Methods and systems for linking chemical emissions with corresponding genetic, medical, and / or pathological conditions.

Abstract

To provide methods and systems for associating discharge of chemicals with corresponding genetic, medical and pathological conditions.SOLUTION: A method comprises: producing an MCD target cell VOC profile by: (a) inducing MCD on non-treated target cell cultures thereby producing post-MCD target cell cultures; and (b) acquiring post-MCD target cell culture VOC emission data; applying the selected treatment to target cell cultures; acquiring VOC emission data of post-treatment target cell cultures; and determining an effect of the selected treatment. The treatment is determined as effective when: (a) concentration values from the VOC emission data of pre-treatment target cell cultures are greater than concentration values from the VOC emission data of post-treatment target cell cultures; and (b) concentration values from the VOC emission data of post-treatment target cell cultures are greater than concentration values from the VOC emission data of pre-treatment target cell cultures.SELECTED DRAWING: Figure 11

Description

[Technical Field]

[0001] Areas of the disclosed technology The disclosed technologies relate, in general, to identifying chemical eliminations in the human body and / or cultures, and more specifically to methods and systems for correlating chemical eliminations in the human body and / or cultures with corresponding genetic, medical, and / or pathological conditions. [Background technology]

[0002] Background of the disclosed technology Metabolic, anabolic, and / or catabolic processes produce chemical compounds. Some of these compounds belong to three groups: volatile organic compounds (VOCs), semi-volatile organic compounds (SVOCs), and volatile sulfur-containing compounds (VSCs). Compounds in these groups typically remain in a gaseous state at room temperature. Monitoring gases associated with metabolic, anabolic, and / or catabolic processes is well known in the art. For example, monitoring oxygen (O2) saturation levels is used to monitor a patient's condition. Similarly, carbon dioxide (CO2) is used as an indicator in a wide range of lung-related diseases.

[0003] The publication, "Summary of Safety and Probable Benefit, Menssana Research, Inc. Hearts Breath Test for Grade 3 Heart Transplant Rejection," focuses on monitoring respiratory VOCs in heart transplant recipients to aid in the diagnosis of grade 3 heart transplant rejection.

[0004] The publication "A Review of the Volatiles From the Healthy Human Body" by de Lacy Costello et al. examines a list of VOCs reported from the bodies of healthy humans. In this list, a total of 1840 VOCs were identified in respiration, saliva, urine, milk, blood, skin secretions, and feces. 872 were found in respiration, 359 in saliva, 154 in blood, 256 in milk, 532 in skin secretions, 279 in urine, and 381 in feces.

[0005] The publication to Ulanowska et al., "The Application of Statistical Methods Using VOCs to Identify Patients with Lung Cancer," focuses on an attempt to determine the grouping of lung cancer biomarkers. For this purpose, respiratory samples were obtained from 137 patients with confirmed lung cancer. These samples were analyzed using the SPME-GC / MS method. Exhaled breath samples were also obtained as a reference group from 143 healthy volunteers with different smoking habits (active smokers, passive smokers, and non-smokers). Statistical methods such as discriminant analysis (DA) and CHAID tree models were used for data processing and evaluation. Ulanowska suggested that chemotherapy for lung cancer can be controlled by utilizing molecular biomarkers such as amino acids, peptides, lipids, and carbohydrates, and that these may be defined as molecules that reflect the pathological state of the organ and may be characteristic pharmacological responses to therapeutic interventions.

[0006] In 2012, Altomare described that respiratory analysis using triple quadrupole gas chromatography-mass spectrometry (hereinafter referred to as "GC-MS / MS") can detect characteristic VOCs in specific medical conditions such as colorectal cancer and melanoma.

[0007] The PCT patent application to Domingues Ortega, WO2014 / 180974, titled "VOC Based, Narcolepsy Diagnostic Method," describes a method for detecting narcolepsy in a patient by obtaining a sample from a subject and detecting the level of at least one VOC in the sample to obtain a VOC profile of the sample. Subsequently, the VOC profile of the sample is compared to a reference VOC profile to determine whether the patient has narcolepsy. [Overview of the project] [Problems that the invention aims to solve]

[0008] Summary of the technology disclosed herein The object of the disclosed technology is to provide novel methods and systems for associating the elimination of chemical substances in at least one of human respiration, bodily fluids, and cell cultures with corresponding expressions of genetic or medical conditions, and for utilizing these associations for diagnosis, and / or for determining the effectiveness of treatment, and / or for determining the choice of treatment. [Means for solving the problem]

[0009] According to embodiments of the disclosed technology, a method is provided for determining the effect of at least one selected treatment administered to a patient for a metabolic, anabolic, catabolic, genetic, and / or medical condition. The method includes the steps of obtaining VOC release data from a pre-treatment target cell culture and creating a pre-treatment target cell VOC profile from the VOC release of the pre-treatment target culture. The method further (a) Inducing massive cell death in untreated target cell cultures, thereby generating post-MCD target cell cultures, and (b) Obtain VOC release data from target cell cultures after MCD. This includes a procedure for creating a VOC profile of MCD-targeted cells.

[0010] The method also includes the steps of applying at least one selected treatment to at least a target cell culture, obtaining VOC emission data of the post-treatment target cell culture for each selected treatment, and determining the effect of the selected treatment. The treatment is (a) when the concentration value of a VOC associated with a pre-treatment target cell VOC profile from the VOC emission data of the pre-treatment target cell culture is greater than the concentration value of the VOC associated with the pre-treatment target cell VOC profile from the VOC emission data of the post-treatment target cell culture, and (b) when the concentration value of a VOC associated with an MCD VOC profile emitted by the post-treatment target cell culture from the VOC emission data of the post-treatment target cell culture is greater than the concentration value of the VOC associated with the MCD VOC profile emitted by the pre-treatment target cell culture from the VOC emission data of the pre-treatment target cell culture, it is determined to be effective.

[0011] According to another aspect of the disclosed technology, a method for determining the effectiveness of a treatment of a metabolic, assimilative, catabolic, genetic and / or medical condition administered to a patient is thus provided. The method includes the steps of obtaining pre-treatment patient VOC emission data of VOCs emitted from at least one of a breath sample and a body fluid sample before at least one selected stage of the treatment, and obtaining VOC emission data of a pre-treatment target cell culture. The method further includes (a) inducing a large amount of cell death in a non-treated target cell culture, thereby generating a post-MCD target cell culture; (b) obtaining VOC emission data of the post-MCD target cell culture; and (c) using a diffusion model to predict the concentration levels of VOCs in the breath and / or body fluid, The method includes a procedure for creating a predicted MCD target cell VOC profile. The method also includes the procedure of applying a selected treatment to a patient, obtaining post-treatment patient VOC release data of VOCs released from at least one respiratory and body fluid sample taken during and / or after at least one selected stage of the treatment, and determining the effectiveness of the selected treatment. A treatment is determined to be effective if the VOC concentration value in the predicted MCD target cell VOC profile during and / or after the selected stage of the treatment from the post-treatment patient VOC release data associated with at least one respiratory and body fluid sample is greater than the VOC concentration value in the predicted MCD target cell VOC profile prior to the selected stage of the treatment from the pre-treatment patient VOC release data associated with at least one respiratory and body fluid sample.

[0012] According to a further aspect of the disclosed technology, a method for determining the effectiveness of a treatment administered to a patient is thus provided. The method includes obtaining at least one of a breath and a bodily fluid sample prior to at least one stage of the treatment, obtaining VOC emission data of VOCs emitted by at least one of the breath sample and the bodily fluid sample obtained prior to at least one stage of the treatment, and identifying a stored Dynamic Differential VOC profile corresponding to the obtained VOC emission data, thereby correlating a pathological condition with the VOC emission data. The method further includes obtaining at least one of a breath and a bodily fluid sample during and / or after at least one stage of the treatment, obtaining VOC emission data of VOCs emitted in at least one of the breath sample and the bodily fluid sample obtained during and / or after at least one stage of the treatment, and classifying the effectiveness of the treatment by comparing at least the concentration values of the VOCs in the identified Dynamic Differential VOC profile obtained prior to at least one stage of the treatment with the concentration values of the VOCs in the identified Dynamic Differential VOC profile obtained during and / or after at least one stage of the treatment. At least one stage of the treatment is classified as successful if the concentration level of the VOCs associated with the target cell VOC profile in the Dynamic Differential VOC profile during and / or after the selected stage of the treatment is reduced compared to the concentration value of the VOCs in the identified Dynamic Differential VOC profile prior to the treatment.

[0013] The disclosed technology will be more fully understood and appreciated from the following detailed description in conjunction with the drawings.

Brief Description of the Drawings

[0014] [Figure 1]This is a schematic example of a system constructed and effective by embodiments of the disclosed technology for associating the excretion of chemicals in at least one of human respiration, bodily fluids, and cell cultures with corresponding expressions of genetic or medical conditions, for utilizing these associations for diagnosis, and / or for determining the effectiveness of treatment, and / or for determining the choice of treatment. [Figure 2A] This is a schematic example of VOC emission data from another embodiment of the disclosed technology. [Figure 2B] This is a schematic example of a differential VOC profile according to another embodiment of the disclosed technology. [Figure 3A] This is a schematic example of an exemplary method for associating VOC releases with corresponding cancer types in a selected population, which will be effective through further embodiments of the disclosed technology. [Figure 3B] This is a schematic example of an exemplary method for associating VOC releases with corresponding cancer types in a selected population, which will be effective through further embodiments of the disclosed technology. [Figure 3C] This is a schematic example of an exemplary method for associating VOC releases with corresponding cancer types in a selected population, which will be effective through further embodiments of the disclosed technology. [Figure 4A] This is a schematic example of a method for associating VOC release with target cells before and after treatment in a selected population, which is effective by another embodiment of the disclosed technology. [Figure 4B] This is a schematic example of a method for associating VOC release with target cells before and after treatment in a selected population, which is effective by another embodiment of the disclosed technology. [Figure 4C] This is a schematic example of a method for associating VOC release with target cells before and after treatment in a selected population, which is effective by another embodiment of the disclosed technology. [Figure 4D]This is a schematic example of a method for associating VOC release with target cells before and after treatment in a selected population, which is effective by another embodiment of the disclosed technology. [Figure 5A] This is a schematic example of a method for relating VOC profiles to pathological conditions arising from pathogens, which will be effective through further embodiments of the disclosed technology. [Figure 5B] This is a schematic example of a method for relating VOC profiles to pathological conditions arising from pathogens, which will be effective through further embodiments of the disclosed technology. [Figure 5C] This is a schematic example of a method for relating VOC profiles to pathological conditions arising from pathogens, which will be effective through further embodiments of the disclosed technology. [Figure 6A] This is a schematic example of a method for determining a dynamic differential VOC profile for selected mismatch repair (MMR) gene activation therapy, according to another embodiment of the disclosed technology. [Figure 6B] This is a schematic example of a method for determining a dynamic differential VOC profile for selected mismatch repair (MMR) gene activation therapy, according to another embodiment of the disclosed technology. [Figure 6C] This is a schematic example of a method for determining a dynamic differential VOC profile for selected mismatch repair (MMR) gene activation therapy, according to another embodiment of the disclosed technology. [Figure 6D] This is a schematic example of a method for determining a dynamic differential VOC profile for selected mismatch repair (MMR) gene activation therapy, according to another embodiment of the disclosed technology. [Figure 7A] This is a schematic example of a method for determining the VOC profile of target cells that have developed treatment-resistant mutations from a selected treatment, which will be effective in further embodiments of the disclosed technology. [Figure 7B]This is a schematic example of a method for determining the VOC profile of target cells that have developed treatment-resistant mutations from a selected treatment, which will be effective in further embodiments of the disclosed technology. [Figure 8A] This is a schematic example of an exemplary method for associating VOC release with corresponding abnormal or pathological cells in individual patients, which is put into effect by another embodiment of the disclosed technology. [Figure 8B] This is a schematic example of an exemplary method for associating VOC release with corresponding abnormal or pathological cells in individual patients, which is put into effect by another embodiment of the disclosed technology. [Figure 9A] This is a schematic example of a method for associating VOC release with corresponding target cells (e.g., oncogenic cells) in individual patients before and after MCD, which will be effective through further embodiments of the disclosed technology. [Figure 9B] This is a schematic example of a method for associating VOC release with corresponding target cells (e.g., oncogenic cells) in individual patients before and after MCD, which will be effective through further embodiments of the disclosed technology. [Figure 10A] This is a schematic example of a method for identifying an individual's personalized treatment-resistant VOC profile for a selected treatment, which is effective by another embodiment of the disclosed technology. [Figure 10B] This is a schematic example of a method for identifying an individual's personalized treatment-resistant VOC profile for a selected treatment, which is effective by another embodiment of the disclosed technology. [Figure 10C] This is a schematic example of a method for identifying an individual's personalized treatment-resistant VOC profile for a selected treatment, which is effective by another embodiment of the disclosed technology. [Figure 11] Here are schematic examples of three exemplary VOC emission data graphs (graps) associated with breast cancer in specific patients, based on further embodiments of the disclosed technology. [Figure 12]This is a schematic example of a method for determining the effectiveness of a treatment, which will be effective in further embodiments of the disclosed technology. [Figure 13A] This is a schematic example of a method for determining the effectiveness of a treatment on an individual, which is effective by another embodiment of the disclosed technology. [Figure 13B] This is a schematic example of a method for determining the effectiveness of a treatment on an individual, which is effective by another embodiment of the disclosed technology. [Figure 13C] This is a schematic example of a method for determining the effectiveness of a treatment on an individual, which is effective by another embodiment of the disclosed technology. [Figure 13D] This is a schematic example of a method for determining the effectiveness of a treatment on an individual, which is effective by another embodiment of the disclosed technology. [Figure 13E] This is a schematic example of a method for determining the effectiveness of a treatment on an individual, which is effective by another embodiment of the disclosed technology. [Figure 14] This is a schematic example of a method for identifying active and inactive mutations in patients and / or cultures, which will be effective through further embodiments of the disclosed technology. [Figure 15A] This is a schematic example of a method for determining the optimal treatment for a patient, which is effective through another embodiment of the disclosed technology. [Figure 15B] This is a schematic example of a method for determining the optimal treatment for a patient, which is effective through another embodiment of the disclosed technology. [Figure 15C] This is a schematic example of a method for determining the optimal treatment for a patient, which is effective through another embodiment of the disclosed technology. [Figure 16] This is a schematic example of the extended Farhi model, which comes into effect through further embodiments of the disclosed technology. [Figure 17A] This is a schematic example of a method for increasing VOC concentrations before sampling and for collecting a quantified amount of air from a selected portion of the lung, which is effective by another embodiment of the disclosed technology. [Figure 17B] This is a schematic example of a method for increasing VOC concentrations before sampling and for collecting a quantified amount of air from a selected portion of the lung, which is effective by another embodiment of the disclosed technology. [Modes for carrying out the invention]

[0015] Detailed description of the embodiment The disclosed technology overcomes the shortcomings of prior art by providing methods and systems for associating the elimination of chemicals such as VOCs, SVOCs, and / or VSCs in at least one of human respiration, bodily fluids, or cell cultures with corresponding expressions of genetic, medical, or pathological conditions. This association may then be used for diagnosis, to determine the effectiveness of treatment, or to select treatment, either for individual patients or for a generally selected population. Genetic and / or medical and / or pathological conditions may include carcinogenic processes at various stages, or conditions caused by pathogens (e.g., bacteria, viruses, fungi, etc.).

[0016] Generally, relating VOC emission data to corresponding genetic, medical, and / or pathological conditions involves obtaining VOC emission data (e.g., from a mass spectrometer) from both healthy and unhealthy patients, either in vivo (e.g., exhaled respiration), in vitro (e.g., cultured cells), or both, and determining VOC profiles associated with genetic, medical, and / or pathological conditions. The VOC profiles may then be stored in a database and used later for various purposes, which will be discussed further below in this specification. The terms “VOC,” “VOC emission data,” and “VOC profile” will also be further elaborated below.

[0017] Furthermore, in this specification, the term “target cell” means any applicable cell that can present a genetic, medical, and / or pathological condition (e.g., cancer cells, Alzheimer’s-infected cells, embryos, or viruses) that causes a genetic, medical, and / or pathological condition, or any applicable cell associated with a genetic, medical, and / or pathological condition originating either inside or outside the body (for example, the papillomavirus can cause cervical cancer, so this virus is applicable in relation to cervical cancer). A target cell may also be a mutation of another target cell. For example, oncogenic cells are target cells. Bacteria, viruses, and fungi may also be target cells. Infected tissue cells may also be target cells.

[0018] The term "cell type" in this specification refers to the type or arc type from which the target cell originates. For example, breast cancer cells originate from cells found only in the breast. Another example is the drug-resistant bacterium string Klebsiella pneumonia / carbapenemase (KPC), which originates from the drug-sensitive source bacterium (arc type) Klebsiella pneumonia (KP).

[0019] The term "culture" refers to at least one type of culture. The term "culture" may also refer to multiple cultures, and further to numerous cell cultures grown using different broth media and broth conditions. For example, KPC can be cultured in Mueller Hinton broth (MB) or triptych soy broth (TSB).

[0020] The term "VOC" refers to any chemical compound or combination of compounds found in vivo and / or in vitro samples (e.g., respiratory samples, urine samples, blood samples, and / or culture samples). For example, the term "VOC" often refers to volatile organic compounds such as hydrocarbons, esters, aldehydes, and ketones, and may further refer to volatile sulfur-containing compounds such as dimethyl sulfide. The term "VOC" may also refer to complex molecular metabolites and / or biological elements, such as proteins, antibodies, enzymes, RNA, and DNA.

[0021] The term "VOC emission data" relates to at least the presence or absence of selected VOCs or multiple VOCs. VOC emission data may further refer to the concentration levels of all or selected VOCs in the sample. VOC emission data may further relate to the mass spectra, ion mobility, and / or retention times (i.e., elution time from the GC column) of selected VOCs or multiple VOCs in the sample. VOC emission data may also relate to the separate or combined total mass spectra (molecular ions and fragments), ion mobility, and / or retention times of all VOCs in the sample. Analytical instruments such as gas chromatography-high field asymmetric waveform ion mobility spectroscopy-mass spectrometry (GC-FAIM-MS) provide GC retention time separation information, mass spectra, and ion mobility information for each detected VOC. VOC emission data may be provided in units such as parts per million (ppm), parts per billion (ppb), and parts per trillion (ppt). VOC emission data may further be provided in terms of count rate, e.g., parts / second, ppm / second. VOC emission data may be represented, for example, in the form of a vector or matrix.

[0022] The term “VOC profile” refers to VOC release data associated with a corresponding metabolic, anabolic, catabolic, genetic, and / or medical condition (e.g., healthy individuals, unhealthy individuals, carcinogenic processes, metabolic processes, cancer types, bacteria, viruses, or fungi). A VOC profile may relate to VOC concentration levels, or, alternatively or in addition, to ratios between selected VOC concentration levels, or to patterns created by some or all of the VOCs appearing in the VOC release data, as further described below. A VOC profile can serve as a template for VOC release associated with a corresponding metabolic, anabolic, catabolic, genetic, and / or medical condition. The term “healthy VOC profile” as used herein refers to a weighted average of VOC release data related to healthy individuals (i.e., from at least one of the following: respiration, body fluids, or cell cultures). Similarly, the term “target cell VOC profile” refers to the weighted average of VOC release data related to the target (i.e., from at least one of the following: respiration, body fluids, or cell cultures). The term “dynamic differential VOC profile” refers to the range of VOC release data for each profile (i.e., healthy profile, target profile), and / or the range between the healthy VOC profile and the target VOC profile. In this specification, the terms VOC release data, healthy VOC profile, target VOC profile, and differential VOC profile may be followed by an adjective describing the relevant term, for example, “MCD pre-target cell VOC profile.”

[0023] Refer now to Figure 1, which is a schematic example of a system, referenced collectively at 100, constructed and effective by embodiments of the disclosed technology, for associating the excretion of chemicals in at least one of human respiration, bodily fluids, and cell cultures with corresponding expressions of genetic or medical conditions, and for utilizing these associations for diagnosis, and / or for determining the effectiveness of treatment, and / or for determining the choice of treatment. The system includes an analyzer 102, a database 104, and a processor 106. The system may further include a user interface 108. The processor 106 is coupled to the analyzer 102, the database 104, and the user interface 108.

[0024] The analyzer 102 may be a mass spectrometer (MS), an ion mobility spectrometer (IMS), a gas chromatograph (GC), various combinations of MS, GC, and IMS, or any other instrument that provides identification and / or quantification of VOC analytes in a sample. The analyzer 102 may be, for example, a triple quadrupole gas chromatography-mass spectrometer (GC-MS / MS), which may include a thermal dissolver and operate in a selected ion monitoring MS mode. Alternatively, the analyzer 102 may be a calibrated proton transfer reaction-time-of-flight mass spectrometer (PTR-TOFMS), a calibrated selective ion flow tube mass spectrometer (SIFT-MS), high-field asymmetric waveform ion mobility spectroscopy (FAIMS), gas chromatograph photoionization detector, or high-field asymmetric waveform ion mobility spectroscopy-time-of-flight mass spectrometer (FAIMS-TOFMS), or gas chromatograph-quadrupole-time-of-flight mass spectrometer (GC-QTOF), or gas chromatograph Orbitrap (e.g., GC-exactive), or gas chromatograph-quadrupole mass spectrometer Orbitrap (GC-Q Exactive) analyzer. The analyzer 102 is used to obtain measurement results for various chemicals, specifically VOCs, in the respiration or body fluids of patient 110. The analyzer 102 is further used to obtain measurement results for various chemicals in cell cultures 112. The analyzer 102 provides raw measurement results to the processor 106.

[0025] Processor 106 associates the excretion of chemical substances with corresponding metabolic, anabolic, catabolic processes, genetic and / or medical and / or pathological conditions, as further detailed below. Processor 106 further utilizes these associations for diagnosis, to determine the effectiveness of treatments, and to select appropriate treatments, as also detailed below.

[0026] Refer here to Figures 2A and 2B, which are schematic examples of VOC emission data, referenced as a whole at 120, and differential VOC profiles, referenced as a whole at 130, respectively, according to another embodiment of the disclosed technology. Referring to Figure 2A, the horizontal axis represents the selected VOC being measured, and the vertical axis represents the concentration level. As mentioned above, the concentration level may be measured in parts-per notation of the VOC molecule (e.g., ppm, ppb, ppt) or as count rate. In general, there are more than 1800 different VOCs that are excreted or found in the respiration and bodily fluids of a typical human. However, not all of these VOCs are necessarily measured.

[0027] Referring to Figure 2B, the dynamic differential VOC profile 130 is defined by the healthy and target VOC profiles determined from the VOC release data of healthy and target cell cultures, the predicted VOC concentration levels present in respiration and / or body fluids, and the VOC concentration levels measured in respiration and / or body fluids. In Figure 2B, symbol "X" 132 T The symbol "X" represents the VOC concentration level measured from healthy cell cultures. H This represents the relative VOC concentration level measured from the target cell culture compared to [a specific value]. (Line 134) T , and line 135 H The square portion 136 above represents the predicted target VOC concentration levels to be observed in the respiration and / or body fluids and / or cell cultures of patients with target cell expression or overexpression. Line 134 H , and line 135 TThe portion of the square 136 below represents the healthy VOC concentration levels expected to be observed in the breath and / or body fluids and / or cell cultures of patients without the expression or overexpression of the target cells. The predicted VOC concentration levels are determined by applying a diffusion model (e.g., Farhi's equation or the modified Farhi model, both further described below) and a cell growth equation to the VOC concentration levels measured from healthy and target cell cultures. The square 136 represents the range of VOC emission data measured from breath and / or body fluids in healthy patients and patients with the expression or overexpression of the target cells. Line 135 T represents the highest concentration level measured in the healthy patient group. Line 135 H represents the lowest concentration level of the VOCs measured in the patient group with the expression or overexpression of the target cells. Line 135 T and line 135 H The range between represents the threshold between the maximum normal VOC expression and the target cell VOC expression of the VOCs in the breath, body fluids or cell cultures. Point 138 H ~138 T The range between is the dynamic range of the VOCs released into the breath and / or body fluids. As shown in FIG. 2A, for example, if the measured concentration level is below the threshold, not all of the measured VOCs are necessarily included in the VOC profile 130.

[0028] Construction of the VOC Profile Database Refer here to Figures 3A, 3B, and 3C, which are schematic examples of exemplary methods for associating VOC releases with corresponding cancer types in a selected population, which will be more effective in further embodiments of the disclosed technology. In step 150, at least one respiratory and bodily fluid (e.g., blood, urine, or sweat) sample is obtained from multiple patients for each selected cancer type. The term “cancer type” refers to the type of cancer (e.g., ovarian, breast, bladder, skin, colon, etc.) and the genetic subtype of the cancer (e.g., HER2+, HER2 triple-negative, etc.). Since the cancer type is known before the sample is obtained, each respiratory and bodily fluid sample is associated with a corresponding cancer type. After step 150, the method proceeds to step 160.

[0029] In step 152, target cell samples are obtained from multiple patients for each selected cancer type. In the examples presented in Figures 3A-3C, the target cells are oncogenic cells of each cell type. These target cell samples are obtained, for example, by a biopsy procedure. After step 152, the method proceeds to step 162.

[0030] In step 154, samples of healthy cells of the same cell type as the target cells are obtained from multiple patients. These healthy cell samples may also be obtained, for example, by a biopsy procedure. After step 154, the method proceeds to step 162.

[0031] In step 156, a sample of control cells of the same type as the target cells is obtained from the control group. The control cells are healthy cells obtained from the control group. The control cells may also be obtained, for example, by a biopsy procedure. After step 156, the method proceeds to step 162.

[0032] In step 158, respiratory and / or body fluid samples are taken from the control group. Following step 158, the method proceeds to step 164.

[0033] In step 160, at least one VOC emission data is obtained from the patient's respiratory and body fluid samples. Referring to Figure 1, the analyzer 102 obtains at least one emission data from the respiratory and body fluids. After step 160, the method proceeds to step 174.

[0034] In step 162, the gene sequences of target cells, healthy cells, and control cells are determined. These gene sequences are then classified according to known oncogenic gene mutations of the selected cancer type. For example, there are currently over 315 known cancer-inducing mutations. These steps are also referred to as molecular classification. After step 162, the method proceeds to step 166.

[0035] In step 164, VOC emission data of VOCs released into respiratory and / or body fluid samples from the control group are acquired. Referring to Figure 1, the analyzer 102 acquires at least one emission data from the respiratory and body fluid samples of the control group. After step 164, the method proceeds to step 176.

[0036] In step 166, target cells, healthy cells, and control cell samples are cultured. After step 166, the method proceeds to step 168.

[0037] In step 168, VOC release data related to the target cell culture, healthy cell culture, and control cell culture are obtained from each respective culture. Referring to Figure 1, the analyzer 102 obtains release data related to the target cell culture, while data for the healthy cell culture and control cell culture are obtained from each respective culture. After step 168, the method proceeds to step 170.

[0038] In step 170, for each gene sequence, target cell VOC profiles, healthy cell VOC profiles, and control cell VOC profiles are determined from target cell culture VOC release data, healthy cell culture VOC release data, and control cell culture VOC release data for each gene mutation. First, the target cell culture VOC release data, healthy cell culture VOC release data, and control cell culture VOC release data are filtered. In general, target and healthy cells may produce VOC artifacts unrelated to gene mutations. Comparing target cell culture VOC release data and healthy cell culture VOC release data with each other and with control cell VOC release data is used to mitigate the impact of unrelated VOC artifacts and to better identify VOCs related to selected gene mutations. For example, in some cases of breast cancer, even healthy cells may have oncogenes and / or be in the process of becoming target cells. In some such cases, the cancer may not be fully active. Nevertheless, the expression of this gene may be a VOC that is not present in the VOC release data from control cell cultures or in the VOC release data from respiratory and body fluid samples obtained from the control group. For this reason, the expression of this gene in healthy cells may be identified. Filtered target cell culture VOC release data is created by comparing the target cell culture VOC release data with both healthy cell culture VOC release data and control cell culture VOC release data. Subsequently, filtered healthy cell culture VOC release data is created by comparing the healthy cell culture VOC release data with both filtered target cell culture VOC release data and control cell culture VOC release data. After this, filtered control cell culture VOC release data is created by comparing the control cell culture VOC release data with filtered healthy cell culture VOC release data. Finally, the target cell VOC profile is determined by comparing the filtered target cell culture VOC release data with filtered healthy cell culture VOC release data and filtered control cell culture VOC release data.The healthy cell VOC profile is determined by comparing filtered healthy cell culture VOC release data with both filtered target cell culture VOC release data and filtered control cell culture VOC release data. The control cell VOC profile is determined by comparing filtered control cell culture VOC release data with filtered healthy cell culture VOC release data. Referring to Figure 1, for each gene sequence, processor 106 determines the target cell VOC profile, healthy cell VOC profile, and control cell VOC profile from the target cell culture VOC release data, healthy cell culture VOC release data, and control cell culture VOC release data for each gene mutation. After step 170, the method proceeds to steps 172 and 180.

[0039] In step 172, the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile are determined by predicting the respiratory and fluid VOC concentration levels from the target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile for each gene mutation. The VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Since the VOC profile is associated with the corresponding gene mutation, the predicted VOC profile is also associated with the corresponding gene mutation. Referring to Figure 1, processor 106 determines the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile. After step 172, the method proceeds to steps 174 and 176.

[0040] In step 174, a dynamic differential VOC profile is created from the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the patient's respiratory and fluid VOC release data. This dynamic differential VOC profile is created by minimizing the error between the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the respiratory and fluid sample VOC release data. Since each predicted target cell VOC profile and the predicted healthy cell VOC profile are associated with the corresponding gene mutation, each dynamic differential VOC profile is also associated with the corresponding gene mutation. Referring to Figure 1, processor 106 determines the dynamic differential VOC profile for each gene mutation. From step 174, the method proceeds to step 178.

[0041] In step 176, a dynamic control VOC profile is created from the predicted control cell VOC profile and the VOC release data from the control group's respiration and body fluid samples. Similar to the dynamic differential VOC profile, the dynamic control VOC profile is created by minimizing the error between the predicted control cell VOC profile and the VOC release data from the control group's respiration and body fluid samples. Referring to Figure 1, processor 106 determines the dynamic control VOC profile for each gene mutation. From step 176, the method proceeds to step 178.

[0042] In step 178, the dynamic differential VOC profile is compared to the dynamic control VOC profile to further identify them. Referring to Figure 1, processor 106 compares the dynamic differential VOC profile to the dynamic control VOC profile to further identify them. From step 178, the method proceeds to step 180.

[0043] In step 180, the VOC profile is saved to the database. Referring to Figure 1, processor 106 saves the dynamic differential VOC profile, the dynamic control VOC profile, and the corresponding cancer type to database 104.

[0044] In some cases, VOC release from target and healthy cells may differ before and after treatment. For example, in oncogenic target cells, mass cell death (MCD) treatment (e.g., radiation therapy, chemotherapy) may be used, and the VOCs released by healthy and target cells may differ before and after MCD treatment. To determine the effect of MCD on patient-released VOCs, MCD is induced in cell cultures in a manner that does not produce VOC artifacts (e.g., by using rapid freezing or ultraviolet-UV light technology), and the VOC releases obtained before and after MCD are then associated with the corresponding target cells for a selected population.

[0045] Refer here to Figures 4A, 4B, 4C, and 4D, which are schematic examples of methods for associating VOC release with target cells before and after treatment in a selected population, which are effective by another embodiment of the disclosed technology. In step 200, respiratory and / or body fluid samples are obtained from multiple patients for each selected target cell type. Following step 200, the method proceeds to step 210.

[0046] In step 202, target cell samples are obtained from multiple patients for each selected target cell type. These target cell samples are obtained, for example, by a biopsy procedure. After step 202, the method proceeds to step 212.

[0047] In step 204, samples of healthy cells of the same cell type as the target cells are obtained from multiple patients. These healthy cell samples may also be obtained by biopsy procedure. After step 204, the method proceeds to step 212.

[0048] In step 206, a sample of control cells of the same type as the target cells is obtained from the control group. These control cell samples may also be obtained by biopsy procedure. After step 206, the method proceeds to step 212.

[0049] In step 208, respiratory and / or body fluid samples are taken from the control group. Following step 208, the method proceeds to step 214.

[0050] In step 210, VOC emission data of VOCs released into respiratory and / or body fluid samples are obtained from multiple patients. Referring to Figure 1, the analyzer 102 obtains at least one emission data from respiratory and body fluid samples from multiple patients. After step 210, the method proceeds to step 236.

[0051] In step 212, the gene sequences of target cells, healthy cells, and control cells are determined, and the gene sequences are classified according to molecular classification (gene classification). After step 212, the method proceeds to step 216.

[0052] In step 214, VOC emission data of VOCs released into respiratory and / or body fluid samples from the control group are acquired. Referring to Figure 1, the analyzer 102 acquires at least one emission data from the respiratory and body fluid samples of the control group. Following step 214, the method proceeds to step 234.

[0053] In step 216, target cell cultures, healthy cell cultures, and control cell cultures are generated by culturing samples of target cells, healthy cells, and control cells. From step 216, the method proceeds to steps 218 and 220.

[0054] In step 218, VOC release data related to the target cell culture, healthy cell culture, and control cell culture are acquired before inducing MCD. Referring to Figure 1, the analyzer 102 acquires VOC release data for target cells, healthy cells, and control cells before inducing MCD. After step 218, the method proceeds to steps 220 and 222.

[0055] In procedure 220, filtered VOC release data from pre-MCD target cell cultures, filtered VOC release data from healthy cell cultures, and filtered VOC release data from control cell cultures are created for each gene sequence of each gene mutation to reduce the impact of irrelevant VOC artifacts and to better identify VOCs associated with selected gene mutations before inducing MCD. To this end, filtered VOC release data from pre-MCD target cell cultures is first created by comparing target cell culture VOC release data with both healthy cell VOC release data and control cell culture VOC release data, all of which are obtained before inducing MCD. Subsequently, filtered VOC release data from healthy cell cultures is created by comparing healthy cell VOC release data (i.e., obtained before inducing MCD) with both filtered VOC release data from pre-MCD target cell cultures and control cell culture VOC release data obtained before inducing MCD (i.e., obtained before inducing MCD). Subsequently, filtered control cell culture VOC release data is determined by comparing the control cell culture VOC release data obtained before inducing MCD with filtered pre-MCD healthy cell culture VOC release data. Referring to Figure 1, processor 106 determines filtered pre-MCD target cell culture VOC release data, filtered pre-MCD healthy cell culture VOC release data, and filtered pre-MCD control cell culture VOC release data for each gene sequence for each gene mutation. After step 220, the method proceeds to steps 228, 230, and 231.

[0056] In step 222, massive cell death is induced in the target cell culture, healthy cell culture, and control cell culture. Preferably, MCD is induced using a method that does not generate VOC artifacts (e.g., by utilizing rapid freezing or UV light technology).

[0057] In step 224, VOC release data related to the target cell culture, healthy cell culture, and control cell culture are obtained from each culture after a large-scale cell death. Note that the cell cultures used before and after MCD are the same cultures. Referring to Figure 1, the analyzer 102 obtains VOC release data for target cells, healthy cells, and control cells after a large-scale cell death. After step 222, the method proceeds to step 226.

[0058] In procedure 226, filtered post-MCD target cell culture VOC release data and filtered post-MCD healthy cell culture VOC release data, as well as filtered post-MCD control cell culture VOC release data, are created for each gene mutation to mitigate the effects of irrelevant VOC artifacts and to better identify VOCs associated with selected gene mutations after MCD induction. First, filtered post-MCD target cell culture VOC release data is created by comparing target cell culture VOC release data obtained after MCD induction with both healthy cell culture VOC release data obtained after MCD induction and control cell culture VOC release data obtained after MCD induction. Subsequently, filtered post-MCD healthy cell culture VOC release data is created by comparing healthy cell culture VOC release data (i.e., obtained after MCD induction) with filtered post-MCD target cell culture VOC release data and control cell culture VOC release data (i.e., obtained after MCD induction). Subsequently, filtered control cell culture VOC release data is determined by comparing the filtered post-MCD healthy cell culture VOC release data (i.e., obtained after inducing MCD) with the control cell culture VOC release data (i.e., obtained after inducing MCD). Referring to Figure 1, processor 106 determines the filtered post-MCD target cell culture VOC release data, filtered post-MCD healthy cell culture VOC release data, and filtered post-MCD control cell culture VOC release data for each gene sequence of each gene mutation. After step 226, the method proceeds to steps 228, 230, and 231.

[0059] In step 228, the MCD pre-target cell VOC profiles and MCD post-target cell VOC profiles are determined from filtered MCD pre-target cell culture VOC release data and filtered MCD post-target cell culture VOC release data. Referring to Figure 1, processor 106 determines the MCD pre-target cell VOC profiles and MCD post-target cell VOC profiles from filtered MCD pre-target cell VOC release data and filtered MCD post-target cell VOC release data. After step 228, the method proceeds to step 232.

[0060] In step 230, the pre-MCD healthy cell VOC profile and the post-MCD healthy cell VOC profile are determined from the filtered pre-MCD healthy cell culture VOC release data and the filtered post-MCD healthy cell culture VOC release data. Referring to Figure 1, processor 106 determines the pre-MCD healthy cell VOC profile and the post-MCD healthy cell VOC profile from the filtered pre-MCD healthy cell culture VOC release data and the filtered post-MCD healthy cell culture VOC release data. After step 230, the method proceeds to step 232.

[0061] In step 231, the pre-MCD control cell VOC profiles and post-MCD control cell VOC profiles are determined from filtered pre-MCD control cell culture VOC release data and filtered post-MCD control cell culture VOC release data. Referring to Figure 1, processor 106 determines the pre-MCD control cell VOC profiles and post-MCD control cell VOC profiles from filtered pre-MCD control cell culture VOC release data and filtered post-MCD control cell culture VOC release data. After step 231, the method proceeds to step 232.

[0062] In procedure 232, the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile are determined for each gene mutation. The predicted target cell VOC profile is determined by predicting the VOC concentration levels in respiration and body fluids from the pre-MCD and post-MCD target cell VOC profiles. The predicted healthy cell VOC profile is determined by predicting the VOC concentration levels in respiration and body fluids from the pre-MCD and post-MCD healthy cell VOC profiles. The predicted control cell VOC profile is determined by predicting the VOC concentration levels in respiration and body fluids from the pre-MCD and post-MCD control cell VOC profiles. The VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile. After step 232, the method proceeds to steps 234, 236, and 240.

[0063] In step 234, a dynamic control cell VOC profile is created from the predicted control cell VOC profile and the VOC release data from the control group's respiration and body fluid samples. Similar to the dynamic differential VOC profile, the dynamic control VOC profile is created by minimizing the error between the predicted control cell VOC profile and the VOC release data from the control group's respiration and body fluid samples. Referring to Figure 1, processor 106 determines the dynamic control VOC profile for each gene mutation. After step 234, the method proceeds to step 238.

[0064] In step 236, a dynamic differential VOC profile is created for each gene mutation from the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the VOC release data from respiration and / or body fluid samples. Referring to Figure 1, processor 106 creates a dynamic differential VOC profile for each gene mutation from the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the VOC release data from respiration and / or body fluid samples. After step 236, the method proceeds to step 238.

[0065] In step 238, the dynamic differential VOC profile is compared to the dynamic control VOC profile to further identify them. Referring to Figure 1, processor 106 compares the dynamic differential VOC profile to the dynamic control VOC profile to further identify them. After step 238, the method proceeds to step 240.

[0066] In step 240, the dynamic differential VOC profile and the dynamic comparative VOC profile are saved to the database. Referring to Figure 1, processor 106 saves the dynamic differential VOC profile and the dynamic comparative VOC profile to database 104.

[0067] According to another embodiment of the disclosed technology, VOC release from the body and / or cultures of a patient or multiple patients may be associated with known pathological conditions arising from pathogens (e.g., normal or pathological bacterial flora) such as bacteria, viruses, or fungi. In some cases (e.g., E. coli), these pathogens may be naturally present in the body, and the pathological condition is characterized by an increase or decrease in the number of such pathogens. The pathological condition may also exhibit different VOC releases before and after treatment as a result of VOC artifacts related to the effect of treatment on other bacteria or pathogens in the body (e.g., VOC artifacts related to the effect of antibiotics on the normal bacterial flora in the gut).

[0068] Refer here to Figures 5A, 5B, and 5C, which are schematic examples of methods for relating VOC profiles to pathological conditions arising from pathogens, which are more effective in further embodiments of the disclosed technology.

[0069] In step 250, at least one respiratory and / or fluid sample is taken from multiple patients with abnormal levels of target cells before the patient receives any treatment related to the target cells (e.g., before the patient receives antibiotics in the case of a bacterial pathogen such as Klebsiella pneumoniae), and at least one respiratory and / or fluid sample is taken from the same multiple patients after the patient has completed the treatment (i.e., after the pathogen is no longer symptomatic and / or no longer present in the patient's culture). It should be noted that the sample taken after a successful treatment is taken after a period including the time required for the effects of the treatment to completely diminish, so that VOC artifacts directly and indirectly related to the treatment are no longer present in the sample. After step 250, the method proceeds to step 258.

[0070] In step 252, at least one respiratory and one bodily fluid sample is obtained from multiple patients with normal levels of target cells. Following step 252, the method proceeds to step 258.

[0071] In step 254, target cell samples are obtained from multiple patients with abnormal levels of target cells before any procedure. Following step 254, the method proceeds to step 260.

[0072] In step 256, target cell samples are obtained from multiple patients with normal levels of target cells. Following step 256, the method proceeds to step 260.

[0073] In procedure 258, VOC release data for VOCs released into at least one respiratory and / or fluid sample from patients with abnormal levels of target cells, taken before and after the procedure, is acquired (i.e., if the procedure is successful). In addition, VOC release data for VOCs released into at least one respiratory and / or fluid sample from patients with normal levels of target cells is also acquired. Referring to Figure 1, analyzer 102 acquires VOC release data for VOCs released into at least one respiratory and / or fluid sample taken before any procedure from patients with abnormal levels of target cells, and VOC release data for VOCs released into at least one respiratory and / or fluid sample taken after the successful treatment related to target cells (samples are taken from the same multiple patients before and after the procedure). Analyzer 102 further acquires VOC release data for VOCs released into at least one respiratory and / or fluid sample from multiple patients with normal levels of target cells. Note that the group of patients with abnormal levels of target cells and the group of patients with normal levels of target cells are two distinct groups. After step 258, the method proceeds to steps 259 and 282.

[0074] In step 259, the respiratory and / or fluid target VOC profile, as well as the respiratory and / or fluid healthy VOC profile, are determined. The respiratory and / or fluid target VOC profile is determined by comparing VOC release data obtained from multiple patients with abnormal levels of target cells before treatment with VOC release data obtained from the same multiple patients with abnormal levels of target cells after successful treatment, and VOC release data obtained from patients with normal levels of target cells. The respiratory and / or fluid healthy VOC profile is determined by comparing respiratory and / or fluid VOC release data from multiple patients with the respiratory and / or fluid target VOC profile. Referring to Figure 1, processor 106 determines the respiratory and / or fluid target VOC profile and the respiratory and / or fluid healthy VOC profile. After step 259, the method proceeds to steps 274 and 276.

[0075] In step 260, the gene sequences of target cells are determined from both patients with abnormal levels of target cells and patients with normal levels of target cells, and the target cells are classified. The gene sequences of target cells from patients with abnormal and normal levels of target cells are determined for molecular classification, and it is determined whether the target cells from patients with abnormal levels of target cells were mutated. After step 260, the method proceeds to step 262.

[0076] In step 262, target cell cultures and normal cell cultures are generated by culturing cells from cell samples from patients with abnormal levels of target cells and from cell samples from patients with normal levels of target cells, respectively. Following step 262, the method proceeds to steps 264 and 266.

[0077] In step 264, VOC release data related to VOCs released by cells in the target and normal cell cultures is acquired before MCD induction. Referring to Figure 1, analyzer 102 acquires VOC release data related to VOCs released by the target and normal cell cultures before MCD induction. After step 264, the method proceeds to steps 266 and 270.

[0078] In step 266, massive cell death is induced in cells in both target and normal cell cultures. As previously described, MCD is induced by a method that does not generate VOC artifacts (e.g., rapid freezing, UV light). Following step 266, the method proceeds to step 268.

[0079] In step 268, VOC release data related to VOCs released by cells in target and normal cell cultures is acquired after MCD induction. Note that the cell culture used to acquire VOC release data after MCD is the same cell culture used to acquire VOCs before MCD. Referring to Figure 1, analyzer 102 acquires VOC release data related to VOCs released by the cell culture after MCD induction. After step 268, the method proceeds to step 272.

[0080] In step 270, target cell culture VOC release data obtained before inducing MCD is compared with normal cell culture VOC release data obtained before inducing MCD to identify them. Target cell culture VOC release data refers to VOC release data from cell cultures originating from patients with abnormal levels of target cells before any treatment. Normal VOC release data refers to VOC release data from cell cultures originating from patients with normal levels of target cells. Referring to Figure 1, processor 106 compares target cell culture VOC release data with normal cell culture VOC release data before inducing MCD to identify them. After step 270, the method proceeds to step 274.

[0081] In step 272, target cell culture VOC release data obtained after induction of MCD is compared with normal cell culture VOC release data obtained after induction of MCD and identified. Referring to Figure 1, processor 106 compares target cell culture VOC release data obtained after induction of MCD with normal cell culture VOC release data obtained after induction of MCD and identifies them. After step 272, the method proceeds to step 276.

[0082] In step 274, pre-MCD target cell VOC profiles and post-MCD target cell VOC profiles are created. These profiles are created by comparing target cell culture VOC release data obtained before induction of MCD with target cell culture VOC release data obtained after induction of MCD, as well as respiratory and / or fluid target VOC profiles. Referring to Figure 1, processor 106 creates pre-MCD target cell VOC profiles and post-MCD target cell VOC profiles by comparing target cell culture VOC release data obtained before induction of MCD with target cell culture VOC release data obtained after induction of MCD, as well as respiratory and / or fluid target VOC profiles. After step 274, the method proceeds to step 278.

[0083] In step 276, pre-MCD normal cell VOC profiles and post-MCD normal cell VOC profiles are created. These profiles are created by comparing normal cell culture VOC release data obtained before induction of MCD with normal cell culture VOC release data obtained after induction of MCD, as well as respiratory and / or fluid-healthy VOC profiles. Referring to Figure 1, processor 106 creates pre-MCD normal cell VOC profiles and post-MCD normal cell VOC profiles by comparing normal cell culture VOC release data obtained before induction of MCD with normal cell culture VOC release data obtained after induction of MCD, as well as respiratory and / or fluid-healthy VOC profiles. After step 276, the method proceeds to step 278.

[0084] In step 278, the predicted target VOC profile and the predicted healthy VOC profile are determined. The predicted target VOC profile is determined by predicting respiratory and fluid concentration levels from the pre-MCD target cell VOC profile and the post-MCD target cell VOC profile. The predicted healthy VOC profile is determined by predicting respiratory and fluid concentration levels from the pre-MCD and post-MCD healthy VOC profiles. VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted target VOC profile and the predicted healthy VOC profile. After step 278, the method proceeds to steps 280 and 282.

[0085] In step 280, abnormal response VOC profiles and normal response VOC profiles are created that relate to the patient's response to target cells by comparing respiration and / or fluid VOC release data from multiple patients with abnormal levels of target cells with respiration and / or fluid VOC release data from multiple patients with normal levels of target cells, predicted target VOC profiles, and predicted healthy VOC profiles. These normal and abnormal response VOC profiles are related to the patient's response to target cells (e.g., immune system, antibody production). Referring to Figure 1, processor 106 creates abnormal response VOC profiles and normal response VOC profiles that relate to the patient's response to target cells by comparing respiration and / or fluid VOC release data from multiple patients with abnormal levels of target cells with respiration and / or fluid VOC release data from multiple patients with normal levels of target cells, predicted target VOC profiles, and predicted healthy VOC profiles. After step 280, the method proceeds to steps 282 and 284.

[0086] In step 282, a dynamic differential VOC profile is created from the predicted target VOC profile, the predicted healthy VOC profile, the abnormal response VOC profile, respiratory and / or fluid VOC release data from multiple patients with abnormal levels of target cells, and respiratory and / or fluid VOC release data from multiple patients with normal levels of target cells. Referring to Figure 1, processor 106 creates the dynamic differential VOC profile. After step 282, the method proceeds to step 284.

[0087] In step 284, the VOC profile is saved to the database. Referring to Figure 1, processor 106 saves the VOC profile to database 104.

[0088] According to another embodiment of the disclosed technology, VOC release from the body of a patient or multiple patients can be associated with the exposure of target cells to MMR activation treatment by determining a dynamic differential VOC profile for each selected mismatch repair gene (MMR) gene activation therapy. Refer here to Figures 6A–6D, which are schematic examples of methods for determining a dynamic differential VOC profile for selected mismatch repair (MMR) gene activation therapy according to another embodiment of the disclosed technology.

[0089] In step 300, for each selected gene, respiratory and / or fluid samples are obtained from multiple patients before applying the MMR gene activation treatment. The genes are selected from those suitable for MMR gene activation therapy. After step 300, the method proceeds to step 302.

[0090] In step 302, VOC emission data of VOCs released into respiratory and / or body fluid samples is acquired. Referring to Figure 1, the analyzer 102 acquires VOC emission data of respiratory and / or body fluid samples. From step 302, the method proceeds to step 348.

[0091] In step 304, target cell samples are obtained from multiple patients for each selected gene. From step 304, the method proceeds to step 308.

[0092] In step 306, for each selected gene, healthy cell samples of the same type as the target cells are obtained from the same multiple patients (i.e., a set of target cells and healthy cells is obtained from each of the multiple patients for each selected gene). From step 306, the method proceeds to step 308.

[0093] In step 308, two target cell culture sets and a healthy cell culture are generated from the target and healthy cell samples. For clarity in the following explanation, the first target cell culture set will be referred to as "Culture Set A," and the second target cell culture set will be referred to as "Culture Set B." From step 308, the method proceeds to step 310.

[0094] In step 310, pre-treatment VOC release data related to cells in two target cell culture sets (i.e., culture set A and culture set B) and a healthy cell culture is acquired. Referring to Figure 1, the analyzer 102 acquires VOC release data related to the two target cell cultures and the healthy cell culture. From step 310, the method proceeds to steps 312, 328, 330, and 331.

[0095] In step 312, the gene sequences of target cells in both target cell culture sets (i.e., in culture set A and culture set B) and healthy cells in the healthy cell culture are validated. When determining the dynamic differential VOC profile for MMR gene activation therapy, the gene sequences of target cells are already known and only validation is required, while healthy cell cultures are gene-sequenced to confirm the absence of pathological steps or conditions, or to determine the level of a step or condition. From step 312, the method proceeds to step 314.

[0096] In step 314, the pre-treatment target cell VOC profile for each target cell and the pre-treatment healthy cell VOC profile for each healthy cell are determined by comparing the target cell culture VOC release data of both target cell culture sets (i.e., culture set A and culture set B) with the healthy cell culture VOC release data of the healthy cell culture. Referring to Figure 1, processor 106 determines the pre-treatment target VOC profile and the pre-treatment healthy VOC profile. From step 314, the method proceeds to steps 316, 318, 320, and 340.

[0097] In step 316, the selected MMR gene activation treatment is applied to cells in the first of the two target cell culture sets (i.e., culture set A). From step 316, the method proceeds to step 322.

[0098] In step 318, a large amount of cell death is induced in the cells of the second of two sets of target cell cultures (i.e., culture set B) that have not been exposed to any treatment, in a manner that does not produce residual VOC artifacts (e.g., by utilizing rapid freezing or ultraviolet light technology on the target cell cultures). From step 318, the method proceeds to step 324.

[0099] In step 320, MMR gene activation therapy is applied to cells in a healthy cell culture. From step 320, the method proceeds to step 326.

[0100] In step 322, post-treatment target cell culture VOC release data related to target cells in the first set of target cell cultures (i.e., culture set A) is obtained after the application of the selected MMR gene activation treatment. Referring to Figure 1, the analyzer 102 obtains post-treatment target cell culture VOC release data related to target cells in culture set A after the application of the MMR gene activation treatment. After step 322, the method proceeds to step 328.

[0101] In step 324, post-MCD target cell culture VOC release data related to target cells in the second set of target cell cultures (i.e., culture set B) is acquired after MCD induction. Referring to Figure 1, analyzer 102 acquires post-MCD target cell culture VOC release data related to target cells in culture set B after MCD induction. After step 324, the method proceeds to step 330.

[0102] In step 326, post-treatment healthy cell culture VOC release data related to healthy cell cultures is obtained after the selected MMR gene activation treatment. Referring to Figure 1, the analyzer 102 obtains post-treatment healthy cell culture VOC release data related to healthy cell cultures after the application of the MMR gene activation treatment. After step 326, the method proceeds to step 332.

[0103] In step 328, an MMR gene activation target VOC profile is created by comparing pre-treatment target cell culture VOC release data obtained from target cells in a first set of target cell cultures (i.e., culture set A) with post-treatment target cell culture VOC release data obtained from target cells in a first set of target cell cultures (i.e., culture set A). The MMR gene activation target VOC profile relates to the VOCs released by the target cell culture when the selected MMR gene activation treatment is applied. Referring to Figure 1, processor 106 creates an MMR gene activation target VOC profile by comparing pre-treatment target cell culture VOC release data obtained from target cells in a first set of target cell cultures with post-treatment target cell culture VOC release data obtained from target cells in a first set of target cell cultures after the selected MMR gene activation therapy has been administered. After step 328, the method proceeds to step 334.

[0104] In step 330, the post-MCD VOC profile is created by comparing pre-treatment target cell culture VOC release data obtained from target cells in a second set of target cell cultures (i.e., culture set B) with post-MCD target cell culture VOC release data obtained from target cells in the second set of target cell cultures (i.e., culture set B) after MCD induction. The post-MCD target cell VOC profile relates to the VOCs released by target cell culture "B" when MCD is induced in a manner that does not generate residual VOCs. Referring to Figure 1, processor 106 creates the post-MCD target cell VOC profile by comparing pre-treatment target cell culture VOC release data obtained from target cells in the second set of target cell cultures before MCD with post-MCD target cell culture VOC release data obtained from target cells in the second set of target cell cultures after MCD induction. After step 330, the method proceeds to step 336.

[0105] In step 332, the MMR gene-activated healthy cell VOC profile is created by comparing the VOC release data from healthy cell cultures before treatment with the VOC release data from healthy cell cultures after treatment. The MMR gene-activated healthy cell VOC profile relates to the VOCs released by healthy cell cultures when the MMR gene activation treatment is applied. Referring to Figure 1, processor 106 creates the MMR gene-activated healthy cell VOC profile by comparing the VOC release data from healthy cell cultures before treatment with the VOC release data from healthy cell cultures after treatment. After step 332, the method proceeds to step 338.

[0106] In step 334, the predicted MMR gene-activated target cell VOC profile is determined by predicting the respiratory and fluid VOC concentration levels from the MMR gene-activated target cell VOC profile. The VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted MMR gene-activated target cell VOC profile. After step 334, the method proceeds to step 342.

[0107] In step 336, the predicted post-MCD target cell VOC profile is determined by predicting the respiratory and fluid VOC concentration levels from the post-MCD target cell VOC profile. The VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted post-MCD target cell VOC profile. After step 336, the method proceeds to step 342.

[0108] In step 338, the predicted MMR gene-activated healthy cell VOC profile is determined by predicting the respiratory and fluid VOC concentration levels from the MMR gene-activated healthy cell VOC profile. The VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted MMR gene-activated healthy cell VOC profile. After step 338, the method proceeds to step 342.

[0109] In step 342, the patient's MMR gene activation treatment is initiated. After step 342, the procedure proceeds to step 344.

[0110] In step 344, at least one respiratory and / or bodily fluid sample is obtained from multiple patients during and / or after at least one selected stage of the selected MMR gene activation treatment. Following step 344, the method proceeds to step 346.

[0111] In step 346, VOC release data of VOCs released into the patient's respiratory and / or body fluid samples is obtained at at least one selected stage of the selected MMR gene activation treatment. Referring to Figure 1, the analyzer 102 obtains VOC release data of VOCs released into the patient's respiratory and / or body fluid samples during and / or after at least one selected stage of the selected MMR gene activation treatment. Following step 346, the method proceeds to step 348.

[0112] In step 348, a dynamic differential VOC profile is created for each selected gene mutation from the predicted MMR gene-activated target cell VOC profile, the predicted MCD target cell VOC profile, the predicted MMR gene-activated healthy cell VOC profile, and respiratory and / or fluid VOC release data from multiple patients before and after treatment. It should be noted that respiratory and / or fluid VOC release data from each patient from samples taken before MMR gene-activation treatment, and respiratory and / or fluid VOC release data from samples taken during or after treatment, should be taken from the same patient. Referring to Figure 1, processor 106 creates a dynamic differential VOC profile from the predicted MMR gene-activated target cell VOC profile, the predicted MCD target cell VOC profile, the predicted MMR gene-activated healthy cell VOC profile, and VOC release data from multiple patients before, during, or after treatment with MMR gene-activation therapy. Following step 348, the method proceeds to step 350.

[0113] In step 350, the VOC profile is saved to the database. Referring to Figure 1, processor 106 saves the VOC profile to database 104.

[0114] Measuring VOCs may also be used to determine whether target cells have developed treatment-resistant mutations or multiple mutations. Furthermore, mutant target cells may develop other mutations when subjected to the same or different treatments. Different treatments may be the same type of treatment with different doses of therapeutic agents (e.g., chemotherapy, radiotherapy, or antibiotics), or different types of treatments. In other words, a treatment may result in several generations of mutations. For example, TP-53 target cells of lung cancer may mutate into KSR target cells when subjected to one treatment. KSR target cells may mutate into another type of off-target cell of lung cancer when subjected to the same or different treatments. For this purpose, VOC release data from target cell cultures known to develop treatment-resistant mutations or multiple mutations are obtained before and after being subjected to the treatment known to induce treatment-resistant mutations or multiple mutations. The pre- and post-treatment cultures are the same culture. VOC release data from target cell cultures induced by one or more treatment-resistant mutations (verified by gene sequencing) are compared to the VOC release data of the culture before the treatment is applied. This step may be repeated multiple times for the same treatment and / or multiple times for different treatments. A complex treatment-resistant target cell VOC profile is created, containing the VOC profile of all generations of mutations or a selected target cell. The term “type” of target cells as used herein relates to target cells that may or may not mutate from other target cells, or target cells that have been treated or not treated in the past. Refer here to Figures 7A and 7B, which are schematic examples of a method for determining the VOC profile of target cells induced by treatment-resistant mutations from a selected treatment, which will come into effect in further embodiments of the disclosed technology.

[0115] In step 400, cell cultures are generated for each of the selected target cell types known to induce treatment-resistant mutations. After step 400, the method proceeds to step 402.

[0116] In step 402, VOC release data related to VOCs released by target cells in the target cell culture is acquired before treatment. Referring to Figure 1, the analyzer 102 acquires VOC release data related to VOCs released by target cells from the target cell culture before treatment. After step 402, the method proceeds to steps 404 and 414.

[0117] In step 404, a treatment known to induce treatment-resistant mutations or multiple mutations is applied to the target cell culture. Following step 404, the method proceeds to step 406.

[0118] In step 406, target cells that have developed treatment resistance are identified in the treated target cell culture, for example, by using a microscopic scan. After step 406, the method proceeds to step 408.

[0119] In step 408, each mutant target cell culture is generated for each identified treatment-resistant mutant target cell (i.e., after the selected treatment has been applied). In other words, new cultures of these treatment-resistant mutant target cells are prepared separately, and the selected treatment can then be applied to the new cultures to identify additional treatment-resistant mutations. After step 408, the method proceeds to step 410.

[0120] In step 410, the target cells in which the treatment-resistant mutations have been identified are genetically sequenced to determine the mutation or multiple mutations (i.e., if present), and to identify the molecular classification of the post-treatment target cells. Following step 410, the method proceeds to step 412.

[0121] In step 412, VOC release data related to mutant target cells is obtained from each muted target cell culture. Referring to Figure 1, the analyzer 102 obtains VOC release data from the mutant target cell culture. After step 412, the method proceeds to step 414.

[0122] In step 414, a VOC profile of treatment-resistant target cells is created by comparing the VOC release data of the pre-treatment target cell culture (i.e., the pre-treatment target cell culture) with the VOC release data of the mutant target cell culture. Referring to Figure 1, processor 106 creates a treatment-resistant VOC profile by comparing the VOC release data of any pre-treatment target cell culture with the target cell culture VOC release data of the mutant target cell culture. After step 414, the method proceeds to step 416.

[0123] In step 416, a complex treatment-resistant VOC profile is determined. The complex treatment-resistant VOC profile contains information from the treatment-resistant target cell VOC profiles of a selected number of mutation generations. Referring to Figure 1, processor 106 determines the complex treatment-resistant VOC profile. After step 416, the method proceeds to step 418.

[0124] In step 418, the predicted complex treatment-resistant VOC profile is determined by predicting respiratory and fluid VOC concentration levels from the complex treatment-resistant VOC profile. VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted complex treatment-resistant VOC profile. Following step 418, the method proceeds to step 420.

[0125] In step 420, the predicted treatment-resistant VOC profile is saved to the database. Referring to Figure 1, processor 106 saves the predicted treatment-resistant VOC profile to database 104.

[0126] It should be noted that the methods described in Figures 7A and 7B may be repeated for a selected number of treatments or mutations or both. Complex treatment-resistant VOC profiles incorporate information from each of these repeats. The saved treatment-resistant VOC profiles described above may be used to identify individual treatment-resistant VOC profiles for each organism.

[0127] Determination of VOC profiles for individual organisms VOC profiles may be determined for individuals in the same way that VOC profiles for a general population are determined. As described above, according to the disclosed technology, VOC emissions from the bodies of one or more patients can be associated with, for example, cellular gene mutations or a combination of mutations resulting in corresponding cancer types. The following is an example of associating VOC emissions from the bodies of individual patients with corresponding cancer types resulting from cellular gene mutations. However, this technology may be applied to cells of any form and type.

[0128] Refer here to Figures 8A and 8B, which are schematic examples of exemplary methods for associating VOC release with corresponding abnormal or pathological cells in individual patients, which are in effect by another embodiment of the disclosed technology. In Figures 8A and 8B, the exemplary pathological cells are cancer-type morphologies.

[0129] In step 450, at least one respiratory and bodily fluid (e.g., blood, urine, or sweat) sample is obtained from the patient for a selected cytogenetic mutation. The cytogenetic mutation may be associated with a cancer type. The term “cancer type” refers to the type of cancer (e.g., ovarian, breast, bladder, skin, colon, etc.) and the genetic subtype of the cancer (e.g., HER2+, HER2 triple-negative, etc.). The cancer type and the genetic mutation causing this cancer are known before the sample is obtained. The obtained respiratory and bodily fluid samples are associated with the cancer type. After step 450, the method proceeds to step 456.

[0130] In step 452, a sample of target cells exhibiting the selected gene mutation is obtained from the patient. In the examples presented in Figures 8A and 8B, the target cells are oncogenic cells of each cancer type exhibiting the respective gene mutation. These target cells are obtained, for example, by a biopsy procedure. After step 452, the method proceeds to step 458.

[0131] In step 454, a sample of healthy cells of the same type as the target cells is obtained from the patient. These healthy cells may also be obtained, for example, by a biopsy procedure. After step 454, the method proceeds to step 458.

[0132] In step 456, at least one VOC emission data is obtained from respiratory and body fluid samples. Referring to Figure 1, the analyzer 102 obtains at least one emission data from respiratory and body fluids. After step 456, the method proceeds to step 468.

[0133] In step 458, target and healthy cells in the cell sample are cultured. After step 458, the method proceeds to step 460.

[0134] In step 460, the gene sequences of cultured target cells and healthy cells are determined. Subsequently, the molecular classification of the cell samples is verified according to known gene mutations. Generally, this classification requires only verification because the gene mutation classification of the patient's target cells is known before the target cells are collected. For example, there are currently over 315 related mutations. This procedure is also called molecular classification. After step 460, the method proceeds to step 462.

[0135] In step 462, VOC release data related to both target cells and healthy cells is acquired. Referring to Figure 1, the analyzer 102 acquires VOC release data related to both target cells and healthy cells. After step 462, the method proceeds to step 464.

[0136] In step 464, the target cell VOC profile for each selected gene mutation (and therefore for each cancer type), and the healthy cell VOC profile for each healthy cell, are determined by comparing the target cell culture VOC release data with the healthy cell culture VOC release data. Referring to Figure 1, processor 106 creates the target cell VOC profile and the healthy cell VOC profile by comparing the target cell culture VOC release data with the healthy cell culture VOC release data. After step 464, the method proceeds to step 466.

[0137] In step 466, the predicted target cell VOC profile is determined by predicting the respiratory and fluid VOC concentration levels from the target cell VOC profile, and the predicted healthy cell VOC profile is determined by predicting the respiratory and fluid VOC concentration levels from the healthy cell VOC profile. The VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in more detail below. Since the target cell VOC profile is associated with a selected gene mutation, the predicted target cell VOC profile is also associated with the same gene mutation. Referring to Figure 1, processor 106 determines the predicted target cell VOC profile and the predicted healthy cell VOC profile from the target cell VOC profile and the healthy cell VOC profile. After step 466, the method proceeds to step 468.

[0138] In step 468, a dynamic differential VOC profile is created from the predicted target cell VOC profile, the predicted healthy cell VOC profile, and respiratory and fluid VOC release data. This dynamic differential VOC profile is created by minimizing the error between the predicted target cell VOC profile and the respiratory and fluid sample VOC release data. Since the predicted target cell VOC profile is associated with the corresponding gene mutation, the dynamic differential VOC profile is also associated with the gene mutation. Referring to Figure 1, processor 106 determines the dynamic differential VOC profile. After step 468, the method proceeds to step 470.

[0139] In step 470, the dynamic differential VOC profile is saved to the database. Referring to Figure 1, processor 106 saves the VOC profile to database 104.

[0140] VOC profiles may be used to determine the effectiveness of treatments in individual patients. However, determining the effectiveness of a treatment requires determining the effect of treatments that induce massive cell death (MCD) (e.g., chemotherapy, radiotherapy) on the VOCs released by the patient. This effect is determined by inducing MCD in cell cultures in a manner that does not produce VOC artifacts (e.g., by using rapid freezing or UV light technology), and by associating VOC release with corresponding target cells in individual patients before and after MCD. Refer here to Figures 9A and 9B, which are schematic examples of methods for associating VOC release with corresponding target cells (e.g., oncogenic cells) in individual patients before and after MCD, which are effective in further embodiments of the disclosed technology.

[0141] In procedure 500, at least one respiratory and bodily fluid (e.g., blood, urine, or sweat) sample is obtained from the patient for selected target cells and / or cellular gene mutations. As above, the target cell type and gene mutation causing the pathological condition are known before the sample is obtained. The obtained respiratory and bodily fluid samples are associated with their target cell type. After procedure 500, the method proceeds to procedure 506.

[0142] In step 502, a target cell sample is obtained from the patient for each selected target cell type. The target cell type may be a target cell exhibiting a selected gene mutation or causing a pathological condition. In the examples presented in Figures 9A and 9B, the target cells are oncogenic cells of each cancer type exhibiting the respective gene mutations. However, the method described in Figures 9A and 9B can be applied to any target cell. These target cells are obtained, for example, by a biopsy procedure. After step 502, the method proceeds to step 508.

[0143] In step 504, a sample of healthy cells of the same type as the target cells is obtained from the patient. These healthy cells may be obtained, for example, by a biopsy procedure. After step 504, the method proceeds to step 508.

[0144] In step 506, at least one VOC emission data is obtained from the respiratory and body fluid samples. Referring to Figure 1, the analyzer 102 obtains at least one emission data from the respiratory and body fluids. After step 506, the method proceeds to step 528.

[0145] In step 508, the gene sequences of target cells and healthy cells are determined. The gene sequences are then classified according to known oncogenic gene mutations of the selected cancer type. This step is also called molecular classification. After step 508, the method proceeds to step 510.

[0146] In step 510, target and healthy cells in the cell sample are cultured. Following step 510, the method proceeds to steps 512 and 514.

[0147] In step 512, VOC release data related to both healthy and target cells is acquired. Referring to Figure 1, the analyzer 102 acquires release data related to both healthy and target cells. After step 512, the method proceeds to step 518.

[0148] In step 514, MCD is induced in the target and healthy cell cultures. Preferably, MCD is induced using a method that does not generate VOC artifacts (e.g., by utilizing rapid freezing or UV light technology). It should be noted that MCD is induced in the same target and healthy cell cultures that were used to obtain pre-MCD VOC release data. After step 514, the method proceeds to step 516.

[0149] In step 516, VOC release data related to the target cell culture and VOC release data related to the healthy cell culture are acquired after MCD has been induced in the target cell and healthy cell cultures. Referring to Figure 1, the analyzer 102 acquires VOC release data from both the target cell culture and the healthy cell culture after MCD. Following step 516, the method proceeds to step 520.

[0150] In step 518, target cell culture VOC release data is compared with healthy cell culture VOC release data prior to MCD to identify them. Referring to Figure 1, processor 106 compares target cell culture VOC release data with healthy cell culture VOC release data prior to MCD to identify them. After step 518, the method proceeds to steps 522 and 524.

[0151] In step 520, target cell culture VOC release data is compared with healthy cell culture VOC release data after MCD to identify them. Referring to Figure 1, processor 106 compares target cell culture VOC release data after MCD with healthy cell culture VOC release data to identify them. After step 520, the method proceeds to steps 522 and 524.

[0152] In step 522, pre-MCD and post-MCD target cell VOC profiles are created by comparing pre-MCD target cell culture VOC release data with post-MCD target cell culture VOC release data. Referring to Figure 1, processor 106 creates pre-MCD and post-MCD target cell VOC profiles by comparing pre-MCD target cell culture VOC release data with post-MCD target cell culture VOC release data. After step 522, the method proceeds to step 526.

[0153] In step 524, pre-MCD healthy cell VOC profiles and post-MCD healthy cell VOC profiles are created by comparing pre-MCD healthy cell culture VOC release data with post-MCD healthy cell culture VOC release data. Referring to Figure 1, processor 106 creates pre-MCD healthy cell VOC profiles and post-MCD healthy cell VOC profiles by comparing pre-MCD healthy cell culture VOC release data with post-MCD healthy cell culture VOC release data. After step 524, the method proceeds to step 526.

[0154] In step 526, the predicted pre-MCD target cell VOC profiles, predicted post-MCD target cell VOC profiles, predicted pre-MCD healthy cell VOC profiles, and predicted post-MCD healthy cell VOC profiles are determined. The predicted pre-MCD target cell VOC profiles and predicted post-MCD target cell VOC profiles are determined by predicting the VOC concentration levels in respiration and body fluids from the pre-MCD target cell VOC profiles and predicted post-MCD healthy cell VOC profiles. The VOC concentration levels are predicted using diffusion models such as the Farhi equation f or a modified Farhi model, both of which are described in further detail below. Since the pre-MCD target cell VOC profile and the post-MCD VOC profile are associated with selected gene mutations, the predicted pre-MCD target cell VOC profile and the predicted post-MCD target cell VOC profile are also associated with the same gene mutations. Referring to Figure 1, processor 106 determines the predicted VOC concentration levels in respiration and body fluids from the pre-MCD target cell VOC profile, the post-MCD target cell VOC profile, the pre-MCD healthy cell VOC profile, and the post-MCD healthy cell VOC profile. After step 526, the method proceeds to step 528.

[0155] In step 528, a dynamic differential VOC profile is created from the predicted pre-MCD target cell VOC profile, the predicted post-MCD target cell VOC profile, the predicted pre-MCD healthy cell VOC profile, the predicted post-MCD healthy cell VOC profile, and respiratory and fluid VOC release data. This dynamic differential VOC profile is created by minimizing the error between the predicted pre-MCD target cell VOC profile, the post-MCD target cell VOC profile, and the respiratory and fluid sample VOC release data. Since the predicted pre-MCD target cell VOC profile and post-MCD target cell VOC profile are associated with corresponding gene mutations, the dynamic differential VOC profile is also associated with those gene mutations. Referring to Figure 1, processor 106 determines the dynamic differential VOC profile. After step 528, the method proceeds to step 530.

[0156] In step 530, the dynamic differential VOC profile is saved to the database. Referring to Figure 1, processor 106 saves the dynamic differential VOC profile to database 104.

[0157] Refer here to Figures 10A, 10B, and 10C, which are schematic examples of a method for identifying an individual treatment-resistant VOC profile for a selected treatment, which is effective in another embodiment of the disclosed technology. In step 550, the individual treatment-resistant VOC profile is determined for an individual according to the method described below herein, in conjunction with, for example, Figures 13A, 13B, 13C, 13D, and 13E below herein. Following step 550, the method proceeds to step 552.

[0158] In step 552, the target cells of the individual to be treated are cultured. After step 552, the method proceeds to step 554.

[0159] In step 554, VOC release data related to the target cell culture is acquired before treatment. Referring to Figure 1, the analyzer 102 acquires VOC release data related to the target cell culture before treatment. After step 554, the method proceeds to steps 556 and 570.

[0160] In step 556, at least one selected treatment is applied to the target cell culture. If the selected treatment or multiple treatments include more than one drug and / or treatment, these selected treatments are applied separately and together to different sets of target cell cultures. Following step 556, the method proceeds to steps 558 and 562.

[0161] In step 558, VOC release data from the target cell culture is obtained after at least one selected treatment has been applied to the target cell culture. Referring to Figure 1, the analyzer 102 obtains VOC release data from the target cell culture after at least one selected treatment. Following step 558, the method proceeds to steps 559 and 568.

[0162] In step 559, pre-treatment target cell VOC profiles and post-treatment target cell VOC profiles are created by separately comparing pre-treatment target cell culture VOC release data with post-treatment target cell culture VOC release data for at least one treatment. Referring to Figure 1, processor 106 creates pre-treatment target cell VOC profiles and post-treatment target cell VOC profiles by comparing pre-treatment target cell culture VOC release data with post-treatment target cell culture VOC release data. After step 559, the method proceeds to step 560.

[0163] In step 560, the identification of treatment-resistant mutations by comparing the pre-treatment target cell VOC profile and the post-treatment target cell VOC profile is compared to a conserved predicted complex treatment-resistant VOC profile (e.g., a profile determined according to the method described earlier herein in conjunction with Figures 7A and 7B). Referring to Figure 1, process 106 compares the pre-treatment target cell VOC profile and the post-treatment target cell VOC profile to a conserved predicted complex treatment-resistant VOC profile in order to identify treatment-resistant mutations. Following step 560, the method proceeds to step 562.

[0164] In step 562, target cells identified as generating treatment-resistant mutations in the corresponding post-treatment target cell VOC profile, and which do not appear in the conserved predicted complex treatment-resistant VOC profile, are cultured separately to generate a new target cell culture. Following step 562, the method proceeds to step 566.

[0165] In step 564, separately cultured target cells exhibiting treatment-resistant mutations (i.e., those identified in the culture but not identified when the post-treatment target cell VOC profile is compared to a conserved, predicted complex treatment-resistant VOC profile) undergo gene sequencing and molecular identification to identify and classify the treatment-resistant mutations. Following step 564, the method proceeds to step 566.

[0166] In step 566, VOC release data from the new target cell culture is acquired. Referring to Figure 1, the analyzer 102 acquires VOC release data from the new target cell culture. After step 566, the method proceeds to step 568.

[0167] In step 568, the new post-treatment target cell VOC profile is determined by comparing the pre-treatment target cell culture VOC release data with the new post-treatment target cell culture VOC release data. Referring to Figure 1, processor 106 determines the new post-treatment target cell VOC profile by comparing the pre-treatment target cell culture VOC release data with the new post-treatment target cell culture VOC release data. After step 568, the method proceeds to step 570.

[0168] In step 570, a complex personalized treatment resistance VOC profile is determined by comparing VOC release data obtained from the target cell culture before treatment with the pre-treatment target cell VOC profile, the post-treatment target cell VOC profile (i.e., related to treatment resistance mutations identified using the conserved predicted complex treatment resistance VOC profile), the new post-treatment target cell VOC profile (i.e., related to mutations identified using gene sequencing and molecular identification), and VOC release data obtained from the new target cell culture (i.e., related to target cells that exhibited treatment resistance). Referring to Figure 1, processor 106 determines the complex personalized treatment resistance VOC profile by comparing VOC release data obtained from the target cell culture before treatment with the pre-treatment target cell VOC profile, the post-treatment target cell VOC profile, the new post-treatment target cell VOC profile, and VOC release data obtained from the new target cell culture. Following step 570, the method proceeds to step 572.

[0169] In step 572, the predicted complex individual treatment-resistant VOC profile is determined by predicting respiratory and fluid VOC concentration levels from the complex individual treatment-resistant VOC profile. VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted complex individual treatment-resistant profile. Following step 572, the method proceeds to step 574.

[0170] In step 574, the dynamic complex treatment resistance VOC profile is determined by using the personalized treatment efficacy VOC profile as a filter for the predicted complex treatment resistance VOC profile, thereby mitigating artifacts unrelated to treatment resistance mutations. Referring to Figure 1, process 106 determines the dynamic complex treatment resistance VOC profile by using the stored personalized treatment efficacy to mitigate artifacts unrelated to treatment resistance mutations in the predicted complex treatment resistance VOC profile. Following step 574, the method proceeds to step 576.

[0171] In step 576, the dynamic, complex, personalized treatment-resistant VOC profile is stored in the database. Referring to Figure 1, processor 106 stores the dynamic, complex, personalized treatment-resistant VOC profile in database 104.

[0172] Uses of saved VOC profiles One application of VOC profiles is to determine whether an individual has oncogenic gene mutations and to further identify which oncogenic gene mutations are active in that individual. For this purpose, respiratory and / or fluid VOC release data is collected from the individual. This VOC release data is then compared to a stored dynamic differential VOC profile (e.g., a dynamic differential VOC profile determined according to the method described in conjunction with Figures 3A, 3B, and 3C). As described above, each of these stored dynamic differential VOC profiles is associated with a respective gene mutation. If a match is detected between the VOC release data and at least one stored dynamic differential VOC profile, the patient is identified as having an oncogenic gene mutation. Furthermore, the gene mutation associated with the dynamic differential VOC profile that best matches the acquired VOC release data is identified as the active gene mutation.

[0173] VOC profiles stored in the database may be used to determine the effectiveness of treatments administered to a patient. Therefore, even during long-term treatments that may include multiple stages (e.g., chemotherapy, radiotherapy, drug therapy), the effectiveness of the treatment may be determined by obtaining VOC emission data from the patient's respiration and / or body fluids before treatment, and by determining which of the dynamic difference or predicted VOC profiles stored in the database matches the obtained VOC emission data. Respiration and / or body fluid VOC emission data are also obtained after at least one selected stage of the selected treatment (i.e., during at least one treatment stage, or at the end of the treatment, or any combination thereof). VOC emission data obtained after at least one selected stage of the selected treatment are also compared with both the stored dynamic difference or predicted VOC profiles and the VOC emission data obtained before treatment to confirm the effectiveness of the treatment administered. If VOC emission data is obtained at more than one stage of treatment, they are compared with each other, with the pre-treatment VOC emission data, and with the stored dynamic difference or predicted VOC profiles. If patient treatment has already begun, the effectiveness of the treatment may be determined by acquiring respiratory and / or fluid VOC emission data from the patient before and after selected stages of treatment. VOC emission data acquired before selected stages of treatment are compared with VOC emission data acquired after selected stages of treatment, and with stored dynamic differential or predicted VOC profiles, to determine the effectiveness of the treatment.

[0174] Furthermore, as mentioned above, MCD is used to determine the VOC profile. As will be further detailed below, the use of MCD in a manner that does not produce VOC artifacts is also used to determine the effectiveness of the treatment. In general, VOCs released by target cells before and after treatment, or after one step of treatment, may differ depending on the patient, the medical condition, and the time of day the VOCs were acquired. For example, VOCs and their concentration levels acquired from a woman during menstruation will differ from VOCs and their concentration levels acquired when the woman is not menstruating. As a further example, a patient may have another medical condition (i.e., a condition other than the one being treated). Therefore, the VOCs released by the target cells of such a patient, and the concentration levels of these VOCs, may differ from those of a patient without another medical condition.

[0175] Since VOCs released by target cells before and after treatment, or after one step of treatment, can vary depending on the patient, the disease state, and the time at which the VOCs are acquired, it is beneficial to obtain information related to the expected VOC release after treatment or one step of treatment (i.e., which VOCs are released and at what concentration). When cells die, their membranes break down. When the membranes break down, VOCs "trapped" within the cells are released. Therefore, if target cells are treated and killed, an increase in the concentration level of "trapped" VOCs within the cells is expected. Inducing MCD in a way that does not produce VOC artifacts in target cells destroys these target cells. Measuring the concentration levels of VOCs released by these destroyed cells, along with the respective genome and medical condition at the time of treatment, provides expected outcomes of treatment for a particular patient. Optimal treatment is expected to destroy all target cells in the patient. Therefore, the concentration values ​​of VOCs associated with MCD (i.e., "trapped" VOCs within the cells) will increase if the treatment is effective. Therefore, comparing the VOC concentration values ​​of target cells obtained after treatment, or one step after treatment, with the VOC concentration values ​​of VOCs obtained after MCD induction provides an indicator of whether the treatment achieved the expected results. As an additional indicator of treatment effectiveness, the VOC concentration values ​​associated with target cells decrease if the treatment is effective. Furthermore, the VOC concentration values ​​associated with healthy cells remain unchanged if the treatment is effective. The treatment is also effective if there is no treatment resistance or mutagenicity, as determined further below.

[0176] Refer here to Figure 11, which is a schematic example of the graph referred to collectively as 580 of three exemplary VOC release data 582, 584, and 586 associated with breast cancer in specific patients, according to further embodiments of the disclosed technology. In the examples presented in Figure 11, VOC release data 582, 584, and 586 relate to specific patients with breast cancer. VOC release data 582 represents the VOC release of selected VOCs from healthy cells after MCD (i.e., either directly to the patient or into a culture, as will be further detailed below). VOC release data 584 represents the VOC release of selected VOCs from target cells before MCD, and VOC release data 586 represents the VOC release of selected VOCs from target cells after MCD. As seen in Figure 11, the VOC referred to as 100 exhibits a higher concentration value in VOC release data 586 (i.e., after MCD) than in VOC release data 584 (i.e., before MCD). Specifically, in the exemplary example shown in Figure 11, the VOC referred to as 100 is 3-methylhexane. Therefore, the optimal treatment in the example of breast cancer in a specific patient related to Graph 580 would involve increasing the concentration values ​​of 3-methylhexane and other VOCs, as shown in Figure 11. Other VOCs measured in Figure 11 include, for example, 2-ethylhexanol, 5-ethyl-3-methyloxane, acetone, ethanol, ethyl acetate, ethylbenzene, isononane, isoprene, nonanal, styrene, toluene, and undecane.

[0177] Refer now to Figure 12, which is a schematic example of a method for determining the effectiveness of a treatment, which will be in effect in further embodiments of the disclosed technology.

[0178] In step 600, respiratory and / or body fluid samples are obtained from the patient at least one step prior to the procedure. After step 600, the method proceeds to step 602.

[0179] In step 602, VOC emission data of VOCs released by the respiratory and / or body fluid samples is acquired at least one step before the procedure. Referring to Figure 1, the analyzer 102 acquires VOC emission data of VOCs released by the respiratory and / or body fluid samples. After step 602, the method proceeds to steps 604 and 610.

[0180] In step 604, the stored dynamic differential VOC profile corresponding to the acquired VOC release data (e.g., as determined in conjunction with Figures 4A-4D, 5A-5C, 6A-6D, and 7A-7B) is identified, thereby associating the pathological condition with the VOC release data. Since the identified, stored dynamic differential VOC profile is associated with the corresponding pathological condition, the acquired VOC release data is also associated with the pathological condition (e.g., the pathological condition arising from the corresponding oncogenic gene mutation or pathogen). Generally, as described above, the stored dynamic differential VOC profile consists of at least one target cell VOC profile and may further consist of several additional VOC profiles. Referring to Figure 1, processor 106 identifies the dynamic differential VOC profile corresponding to the acquired VOC release data. After step 604, the method proceeds to step 606.

[0181] In step 606, respiratory and / or fluid samples are taken during and / or after at least one selected stage of the selected procedure. Following step 606, the method proceeds to step 608.

[0182] In step 608, VOC emission data of VOCs released into respiratory and / or body fluid samples are acquired during and / or after at least one selected stage of the selected procedure. Referring to Figure 1, the analyzer 102 acquires VOC emission data of VOCs released into respiratory and / or body fluid samples after the procedure. Following step 608, the method proceeds to step 610.

[0183] In step 610, the effectiveness of the treatment is classified by determining, at least, the VOC concentration values ​​in the identified dynamic differential VOC profile obtained at least one step before the treatment, together with the VOC concentration values ​​in the identified dynamic differential VOC profile obtained during and / or after at least one step of the treatment. For example, if the VOC concentration values ​​in the identified dynamic differential VOC profile during and / or after the treatment are reduced compared to the VOC concentration values ​​in the identified dynamic differential VOC profile before the treatment, the treatment may be classified as successful. Otherwise, the treatment may be classified as unsuccessful. To detect whether new mutations have occurred, VOC emission data obtained before the treatment, and during and / or after the treatment, are compared with other dynamic differential VOC profiles stored in the database. If no new mutations have occurred (i.e., no other dynamic differential profiles are identified in the database), the treatment may be considered successful. Referring to Figure 1, the processor 106 determines the effectiveness of the treatment.

[0184] As described above, VOC profiles may be used to determine the effectiveness of a selected treatment at a selected stage of the treatment, and to detect the development of mutations that could render the treatment ineffective. For example, these VOC profiles may be used to determine the effectiveness of chemotherapy. Furthermore, in the case of cancer treatment, VOC profiles may be used to determine whether cells mutated into a different cancer subtype during the treatment, thereby rendering the treatment ineffective.

[0185] Refer here to Figures 13A–13E, which are schematic examples of methods for determining the effectiveness of a treatment for an individual, which are also effective in other embodiments of the disclosed technology.

[0186] In step 650, at least one respiratory sample and one body fluid sample are taken from the patient before applying the selected procedure. Following step 650, the method proceeds to step 652.

[0187] In step 652, at least one VOC emission data from respiratory and / or fluid samples is acquired prior to at least one selected stage of the selected procedure. The VOC emission data from respiratory and / or fluid samples acquired prior to at least one selected stage of the selected procedure are referred to as pre-procedure patient VOC emission data. Referring to Figure 1, the analyzer 102 acquires at least one emission data from respiratory and / or fluid samples. From step 652, the method proceeds to step 702.

[0188] In step 654, the target cell sample is obtained from the patient. From step 654, the method proceeds to step 658.

[0189] In step 656, a sample of healthy cells of the same type as the target cells is obtained from the patient. From step 656, the method proceeds to step 658.

[0190] In step 658, two sets of target cell cultures (i.e., target cell culture "A" and target cell culture "B") and two sets of healthy cell cultures (i.e., healthy cell culture "A" and healthy cell culture "B") are generated from the target cell sample and the healthy cell sample, respectively. Each set contains at least one culture. From step 658, the method proceeds to step 660.

[0191] In step 660, pre-treatment target cell culture VOC release data related to both target cell culture sets (i.e., target cell culture set "A" and target cell culture set "B") and pre-treatment healthy cell culture VOC release data related to both healthy cell culture sets (i.e., healthy cell culture set "A" and healthy cell culture set "B"). Referring to Figure 1, the analyzer 102 acquires VOC release data related to both target cell cultures and both healthy cell cultures. From step 660, the method proceeds to steps 662, 680, 682, 684, 686 and 698.

[0192] In step 662, the pre-treatment target cell VOC profile and the pre-treatment healthy cell VOC profile are determined by comparing the pre-treatment target cell culture VOC release data of both sets of target cell cultures (i.e., target cell culture set "A" and target cell culture set "B") with the pre-treatment healthy cell culture VOC release data of both sets of healthy cell cultures (i.e., healthy cell culture set "A" and healthy cell culture set "B"). Referring to Figure 1, processor 106 determines the pre-treatment target cell VOC profile and the pre-treatment healthy cell VOC profile. From step 662, the method proceeds to steps 680 and 698.

[0193] In step 664, the selected treatment is applied to the target cells in the first set of target cell cultures (for example, target cell culture set "A"). From step 664, the method proceeds to step 672.

[0194] In step 666, a large amount of cell death is induced in the target cells in a second set of target cell cultures (i.e., target cell culture set "B") in a manner that does not produce residual VOC artifacts (for example, by using rapid freezing or ultraviolet light techniques on the target cell cultures). From step 666, the method proceeds to step 674.

[0195] In step 668, the selected treatment is applied to healthy cells in the first set of healthy cell cultures (i.e., healthy cell culture set "A"). From step 668, the method proceeds to step 676.

[0196] In step 670, a large amount of cell death is induced in healthy cells in a second set of healthy cell cultures (e.g., healthy cell culture set "B") in a manner that does not produce residual VOC artifacts (e.g., by using rapid freezing or ultraviolet light technology on the healthy cell cultures). From step 670, the method proceeds to step 678.

[0197] In step 672, post-treatment target cell culture VOC release data related to target cells in the first set of target cell cultures (i.e., target cell culture set "A") is acquired after the treatment is applied. Referring to Figure 1, the analyzer 102 acquires post-treatment target cell culture VOC release data related to target cells in the first set of target cell cultures after the treatment is applied. Following step 672, the method proceeds to steps 680 and 698.

[0198] In step 674, post-MCD target cell culture VOC release data related to target cells in the second target cell culture (i.e., target cell culture set "B") is acquired after MCD induction. Referring to Figure 1, the analyzer 102 acquires post-MCD target cell culture VOC release data related to target cells in the second target cell culture after MCD induction. After step 674, the method proceeds to step 682.

[0199] In step 676, post-treatment healthy cell culture VOC release data related to cells in the first set of healthy cell cultures (i.e., healthy cell culture set "A") is acquired after the treatment is applied. Referring to Figure 1, the analyzer 102 acquires post-treatment target cell culture VOC release data related to healthy cells in the first set of healthy cell cultures after the treatment is applied. After step 676, the method proceeds to step 684.

[0200] In step 678, post-MCD healthy cell culture VOC release data related to cells in the second set of healthy cell cultures (i.e., healthy cell culture set "B") is acquired after MCD induction. Referring to Figure 1, the analyzer 102 acquires post-MCD healthy cell culture VOC release data related to healthy cells in the second set of healthy cell cultures after MCD induction. After step 678, the method proceeds to step 686.

[0201] In step 680, the treatment-inducible MCD target cell VOC profile is created by comparing pre-treatment target cell culture VOC release data of target cells in a first set of target cell cultures (i.e., target cell culture set "A") with post-treatment target cell culture VOC release data of target cells in the first set of target cell cultures (i.e., target cell culture set "A"). The treatment-inducible MCD target cell VOC profile relates to the VOCs released by target cell culture set "A" when MCD is induced by the selected treatment. Referring to Figure 1, processor 106 creates the treatment-inducible MCD target cell VOC profile by comparing pre-treatment target cell culture VOC release data of target cells in a first set of target cell cultures with post-treatment target cell culture VOC release data of target cells in a first set of target cell cultures. After step 680, the method proceeds to steps 688 and 698.

[0202] In step 682, the MCD target cell VOC profile is created by comparing the pre-treatment target cell culture VOC release data of target cells in a second set of target cell cultures (i.e., target cell culture set "B") with the post-MCD target cell culture VOC release data of target cells in the second set of target cell cultures (i.e., target cell culture set "B"). The MCD target cell VOC profile relates to the VOCs released by target cell culture "B" when MCD is induced in a manner that does not produce residual VOC artifacts (e.g., by utilizing rapid freezing technology or ultraviolet-ultraviolet light technology). Referring to Figure 1, processor 106 creates the MCD target cell VOC profile by comparing the pre-treatment target cell culture VOC release data of target cells in the second set of target cell cultures with the post-MCD target cell culture VOC release data of target cells in the second set of target cell cultures. Following step 682, the method proceeds to steps 690 and 698.

[0203] In procedure 684, the treatment-induced MCD healthy cell VOC profile is created by comparing the pre-treatment healthy cell culture VOC release data of healthy cells in the first set of healthy cell cultures (i.e., healthy cell culture set "A") with the post-treatment healthy cell culture VOC release data of healthy cells in the first set of healthy cell cultures (i.e., healthy cell culture set "A"). The treatment-induced MCD healthy cell VOC profile relates to the VOCs released by healthy cell culture "A" when MCD is induced by the selected treatment. Referring to Figure 1, processor 106 creates the treatment-induced MCD healthy cell VOC profile by comparing the pre-treatment healthy cell culture VOC release data of healthy cells in the first set of healthy cell cultures with the post-treatment healthy cell culture VOC release data of healthy cells in the first set of healthy cell cultures.

[0204] In step 686, the MCD healthy cell VOC profile is created by comparing the pre-treatment healthy cell culture VOC release data of healthy cells in a second set of healthy cell cultures (i.e., healthy cell culture set "B") with the post-MCD healthy cell culture VOC release data of healthy cells in the second set of healthy cell cultures (i.e., healthy cell culture set "B"). The MCD healthy cell VOC profile relates to the VOCs released by healthy cell culture "B" when MCD is induced in a manner that does not produce residual VOC artifacts (e.g., by using rapid freezing technology or ultraviolet-UV light technology). Referring to Figure 1, processor 106 creates the MCD healthy cell VOC profile by comparing the pre-treatment healthy cell culture VOC release data of healthy cells in a second set of healthy cell cultures with the post-MCD healthy cell culture VOC release data of healthy cells in a second set of healthy cell cultures. After step 686, the method proceeds to step 694.

[0205] In step 688, the predicted treatment-inducible MCD target cell VOC profile is determined by predicting the respiratory and / or fluid VOC concentration levels from the treatment-inducible MCD target cell VOC profile. The VOC concentration levels are predicted by utilizing diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted treatment-inducible MCD target cell VOC profile. Following step 688, the method proceeds to step 696.

[0206] In step 690, the predicted MCD target cell VOC profile is determined by predicting the VOC concentration levels in respiration and / or body fluids from the MCD target cell VOC profile. The VOC concentration levels are predicted by utilizing diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted MCD target cell VOC profile. After step 690, the method proceeds to step 696.

[0207] In step 692, the predicted treatment-induced MCD healthy cell VOC profile is determined by predicting the respiratory and / or fluid VOC concentration levels from the treatment-induced MCD healthy cell VOC profile. The VOC concentration levels are predicted by utilizing diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted treatment-induced MCD healthy cell VOC profile. Following step 692, the method proceeds to step 696.

[0208] In step 694, the predicted healthy MCD VOC profile is determined by predicting the respiratory and / or fluid VOC concentration levels from the healthy MCD cell VOC profile. The VOC concentration levels are predicted using diffusion models such as the Farhi equation or a modified Farhi model, both of which are described in further detail below. Referring to Figure 1, processor 106 determines the predicted healthy MCD cell VOC profile. Following step 694, the method proceeds to step 696.

[0209] In step 696, the predicted treatment effect differential VOC profile is determined from the pre-treatment target cell VOC profile, the pre-treatment healthy cell VOC profile, the predicted treatment-inducible MCD healthy cell VOC profile, the predicted treatment-inducible MCD target cell VOC profile, the predicted MCD healthy cell VOC profile, and the predicted MCD target cell VOC profile. Referring to Figure 1, processor 106 determines the predicted treatment effect differential VOC profile. From step 696, the method proceeds to step 702.

[0210] In procedure 698, the dynamic treatment resistance differential VOC profile and the mutation rate differential VOC profile for at least one mutation and / or pathogen are determined. First, the post-treatment target cell culture VOC release data of target cells in target cell culture set "A" is filtered by the MCD target cell VOC profile and the treatment-inducible MCD target cell VOC profile to reduce cell death-related VOCs. The filtered post-treatment target cell culture VOC release data is compared with the pre-treatment target cell VOC profile to determine the dynamic treatment resistance profile. The VOC concentration levels in this treatment resistance profile are an indicator of the portion of cells that survive the treatment and are not mutated.

[0211] The filtered post-treatment target cell culture VOC release data is further filtered by the pre-treatment target cell VOC profile. The twice-filtered post-treatment target cell culture VOC release data is compared with the stored dynamic differential VOC profile (e.g., as determined herein in conjunction with Figures 3A-3C, 5A-5C, and 6A-6D) to determine the mutation rate differential VOC profile or multiple profiles for various mutations and / or pathogens. The VOC concentration levels in these mutation rate differential VOC profiles are indicators of the mutation rate of various mutations and pathogens (i.e., the portion of cells that mutated during treatment and the number of new mutations that appeared after treatment). Referring to Figure 1, processor 106 determines the dynamic treatment resistance profile and mutation rate differential VOC profile or multiple profiles for various mutations and / or pathogens. After step 698, the method proceeds to step 710.

[0212] In step 702, the dynamic treatment effect VOC profile is determined from the predicted treatment effect difference VOC profile and the VOC release data of VOCs released into the respiratory and / or body fluid samples. Referring to Figure 1, the processor 106 determines the dynamic treatment effect VOC profile. After step 702, the method proceeds to step 704.

[0213] In step 704, the selected treatment for the patient is initiated. After step 704, the method proceeds to step 706.

[0214] In step 706, at least one respiratory sample and / or body fluid sample is taken from the patient during and / or after at least one selected stage of the selected procedure. The respiratory sample and / or body fluid sample taken from the patient during and / or after at least one selected stage of the selected procedure are referred to herein as post-procedure patient VOC emission data. Following step 706, the method proceeds to step 708.

[0215] Step 708 is post-treatment patient VOC emission data of VOCs released into respiratory and / or body fluid samples acquired during and / or after at least one selected stage of the selected treatment. Referring to Figure 1, analyzer 102 acquires VOC emission data of VOCs released into respiratory and / or body fluid samples during and / or after at least one selected stage of the selected treatment. Following step 708, the method proceeds to step 710.

[0216] In procedure 710, the effectiveness of at least one selected stage of a selected treatment administered to an individual patient (i.e., the effectiveness of the individualized treatment) is determined from at least VOC emission data of respiratory and / or body fluid samples taken before at least one selected stage of the selected treatment, and VOC emission data of respiratory and / or body fluid samples taken during and / or after at least one selected stage of the selected treatment (i.e., pre-treatment patient VOC emission data and post-treatment patient VOC emission data). To determine the effectiveness of a selected stage of the selected treatment, the following is determined: • VOC concentration values ​​in the pre-treatment target cell VOC profile prior to the selected stage of the selected treatment, from pre-treatment patient VOC release data. • VOC concentration values ​​in the pre-treatment target cell VOC profile during and / or after the selected stage of the selected treatment, from post-treatment patient VOC release data. • VOC concentration values ​​in the predicted treatment-inducible MCD target cell VOC profile prior to the selected stage of the selected treatment, based on pre-treatment patient VOC release data. • VOC concentration values ​​in the predicted treatment-inducible MCD target cell VOC profile during and / or after selected stages of the selected treatment, from post-treatment patient VOC release data. • VOC concentration values ​​in the predicted MCD target cell VOC profile prior to the selected stage of the selected treatment, based on pre-treatment patient VOC release data. • VOC concentration values ​​in the predicted MCD target cell VOC profile during and / or after selected stages of the selected treatment, based on post-treatment patient VOC release data.

[0217] A selected stage of a selected treatment is determined to be effective if the VOC concentration value in the predicted MCD target cell VOC profile during and / or after the selected stage of the treatment, based on treatment patient VOC release data associated with at least one of respiratory and body fluid samples, is greater than the VOC concentration value in the predicted MCD target cell VOC profile prior to the selected stage of the treatment, based on pre-treatment patient VOC release data associated with at least one of respiratory and body fluid samples.

[0218] The treatment is further determined to be effective if the concentration value of VOCs from pre-treatment target cell cultures, associated with the pre-treatment target cell VOC profile, is greater than the concentration value of VOCs from post-treatment target cell cultures, associated with the pre-treatment target cell VOC profile.

[0219] The treatment is further determined to be effective if the VOC concentration value associated with the predicted treatment-inducible MCD target cell VOC profile from post-treatment patient VOC release data associated with at least one of the respiratory and body fluid samples is greater than the VOC concentration value associated with the predicted treatment-inducible MCD target cell VOC profile from pre-treatment patient VOC release data associated with at least one of the respiratory and body fluid samples.

[0220] Furthermore, in order to determine the effectiveness of the selected stage of the selected treatment, the following will also be determined: • VOC concentration values ​​in the pre-treatment healthy cell VOC profile related to healthy cells prior to the selected stage of the selected treatment, from pre-treatment patient VOC emission data. • VOC concentration values ​​in the pre-treatment healthy cell VOC profile during and / or after selected stages of the selected treatment, determined from post-treatment patient VOC release data. • VOC concentration values ​​in the predicted treatment-induced MCD healthy cell VOC profile prior to the selected stage of the selected treatment, based on pre-treatment patient VOC release data. • VOC concentration values ​​in the predicted treatment-induced MCD healthy cell VOC profile during and / or after selected stages of the selected treatment, from post-treatment patient VOC release data. • VOC concentration values ​​in the predicted MCD healthy cell VOC profile prior to the selected stage of the selected treatment, based on pre-treatment patient VOC release data. • VOC concentration values ​​in the predicted MCD healthy cell VOC profile during and / or after selected stages of the selected treatment, based on post-treatment patient VOC release data.

[0221] The treatment is determined to be effective if the concentration values ​​from post-treatment patient VOC emission data associated with at least one of the respiratory and fluid samples of the VOC in the predicted treatment-inducible MCD healthy VOC remain unchanged from the concentration values ​​from pre-treatment patient VOC emission data associated with at least one of the respiratory and fluid samples of the VOC in the predicted treatment-inducible MCD healthy VOC (for example, if the difference between those values ​​is within a predetermined threshold).

[0222] The effectiveness of the selected stage of the selected treatment may further be determined from the VOC concentration values ​​in the dynamic patient treatment resistance differential VOC profile and the mutational differential VOC profile before, during, and / or after the selected stage of the selected treatment. The VOC concentration values ​​in the pre-treatment dynamic patient treatment resistance differential VOC profile and the mutational differential VOC profile are determined from pre-treatment patient VOC release data.

[0223] The VOC concentration values ​​in the dynamic patient treatment resistance differential VOC profile and the mutation rate differential VOC profile during and / or after the selected stages of the selected treatment are determined from post-treatment patient VOC release data. The selected stage of the selected treatment is determined to be effective if the VOC concentration values ​​in the dynamic patient treatment resistance differential VOC profile remain unchanged before, during, and / or after the selected stage of the selected treatment. The selected stage of the selected treatment is also determined to be effective if the VOC concentration values ​​in the mutation rate differential VOC profile during and / or after at least one selected stage of the selected treatment remain unchanged compared to the VOC concentration values ​​before the selected stage of the selected treatment.

[0224] Furthermore, the selected stage of the selected treatment is determined to be effective if no new mutations are identified. New mutations or multiple mutations are identified by comparing post-treatment patient VOC release data with stored dynamic differential VOC profiles. Before attempting to identify new mutations or multiple mutations, post-treatment patient VOC release data is filtered by pre-treatment target cell VOC profiles, treatment-inducible MCD target cell VOC profiles, and MCD target cell VOC profiles, thereby reducing VOC-related information associated with pre-treatment target cell VOC profiles, treatment-inducible MCD target cell VOC profiles, and MCD target cell VOC profiles.

[0225] Referring to Figure 1, the processor 106 determines the effectiveness of at least one selected stage of the selected treatment administered to the individual patient.

[0226] Another use of stored VOC profiles is to identify active and inactive mutations in patients and / or cultures. Refer here to Figure 14, which is a schematic example of a method for identifying active and inactive mutations in patients and / or cultures, which will come into effect in further embodiments of the disclosed technology.

[0227] In step 750, at least one respiratory sample and / or body fluid sample is obtained from the patient. Following step 750, the method proceeds to step 752.

[0228] In step 752, VOC emission data of VOCs released into respiratory and / or body fluid samples is acquired. Referring to Figure 1, the analyzer 102 acquires VOC emission data of VOCs released into respiratory and / or body fluid samples. After step 752, the method proceeds to step 766.

[0229] In step 754, target and healthy cell samples are obtained from the patient. Following step 754, the method proceeds to step 756.

[0230] In step 756, target cells and healthy cells are genetically sequenced, molecularly identified, and genetic mutations are identified. After step 756, the method proceeds to step 758.

[0231] In step 758, target and healthy cell samples are cultured. After step 758, the method proceeds to step 760.

[0232] In step 760, VOC release data related to VOCs released by the target and healthy cell cultures is acquired. Referring to Figure 1, the analyzer 102 acquires VOC release data related to VOCs released by the target and healthy cell cultures. After step 760, the method proceeds to step 762.

[0233] In step 762, the target cell VOC profile is determined by comparing VOC release data from the target cell culture with VOC release data from a healthy cell culture. Referring to Figure 1, processor 106 determines the target cell VOC profile by comparing VOC release data from the target cell culture with that of a healthy cell culture. After step 762, the method proceeds to step 764.

[0234] In step 764, active mutations or multiple mutations in the target cell culture are determined by comparing the target cell VOC profile with a stored dynamic differential VOC profile determined as described earlier in this specification, in conjunction with Figures 3A–3C, 5A–5C, 7A–7B, 8A–8B, and 10A–10C. Referring to Figure 1, process 106 determines active mutations or multiple mutations in the target cell culture from a list of mutations received by gene sequencing by comparing the target cell VOC profile with VOC profiles in database 104. Following step 764, the method proceeds to step 766.

[0235] In procedure 766, the patient's active mutation or multiple mutations are determined by comparing the patient's respiratory and / or fluid VOC release data with the identified dynamic differential VOC profile or multiple profiles (i.e., the identified dynamic differential VOC profile or multiple profiles relate to the dynamic differential VOC profile identified by comparing the target cell VOC profile with the stored dynamic differential VOC profile). Furthermore, the respiratory and / or fluid VOC release data is filtered by the target cell VOC profile. In addition, the filtered respiratory and / or fluid VOC release data is compared with the stored dynamic differential VOC profile determined as described earlier in this specification, in conjunction with Figures 3A-3C, 5A-5C, 7A-7B, 8A-8B and 10A-10C. Referring to Figure 1, processor 106 determines the active mutation or multiple mutations in the patient.

[0236] Another use of the stored VOC profile is to determine the optimal treatment for a patient. Refer here to Figures 15A and 15B, which are schematic examples of a method for determining the optimal treatment for a patient, which is in effect by another embodiment of the disclosed technology.

[0237] In step 800, target and healthy cell samples are obtained from the patient. Following step 800, the method proceeds to step 802.

[0238] In step 802, two sets of target cell cultures (i.e., target cell culture "A" and target cell culture "B") and two sets of healthy cell cultures (i.e., healthy cell culture "A" and healthy cell culture "B") are generated from the target cell sample and the healthy cell sample, respectively. Each set contains at least one culture. After step 802, the method proceeds to step 804.

[0239] In step 804, pre-treatment target cell culture VOC release data relating to VOCs released by target cells in both sets of target cell cultures (i.e., target cell culture set "A" and target cell culture set "B") and pre-treatment healthy cell culture VOC release data relating to VOCs released by healthy cells in both sets of healthy cell cultures (i.e., healthy cell culture set "A" and healthy cell culture set "B"). Referring to Figure 1, the analyzer 102 acquires pre-treatment target cell culture VOC release data relating to VOCs released by target cells from target cells in both sets of target cell cultures and pre-treatment healthy cell culture VOC release data relating to VOCs released by healthy cells from healthy cells in both sets of healthy cell cultures. After step 804, the method proceeds to steps 806, 826, 828 and 830.

[0240] In step 806, pre-treatment target cell VOC profiles and pre-treatment healthy cell VOC profiles are created by comparing pre-treatment target cell culture VOC release data from both target cell culture sets (i.e., target cell culture set "A" and target cell culture set "B") with pre-treatment healthy cell culture VOC release data from both healthy cell culture sets (i.e., healthy cell culture set "A" and healthy cell culture set "B"). Referring to Figure 1, processor 106 determines pre-treatment target cell VOC profiles and pre-treatment healthy cell VOC profiles by comparing target cell culture VOC release data from target cells in both target cell culture sets with healthy cell culture VOC release data from healthy cells in both healthy cell culture sets. After step 806, the method proceeds to steps 808, 810, 820, 826, 828, and 830.

[0241] In step 808, a large amount of cell death is induced in the target cells in the first set of target cell cultures (i.e., target cell culture set "A") using a method that does not produce VOC artifacts (e.g., by using rapid freezing or UV light technology). From step 808, the method proceeds to step 812.

[0242] In step 810, a large amount of cell death is induced in healthy cells within the first set of healthy cell cultures (i.e., healthy cell culture set "A") in a manner that does not produce VOC artifacts (e.g., by using rapid freezing or UV light technology) and does not produce residual VOC artifacts. From step 810, the method proceeds to step 814.

[0243] In step 812, post-MCD target cell culture VOC release data related to target cells in the first set of target cell cultures (i.e., target cell culture set "A") is acquired. Referring to Figure 1, after induction of MCD, the analyzer 102 acquires post-MCD target cell culture VOC release data related to target cells in target cell culture "A". After step 812, the method proceeds to step 816.

[0244] In step 814, post-MCD healthy cell culture VOC release data related to healthy cells in the first set of healthy cell cultures (i.e., healthy cell culture set "A") is acquired. Referring to Figure 1, after induction of MCD, the analyzer 102 acquires post-MCD healthy cell culture VOC release data related to healthy cells in healthy cell culture "A". After step 814, the method proceeds to step 818.

[0245] In step 816, the MCD target cell VOC profile is created by comparing pre-treatment target cell culture VOC release data with post-MCD target cell culture VOC release data. The MCD target cell VOC profile relates to the VOCs released by target cells in a first set of target cell cultures (i.e., target cell culture set "A") when MCD is induced in a manner that does not produce residual VOC artifacts (e.g., by utilizing rapid freezing or UV light technology). Referring to Figure 1, processor 106 creates the MCD target cell VOC profile by comparing pre-treatment target cell culture VOC release data of target cells in the first set of target cell cultures with post-MCD target cell culture VOC release data of target cells in the first set of target cell cultures. Following step 816, the method proceeds to steps 826 and 828.

[0246] In step 818, the MCD healthy cell VOC profile is created by comparing the pre-treatment healthy cell culture VOC emission data of the healthy cells in the first set of healthy cell cultures (i.e., healthy cell culture set "A") with the post-MCD healthy cell culture VOC emission data of the healthy cells in the first set of healthy cell cultures (i.e., healthy cell culture set "A"). The MCD healthy cell VOC profile relates to the VOCs emitted by healthy cell culture "A" when MCD is induced in a way that does not generate residual VOC artifacts (e.g., by using rapid freezing technology or UV light technology). Referring to FIG. 1, the processor 106 creates the MCD healthy cell VOC profile by comparing the pre-treatment healthy cell culture VOC emission data of the healthy cells in the first set of healthy cell cultures with the post-MCD healthy cell culture VOC emission data of the healthy cells in the first set of healthy cell cultures. After step 818, the method proceeds to step 830.

[0247] In step 820, at least one selected treatment is applied to the target cells in the second set of target cell cultures (i.e., target cell culture set "B") and the healthy cells in the second set of healthy cell cultures (i.e., healthy cell culture set "B"). If more than one treatment is applied, the treatments are used as comparative references to each other. From step 820, the method proceeds to step 824.

[0248] In step 822, for each selected treatment, post-treatment target cell culture VOC emission data related to the target cells in the second set of target cell cultures (i.e., target cell culture set "B") is obtained. Referring to FIG. 1, the analyzer 102 obtains the post-treatment target cell culture VOC emission data related to the target cells in the second set of target cell cultures for each selected treatment. After step 822, the method proceeds to steps 826 and 828.

[0249] In step 824, for each selected treatment, post-treatment healthy cell culture VOC release data related to healthy cells in the second set of healthy cell cultures (i.e., healthy cell culture set "B") is acquired after the application of the selected treatment. Referring to Figure 1, the analyzer 102 acquires post-treatment healthy cell culture VOC release data related to healthy cells in the second set of healthy cell cultures for each selected treatment after the application of the selected treatment. After step 824, the method proceeds to step 830.

[0250] In procedure 826, treatment resistance and target cell mutation rates are determined for each selected treatment. Both treatment resistance and target cell mutation rates are used to determine whether the selected treatment is effective or not (i.e., the treatment effect). First, the VOC concentration values ​​in the pre-treatment target cell VOC profile are determined from the pre-treatment target cell culture VOC release data related to target cell culture "B" (i.e., the relevant VOC concentration levels before treatment are determined). Then, the post-treatment target cell culture VOC release data is filtered with the MCD target cell VOC profile to reduce cell death-related VOCs. The MCD concentration values ​​in the pre-treatment target cell VOC profile are determined from the filtered post-treatment target cell culture VOC release data (i.e., the relevant VOC concentration levels during and / or after treatment are determined). The treatment resistance of the selected treatment is determined by comparing the VOC concentration values ​​in the pre-treatment target cell VOC profile before the selected treatment with the VOC concentration values ​​in the pre-treatment target cell VOC profile during and / or after the selected treatment. The variation in the concentration values ​​of the relevant VOCs is an indicator of the portion of cells that survived the selected treatment and did not mutate. The selected treatment is determined to be effective if the VOC concentration value in the pre-treatment target cell VOC profile is reduced. The selected treatment may be considered most effective if the VOC concentration value in the pre-treatment target cell VOC profile is zero.

[0251] The filtered post-treatment target cell culture VOC release data is further filtered by the pre-treatment target cell VOC profile. Mutagenesis capacity is determined by comparing the twice-filtered post-treatment target cell culture VOC release data with a preserved dynamic differential VOC profile (e.g., as determined earlier in this specification in conjunction with Figures 3A-3C, 5A-5C, and 6A-6D), thereby identifying the preserved dynamic differential VOC profile or multiple profiles corresponding to the twice-filtered post-treatment target cell culture VOC release data. The VOC concentration values ​​in the identified preserved dynamic differential VOC profile or multiple profiles are then determined from the twice-filtered post-treatment target cell culture VOC release data. The concentration levels of the identified preserved dynamic differential VOC profiles are indicators of the mutation rates of various mutations and pathogens (i.e., the portion of cells that mutated during the selected treatment, and the number of new mutations that appeared after the selected treatment). The selected treatment is determined to be most effective if no new preserved dynamic differential VOC profiles are identified from the twice-filtered post-treatment target cell culture VOC release data. If more than one selected treatment is available, the optimal treatment may be determined by selecting the treatment that exhibits the greatest variation in VOC concentration levels in the pre-treatment target cell VOC profile, showing a reduction in the pre-treatment target cell VOC profile in the post-treatment target cell culture VOC release data (i.e., optimal efficiency is shown when the pre-treatment target cell VOC profile is no longer identified in the post-treatment target cell culture VOC release data). If the stored dynamic fibrillation VOC profiles are not identified in the post-treatment target cell culture VOC release data, or if they are identified in all selected treatments, the treatment that exhibits the smallest variation in VOC concentration levels in the identified stored dynamic differential VOC profile in the post-treatment target cell culture VOC release data may be determined as the optimal treatment. Referring to Figure 1, processor 106 determines the treatment resistance and mutation rate. After step 826, the method proceeds to step 832.

[0252] In step 828, the effect of each selected treatment on the target cells is determined. To determine the effect of the selected treatments, the VOC concentration values ​​in the pre-treatment target cell VOC profile and the VOC concentration values ​​in the pre-treatment MCD target cell VOC profile are determined. For this purpose, the VOC concentration level in the pre-treatment target cell VOC profile is determined from the pre-treatment target cell culture VOC release data obtained from target cell culture "B". Furthermore, the VOC concentration level in the MCD target cell VOC profile is determined from the pre-treatment target cell culture VOC release data obtained from target cell culture "A". In addition, the VOC concentration value in the pre-treatment target cell VOC profile is determined from the post-treatment target cell culture VOC release data obtained from target cell culture "B", and the VOC concentration value in the MCD target cell VOC profile is determined from the post-treatment target cell culture VOC release data obtained from target cell culture "A". The selected treatments are, (a) When the concentration value of VOCs associated with the pre-treatment target cell VOC profile, based on VOC release data from the pre-treatment target cell culture, is greater than the concentration value of VOCs associated with the pre-treatment target cell VOC profile, based on VOC release data from the post-treatment target cell culture; and (b) If the concentration of VOCs associated with the MCD target cell VOC profile released by the post-treatment target cell culture, based on the VOC release data of the post-treatment target cell culture, is greater than the concentration of VOCs associated with the MCD target cell VOC profile, based on the VOC release data of the pre-treatment target cell culture, It is determined to be effective.

[0253] In another example, the ratio of the concentration of VOCs associated with the MCD VOC profile released by the post-treatment target cell culture from the VOC release data of the post-treatment target cell culture to the concentration of VOCs associated with the pre-treatment target cell VOC profile from the VOC release data of the post-treatment target cell culture is greater than the ratio of the concentration of VOCs associated with the MCD VOC profile from the VOC release data of the pre-treatment target cell culture to the concentration of VOCs associated with the pre-treatment target cell VOC profile from the VOC release data of the pre-treatment target cell culture.

[0254] If more than one selected treatment is available, the optimal treatment is determined from the selected treatments by determining the treatment that shows the greatest increase in VOC concentration levels in the MCD target cell VOC profile and the greatest decrease in VOC concentration levels in the pre-treatment target cell VOC profile. Referring to Figure 1, processor 106 determines the effect of the selected treatment. After step 828, the method proceeds to step 832.

[0255] In procedure 830, the effect of each selected treatment on healthy cells is determined. To determine the effect of the selected treatment, the VOC concentration values ​​in the pre-treatment healthy cell VOC profile and the MCD healthy cell VOC profile before and after the selected treatment are determined. For this purpose, the VOC concentration value in the pre-treatment healthy cell VOC profile is determined from the pre-treatment healthy cell culture VOC release data obtained from healthy cell culture "B". Furthermore, the VOC concentration value in the MCD healthy cell VOC profile is determined from the pre-treatment healthy cell culture VOC release data obtained from healthy cell culture "A". In addition, the VOC concentration value in the pre-treatment healthy cell VOC profile is determined from the post-treatment healthy cell culture VOC release data obtained from healthy cell culture "B", and the VOC concentration value in the MCD healthy cell VOC profile is determined from the post-treatment healthy cell culture VOC release data obtained from healthy cell culture "A". The selected treatment is, (a) When the concentration values ​​of VOCs in the pre-treatment healthy cell VOC profile from post-treatment healthy cell culture VOC release data remain unchanged compared to the concentration values ​​of VOCs associated with the pre-treatment healthy cell VOC profile from pre-treatment healthy cell culture VOC release data; and (b) If the concentration values ​​of VOCs associated with the MCD healthy cell VOC profile from post-treatment healthy cell culture VOC release data remain unchanged compared to the concentration values ​​of VOCs in the MCD healthy cell VOC profile from pre-treatment healthy cell culture VOC release data, It is determined to be effective.

[0256] In another example, the concentration values ​​of VOCs associated with the MCD healthy cell VOC profile from post-treatment healthy cell culture VOC release data, and the concentration values ​​of VOCs in the pre-treatment healthy cell VOC profile from post-treatment healthy cell culture VOC release data, are equal to the concentration values ​​of VOCs in the MCD healthy cell VOC profile from pre-treatment healthy cell culture VOC release data, and the concentration values ​​of VOCs associated with the pre-treatment healthy cell VOC profile from pre-treatment healthy cell culture VOC release data.

[0257] If more than one selected treatment is available, the optimal treatment is determined from the selected treatments by determining the treatment that exhibited the least variation in the MCD healthy cell VOC profile and the VOC concentration levels in the pre-treatment healthy cell VOC profile. Referring to Figure 1, processor 106 determines the effect of the selected treatment. After step 830, the method proceeds to step 832.

[0258] In procedure 832, the compound treatment response is determined for each selected treatment based on four treatment effect parameters (i.e., the effect of the treatment on target cells, the effect of the treatment on healthy cells, treatment resistance, and mutation rate potential). If the selected treatment is determined to be effective in all four of these parameters, the compound treatment response is considered positive, and this treatment may be considered optimal. If more than one selected treatment is available, the treatment with the best positive response in all four treatment effect parameters may be determined to be the optimal treatment among the selected treatments. Referring to Figure 1, process 106 determines the compound treatment response to each selected treatment based on the four treatment effect parameters.

[0259] Diffusion model As described above, VOC concentration levels in respiration and body fluids are predicted by utilizing a diffusion model of VOCs from target and healthy cells to body fluids and respiration. Specifically, a dynamic differential VOC profile is determined from the predicted target cell VOC profile, the predicted healthy cell VOC profile, and, in some cases, additional predicted profiles. The various predicted profiles are determined by predicting the VOC concentration levels of the relevant VOCs in respiration and body fluid VOC release data based on the metabolic rates and production rates of target, healthy, and control cells in vitro. During the determination of the dynamic differential VOC profile, respiration and body fluid sample VOC release data are also used to minimize the possible tolerance between the various predicted profiles and the actual results. One such model relating alveolar VOC concentrations (i.e., during respiration) to their basic blood concentrations is the Farhi equation, which takes the following form:

number

[0260] The standard Farhi equation, as explained earlier, only considers possible VOC concentrations within the alveolar (i.e., lower lung) compartment. This model can lead to erroneous results.

[0261] The model described herein extends the standard Farhi equations (from a two-part model to a three-part model, i.e., an extended Farhi model). Referring here to Figure 16, it is a schematic example of the extended Farhi, referred to as 840 as a whole, which will come into effect in further embodiments of the disclosed art. As shown in Figure 16, the extended Farhi model 840 includes three parts: the bronchial part 842, the alveolar part 844, and the body part 846. The bronchial part 842 and the alveolar part 844 relate to the lungs. The body part 844 (i.e., metabolism and production) combines the blood and tissue parts (i.e., any form of target cells, healthy cells, or control cells in the body) into one part of an effective volume V(~). The blood and tissue parts of the body are assumed to be in equilibrium, and therefore

number

[0262] The use of the three - compartment model may be employed to create a more accurate dynamic differential VOC profile. The three - compartment model incorporates the effects of the upper airway (bronchi) and exhaled VOC concentrations to address known issues associated with the standard Farhi equations. The three - compartment model detailed below in this specification also accounts for the influence of inhaled VOCs (environmental pollution sources) on exhaled breath concentrations for VOCs with higher Henry's constants. The bronchial compartment 842 is considered a distinct compartment separated into a gas phase and a mucosa that is assumed to inherit the physical properties of water and act as a reservoir. A portion of the VOCs dissolved in this layer migrates to the bronchial environment, whereby it is presumed that the major fraction of the associated venous drainage pathway converges into the pulmonary vein via the post - capillary anastomosis. The amount of VOCs transported to the bronchial compartment 842 at time "t" via exhalation and inhalation is therefore,

Number

Number

Number

[0263] The decrease in solubility in mucosa with increasing temperature can be described over the ambient temperature range using a Van't Hoff-type equation:

number

number

[0264] The exchange between bronchial segment 842 and alveolar segment 844 is modeled as a diffusion process:

number

[0265] The total material balance for bronchial category 842 is:

number

[0266] The material balance for alveolar segment 844 is:

number

number

[0267] Combining the three linear difference equations (9), (10), and (11), we get VOC(m tot The total mass change of ) is obtained:

number

[0268] VOC filter In the above, all VOC release data obtained from respiration and / or body fluids and / or cell cultures, as well as comparisons between them, may be used to detect common VOCs among them. These common VOCs may be used to define a VOC filter. Such a VOC filter may be used to filter out common VOCs when obtaining new VOC release data from respiration and / or body fluids and / or cell cultures. Such a VOC filter may also be used to filter out common VOCs in a VOC profile. The filter is constructed by comparing healthy VOC release data from healthy cells of the same type from multiple patients and identifying VOCs associated with the normal activity of these healthy cells. A diffusion equation (e.g., the Farhi equation discussed earlier) is used to generate a range of VOC concentration levels associated with normal activity. These VOC concentration levels associated with normal activity are used to identify abnormal concentrations by filtering out these VOCs from the VOC release data.

[0269] Comparison of VOC profiles In the embodiments described above, VOC profiles are compared with each other. According to one alternative method, VOC profiles are compared by comparing the polygons defined by the VOC profiles. The VOC profiles may be considered as a two-dimensional Euclidean space where the horizontal axis is defined by the VOC and the vertical axis is defined by the concentration level. In the VOC profile space, a polygon is defined by a reference point and the peak value of the selected VOC. The reference point may be the zero coordinate of the two-dimensional space. When comparing two VOC profiles, the same polygon or more polygons are defined in the two VOC profiles (i.e., using the same reference point and the same selected VOC). These two polygons are then compared, for example, by the turning function between the two polygons. p They are compared to each other by using distance.

[0270] Increase in VOC concentration When determining VOC emission data in respiratory samples, it may be desirable to increase the VOCs in the lungs before collecting the breath. For this purpose, the patient is asked to exhale to their maximum capacity before sample collection. The patient then inhales, holds their breath for a predetermined period (e.g., 5, 10, 20, 30 seconds, 1 minute), and exhales the respiratory sample into the collector. The duration of breath-holding for each patient is measured to correlate respiratory samples obtained with different breath-holding durations.

[0271] Furthermore, the previously detailed methods for increasing the VOC concentration levels during respiration before sampling may be combined with methods known in the art for sampling a quantified amount of air exhaled from a selected part of the lung (e.g., bronchi, alveoli, or whole lung), thereby raising the VOC concentration levels of VOCs typically found during respiration below the detection limit of an analyzer to levels at which they can be detected, identified, and quantified. Refer here to Figures 17A and 17B, which are schematic examples of methods for increasing VOC concentrations before sampling and for sampling a quantified amount of air from a selected part of the lung, which are effective in other embodiments of the disclosed technology.

[0272] In step 850, the part of the lung from which air is drawn and the volume of air to be drawn are selected. After step 850, the method proceeds to step 852.

[0273] In step 852, the inhalation flow rate, exhalation rate, and carbon dioxide (CO2) concentration level are measured while the patient is breathing normally over a period of time (e.g., several breaths). The inhalation and exhalation flow rates, as well as the CO2 concentration, may be measured using a spirometer that includes a flow meter and a CO2 sensor. After step 852, the method proceeds to step 854.

[0274] In step 854, the inhalation flow rate, exhalation flow rate, and CO2 concentration level are measured when the patient exhales and inhales to their maximum capacity. Following step 854, the method proceeds to step 856.

[0275] In step 856, a breathing pattern is determined that distinguishes between the bronchial portion and the alveolar portion of the exhaled breath. Following step 856, the method proceeds to steps 858 and 860.

[0276] In step 858, a respiratory sample is taken when the patient exhales to maximum capacity, inhales to maximum capacity, and then exhales to maximum capacity after holding their breath for at least a predetermined period. Each respiratory sample is associated with a specific time point in the determined breathing pattern. Following step 858, the method proceeds to step 860.

[0277] In step 860, a respiratory sample corresponding to a selected portion of the lung is selected according to the respiratory pattern and the associated time of the respiratory sample. Following step 858, the method proceeds to step 862.

[0278] In step 862, VOC emission data is obtained from the selected respiratory sample. Referring to Figure 1, the analyzer 102 obtains VOC emission data from the selected respiratory sample. After step 862, the method proceeds to step 864.

[0279] In procedure 864, the duration of respiratory arrest in the patient is measured, and adjustments (i.e., standardization) that may be required to correlate different respiratory arrest durations with the resulting increases in concentration levels in the acquired samples are determined.

[0280] In some cases, the selected volume of air cannot be obtained in a single repetition. Therefore, procedure 858 may be repeated until the selected volume of air is obtained, with the patient holding their breath for the same duration in each repetition.

[0281] Those skilled in the art will notice that the disclosed technology is not limited to what is indicated and described in detail further herein. Rather, the scope of the disclosed technology is defined solely by the following claims.

Claims

1. A method for associating volatile organic compound (VOC) release with at least one target cell type before and after treatment in a selected population, comprising the following steps: For each selected target cell type, a procedure (212) to determine the gene sequence of at least one of the target cell samples (202) obtained from multiple patients; A procedure (212) for determining the gene sequence of at least one healthy cell sample of healthy cells collected (204) from the plurality of patients with respect to each of the selected target cell types, wherein the healthy cell sample is of the same cell type as the target cell sample; A procedure (212) for determining the gene sequence of at least one control cell sample of control cells taken from a control group (206) with respect to each selected target cell type, wherein the control cell sample is of the same cell type as the target cell sample; A procedure (212) for classifying each of the at least one gene sequences according to molecular classification; A procedure (216) for culturing the target cell sample, the healthy cell sample, and the control cell sample to produce target cell cultures, healthy cell cultures, and control cell cultures, respectively; A procedure (218) for obtaining VOC release data for the target cell culture, the healthy cell culture, and the control cell culture before inducing mass cell death (MCD); A procedure (220) for generating filtered VOC release data from pre-MCD target cell cultures and filtered VOC release data from healthy pre-MCD cell cultures with respect to at least one gene sequence of each of the molecular classifications; Procedure (220) for generating filtered VOC release data from pre-MCD control cell cultures; A procedure for inducing MCD in the target cell culture, the healthy cell culture, and the control cell culture (222); Procedure for obtaining VOC release data from target cell cultures after MCD, VOC release data from healthy cell cultures after MCD, and VOC release data from control cell cultures after MCD (224); A procedure (226) for generating filtered post-MCD target cell culture VOC release data and filtered post-MCD healthy cell culture VOC release data with respect to at least one gene sequence of each of the molecular classifications; Procedure for generating filtered VOC release data from post-MCD control cell cultures (226); A procedure (228) for determining the VOC profile of pre-MCD target cells and post-MCD target cells from the filtered VOC release data of pre-MCD target cell cultures and the filtered VOC release data of post-MCD target cell cultures; Procedure (230) for determining the pre-MCD healthy cell VOC profile and the post-MCD healthy cell VOC profile from the filtered pre-MCD healthy cell culture VOC release data and the filtered post-MCD healthy cell culture VOC release data; A procedure (231) for determining the pre-MCD control cell VOC profile and the post-MCD control cell VOC profile from the filtered pre-MCD control cell culture VOC release data and the filtered post-MCD control cell culture VOC release data; A procedure (232) for determining the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile for each of the aforementioned molecular classifications, with respect to at least one gene sequence; A procedure (214) for obtaining VOC emission data of VOCs released from at least one sample of respiration and at least one body fluid collected from a control group (208); A procedure (234) for generating a dynamic control cell VOC profile from the predicted control cell VOC profile and the VOC release data of the VOC released in at least one sample of the control group; A procedure (210) for obtaining VOC emission data of VOCs released from at least one sample of respiration and at least one body fluid collected (200) from multiple patients (200) in the selected population with respect to each of the selected target cell types; Procedure (236) for generating a dynamic differential VOC profile, wherein the dynamic differential VOC profile is generated with respect to each of the at least one gene sequences of the molecular classification from the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the VOC emission data of the at least one sample; A procedure (238) for comparing the dynamic differential VOC profile with the dynamic control cell VOC profile; and Procedure (240) for saving the dynamic differential VOC profile and the dynamic control cell VOC profile in a database. A method that includes this.

2. The method according to claim 1, wherein the molecular classification classifies each of the at least one gene sequences as a gene mutation according to the cancer type.

3. The method according to claim 1, wherein the procedure for generating the filtered pre-MCD target cell culture VOC release data, the filtered pre-MCD healthy cell culture VOC release data, and the filtered pre-MCD control cell culture VOC release data reduces the influence of irrelevant VOC artifacts and thereby better identifies VOCs associated with the at least one gene sequence before the procedure for inducing the MCD.

4. The procedure for generating the filtered VOC release data from pre-MCD target cell cultures, the filtered VOC release data from healthy pre-MCD cell cultures, and the filtered VOC release data from pre-MCD control cell cultures is as follows: A procedure for comparing the VOC release data of the target cell culture with both the filtered VOC release data of healthy cell cultures before MCD and the filtered VOC release data of control cell cultures before MCD; A procedure for comparing the VOC release data of healthy cell cultures with both the filtered VOC release data of pre-target cell cultures and the filtered VOC release data of pre-control cell cultures; A procedure for comparing the VOC release data of the control cell culture with the filtered VOC release data of the pre-MCD healthy cell culture. The method according to claim 1, including the method described in claim 1.

5. The method according to claim 1, wherein the procedure for inducing the MCD includes a method that does not generate VOC artifacts.

6. The method according to claim 5, wherein the method is selected from the group consisting of rapid freezing technology and UV light technology.

7. The method according to claim 1, wherein the VOC release data related to the target cell culture, the healthy cell culture, and the control cell culture, as well as the post-MCD target cell culture VOC release data, the post-MCD healthy cell culture VOC release data, and the post-MCD control cell culture VOC release data, are derived from the same target cell culture, healthy cell culture, and control cell culture.

8. The method according to claim 1, wherein the procedure for generating the filtered post-MCD target cell culture VOC release data, the filtered post-MCD healthy cell culture VOC release data, and the filtered post-MCD control cell culture VOC release data reduces the influence of irrelevant VOC artifacts and thereby better identifies VOCs associated with the at least one gene sequence after the procedure for introducing the MCD.

9. The procedure for producing the filtered post-MCD target cell culture VOC release data, the filtered post-MCD healthy cell culture VOC release data, and the filtered post-MCD control cell culture VOC release data is as follows: A procedure for comparing the VOC release data of the target cell culture with both the VOC release data of healthy cell cultures after MCD and the VOC release data of control cell cultures after MCD; A procedure for comparing the VOC release data of the healthy cell culture with both the VOC release data of the post-MCD target cell culture and the VOC release data of the post-MCD control cell culture; and Procedure for comparing the VOC release data of the control cell culture with the VOC release data of healthy cell cultures after MCD. The method according to claim 1, including the method described in claim 1.

10. The procedure for determining the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile is as follows: A procedure for predicting the VOC concentration level in at least one sample from the VOC profiles of pre-MCD target cells and post-MCD target cells; A procedure for predicting the VOC concentration level in at least one sample from the pre-MCD healthy cell VOC profile and the post-MCD healthy cell VOC profile; and A procedure for predicting the VOC concentration level in at least one sample from the VOC profile of a control cell before MCD and the VOC profile of a control cell after MCD. The method according to claim 1, including the method described in claim 1.

11. The method according to claim 10, wherein the VOC concentration level is predicted by using a diffusion model.

12. The method according to claim 11, wherein the diffusion model is selected from the group consisting of Farhi's equation and a modified Farhi model.

13. The method according to claim 1, wherein the step of generating the dynamic control cell VOC profile includes a step of minimizing the error between the predicted control cell VOC profile and the VOC emission data in the at least one sample of the control group.

14. The method according to claim 1, further comprising the step of associating the VOC emission data with at least one known medical condition.

15. The method according to claim 14, further comprising a step of comparing VOC emission data before and after treatment of the aforementioned medical condition.

16. A method for associating volatile organic compound (VOC) emissions with selected cancers in a selected population, comprising the following steps: A procedure (162) to determine the gene sequence of at least one of the target cell samples (152) of target cells collected from multiple patients of a selected cancer type in the aforementioned selected population; A procedure (162) for determining the gene sequence of at least one healthy cell sample of healthy cells collected (154) from the aforementioned multiple patients, wherein the healthy cell sample is of the same cell type as the target cell sample; A procedure (162) for determining the gene sequence of at least one control cell sample of control cells collected (156) from a control group, wherein the control cell sample is of the same cell type as the target cell sample; A procedure (162) for classifying each of the at least one gene sequences based on at least one gene mutation of the selected cancer type; A procedure (166) for culturing one of the target cell sample, the healthy cell sample, and the control cell sample to produce a target cell culture, a healthy cell culture, and a control cell culture, respectively; Procedure for obtaining VOC release data for the target cell culture, the healthy cell culture, and the control cell culture (168); A procedure (170) for generating a target cell VOC profile, a healthy cell VOC profile, and a control cell VOC profile, respectively, from the target cell culture VOC release data, the healthy cell culture VOC release data, and the control cell culture VOC release data, for each of the at least one gene mutations; A procedure (164) for obtaining VOC emission data of VOCs released from respiration and at least one sample of at least one body fluid collected from a control group (158); A procedure (172) for determining the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the predicted control cell VOC profile for each of the at least one gene mutations of the selected cancer type by predicting the VOC concentration level in the at least one sample obtained from the target cell VOC profile, the healthy cell VOC profile, and the control cell VOC profile, respectively; A procedure (176) for generating a dynamic control cell VOC profile from the predicted control cell VOC profile and the VOC release data of the VOC released in at least one sample of the control group; A procedure (174) for generating a dynamic differential VOC profile for each of the at least one gene mutations of the selected cancer type from the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the VOC release data of the at least one sample; A procedure for comparing the dynamic differential VOC profile with the dynamic control cell VOC profile (178); and Procedure for saving the dynamic differential VOC profile and the dynamic control cell VOC profile to a database (180) A method that includes this.

17. A procedure for filtering VOC release data from the target cell culture; Procedure for filtering VOC release data from the healthy cell culture; and Procedure for filtering VOC release data from the aforementioned control cell culture The method according to claim 16, further comprising:

18. The filtering procedure described above is further divided into the following sub-procedures: A procedure for comparing the VOC release data of the target cell culture with both the VOC release data of the healthy cell culture and the VOC release data of the control cell culture; A procedure for comparing the VOC release data of the healthy cell culture with both the VOC release data of the target cell culture and the VOC release data of the control cell culture; Procedure for comparing the VOC release data of the control cell culture with the VOC release data of the healthy cell culture. The method according to claim 17, including the method described in claim 17.

19. The procedure for generating the target cell VOC profile, the healthy cell VOC profile, and the control cell VOC profile is comprised of the following sub-procedures: A procedure for comparing the filtered target cell culture VOC release data with both the filtered healthy cell culture VOC release data and the filtered control cell culture VOC release data; A procedure for comparing the filtered VOC release data from healthy cell cultures with both the filtered VOC release data from target cell cultures and the filtered VOC release data from control cell cultures; and A procedure for comparing the filtered VOC release data from control cell cultures with the filtered VOC release data from healthy cell cultures. The method according to claim 17, including the method described in claim 17.

20. The method according to claim 16, wherein the VOC concentration level is predicted by using a diffusion model.

21. The method according to claim 20, wherein the diffusion model is selected from the group consisting of Farhi's equation and a modified Farhi model.

22. The method according to claim 16, wherein the step of generating the dynamic control cell VOC profile includes a step of minimizing the error between the predicted control cell VOC profile and the VOC emission data of the at least one sample of the control group.

23. The method according to claim 16, wherein the step of generating the dynamic differential VOC profile includes a step of minimizing the error between the predicted target cell VOC profile, the predicted healthy cell VOC profile, and the VOC emission data of the at least one sample.

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