JP2006501906A - Method and apparatus for non-invasive analysis of metabolic processes - Google Patents

Abstract

The present invention relates to methods and arrangements for non-invasive analysis of control and control processes in human and animal metabolism in order to be able to draw conclusions about specific diseases from changes in individual metabolic parameters. The method involves prophylactic analysis and specific physical and mental stress for therapeutic control of individual clinical features for early detection of cancer, inflammatory diseases, and determination of antioxidant needs. It can also be used for regular checkups of professional occupation groups. In accordance with the present invention, bioactive substances that are metabolically relevant to describe the control and control processes in the human body and that have autofluorescent properties are selected from the native fluorescence spectrum in the wavelength range between 287 nm and 640 nm and Linked in chemical and biophysical models. The fluorescence spectrum consists of a light source (5), a fiber optic cable (1) for supplying stimulation light to the measurement site, a fiber optic cable (2) for branching fluorescence to the spectrometer (6), and a quantification computer (7). It is detected by an optical measurement path consisting of

Description

  The present invention relates to methods and arrangements for non-invasive analysis of control and control processes in human and animal metabolism in order to be able to draw conclusions about specific diseases from changes in individual metabolic parameters . The method includes prophylactic analysis for therapeutic control of individual clinical features for early detection of cancer, inflammatory diseases, and determination of the need for antioxidants, and specific physical and mental stress It can also be used for regular check-ups of professional groups with

  For several years, fluorescence spectrum analysis has been known as a highly accurate and very specific method in biomedical analysis as a basic biology study and diagnostic aid for transport processes through biological membranes, And now it is in a developmental period that is making steady progress. The basis of this measurement method is knowledge of the properties of artificial fluorophores and knowledge of the excitation and emission wavelengths of autofluorphores. Tryptophan, adenosine triphosphate (ATP), guanosine triphosphate (GTP), nicotinamide adenine dinucleotide phosphate (NADP), reduced nicotinamide adenine dinucleotide (NADH), kynurenine, flavin adenine dinucleotide (FAD) and Many parameters related to metabolism, such as thromboxane, have so-called autofluorescence.

  The measurement of autofluorescence has the advantage that the metabolism does not need to be supplied with non-physiological substances. For example, in patent DE 3542167 A1, the change in the autofluorescence of ascorbic acid during the oxidation process is used to measure eye lens opacity in a non-invasive manner.

Further work is DE 69591815 T2, which uses NADH's high native fluorescence for the detection of melanoma.
In Patent DE 3210593 A1, NADH autofluorescence is used to measure the redox state of organs in an invasive method using an endoscope.

  In patent DE 19535114 A1, in the emission range of 520 to 600 nm, various autofluorescences of biological tissues are used for diagnosis of cancerous tissues without reference to specific biologically relevant substances. Yes. In the case of this method as well, the measurement device must be brought invasively to the measurement site by means of an endoscope.

  After complex experiments, fluorescence spectroscopic behaviors of living tissues and organs are described in US Pat. No. 5983125, US 5769081, US 5369496, US 6091985, US 6080584, US 6346101 B1, US. No. 2002/0002337 A1, US Pat. No. 5,943,113 and US Pat. No. 6,205,353 B1 were described in connection with preventive cancer diagnosis. For this analysis, the intensity of individual substances such as tryptophan, NADH and flavins and the maximum fluorescence intensity in the wavelength range from 320 to 580 nm were considered. In addition, the results of Fourier analysis were considered.

  In their analysis, the use of the maximum fluorescence intensity in the wavelength range from 320 nm to 600 nm or the absolute fluorescence intensity of related metabolic parameters such as NADH, tryptophan, FAD and kynurenine, as well as the ratio of two substances such as NADH and kynurenine. The disadvantage is that it does not allow a clear separation between “healthy” and “cancerous”. Thus, for example, a low ratio between the strengths of NADH and kynurenine is not only unique to cancer, but all inflammatory diseases exhibit a similar ratio. This is not strange because many cancerous diseases involve inflammation.

  A further disadvantage of the invasive method described above is that the stress load of the measurement process gives a false instantaneous picture of the metabolism and makes no statement about the metabolic control process. Such a statement is possible only by measuring without stress, which can be repeated at short intervals, or in a specific way with respect to the time before and after stress loading.

  The problem underlying the present invention is a method that enables the control of human and animal metabolism and the description of the control process in order to be able to conclude a specific clinical picture in the case of changes in the following processes and The provision of equipment. The method is intended to allow the actual measurement process to be repeated non-invasively and rapidly so as not to cause stress loading due to the measurement process.

According to the invention, the problem is solved by the features disclosed in the claims.
An advantage of the present invention is the non-invasive measurement of the fluorescence spectrum and the lack of stress guaranteed thereby. This measurement method makes it possible to make repeated measurements at very short intervals and therefore to recognize regulatory processes in metabolism. By changing these control processes under specific stress conditions, it is possible to draw conclusions about the pathological changes of the organism.

In the following, the present invention will be described in detail by means of embodiments.
Using the optical measurement section according to FIG. 1 consisting of a fiber optic cable 1 for supplying excitation light and a fiber optic cable 2 with a collimator for branching the measurement signal, [ ¹ ] is the appropriate part of the body, preferably the thumb and Placed in the crease between the index finger. Both fiber optic cables 1, 2 are located in an ergonomically made carrier, preferably the handpiece 4, and their outlets are preferably located perpendicular to each other. A light source 5 consisting of a laser with a downstream monochromator or filter or a control xenon flash lamp emits light for excitation of autofluorescence and is directed via a fiber optic cable 1 to a measurement site. The wavelength of the excitation light is preferably 287 nm, 305 nm, 326 nm and 337 nm.

  The fluorescence emitted at the measurement site by excitation is collected by the collimator 3, connected into the fiber optic cable 2, and directed to the spectrometer 6. The spectrometer can also have both a CCD line sensor and a photomultiplier tube with a downstream acousto-optic monochromator as a transducer unit. The optical spectrum converted into an electrical signal by the spectrometer 6 is now stored in the corresponding computer structure 7.

  A fluorescence spectrum stored in the computer, consisting of recording wavelengths in the range from 287 nm to 600 nm and the corresponding fluorescence intensity, is prepared for analysis in a suitable table format.

FIG. 2 shows an example of such a native spectrum.
A combination of numerical values (wavelength and fluorescence intensity) for metabolically related biologically active substances such as ATP, GTP, tryptophan, orotic acid, NADP, NADH, FAD, etc. is selected from these tables. The excitation and emission wavelengths of these materials were determined by a composite pilot test. Since different skin structures and skin components do not allow the use of absolute values, further analysis can only be performed with relative values. It is therefore necessary to determine numerical combinations of relevant bioactive substances and correlate them with biophysical and biochemical models. These models include substances that react with each other during the metabolic process, transform with each other, and / or influence each other with their concentration and reactivity.

  FIG. 3 shows a graphical illustration of the results of a simple biochemical model that is used as the first stage of diagnosis and consists of a combination of NADH, kynurenine, FAD, NADP and thromboxane. This illustration proves that even the use of five metabolism-related substances is not sufficient to separate cancer disease from inflammatory disease. The first selection stage is only suitable for distinguishing between “sick” and “healthy”. Subsequently, further selection steps are performed to distinguish between inflammatory and cancer diseases and also to detect the distinction among inflammatory diseases.

FIG. 4 shows the separation between cancer diseases and treated cancer diseases and inflammatory diseases.
Healthy probands and patients with spectrum analysis by selection of diseases using biophysical and biochemical models based on known bioactive substances, without using known numerical combinations (wavelength and intensity) of bioactive substances Simultaneously with an analysis using a self-learning system that searches for differences in the spectrum of

  FIG. 5 shows additional selections at wavelengths of 509 nm and 495 nm, where the luminescent material is not known so far, but shows the successful use of this selection.

FIG. 6 is a block diagram of a measurement reading record. It is a figure which shows a native fluorescence spectrum. It is a figure which shows the result of the simple biochemical model as a selection step. It is a figure which shows the result of isolation | separation of a cancer disease and an inflammatory disease. FIG. 6 shows selection using emission wavelengths 509 nm and 495 nm.

Explanation of symbols

1. 1. Fiber optical cable for supplying excitation light 2. Fiber optical cable for fluorescence branching Collimator 4. Handpiece 5. Light source 6. Spectrometer 7. Computer structure

Claims (10)

  For the process of dialysis and apheresis for therapeutic control, for routine screening of professional and sports professionals with high levels of physical and mental stress, for disease diagnosis and prophylactic testing And non-invasive and / or invasive analytical methods of control and control processes in human and animal metabolism for the determination of the need for antioxidants, which interact with each other during metabolic processes Metabolites related to each other that are converted to each other and / or interact with each other in their concentration and reactivity and exhibit autofluorescence (endogenous), in their fluorescence intensity and thus indirectly in their concentration Measured, placed in mutual mathematical relationship according to biochemical requirements, and compared to a symptom-specific model that defines the metabolic state of the disease Wherein the door.   The sign-specific model corresponds to each metabolic state of the clinical picture and is calculated from the fluorescence intensity by a mathematical combination such as quotient, product, sum, subtraction, or more complex formulas ( 2. Method according to claim 1, characterized in that it consists of at least 6) calculated values.   The fluorescence intensity is in the wavelength range from 287 nm to 800 nm, preferably from 340 nm for metabolic related substances whose emission wavelengths are known, preferably ATP, GTP, FAD, NADH, NADP, kynurenine, orotic acid, thromboxane and tryptophan. Method according to claims 1 and 2, characterized in that it is measured at 600 nm.   Method according to claims 1-3, characterized in that the measurement of the fluorescence intensity is carried out at specific times and / or at specific intervals, so that control and control processes are detected by these process measurements.   The patient is afflicted with mental or physiological stress at a specific point in the measurement, the fluorescence intensity is measured several times before and after the stress load and the control process in the metabolism is determined. Method according to 1-4.   A bioactive substance exhibiting autofluorescence is stimulated for light emission by light having an excitation wavelength of 287 nm to 340 nm, preferably 340 nm, between cells and between the cells. By the method.   Device according to claims 1 to 6, characterized in that the place stimulated to fluoresce is located in the earlobe, hand and nostril, preferably in the crease between the thumb and forefinger.   Device for carrying out the method according to the preceding claim, wherein the monochromatic light required for excitation is emitted by a light source (5), preferably a laser or xenon flash lamp with a light filter, and a fiber optic cable (1) A device characterized by being directed to the site of measurement via   The emitted light of the autofluorophore is directed and measured from the measurement site via a fiber optic cable (2) to a spectrometer (6) comprising a CCD line sensor or acousto-optic monochromator and photomultiplier tube 9. Device according to claim 8, characterized in that, after digitizing the numerical values, the emission intensity is analyzed by means of a suitable computer structure (7).   Device according to claims 8 to 9, characterized in that the analysis in the structure by means of a computer is performed by a mathematical model of a biological control system and / or a self-learning system.

Images