werkhaven, "Endogenous porphyrin fluorescence in tumors," Lasers Surg. Med. 7, 467-472 (1987).
[CrossRef] [PubMed]

P. Jichlinski, M. Forrer, J. Mizeret, T. Glanzmann, D. Braichotte, G. Wagnières, G. Zimmer, L. Gulillou, F. Schmidlin, P. Graber, H. van den Bergh, and H. J. Leisinger, "Clinical evaluation of a method for detecting superficial transitional cell carcinoma of the bladder by light-induced fluorescence of protoporphyrin IX following topical application of 5-aminolevulinic acid," Lasers Surg. Med. 20, 402-408 (1997).
[CrossRef] [PubMed]

Nature (1)

R. Aebersold and M. Mann, "Mass spectroscopy-based proteomics," Nature (London) 422, 198-207 (2003).
[CrossRef] [PubMed]

Other (8)

A. A. Pineda, ed., Selective Plasma Component Removal (Futura, 1984), p. 156.

B. F. Feldman, J. G. Zinkl, and N. C. Jain, eds., Schalm's Veterinary Hematology, 5th ed. (Lippincott Williams & Wilkins, 2000), pp. 565-570 and 899-903.

S. C. Gad, ed., Drug Discovery Handbook (Wiley-Interscience, 2005), pp. 81-83.

R. S. Cotran, V. Kumar, and T. Collins, eds., Robbins Pathological Basis of Disease (Saunders, 1999), pp. 260-327.

R. Richards-Kortum, R. Drezek, K. Basen-Engquist, S. B. Cantor, U. Utzinger, C. Brookner, and M. Follen, "Cervical dysplasia diagnosis with fluorescence spectroscopy," in Handbook of Biomedical Fluorescence, M. A. Mycek and B. W. Poque, eds. (Marcel Dekker, 2003), pp. 265-314.

G. Wagnières, A. McWilliams, and S. Lam, "Lung cancer imaging with fluorescence endoscopy," in Handbook of Biomedical Fluorescence, M. A. Mycek and B. W. Poque, eds. (Marcel Dekker, 2003), pp. 361-396.

S. Welle, Human Protein Metabolism (Springer, 1999), pp. 204-204.

R. J. Henry, D. C. Cannon, and J. W. Winkelman, eds., Clinical Chemistry Principles and Techniques, 2nd ed. (Harper & Row, 1974), pp. 449, 1071-1072, 1117, and 1239.

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Figures (14)

Fig. 1
Fig. 1

Average UV absorbance spectra for diluted serum from dog populations grouped by health status. On the average, the serum absorbance of geriatric patients and those with cancer was higher than that of healthy dogs across the wavelength range from 250 to 500   nm ; however, there was high variability (standard deviations indicated by error bars) within the cancer and geriatric populations. The absorbance spectra shown are through samples that have been diluted in water to 1 / 60 of their original concentrations.

Fig. 2
Fig. 2

Absorbance spectra of reference solutions for primary serum proteins. The spectral gap between 370 and 380   nm is a low signal region between two light sources.

Fig. 3
Fig. 3

Fluorescence spectra of reference solutions for primary serum proteins. Compared to albumin, globulins exhibited enhanced fluorescence near 300   nm , while hemoglobin showed negligible fluorescence.

Fig. 4
Fig. 4

Linear absorbance of serum components. The absorbance peak amplitude at the indicated wavelengths was proportional to that protein's concentration

Fig. 5
Fig. 5

Scattering contributions to serum absorbance spectra. The absorbance spectra of some samples exhibited a significant background, which increased towards shorter wavelengths. The dashed curve in this plot shows the smoothed scattering background, Scat ( λ ) , after normalizing the spectra so that Scat ( λ = 350   nm ) is 1.

Fig. 6
Fig. 6

Fluorescence of serum components saturates at higher concentrations due to reabsorbance.

Fig. 7
Fig. 7

Fitting typical serum spectra. Most all canine serum spectra were easily modeled as a linear combination of reference absorbance spectra and normalized fluorescence spectra from albumin, globulins, and hemoglobin.

Fig. 8
Fig. 8

Accurate spectroscopic estimates of albumin concentration in mixtures.

Fig. 9
Fig. 9

Accurate spectroscopic estimates of globulins in mixtures. Standard chemical blood tests (labeled “Globulins Chemical”) underestimate globulin levels.

Fig. 10
Fig. 10

Very accurate spectroscopic estimates of total protein (albumin + globulins) in mixtures, compared to less chemical test methods used in standard blood tests.

Fig. 11
Fig. 11

Sensitive spectroscopic estimates of trace hemoglobin levels in mixtures. Standard chemical blood tests (labeled “Hemoglobin Chemical”) did not accurately measure such low hemoglobin levels.

Fig. 12
Fig. 12

Identifying selected low-risk patients based on globulins and albumin levels in serum. The “safe” subset of healthy patients could be excluded from the high-risk population based on their albumin and globulin levels, regardless of their serum hemoglobin and total protein levels.

Fig. 13
Fig. 13

Screening for cancer based on hemoglobin and total protein levels in serum. Most of the cancer patients had lower serum hemoglobin levels than the healthy patients, as shown in the “suspect” compositional region.

Fig. 14
Fig. 14

Tracking temporal changes in serum composition for a dog with cancer. During the course of treatment following the Wisconsin protocol, fluctuations in (a) serum protein composition and (b) hemoglobin levels were observed using the optical methods described in this paper. The cause of the hemoglobin spike after 10 weeks of chemotherapy is unknown, but this dog's medical history is outlined in Table 2.

Tables (2)

Tables Icon

Table 1 Correlation Coefficients between Canine Serum Protein Concentrations and Healthy Group a

Tables Icon

Table 2 Medical History of a Dog Treated for Lymphoma

Equations (4)

Equations on this page are rendered with MathJax. Learn more.

A b s fit ( λ ) = A 1.0 A l b u ( λ ) + G 1.0 G l o b ( λ ) + H 0.1 H e m ( λ ) + S × S c a t ( λ ) .
E a b s [ i = 1 n { ( A b s ( λ i ) A b s fit ( λ i ) ) 2 } ] 1 / 2 / i = 1 n { A b s ( λ i ) } .
P F a l b u = 180 A 1 + 1.2 A 1.2 , P F g l o b = 368 G 1 + 1.8 G 1.2 ,
F fit ( λ ) = P F a l b u F a l b u ( λ ) + P F g l o b F g l o b ( λ ) P F a l b u + P F g l o b .

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