Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2000-10-13
2003-04-15
Horlick, Kenneth R. (Department: 1634)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C536S023100, C536S024300, C536S024330, C435S091100, C435S091200
Reexamination Certificate
active
06548252
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to improvements in diagnostic techniques, and more particularly to diagnostic techniques for detecting and identifying DNA or RNA damage. The invention has particular utility in the detection of cervical cancer and will be described in connection with such utility, although other utilities are contemplated, as will be discussed below in detail, including detection of other cancers and other disease conditions, as well as health conditions brought out by exposure to environmental insults such as radiation, metals, smoke and solvents.
BACKGROUND OF THE INVENTION
The current gold standard for detecting cervical cancer in women is the so-called “PAP Smear.” However, the reliability of PAP Smear testing, which relies upon a technician's observations, under the microscope, of cellular morphology, may be compromised by technician fatigue and/or subjectivity. Even very experienced technicians may misread a slide. If a false-positive is “called” or the slide results in an “uncertain” call, the physical may err on the side of the patient's safety, and call for a hysterectomy (uterus removal) or total hysterectomy (uterus and ovary removal). This results in the patient taking a lifetime dose of hormone replacement medications to keep the body in balance. Of course, the failure to identify “a pre-cancerous condition” could lead to an even more disastrous result to a patient.
OBJECTIVES OF THE INVENTION
It is thus a primary object of the invention to overcome the aforesaid and other disadvantages of the prior art. Another object of the invention is to provide an analytical technique for detecting and diagnosing disease conditions, as well as health conditions due to exposure to environmental conditions, by detecting and identifying DNA or RNA damage markers. Another more object of the invention is to provide a reliable, totally objective analytical technique for detecting cancer.
BRIEF DESCRIPTION OF THE INVENTION
In order to effect the foregoing and other objects, the present invention provides an analytical technique for detecting cancer or other disease and/or health conditions based on measurement of free levels of nucleotide excision products resulting from DNA or RNA damage, such as 8OH2′dG-8 hydroxy 2′ deoxyguanosine; O6MG—O-6-methylguanine; 2′dG—2′deoxyguanosine; 7MG—7-methylguanine; 8NG—8-nitroguanine; 8OHG—8-hydroxyguanine; 8OH2′dA—8-hydroxy-2-deoxyadenosine; 8OHA—8-hydroxyadenine; 5OH2′dCy—5-hydroxy 2′deoxycytidine; 5OHU—5-hydroxyuracil; 3NT—3-nitrotyrosine; or 3 CIT—3-chlorotyrosine in biological samples from selected areas of the body. More particularly, the present invention is based on the hypothesis that specific areas of the body are semi-isolated in-situ biochemical environments for nucleotide excision products such as 8OH2′dG or other nucleotide excision products, and that the levels of such free 8OH2′dG or other nucleotide excision products in the semi-isolated environment magnify the combined defects of DNA or RNA damage and repair mechanisms. By way of specific example; the effective damage and repair rate increases in DNA in cancer, or pre-cancerous cells, has been found to be magnified by accumulation in the extra-cellular matrix in selected areas of the body of 8OH2′dG. For example, as applied to cervical cancer, the cervix has been found to be a semi-isolated in-situ biochemical environment which may be accessed through cervico vaginal lavage sampling. Thus, an objective analytical technique for determination of cervical cancer is provided.
REFERENCES:
patent: PCT/US98/22275 (1998-10-01), None
Kristal et al. Simulateneous Analysis of the Majority of Low-Molecular-Weight, Redox-Active Compounds from mitochondria. Analytical Biochemistry vol. 263: Oct. 18-25, 1998.*
Kasai H. Analysis of a form of oxidative DNA damage, 8-hydroxy-2'deoxyguanosine, as a marker of cellular oxidative stress during carcinogenesis. Mutation Research, vol. 387, pp. 147-163, Dec. 1997.*
Ames B. Endogeneous Oxidative DNA Damage, aging and Cancer, Free Rad. Res. Communications, vol. 7, No. 3-6, pp. 121-128, Jun. 1989.*
Kasai H. Analysis of a form of oxidative DNA damage, 8-hydroxy-2'deoxyguanosine, as a marker of cellular oxidative stress during carcinogenesis. Mutation Research, vol. 387, pp. 147-163, Dec. 1997.*
“A Carbon Column-based Liquid Chromatography Electrochemical Approach to Routine 8-Hydroxy-2'Deoxyguanosine Measurements in Urine and Other Biologic Matrices: A One-Year Evaluation of Methods” by Mikhail Bogdanov, et al Free Radical Biology & Medicine, vol. 27, Nos. 5/6, pp 647-666, 1999.
“Simultaneous Analysis of the Majority of Low-Molecular-Weight, Redox-Active Compounds from Mitochondria”, by Bruce Kristal, et al, Analytical Biochemistry 263, pp. 18-25 (1998) Artticle No. AB982831.
ESA, Inc.
Hayes & Soloway P.C.
Horlick Kenneth R.
Wilder Cynthia
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