METHOD FOR DIAGNOSTICS OF PATHOLOGY IN BIOLOGOCAL OBJECTS

Abstract

A method of the present invention is used for diagnostics of pathology in biological objects. The method includes the steps of analyzing a first biological element obtained from a human to identify ratio between first isotopes and second isotopes wherein weight of the first isotopes differs from weight of the second isotopes. The method further includes the step of analyzing information about natural distribution of the first isotopes and the second isotopes in a second biological element. The method includes the step of comparing the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element thereby changing the ratio of the first isotopes and the second isotopes of the first biological element to match the ratio of the first isotopes and the second isotopes of the second biological element.

Description

RELATED APPLICATIONS

This application claims priority to U.S. provisional application Ser. No. 62/123,900 filed on Dec. 1, 2014 and entitled “SYSTEM, APPARATUS, METHODS AND COMPOSITIONS FOR THE TREATMENT OF GROUP OF DISEASES INVOLVING ABNORMAL CELL GROWTH AND OTHER HEALTH ABNORMALITIES OF MAMMALS”, which is incorporated herewith in by reference in its entirety.

FIELD OF THE INVENTION

The invention relates to biology and medicine, and more particularly to a method to estimate the internal state of a biological object by detecting pathology its body, in particular, for screening with the purpose of detecting and monitoring a human body.

BACKGROUND OF THE INVENTION

People say that nothing is eternal under the Moon. It is true. There are plenty of reasons for something in live to go wrong in such a complex hierarchical systems, as, for example, a biological organism. In terms of automatic control theory, “mistakes” may occur on any particular level of said hierarchy causing breakage of negative feedback with integral (source of energy) or differential components or both. In technical systems, diagnostics allows to quickly establish the source of problem and eliminate it by simple substitution of broken part by the spare one.

In case of complex biological objects, diagnostics usually results in formulation of the set of symptoms with subsequent classification of pathological conditions. Fixing problems of deviation from the normal state here is not as easy as in manmade systems. Modern medicine is able to compensate excess or lack of certain chemicals (hormones, for example), to induce necessary changes in functions or organs and tissues. At the same time, it is still impossible to predict all the consequences of treatment whether it is pharmaceutical or surgical.

Proper diagnostics in fact is a key to successful treatment of diseases, given it provides information not only about how certain illness expresses itself, but also allows for exact determination of the level and place where deviation from the normal state had taken place. History of all components of medicine such as treatment, prevention and diagnostics can be explained in terms of going deeper and deeper in defining and treating source of pathology from body as a whole to organs, then to tissues, cells and nowadays—to molecules and macromolecules like proteins, amino acids and even RNA and DNA. Mutations accumulated in DNA both inherited and induced by pathogens believed to be the real reason behind most serious irreversible degenerative disorders, including cancer.

Random mutations in the genetic material of somatic cells accumulated with time considered as a main reason of aging. Genetic diseases caused by changes in DNA sequences of critical genes lead to the changes in protein composition of affected cells (tumor cells for example). That is why modern diagnostics depend heavily on development and detection of unique molecular markers. If we refer to FDA definition of biological marker it explicitly states that a biological marker is a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or biological responses to a therapeutic intervention.

Biomarkers presence can be, as in the case of cancer, produced by abnormal tissue/tumor or a response to the pathology-affected tissue. Examples of tumor markers are PSA and PAP for prostate cancer, CA 15-3 or CA 27-29 for breast cancer, MCA for breast metastatic cancer, HER 2 for primary or metastatic cancer, neuron specific enolase (NSE) for neuroblastoma and small cell lung cancer. Due to complexity of cancer the efficiency of single biomarkers is not sufficient. Much better chances for successful diagnostics present development of so-called gene or protein signatures, where multiple genes or proteins selected based on statistical correlation with disease. The use of the biomarkers is the base for new biosensors development including multichannel biosensors for simultaneous analysis of samples. Known methods of diagnostics discussed in details in “Biosensors and Molecular Technologies for Cancer Diagnostics” published by Keith E. Herold and Avraham Rasooly, Taylor & Francis 2012, Print ISBN: 978-1-4398-4165-5.

In addition, traditional methods of diagnostics, like MRI are also playing important role allowing to precisely pinpoint cancerous tissues in the patient's body. It used for early detection of metastases in soft tissues and inside of bones. Current diagnostics methods are crucial for successful treatment. MRI is extremely important for surgical procedures and protein signatures define cells targeted by chemotherapy or an immunotherapy. All three methods: the surgery, the chemotherapy and the immunotherapy each in its own manner supposed to eliminate malignant tumors. Last two, i.e. the chemotherapy and the immunotherapy, by killing cancer cells. Therefore, that the essence of fighting disease can be reduced to find and remove or find and kill strategies.

The problem is in “collateral damage” and often life threatening side effects at quite low efficiency. Surgical tumor removal creates huge risk of cancer spreading all over the organism. Targeting cells with certain protein signature often becomes obsolete as soon as additional mutation happens, this time in cancer cells itself. One can only hope that both the chemotherapy and the immunotherapy somehow will become less harmful and more efficient. At the same time, not only for cancer, but also for all types of degenerative chronic diseases the following statement is true: diagnostics defines outcome of treatment.

When treatment on the level of macromolecule is required, then diagnostics on the level of tissue or organ is of little help. The same is correct for treatment on the level of molecules/macromolecules. In case of irreversible damage on the level of chemical bonds, fixing will require manipulations with atoms. What is the first, elementary step of pathological change caused by any pathogen without exception? Life scientists would start answering with the reference to processes in the cells.

The fact is that we can completely ignore any pathogens as long as not a single chemical bond in the organism is damaged. It means that the interaction between pathogen and organism did not take place. When it does, a lot depends on the type of chemical bond that was destroyed and what have happened in the result. When damages caused by pathogens are limited to a certain chemical bond, they are repaired by internal mechanisms the same way as reparation of mistakes during mitosis happens. The problem is that damage sometime goes deeper than chemical bonds level—to the nucleus. Once it happens, there is no way back, and change becomes irreversible contributing to the unavoidable finale whether it is about illness or aging.

The art is replete with various compositions and methods for diagnostics. U.S. Pat. No. 7,214,194 to Klyukin teaches a medical equipment designed for the temperature diagnostics of biological object pathologies. The method comprises dynamical measuring a body temperature in a series of points. Based on the difference temperature field on the surface of a body, zones are determined inside the biological object that are identified with pathology loci. A temperature is each point is estimated with taking into account a type of skin of the biological object, a type of a temperature sensor and the form of the sensor temperature change curve in process of measurement. The device comprises temperature measurement units, temperature sensors being connected to the temperature measurement units, a RAM unit, a program realization unit, a comparator unit, a programming unit, and a visualization unit. The invention allows to increase speed and reliability of exposing pathology loci.

Another prior art reference, U.S. Pat. No. 8,822,224 to Markin, for example, teaches a method for automatic evaluation, processing and/or testing of an anatomic pathology specimen is disclosed. The specimen is placed into a primary or secondary container labeled with a unique identification code, placed into a specimen carrier, and the carrier marked with an identification code which uniquely identifies the specimen and, by virtue of the identification code, the evaluation, processing and/or tests to be conducted thereon. The identification code may be in the form of a bar code, an RFID tag or similar device or any other identification that is either human read able, machine readable or electronically transferred. The specimen contained within the specimen container or within the specimen carrier is entered into the anatomic pathology, histology or molecular diagnostics LAS at a receiving station, which reads the identification code.

Bur let us consider elementary act of irreversible pathology, and why it happens on sub-atomic level. It is not that an effect of perturbation or ionization of electron subsystem on parameters of nucleus is unknown. It was demonstrated that atom ionization and other perturbations in electron shells (caused, for example, by magnetic fields) not only changes the decay periods of unstable nuclei, but also alters decay schemes and modifies the stability condition”. That is for unstable nuclei and at full ionization, which is definitely not the case for the subjects of life science.

Using data with ionization potentials, one can show additional arguments for interconnection between electron states and atoms stability, but again for heavy atoms. For “normal” s-, p-, d-elements it is unreasonable to expect any effect of ionization, let alone excitation of electron state on parameters of nuclei, for example isotope composition. Small change in energy and character of electron state would never lead to a nuclear reaction. At the same time we believe, that interaction between electrons, protons and neutrons goes through the polarization potential field for any and not only heavy atoms.

We have to remember that this interaction occurs two levels higher than electromagnetic interaction and therefore it is very important but not necessarily in terms of the interaction energy, but rather in terms of long-range symmetries. The excess of neutrons over protons in the atoms is directly proportional to the sum of d- and f-electrons. It does not mean that change in the number of d- or f-electrons will cause correspondently change in number of neutrons. It can be an indication of the fact that when number of d- or f-electrons or just symmetry of external electron configuration is changed the atom would be “better off” (more stable) with different number of neutrons in the nucleus being in fact a different isotope of the same chemical element. The opposite situation may happen as well when change in isotope composition requires subsequent change in the number or symmetry of external electrons. In inorganic materials or in this regards in any non-live matter these kinds of adjustments are unrealistic.

In live organisms situation is completely different. Due to metabolism, there is a constant flow of materials including ions under discussion with natural isotope distribution. In results of the changes in electron configuration of d- and f-electrons containing elements due to ionization or interaction with free radical or any other pathogen, the adjustment of isotope composition happens. It occurs not by changes in the nucleus of the atom affected by external agents, but by simple substitution of said atom on different isotope more suitable for the electron configuration formed in result of external action thanks to the constant supply of all kind of elements feeding live organism.

One more time, change in electron configuration of d- and f-electron containing elements that are part of live organism may require change in isotope composition of said elements and vise-versa: change in their isotope composition may cause the change not only external electron configuration but also both short—and long—range order or symmetry. In general, as we have demonstrated above the same should be true not only for elements with d- and f-electrons, but for s- and p-elements as well, only expressed in much less degree. Exactly this interconnection in what we consider an elementary act of irreversible pathology gives us the hope that even without being able to control parameters of vector and polarization potentials, manipulation with isotope composition gives a solid chance to transform pathology-affected organism back into the normal state.

The human being is a stable hierarchical system based on a number vector functions with a complex system of negative feedback connections uniting all of its differential and integral components starting from electrons, protons and neutrons and ending up with mind/intellect. Disease depends on severity or stage and expressed on different levels of said hierarchical system. To deal with illness we can “from above or below” meaning from integral or differential levels. Most serious pathologies like progressive degenerative diseases (including cancer) and aging quite often considered as results of accumulated mutations. There is an opinion that mutations accumulated in healthy cells cause aging, and mutations accumulated in adult stem cells cause cancer.

Both situations can be characterized by avalanche like destruction of negative feedback connections leading to a situation when whole hierarchical system losing its stability and disintegrating. Any pathological change in organism starts from excitation or breakage of chemical bond. Not every chemical bond in organism (macromolecule) is equally important. Due to the nature of human organism, the vital chemical bonds are ones created with d-electrons containing metals. Not every carcinogen cause mutation and not every mutation leads to cancer is just a reflection of existence of the above-mentioned vital and not vital chemical bonds. When ultraviolet radiation or free radical from cigarette smoke or wine affects skin or epithelial tissue, it has very little chance to destroy chemical bond under discussion. In metal containing amino acids or proteins, concentration of transition metals is negligibly low. In many cases about one d-electron-containing element per thousands of other atoms. These explain seemingly random character with which cancer choosing its victims. Once pathogen becomes lucky enough to hit and destroy vital chemical bond, organism will try to fix the problem of broken negative feedback connection and jeopardized stability by supplying heavier isotope of the same d-electron containing element.

Once it takes the place of a light one, irreversible damage is done and fixed. The same effect is achievable with s- and p-elements but it requires much higher concentration of elements (one or two orders of magnitude more).

The incidence of cancer continues to climb as the general population ages, as new cancers develop, and as susceptible populations (e.g., people infected with AIDS or excessively exposed to sunlight) grow. A tremendous demand therefore exists for new methods and compositions that can be used to diagnose early stages of cancer and other pathologies like progressive degenerative diseases.

SUMMARY OF THE INVENTION

A system for diagnostics of pathology in biological objects. The system includes a first device adaptable to receive and analyze information from a first biological element obtained from a human to identify ratio between first isotopes and second isotopes wherein weight of the first isotopes differs from weight of the second isotopes in order to generate a first set of data. The system includes a second device cooperable with said first device, said second device adaptable to receive and analyze information about natural distribution of the first isotopes and the second isotopes in a second biological element in order to generate a second set of data. The system further includes a controller (a central processing unit) having a comparative software. The controller configured to receive said first set of data and said second set of data with said comparative software comparing the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element thereby generating a third set of data allowing to change the ratio of the first isotopes and the second isotopes of the first biological element to match the ratio of the first isotopes and the second isotopes of the second biological element.

A method for diagnostics of pathology in biological objects of the present invention begins with analyzing a first biological element obtained from a human to identify ratio between first isotopes and second isotopes wherein weight of the first isotopes differs from weight of the second isotopes. Then, information about natural distribution of the first isotopes and the second isotopes in a second biological element is analyzing.

A comparison analysis is conducted to compare the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element thereby changing the ratio of the first isotopes and the second isotopes of the first biological element to match the ratio of the first isotopes and the second isotopes of the second biological element. The first isotopes are light isotopes and the second isotopes are heavy isotopes. The first biological element is a tissue from a human body being diagnosed wherein the second biological element is a tissue from a human body without any pathology.

The method for diagnostics of the present invention uses a mass-spectrometry to identify the amount and type of chemicals presented in the first biological element by measuring the mass-to-charge ratio and abundance of gas-phase ions. The method for diagnostics also uses MRI on selected NMR active nuclei, wherein the step of using MRI on selected NMR active nuclei is further defined by isotopes K-39, K-40, K-41, Mg-25, and Zn-67. The method for diagnostics is further defined by studying isotope distribution of chemical elements of potassium, magnesium and zinc. The comparative analysis is performed on the samples of body fluids, wherein body fluids are samples of blood and/or urine. The pathology is a chronicle degenerative disorder such as a cancer, results of infectious diseases, and local aging of certain tissue part of a body.

An advantage of the present invention is to provide a method to improve health, cure abnormalities and degenerative disease and achieve therapeutic effect on mammals.

Another advantage of the present invention is to provide a method of to amplify therapeutic effect on cancer tissue and to protect healthy tissue from chemotherapy side effects and from immunotherapy side effect.

Still another advantage of the present invention is to provide a method to allow administering the isotope selective ingredient to cause fast and significant reduction in degree of malignancy and to induce changes of cells phenotype form malignant phenotype to a benign or normal phenotype.

These and other objects of the present invention will become readily apparent upon further review of the following specification and drawings.

Other objects, features, and advantages of the present invention will become apparent upon consideration of the following detailed description of a preferred embodiment thereof, when taken in conjunction with the appended drawing.

BRIEF DESCRIPTION OF THE DRAWINGS

Other advantages of the present invention will be readily appreciated as the same becomes better understood by reference to the following detailed description when considered in connection with the accompanying drawings wherein:

FIG. 1 illustrates a graph with excess of neutrons over protons (OY) versus sum of d- and f-electrons (OX);

FIG. 2 illustrates a mass spectrum representing recorded m/z relationship graphically;

FIG. 3 illustrates a method of direct imaging of isotope K³⁹ on the sample surface;

FIG. 4 illustrates a character of homogeneity of potassium isotope distribution obtained by producing a profilogram;

FIG. 5 illustrates an isotope distribution in young and old biological tissue;

FIG. 6 illustrates a mass spectrum of the young tissue sample in a range of 1 to 50 amu;

FIG. 7 illustrates a mass spectrum of the old tissue sample in a range of 1 to 50 amu;

FIG. 8 illustrates a mass spectrum of the young tissue sample in a range of 50 to 100 amu;

FIG. 9 illustrates a mass spectrum of the old tissue sample in a range of 50 to 100 amu;

FIG. 10 illustrates a diagram of deviation of the isotopic composition of young and old tissues from the natural isotope distribution;

FIG. 11 illustrates a comparative assessment of changes in the concentration of heavy and light isotopes in biological tissues of different ages;

FIG. 12 illustrates an isotope distribution in samples 12 and 14;

FIG. 13 illustrates an isotope distribution in samples 18 and 20;

FIG. 14 illustrates a diagram of deviations of the isotopic composition in normal and pathological tissues of an adult from the natural isotope distribution obtained as a result of analysis of samples 12 and 14;

FIG. 15 illustrates a comparative assessment of changes in the concentration of heavy isotopes in samples 12 and 14;

FIG. 16 illustrates a diagram of deviations of the isotopic composition in normal and pathological tissues of an adult from the natural isotope distribution obtained as a result of analysis of samples 18 and 20;

FIG. 17 illustrates a comparative assessment of changes in the concentration of heavy isotopes in samples 18 and 20;

FIG. 18 illustrates a quantification of isotope content in the samples of fungus and cortex and comparison of the obtained results with natural distribution of isotopes;

FIG. 19 illustrates a diagram of deviation of the isotopic composition in the samples of fungus and cortex from the natural isotope distribution; and

FIG. 20 illustrates a system of the present invention for diagnostics of pathology in biological objects.

DESCRIPTION OF THE INVENTION

A system for diagnostics of pathology in biological objects is generally shown at 10 in FIG. 20. The system 10 includes a first device 12 adaptable to receive and analyze information from a first biological element 14, affected by pathology, such as human tissue, fluids, obtained from a human 16 to identify ratio between first isotopes and second isotopes wherein weight of the first isotopes differs from weight of the second isotopes in order to generate a first set of data 18. The system 10 includes a second device 20 cooperable with the first device 12. The second device 20 is adaptable to receive and analyze information about natural distribution of the first isotopes and the second isotopes in a second biological element 22 in order to generate a second set of data 24.

The system 10 further includes a controller 26 (a central processing unit) having a comparative software 28. The controller 26 configured to receive the first set of data 18 and the second set of data 24 with the comparative software 28 comparing the ratio of the first isotopes and the second isotopes of the first biological element 14 with the ratio of the first isotopes and the second isotopes of the second biological element 22 thereby generating a third set of data 30 allowing to change the ratio of the first isotopes and the second isotopes of the first biological element 14 to match the ratio of the first isotopes and the second isotopes of the second biological element 22.

A method for diagnostics of pathology in biological objects of the present invention begins with analyzing a first biological element obtained from a human to identify ratio between first isotopes and second isotopes wherein weight of the first isotopes differs from weight of the second isotopes. Then, information about natural distribution of the first isotopes and the second isotopes in a second biological element is analyzing. A comparison analysis is conducted to compare the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element thereby changing the ratio of the first isotopes and the second isotopes of the first biological element to match the ratio of the first isotopes and the second isotopes of the second biological element. The first isotopes are light isotopes and the second isotopes are heavy isotopes. The first biological element is a tissue from a human body being diagnosed wherein the second biological element is a tissue from a human body without any pathology. The method for diagnostics of the present invention uses a mass-spectrometry to identify the amount and type of chemicals presented in the first biological element by measuring the mass-to-charge ratio and abundance of gas-phase ions. The method for diagnostics also uses MRI on selected NMR active nuclei, wherein the step of using MRI on selected NMR active nuclei is further defined by isotopes K-39, K-40, K-41, Mg-25, and Zn-67. The method for diagnostics is further defined by studying isotope distribution of chemical elements of potassium, magnesium and zinc. The comparative analysis is performed on the samples of body fluids, wherein body fluids are samples of blood and/or urine. The pathology is a chronicle degenerative disorder such as is a cancer, result of infectious diseases, and local aging of certain tissue part of a body.

Alluding and supporting of the above, isotope composition is the main difference between healthy and disease affected cells, as well as between “young” (stem cells, umbilical cord etc.), normal and “old” cells. Disease progression and aging both related to accumulation of heavy isotopes and depletion of light isotopes in cells and already damaged tissue. People say that age is just a number, which of course is wrong. In fact, age is a function of a ratio of two numbers—concentrations of light and heavy isotopes of d-electrons containing metals, for example such as zinc, i.e. not transition metals, but d-electrons containing metals. The only way to transform cells back to normal or young one is shifting isotope distribution back to the natural one and even better—increasing concentration of light isotopes and completely excluding heavy isotopes. Mammals, i.e. humans and animals, are not isotope selective. To stay alive and healthy they need to follow very special diet, rich with light isotopes.

Although we have to admit that system build with heavy isotopes only will have more neutrons and therefore more negative feedback connections being better fitted into the environment and hence much smarter compare to one, made with only light isotopes. Most elements in nature is a mixture of isotopes. Isotopes are atoms of the same chemical element with different masses. The difference in masses is due to the different numbers of neutrons at the same numbers of protons and electrons. In terms of concentrations, the main elements of body tissue are C, H, N, O, P, and S. We have found that in terms of pathologies, the elements with electron structure containing d-electrons and possibly f-electrons are extremely important and especially their isotope composition. In accordance to modem science, one would never expect any deviations from natural distribution of isotopes for a given element. It does not matter, which object of nature this element is belong to, and where it is in our universe.

For example, potassium is a mixture of two stable isotopes K-39 (93.3%), K-41 (6.7%) and long-lived radioisotope of K-40 (0.012%). Substitution of one isotope on another may result in the isotope effect—kinetic or magnetic isotope effect, isotopic shift or small variation in the temperature of superconductive transition. Isotope effect should not cause any dramatic changes in chemical properties or structure of materials. This is correct for non live matter but not for microbes and not for animals, not even for viruses or plants. The results and experimental conditions of detailed study of isotope distribution peculiarities in healthy and pathology affected biological tissues, as well as in “young” and “old” tissues are described below.

At present stage, all results obtained are on the level of tissue and are not protein or amino acid specific. This direction is a subject for further study that should bring extremely fine results. It is like studying physical properties of multiphased polycrystalline compounds compared to singlephased monocrystalline material. Never the less, the results are very important and allow for new ways of diagnostics, treatment and prevention of irreversible pathologies including cancer and ageing. Although samples represent different types of cancer or “young” and “old” tissues, the conclusions made have much broader significance, as many pathologies can be discussed in terms of extreme local ageing effects. We also believe that notion of immune system should be used not only to describe body's defense against microbes, viruses and other invaders, but also the ability to repair mutation—threatening damage to the most important macromolecules.

This function is crucial for irreversible disease prevention and as proved by our data requires uninterrupted critical “spare parts” supply. The “spare parts” are set of lightest isotopes of vitally important metals. Mass spectrometry data shows that in healthy tissue of not too young and too old person there is so-called natural distribution of isotopes in complete correspondence with all textbooks. In the tissue from the body of 77 years old, there is a shift in isotope distribution to the side of heavy isotopes. Concentration of light isotopes in a number of elements is significantly decreased and concentration of heavy isotopes is correspondently increased.

Shift of isotope distribution in favor of heavy isotopes was also observed in samples affected by a cancer tumor. There is correlation between severity of disease and concentration of heavy isotopes in the tissue. In the sample taken from malignant tumor with well expressed metastases, concentration of heavy isotopes was much higher and for light isotopes much lower than in the tissue of malignant tumor without metastases. In contrast to the “old” and cancer affected samples, in “young” tissue the shift of isotope distribution in the favor of light isotopes was detected along with increased concentration of light isotopes and significant decrease in concentration of heavy ones.

Here below in examples that we have presented most conservative data, but it worse to mention that the difference in concentrations of heavy and light isotopes in healthy and cancer tissues of the same patient depends on progress of the disease and can be higher than tenfold. It is important to notice that as much as light isotopes are spare parts to fix broken chemical bonds back to normal, the heavy isotopes are also spare parts without which dangerous damage to the chemical bonds cannot hold on in time or stabilized and eventually is repaired. It is logical to expect that in right amount light isotopes should be able to rejuvenate biological tissue and transform cells affected by pathology back to normal state. At the same time, one can consider heavy isotopes of the same elements as an efficient and multipotent pathogen or kind of a poison.

At certain concentration, they can cause most of the broken chemical bond to become a source of irreversible pathology. Concentration of light isotopes is also a key to successful treatment of various illnesses. To achieve positive result the amount of “cure” should be much higher than amount of “poison”. A lot depends on the damage inflicted by disease. That is why it is inappropriate to speak of one universal dose of “cure” for any disease at any stage. The target is to prevent access of heavy isotopes and provide right amount of light isotopes supply during the treatment time. Otherwise, results will by inconclusive and irreproducible creating uncertainty and confusion.

Let us take two examples to demonstrate what happens: one from biotechnology and another from pharmaceutics. First, we will discuss stem cells treatment. It is proclaimed as possible universal cure and sometime it really helps to some patients with some illnesses to a few and not always, and no explanation why. Let us put aside problems related to the reaction of immune system on the alien stem cells infusion. Then stem cells should be able to help healthy organism to become stronger due to the regenerative ability and theoretical possibility to differentiate and substitute damaged cells of various organs. It is necessary to add-given there is sufficient supply of key light isotopes to the tissue or organ under discussion. Otherwise, the lack of light isotopes and excess of heavy ones may cause transformation of stem cells into cancer cells. Stem cells are definitely “young”. Therefore, their positive effect can be attributed to the fact that they contain higher that natural concentration of light isotopes. Here we speak of ratio between light and heavy isotopes. Absolute amount of light isotopes in stem cells is very low.

In general, stem cells can be considered as source and carriers of very low quantities of light isotopes. It is better than nothing and sometime even helpful, but it is difficult to predict when. Second example is birch tree mushrooms—“chaga”. It is believed to be a “virtual cocktail of antioxidants and phytonutrients” able to bust immune system, reduce inflammation and eliminate cancer. It was used for many centuries as folk medicine with universal properties. Nobel Prize winner Alexander Solzhenitsyn believed he was cured from cancer with chaga and described the story in his famous book “The Cancer Ward”. Yet, there is no solid data or statistics to support the legend. They say taking chaga tea over a long period is extremely beneficial. At the same time, nobody dares even to speculate on probability to cure any disease, let alone cancer, with chaga in less than a lifetime.

To figure out the nature of legendary claims on chaga properties we have conducted mass-spectrometry study of samples prepared from chaga mushroom, surrounding bark and birch rings (inner volume of the tree). Results should help to understand real reasons of biological activity for not only chaga, but other folk recipes as well. Chaga mushroom appeared to have a really unique and unheard of feature—isotope selectivity. Like in the young human tissue, the isotope distribution is different from the natural one with much higher concentration of light isotopes and lower concentration of heavy isotopes in chaga samples, reverse picture (like in cancer affected tissues) in surrounding bark and natural distribution deep inside birch rings/trunk. They believe that birch tree host gradually dies of due to the mushroom consuming all nutrients from the bark and trunk.

At the same time, nothing is preventing additional supply from the root system. The real problem is the constant excess of heavy isotopes over light ones in the bark and trunk around the mushroom. That is what kills healthy tissue. Chaga contains up to 30% higher concentration of light isotopes of vitally important metals like K, Zn, Mg and Rb. There is a significant shift in isotope distribution but heavy isotopes are still present although in a smaller amounts. We have also discovered that light isotopes spread not equally over the samples. It means that there are areas in mushroom cross-section with no light isotopes at all.

Hence, seemingly random biological activity of chaga extracts can be explain by competition between effects of higher than normal amount of light isotopes and less than normal but still significant presence of heavy isotopes. Hundreds to thousands times smaller quantities of light isotopes compared to the daily intake of vital elements. Non-homogeneous distribution of chemical elements in the bulk of mushroom. It means that quite often extracts are made from chaga's parts that do not contain any useful elements at all. Chaga's extract consumer should be lucky enough to have multiple random events to coincide to get theoretically possible therapeutic effect. Isotope selectivity obviously is not so rare effect. It just never been studied in biological objects in current scientific paradigm. It is reasonable to suggest that it is a common feature in many plants, especially those used for treatment in both traditional and alternative medicine.

There is high probability that spermatozoid and egg in ovary are also isotope selective. Human embryo seems also has this property but at some stage of the development it is suppressed and lost. Experimental results prove that peculiarities of isotope distribution in biological tissues are perfect indicators of the balance between youth and aging, perfect health and irreversible pathological change. Therefore, new method of diagnostics is based on evaluation of the isotopes distribution for the vital chemical elements like K, Mg, Zn, Mo, Si, Se etc. in the samples of biological tissue taken for the analysis. The key parameter to measure is the ratio between concentrations of light and heavy isotopes of the same chemical elements in the samples.

When said ratio is in favor of heavy isotopes then it has to be considered as an indication of dangerous irreversible pathology or extreme local adding. Combined with detection of additional chemical elements compared to ones present in normal tissues, it is a strong indication of malignant tumor and metastases. Diagnostics based on isotopes distribution analysis can be easily performed by those skilled in the art of standard mass-spectrometry.

Mass-spectrometry requires samples preparations. It makes sense to create MRI systems based on spectroscopy of magnetic nuclei of chemical elements. Potassium has three NMR active nuclei: K-39, K-40 and K-41, Magnesium has Mg-25. The only NMR active nucleus of zinc is Zn-67. Therefore, the creation and use of MRI systems based but not limited to K-40 or K-41, M-25 and Zn-67 should help with early non-invasive diagnostics of slightest pathological changes in human organism.

Referring to the above, our first Example covers an analysis of cancer affected and healthy tissues and “old” and “young” tissues. Experimental samples obtained by surgery or filtration depends on the objects of investigation (blood, lymph etc.). The samples having a mass of about 1 gram were quick-frozen by immersion in liquid nitrogen. To reach high rate of freezing small portions of samples have been used. In ultrafast cooling at a rate of about one hundred degrees in 1 sec., water was transformed into amorphous ice without volumetric expansion. After that, the amorphous ice was removed from the sample by means of vacuum freeze drying at a low temperature. The ice evaporates and the mineral particles and organic components of the sample remain in the same position in which they were in the initial wet sample, and thus the structure and chemical composition of the sample is preserved.

To reduce the drying time of the sample during sublimation, dry nitrogen was fed automatically to the vacuum chamber. This technique increases thermal conductivity of the sample and therefore the convection heat supply accelerates the process of sublimation. The gas feed rate to the vacuum chamber did not exceed 0.1 I/min. The samples drying time under these conditions was 10 hours. At the final stage of preparation of the samples an additional drying under the high vacuum was done. At almost complete (99%) dehydration under the high vacuum, samples were heated to a temperature of 35-40° C. and then have been aged isothermally for about 1 hour under these conditions. Method of secondary ion mass spectrometry with Cameca IMS-4F mass spectrometer was used. Mass spectrometry in the biological samples was performed based on the m/z ratio analysis in the pulse counting mode as well as using results displayed as a mass-spectrum as shown in FIG. _——— or as a profilogram that characterizes distribution of the detected masses along the sample plane as shown in FIG. _——— and in depth as shown in FIG. _———.

The sample prepared for the experiment was placed in a sample holder. After evacuating the system, the probe was calibrated and its mode was stabilized. To prevent the destructive action of the ion beam on the sample and to reduce the amount of static electricity caused by ionization, the analysis was carried out at very low rates of the sample sputtering (less than 10⁻⁴ of the monolayer per second).

A subsequent analysis of the masses obtained in result of the ion beam bombardment of the sample surface was based on the interaction of secondary ions ejected from the sample surface with the electric and magnetic fields of the detectors. Utilization of a double-focusing spectrometer with combination of electric and magnetic fields controls made it possible to maximize the sensitivity of the instrument. For such multistage magnetic spectrometers a background signal resulting from the residuals of the main peaks of the matrix material (wall scattering, on the gas atoms, etc.) can be reduced to a level of less than 10⁻⁹ for the general background and only 10⁻⁶ for all masses close to the main peak.

To minimize the amount of gases adsorbed on the sample surface (H₂, N₂, O₂, H₂O, CO₂ and CO) the measurements were performed in an ultrahigh vacuum free of hydrocarbons using cryogenic and getter pumping near the sample. To reduce the formation of positive charge on the surface of the test sample due to electron ionization, the latter was irradiated with electrons emitted by a thermal cathode located nearby. In addition, an electro-conductive additive to the sample increased samples conductivity.

Sample dried by sublimation was placed on a metal mesh with a mesh size of 25×25 mm using a pressing technique with the addition of electro-conductive highly dispersed carbon black. The quantitative content of the electro-conductive additive was 0.1% of the sample weight to drain off accumulated charge on the metal mesh. For the measurement, several methods were applied using Cameca IMS-4F instrument: a) mass spectrometry, b) the method of direct imaging of isotope distribution on the sample surface, c) the method of profilograms, which made it possible to assess a degree of homogeneity of the isotope distribution in the depth of the sample as shown in FIGS. _———. Description of the samples is presented in FIG. _———. The experimental results are presented in FIG. _———. Two right columns in FIG. _——— characterize different isotopes concentration in the examined tissues. For visual presentation of a graph shown in FIG. _——— was built.

A vertical line in the middle part corresponds to the natural distribution for each of the detected isotopes. Horizontal yellow and green diagrams characterize deviations from the natural content for each of the said isotopes. Therefore the left side of the graph (with shaded diagram strips) corresponds to an increase in the concentration of isotopes and is represented mainly by heavy isotopes, while the right side (with unshaded diagram strips) on the contrary—a decrease in the percentage of isotopes in relation to the natural content primarily due to light isotopes.

FIG. _——— illustrates the character of distribution of isotopes in the “young” and “old” tissues. We analyzed the isotopic ratios in pathological (cancer) and healthy tissues of the following chemical elements: magnesium, silicon, sulfur, chlorine, potassium, calcium, chromium, iron, nickel, copper, zinc, bromine, rubidium, molybdenum and silver. The following types of biological tissues were studied: biological tissues containing cancerous tumor cells (sample 20 and sample 12) where sample 20 had metastases. Normal biological tissues (sample 12 and sample 18). Within each pair of samples (tumor and normal tissue were divided into the following pairs: 12-14 and 18-20), the pairs of samples belonged to the same biological organism. Isotope distribution was studied on the following pairs of samples as shown in FIG. _———. Samples 12 and 14, where sample 12 was taken from the central area of the tumor and sample 14 from normal tissue. Samples 18 and 20, where sample 18 was taken from normal tissue and sample 20 from tissue affected by the tumor. Distribution of isotopes detected in samples 12 and 14 is shown as shown in FIG. _——— and in samples 18 and 20 as shown in FIG. _———. Comparative assessment of the experimental results on samples 12 and 14 is shown in shown in FIG. _——— and on samples 18 and 20 as shown in FIG. _———.

Analysis of the obtained data evidences the following pattern of fractionation of the isotopic composition in the tissues affected by the tumor. Tissue samples 20 and 12 characterizing tumor tissues had the following factors of similarity and differences. The similarity consisted in their belonging to the tumor tissues and the difference was in the presence of metastases of poorly differentiated transitional cell carcinoma in sample 20, which characterized this biological sample as an object with a more pronounced degree of damage by cancer.

When analyzing the results of obtained on samples 12 and 14, it can be found that of 20 isotopes detected in these samples represented by magnesium, silicon, potassium, iron, copper, zinc, bromine, rubidium, and silver, the major part, in the amount of 11 isotopes, is characterized as heavy isotopes primarily concentrated in the tissue affected by the tumor in the “cancer tissue” as shown in FIG. _———, these isotopes are highlighted in red. This is in correspondence with the data as shown in FIG. _———. In FIG. _——— the content of heavy isotopes in the normal and cancer tissues is shown in red, and the analysis of these results reveals that heavy isotopes is observed in the normal tissue as well.

However, the general trend is accumulation of heavy isotopes in the tissue affected by the tumor. Now let us consider the results obtained on samples 18 and 20. Sample 18 was represented by healthy tissue and sample 20 was a metastasis of poorly differentiated transitional cell carcinoma with extensive necrosis. It should be noted that compared with samples 12 and 14 these samples contained a much larger number of both the elements and isotopes. This can be explained by the man-caused factors of existence of a biological organism as well as by certain selective conditions of accumulation of some elements and their isotopes in a pathological tissue.

We have determined that a pair of samples 18 and 20 contained new elements, such as molybdenum, nickel, chromium, chlorine and sulfur, and the list of isotopes found in samples 12 and 14 was considerably expanded and included such isotopes as Zn₆₈, Zn₆₄, Fe₅₇. If we describe the results obtained on samples 18 and 20 statistically, the following can be concluded. Of the 30 isotopes identified in this pair of samples (which are represented by 11 elements such as magnesium, silicon, sulfur, chlorine, potassium, chromium, iron, nickel, zinc, rubidium, molybdenum) 15 isotopes can be characterized as heavy.

According to FIG. _——— which reflect the results of examination of samples 18 and 20, one can see a predominant increase in the number of heavy isotopes in the tumor tissue affected by metastases as compared to their content in normal tissue. Quantitative assessment of the nature of distribution of light isotopes is bidirectional and consists in the following. A slight increase in the content of light isotopes of such elements as magnesium, sulfur, chlorine, nickel and molybdenum in the healthy tissue relative to their natural distribution (shown in blue) was observed. As it was noted earlier (in the examination of young and old tissues as well as of samples 12 and 14), an increase in the concentration of heavy isotopes is accompanied by a decrease in the concentration of light isotopes. This is also true for samples 18 and 20. It is important to note a few more facts that characterize the correlation between the concentration of heavy isotopes and a degree of disease of biological tissue. Sample 20 was taken from metastases of poorly differentiated transitional cell carcinoma with extensive necrosis and sample 12 of renal cell carcinoma. From a medical point of view, a metastasis is a recurrence of cancer and it is more dangerous and serious complication for the life of a patient than the primary tumor. The poorly differentiated cancers have the most adverse outcomes. This happens because the tumor overcomes protective barriers and cancer cells get in the lymph and blood stream.

For convenience, heavy isotopes in as shown in FIG. _———. Quantitative assessment of the isotopes detected in samples 12 and 20 indicates an increase in their number (in sample 20) along with an increase in the degree cancer aggressiveness. The range of deviation of isotope concentrations from the one in natural isotope distribution is much higher in sample 20. The maximum deviation from the natural content found on silicon isotopes in sample 20 with an increase in Si₂₉ by 11.6%, while the content of Si₂₈ isotope reduced by 20.6%. Such a difference is expressed less clearly in the pair of samples 12 and 14. The largest increase here was observed in the content of Rb₈₇ isotope which amounted to +8.35% in the tissue affected by tumor, while the largest decrease in the isotopic concentration was found in Mg₂₄ isotope which was −8.1%. This trend is observed on other elements as well. However, a shift in the distribution towards heavy isotopes is considerably stronger in the pair of samples 18 and 20. Biological tissue affected by cancer cells and metastases has a higher concentration of heavy isotopes while the bordering normal tissue has a lower concentration of light isotopes.

Our second example covers a mass spectrometry of isotope composition of fungus and cortex samples. Analysis of the content of trace quantities of isotopes requires prior sample preparation as elements in most objects are in a bound state. They form quite strong organic complexes that prevent accurate and reproducible determination of their content. Therefore, prior to any analysis it is necessary to destroy the organic portion of the sample. Preparation of the samples of fungus and cortex for the analysis was carried out using the dry ashing method. The dry ashing method involves sequential heating of a substance to the ashing temperature in oxygen in a closed system.

Dry ashing of the samples of fungus and cortex was performed in a ceramic crucible placed in a muffle furnace. For the destruction of the organic bond of substances the same quantities of the samples weighed for the analysis (6 grams each) were used. The samples in crucibles were placed in a muffle furnace without protective atmosphere (i.e. in air) at the temperature of 150° C. At this temperature, the samples were heated for one hour and then they were dried step by step by raising the temperature of the furnace by 50° C. every hour. The temperature of the sample was raised in such a mode up to 350° C. (for 4 hours), whereupon it was raised to 360° C.

The samples were then held at this temperature for 20 minutes. Then crucibles with ash were extracted from the furnace and were cooled under the atmospheric conditions to the room temperature. With the same initial masses of fungus and cortex, after completion of the dry ashing procedure the fungus weighed 112 mg while the cortex weighed only 32 mg. The solid residues of the samples of fungus and cortex were pressed in a nickel metal mesh with a mesh size of 50×50 microns. The metal mesh helped to keep the sample material fixed and additionally served as the current collector reducing the amount of static electricity caused by ionization by the ion beam. Information on the isotopic composition of the samples was obtained from the surface area 200×200 mm in size.

Analysis of the samples of fungus and cortex was performed with respect to the following detected elements: magnesium, silicon, potassium, calcium, titanium, iron, copper, zinc, rubidium, strontium. The total number of isotopes found in the samples was 36 isotopes in the 10 detected elements. Quantitative assessment of each isotope was obtained based on the primary analysis of the experimental data characterizing the relationship of mass and impulse response for each of the isotopes. FIGS. _——— show the results of quantitative assessment of the content of isotopes in fungus and birch cortex, and the diagram in FIG. _——— makes it possible to visually compare the quantitative characteristics of isotopic distribution in fungus and the birch cortex and rings.

The following conclusions were made based on the results of our study: In samples from the interior of the tree (birch ring), the natural distribution of isotopes was observed. In the samples prepared from the fungus, the isotopic distribution shifted toward the light isotopes. The concentration of light isotopes was considerably higher and the concentration of heavy isotopes was considerably lower than in the natural distribution. In the samples prepared from the cortex, surrounding the fungus the isotopic distribution shifted towards an increase in a portion of heavy isotopes and a reduction in the concentration of light isotopes. It should be noted, that the fungus itself is mechanically strong while the cortex around it is in fact quasi-dead tissue with no mechanical strength.

Isotope composition is the main difference between healthy and disease affected cells, as well as between “young” (stem cells, umbilical cord etc.), normal and “old” cells. Disease progression and aging both related to accumulation of heavy isotopes and depletion of light isotopes in cells and already damaged tissue. The only way to transform cells back to normal or young one is shifting isotope distribution back to the natural one and even better—increasing concentration of light isotopes and completely excluding heavy isotopes.

We have to repeat that mammals are not isotope selective. To stay alive and healthy they need to follow very special diet, rich with light isotopes. Violations (heavy isotopes consumption) should be qualified as sin and be punishable. Although we have to admit that system build with heavy isotopes only will have more neutrons and therefore more negative feedback connections being better fitted into the environment and hence much smarter compare to one, made with only light isotopes.

The basis for nuclear magnetic resonance is the observation that many atomic nuclei spin about an axis and generate their own magnetic field, or magnetic moment. For reasons that are outside the scope of this text, only those nuclei with an odd number of protons and/or neutrons have a magnetic moment. Several common nuclei, including hydrogen (¹H), the ¹³C isotope of carbon, the ¹⁹F isotope of fluorine, and the ³¹P isotope of phosphorus, all have magnetic moments and therefore can be observed by NMR—they are, in other words, NMR-active. Other nuclei—such as the common ¹²C and ¹⁶O isotopes of carbon and oxygen—do not have magnetic moments, and are essentially invisible in NMR. Other nuclei such as deuterium (²H) and nitrogen (¹⁴N) have magnetic moments and are NMR-active, but the nature of their magnetic moments is such that these nuclei are more difficult to analyze by NMR.

In practice it is ¹H, ¹³C, ¹⁹F, and ³¹P that are most often observed by NMR spectroscopy. In this chapter, we will develop our understanding of the principles behind NMR spectroscopy by focusing our attention first on the detection of protons in ¹H-NMR experiments (in discussion about NMR, the terms ‘hydrogen’ and ‘proton’ are used interchangeably). Much of what we learn, however, will also apply to the detection and analysis of other NMR-active nuclei, and later in the chapter we will shift our attention to NMR experiments involving ¹³C and ³¹P atoms.

Nuclear magnetic resonance spectroscopy technique used in the present method, most commonly known as NMR spectroscopy, is a research technique that exploits themagnetic properties of certain atomic nuclei. It determines the physical and chemical properties of atoms or the molecules in which they are contained. It relies on the phenomenon of nuclear magnetic resonance and can provide detailed information about the structure, dynamics, reaction state, and chemical environment of molecules. The intramolecular magnetic field around an atom in a molecule changes the resonance frequency, thus giving access to details of the electronic structure of a molecule.

Most frequently, NMR spectroscopy is used by chemists and biochemists to investigate the properties of organic molecules, although it is applicable to any kind of sample that contains nuclei possessing spin. Suitable samples range from small compounds analyzed with 1-dimensional proton or carbon-13 NMR spectroscopy to large proteins or nucleic acids using 3 or 4-dimensional techniques. The impact of NMR spectroscopy on the sciences has been substantial because of the range of information and the diversity of samples, including solutions and solids.

While the invention has been described with reference to an exemplary embodiment, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from the essential scope thereof. Therefore, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention will include all embodiments falling within the scope of the appended claims.

Claims

1. A method for diagnostics of pathology in biological objects, said method comprising the steps of:

analyzing a first biological element obtained from a human to identify ratio between first isotopes and second isotopes wherein weight of the first isotopes differs from weight of the second isotopes;

analyzing information about natural distribution of the first isotopes and the second isotopes in a second biological element; and

comparing the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element thereby changing the ratio of the first isotopes and the second isotopes of the first biological element to match the ratio of the first isotopes and the second isotopes of the second biological element.

2. The method for diagnostics as set forth in claim 1, wherein the first isotopes are light isotopes and the second isotopes are heavy isotopes.

3. The method for diagnostics as set forth in claim 1, wherein the first biological element is a tissue from a human body being diagnosed.

4. The method for diagnostics as set forth in claim 1, wherein the second biological element is a tissue from a human body without any pathology.

5. The method for diagnostics as set forth in claim 1, wherein the step of comparing the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element is further define by using a mass-spectrometry to identify the amount and type of chemicals presented in the first biological element by measuring the mass-to-charge ratio and abundance of gas-phase ions.

6. The method for diagnostics as set forth in claim 1, wherein the step of comparing the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element is further define by using MRI on selected NMR active nuclei.

7. The method for diagnostics as set forth in claim 1, wherein the step of using MRI on selected NMR active nuclei is further defined by isotopes K-39, K-40, K-41, Mg-25, and Zn-67.

8. The method for diagnostics as set forth in claim 1, wherein the step of comparing the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element is further define by studying isotope distribution of chemical elements of potassium, magnesium and zinc.

9. The method for diagnostics as set forth in claim 1, wherein comparative analysis is performed on the samples of body fluids.

10. The method for diagnostics as set forth in claim 1, wherein body fluids are samples of blood and/or urine.

11. The method for diagnostics as set forth in claim 1, wherein the pathology is a chronicle degenerative disorder.

12. The method for diagnostics as set forth in claim 1, wherein the degenerative disorder is a cancer.

13. The method for diagnostics as set forth in claim 1, wherein the pathology is a result of infectious diseases.

14. The method for diagnostics as set forth in claim 1, wherein the pathology is local aging of certain tissue part of a body.

15. A system for diagnostics of pathology in biological objects, said system comprising:

a first device adaptable to receive and analyze information from a first biological element obtained from a human to identify ratio between first isotopes and second isotopes wherein weight of the first isotopes differs from weight of the second isotopes in order to generate a first set of data;

a second device cooperable with said first device, said second device adaptable to receive and analyze information about natural distribution of the first isotopes and the second isotopes in a second biological element in order to generate a second set of data; and

a controller having a comparative software, said controller configured to receive said first set of data and said second set of data with said comparative software comparing the ratio of the first isotopes and the second isotopes of the first biological element with the ratio of the first isotopes and the second isotopes of the second biological element thereby generating a third set of data allowing to change the ratio of the first isotopes and the second isotopes of the first biological element to match the ratio of the first isotopes and the second isotopes of the second biological element.