Apparatus and methods for performing in-vivo measurements of enzyme activity

• US 4,071,020 A
• Filed: 06/03/1976
• Issued: 01/31/1978
• Est. Priority Date: 06/03/1976
• Status: Expired due to Term

• Current Assignee: Pharmacontrol Corporation (14605, Inc.)
• Original Assignee: Xienta, Inc.
• Inventors: Pugliese, Peter
• Primary Examiner(s): Padgett, Benjamin R.
• Assistant Examiner(s): Nucker, Christine M.
• Application Number:
  US05/692,511
• Time in Patent Office: 607 Days
• Field of Search: 128/2 A, 128/2 E, 128/2 W, 128/2.1 E, 195/103.5 R, 195/103.5 C, 424/7, 424/9, 23/253 TP
• US Class Current: 424/9.6
• CPC Class Codes: A61B 5/0071 by measuring fluorescence emission
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: A61B 5/1455 using optical sensors, e.g. spectral photometrical oximeters
• CPC Class Codes: A61B 5/441 Skin evaluation, e.g. for skin disorder diagnosis
• CPC Class Codes: A61B 5/6835 Supports or holders, e.g., articulated arms
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53

Abstract

There are disclosed apparatus and methods for performing in-vivo measurements of enzyme kinetics on the epidermis and mucous membranes of a subject. One or more reactants are placed on a predetermined area of the skin in order to react with the enzyme or other biological substance of interest. At least one physical characteristic of one or more of the reaction products formed is measured. Changes in enzymatic or related activity in response to various stimuli can be observed by continuously monitoring the physical characteristic of the reaction product while certain controls are varied, thereby providing dynamic information not previously obtainable.

Claims

1. Apparatus for performing an in-vivo measurement of a substrate, enzyme, or other biochemical agent on the skin or mucous membranes of an accessible body surface of a patient comprising means for reacting at least one chemical with said substrate, enzyme or other biochemical agent on a predetermined area of the skin or mucous membranes of the patient to produce at least one reaction product, and means for measuring a physical characteristic of said at least one reaction product.

2. Apparatus in accordance with claim 1 wherein said measuring means measures said physical characteristic while said at least one reaction product is on the skin or mucous membranes of the patient.

3. Apparatus in accordance with claim 2 wherein said measuring means includes first means for irradiating said at least one reaction product while on the skin or mucous membranes of the patient and second means for measuring a wavelength-dependent physical characteristic thereof.

4. Apparatus in accordance with claim 3 wherein said second means measures the intensity of the fluorescence of said at least one reaction product while it is being irradiated by said first means.

5. Apparatus in accordance with claim 1 wherein said physical characteristic is wavelength-dependent.

6. Apparatus in accordance with claim 5 wherein said measuring means includes first means for irradiating said at least one reaction product and second means for measuring the intensity of the fluorescence of said at least one reaction product while it is being irradiated by said first means.

7. A method for performing an in-vivo measurement of a substrate, enzyme, or other biochemical agent on the skin or mucous membranes of an accessible body surface of a patient comprising the steps of reacting at least one chemical with said substrate, enzyme or other biochemical agent on a predetermined area of the skin or mucous membranes of the patient to produce at least one reaction product, and measuring a physical characteristic of said at least one reaction product.

8. A method in accordance with claim 7 wherein said physical characteristic is measured while said at least one reaction product is on the skin or mucous membranes of the patient.

9. A method in accordance with claim 8 wherein said measuring step includes a first sub-step of irradiating said at least one reaction product while on the skin or mucous membranes of the patient and a second sub-step of measuring a wavelength-dependent physical characteristic thereof.

10. A method in accordance with claim 9 wherein in said second sub-step the intensity of the fluorescence of said at least one reaction product is measured while it is being irradiated.

11. A method in accordance with claim 8 further including the step of recording continuously the measurements taken over an interval of at least several minutes.

12. A method in accordance with claim 7 wherein said physical characteristic is wavelength-dependent.

13. A method in accordance with claim 12 wherein said measuring step includes a first sub-step of irradiating said at least one reaction product and a second sub-step of measuring the intensity of the fluorescence of said at least one reaction product while it is being irradiated.

14. A method in accordance with claim 13 wherein said at least one chemical includes nicotinamide adenine dinucleotide phosphate, and said at least one reaction product includes reduced nicotinamide adenine dinucleotide phosphate, and wherein in said first sub-step said at least one reaction product is irradiated by ultraviolet light at a wavelength in the range 350-365 millimicrons, and in said second sub-step the fluorescence of said at least one reaction product is measured at a wavelength of approximately 460 millimicrons.

15. A method in accordance with claim 14 wherein said at least one chemical also includes glucose-6-phosphate, and said enzyme or other biochemical agent is glucose-6-phosphate dehydrogenase.

16. A method in accordance with claim 14 wherein said at least one chemical also includes glucose-6-phosphate dehydrogenase, and said enzyme or other biochemical agent is glucose-6-phosphate.

17. A method in accordance with claim 7 further including the step of recording continuously the measurements taken over an interval of at least several minutes.