ELECTROCHEMICAL MEASURING OF REACTIVE OXYGEN SPECIES (ROS) LEVELS IN PERIPHERAL BLOOD TO DETECT RATIO OF LOW-DENSITY NEUTROPHILS (LDNS) TO HIGH-DENSITY NEUTROPHILS (HDNS), SUITABLE TO ALARM PRESENCE OF CANCER IN SUSPICIOUS CASES
20230140879 · 2023-05-11
Assignee
Inventors
Cpc classification
International classification
Abstract
A system for real-time detecting cancer by analyzing unprocessed blood. The system includes a sensor, an electrical stimulator-analyzer connected to the sensor, and a processing unit connected to the electrical stimulator-analyzer. The sensor includes three electrodes including a working electrode, a counter electrode, and a reference electrode configured to be put in contact with an unprocessed blood sample. The processing unit is configured to perform a method. The method includes applying a set of voltages between the reference electrode and the working electrode utilizing the stimulator-analyzer, measuring a produced set of currents between the counter electrode and the working electrode utilizing the stimulator-analyzer, measuring a level of a ratio of low-density neutrophils (LDNs) to high-density neutrophils (HDNs) in the unprocessed blood sample by measuring a maximum current of the measured set of currents, and detecting a cancer disease if the measured maximum current is less than a threshold value.
Claims
1. A system for real-time detecting cancer by analyzing unprocessed blood, comprising: a sensor, comprising: a substrate; and three electrodes formed on the substrate, the three electrodes comprising a working electrode, a counter electrode, and a reference electrode, each respective electrode comprising: an electrically conductive layer deposited on the substrate, the electrically conductive layer comprising a proximal end and a circular distal end; and an array of multi-walled carbon nanotubes (VAMWCNTs) grown on the circular distal end, the array of VAMWCNTs configured to be put in contact with an unprocessed blood sample dropped on surface of the sensor, the unprocessed blood sample being drawn from a suspected person to have cancer; an electrical stimulator-analyzer device electrically connected to the sensor at the respective proximal end of each respective electrode of the three electrodes, the stimulator-analyzer device configured to: apply a set of voltages in a sweeping range from −0.8 V to +0.8 V to the sensor comprising the VAMWCNTs being in contact with the unprocessed blood sample; and measure a produced set of electrical currents of the sensor responsive to the applied set of voltages; and a processing unit electrically connected to the electrical stimulator-analyzer device, the processing unit comprising: a memory having processor-readable instructions stored therein; and a processor configured to access the memory and execute the processor-readable instructions, which, when executed by the processor configures the processor to perform a method, the method comprising: applying, utilizing the stimulator-analyzer device, the set of voltages in the sweeping range from −0.8 V to +0.8 V between the reference electrode and the working electrode; measuring, utilizing the stimulator-analyzer device, the produced set of electrical currents between the counter electrode and the working electrode; measuring a level of a ratio of low-density neutrophils (LDNs) to high-density neutrophils (HDNs) in the unprocessed blood sample by measuring a level of reactive oxygen species (ROS) in the unprocessed blood sample, measuring the level of ROS in the unprocessed blood sample comprising measuring a maximum electrical current of the measured set of electrical currents; and detecting cancer status of the suspected person, comprising: detecting a cancer disease in suspected person's body if the measured ratio of LDNs to HDNs in the unprocessed blood sample is more than 1 by detecting the measured maximum electrical current of the measured set of electrical currents being less than a first threshold electrical current value; or detecting no cancer disease in the suspected person's body if the measured ratio of LDNs to HDNs in the unprocessed blood sample is less than 1 by detecting the measured maximum electrical current of the measured set of electrical currents being more than a second threshold electrical current value.
2. The system of claim 1, wherein detecting the cancer disease in the suspected person's body comprises one of: detecting the cancer disease in the suspected person's body if the suspected person's body is an adult responsive to the measured maximum electrical current of the measured set of electrical currents being less than 300 μA corresponding to a ratio of LDNs to HDNs in the unprocessed blood sample being more than 1; or detecting the cancer disease in the suspected person's body if the suspected person's body is a child responsive to the measured maximum electrical current of the measured set of electrical currents being less than 100 μA corresponding to a ratio of LDNs to HDNs in the unprocessed blood sample being more than 1.
3. The system of claim 1, wherein detecting no cancer disease in the suspected person's body comprises one of: detecting no cancer disease in the suspected person's body if the suspected person's body is an adult responsive to the measured maximum electrical current of the measured set of electrical currents being more than 450 μA corresponding to a ratio of LDNs to HDNs in the unprocessed blood sample being less than 1; or detecting no cancer disease in the suspected person's body if the suspected person's body is a child responsive to the measured maximum electrical current of the measured set of electrical currents being more than 300 μA corresponding to a ratio of LDNs to HDNs in the unprocessed blood sample being less than 1.
4. The system of claim 1, wherein the method further comprises determining the first threshold electrical current value and the second threshold electrical current value, comprising: generating a first dataset of a plurality of unprocessed blood samples associated with a plurality of cancer patients, comprising: measuring a first set of electrical current peaks of the unprocessed blood samples associated with the plurality of cancer patients; measuring a first set of ratio of LDNs to HDNs in the unprocessed blood samples associated with the plurality of cancer patients utilizing a cell counter; and assigning each measured ratio of LDNs to HDNs of the first set of ratio of LDNs to HDNs to the respective measured electrical current peak of the first set of electrical current peaks; generating a second dataset of a plurality of unprocessed blood samples associated with a plurality of healthy persons, comprising: measuring a second set of electrical current peaks of the unprocessed blood samples associated with the plurality of healthy persons; measuring a second set of ratio of LDNs to HDNs in the unprocessed blood samples associated with the plurality of healthy persons utilizing a cell counter; and assigning each measured ratio of LDNs to HDNs of the second set of ratio of LDNs to HDNs to the respective measured electrical current peak of the second set of electrical current peaks; and determining the first threshold electrical current value and the second threshold electrical current value, comprising: determining the first threshold electrical current value equal to a maximum electrical current peak among the first set of electrical current peaks; and determining the second threshold electrical current value equal to a minimum electrical current peak among the second set of electrical current peaks.
5. The system of claim 1, wherein the method is conducted in less than 30 seconds.
6. The system of claim 1, wherein the circular distal end has a diameter in a range of 0.5 mm to 3 mm.
7. The system of claim 1, wherein the three respective circular distal ends of the three electrodes are placed apart from each other by a distance between 1 mm and 5 mm.
8. The system of claim 1, wherein the substrate comprises a first layer of silicon dioxide deposited on a layer of silicon.
9. The system of claim 8, wherein the sensor further comprises a second layer of silicon dioxide deposited on surface of sensor except surface of the circular distal end and the proximal end of each respective electrode of the three electrodes.
10. The system of claim 1, wherein the electrically conductive layer comprises a layer of at least one of nickel, gold, and combinations thereof.
11. The system of claim 1, wherein the electrically conductive layer has a thickness in a range of 5 nm to 20 nm.
12. The system of claim 1, wherein: the array of VAMWCNTs comprises VAMWCNTs with a length in a range of 2.5 μm to 5 μm; and the array of VAMWCNTs comprises VAMWCNTs with a diameter in a range of 50 nm to 70 nm.
13. A method for real-time detecting cancer, comprising: putting an unprocessed blood sample in contact with sensing parts of a set of a working electrode, a counter electrode, and a reference electrode of a sensor, the unprocessed blood sample drawn from a suspected person to have cancer; applying a set of voltages in a sweeping range from −0.8 V to +0.8 V between the reference electrode and the working electrode; measuring, utilizing one or more processors, a produced set of electrical currents between the counter electrode and the working electrode versus the applied set of voltages; measuring, utilizing one or more processors, a level of ratio of low-density neutrophils (LDNs) to high-density neutrophils (HDNs) in the unprocessed blood sample, comprising: measuring a level of reactive oxygen species (ROS) in the unprocessed blood sample by measuring a maximum electrical current of the measured set of electrical currents; and determining the level of ratio of LDNs to HDNs in the unprocessed blood sample, comprising: detecting the level of ratio of LDNs to HDNs in the unprocessed blood sample is more than 1 if the measured maximum electrical current of the measured set of electrical currents is less than a first threshold electrical current value; and detecting the level of ratio of LDNs to HDNs in the unprocessed blood sample is less than 1 if the measured maximum electrical current of the measured set of electrical currents is more than a second threshold electrical current value; and detecting, utilizing one or more processors, cancer status of the suspected person, comprising one of: detecting a cancer disease in the suspected person's body if the level of ratio of LDNs to HDNs in the unprocessed blood sample is more than one; or detecting no cancer disease in the suspected person's body if the level of ratio of LDNs to HDNs in the unprocessed blood sample is less than one.
14. The method of claim 13, further comprising generating the first threshold electrical current value and the second threshold electrical current value, comprising: generating a first dataset of a plurality of unprocessed blood samples associated with a plurality of cancer patients, comprising: measuring a first set of electrical current peaks of the unprocessed blood samples associated with the plurality of cancer patients; measuring a first set of ratio of LDNs to HDNs in the unprocessed blood samples associated with the plurality of cancer patients utilizing a cell counter; and assigning each measured ratio of LDNs to HDNs of the first set of ratio of LDNs to HDNs to the respective measured electrical current peak of the first set of electrical current peaks; generating a second dataset of a plurality of unprocessed blood samples associated with a plurality of healthy persons, comprising: measuring a second set of electrical current peaks of the unprocessed blood samples associated with the plurality of healthy persons; measuring a second set of ratio of LDNs to HDNs in the unprocessed blood samples associated with the plurality of healthy persons utilizing a cell counter; and assigning each measured ratio of LDNs to HDNs of the second set of ratio of LDNs to HDNs to the respective measured electrical current peak of the second set of electrical current peaks; and determining the first threshold electrical current value and the second threshold electrical current value, comprising: determining the first threshold electrical current value equal to a maximum electrical current peak among the first set of electrical current peaks; and determining the second threshold electrical current value equal to a minimum electrical current peak among the second set of electrical current peaks.
15. The method of claim 13, wherein: the first threshold electrical current value is 300 μA for an adult person; the first threshold electrical current value is 100 μA for a child; the second threshold electrical current value is 450 μA for an adult person; and the second threshold electrical current value is 300 μA for a child.
16. The method of claim 13, wherein the method is conducted in less than 30 seconds.
17. The method of claim 13, further comprising acquiring the unprocessed blood sample from the suspected person to have cancer.
18. The method of claim 13, wherein putting the unprocessed blood sample in contact with the sensing parts of the set of the working electrode, the counter electrode, and the reference electrode of the sensor comprises putting the unprocessed blood sample in contact with three respective arrays of multi-walled carbon nanotubes (VAMWCNTs) grown on the sensing parts of the set of the working electrode, the counter electrode, and the reference electrode by dropping the unprocessed blood sample on surface of the sensor.
19. The method of claim 13, wherein applying the set of voltages in the sweeping range from −0.8 V to +0.8 V between the reference electrode and the working electrode comprises: connecting respective proximal ends of the set of the working electrode, the counter electrode, and the reference electrode of the sensor to an electrical stimulator-analyzer device; and applying the set of voltages in a sweeping range from −0.8 V to +0.8 V to the sensor using the electrical stimulator-analyzer device.
20. The method of claim 19, wherein measuring the produced set of electrical currents between the counter electrode and the working electrode versus the applied set of voltages is done utilizing the electrical stimulator-analyzer device.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0027] The drawing figures depict one or more embodiments in accord with the present teachings, by way of example only, not by way of limitation. In the figures, like reference numerals refer to the same or similar elements.
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DETAILED DESCRIPTION
[0048] In the following detailed description, numerous specific details are set forth by way of examples in order to provide a thorough understanding of the relevant teachings. However, it should be apparent that the present teachings may be practiced without such details. In other instances, well known methods, procedures, components, and/or circuitry have been described at a relatively high-level, without detail, in order to avoid unnecessarily obscuring aspects of the present teachings. The following detailed description is presented to enable a person skilled in the art to make and use the methods and devices disclosed in exemplary embodiments of the present disclosure. For purposes of explanation, specific nomenclature is set forth to provide a thorough understanding of the present disclosure. However, it will be apparent to one skilled in the art that these specific details are not required to practice the disclosed exemplary embodiments. Descriptions of specific exemplary embodiments are provided only as representative examples. Various modifications to the exemplary embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be applied to other embodiments and applications without departing from the scope of the present disclosure. The present disclosure is not intended to be limited to the embodiments shown, but is to be accorded the widest possible scope consistent with the principles and features disclosed herein.
[0049] In cancer patients with solid tumors, neutrophils can paradoxically mediate a broad range of anti- and pro-tumor activities from cancer cell killing (done by HDNs) to tumor cell proliferation, angiogenesis, metastasis, and organizing other immune responses (done by LDNs). Neutrophil density changes due to their increased granularity and variations in cell size during development. Immature neutrophils are typically found in low density (LD) fractions, whereas mature neutrophils are detected in normal/high density (N/HD) fractions. LDNs are either immature with a banded/ring nucleus or mature with segmented nuclei. All LDNs are pro-tumor that present immunosuppressive properties. HDNs (or NDNs) are mature with segmented nuclei which are antitumor. It should be noted that TGF-β could also mediate transition of HDN to LDN (displaying plasticity) in the murine model.
[0050] An increased number of immature neutrophils in peripheral blood and tissues is a consequence of cancer development in human patients. LDNs may be detectable in peripheral blood of patients with lung cancer, breast cancer, and ovarian cancer. This increase may result from promotion of immature neutrophil production and release due to increased systemic chemokines, (e.g., granulocyte colony-stimulating (G-CSF)). It should be noted that tumors produce granulocyte colony-stimulating factor (G-CSF). G-CSF reduces chemokine receptor type 4 (CXCR4) expression in human myeloid lineage cells, resulting in decreasing their response to the bone marrow retention signal stromal cell-derived factor 1 (SDF-1).
[0051] Immature neutrophils may have both anti- and pro-tumor properties. These include altered localization resulting from reduced surface chemotactic receptor expression (e.g., CXCR2) and less segmented nuclear morphology compared to mature ones which reduce their immunological functions due to their reduced ROS production and altered cell surface receptor expression. Also, immature neutrophils are unable to kill tumor cell via FcγRI receptors. These differences in properties and functions of immature neutrophils could lead to their negative effect on cancer therapy. ROS production is crucial in several neutrophil effector mechanisms including their microbicidal, phagocytic, suppressive capacity and neutrophil anti, pro-tumor functions. Reduced ROS production may be the main deficiency of immature neutrophils against cancer cells.
[0052] Herein, an exemplary real-time electrochemical diagnostic sensor, system, and method for detecting cancer by detecting a level of a ratio of LDNs/HDNs in a blood sample via detecting reactive oxygen species (ROS) levels in a small amount (i.e., about 1 cc) of peripheral blood is disclosed. An exemplary sensor may include three disks like electrodes (working electrode (WE), counter electrode (CE), and reference electrode (RE)) covered by Multi-Wall Carbon Nanotubes (MWCNTs) as a ROS detecting agent in a solution. A cancerous status of a suspected person may be determined utilizing exemplary sensor, system, and/or method in a time period of less than about 30 seconds. An exemplary method and system may be utilized for a rapid detection of cancer via a simple procedure using a small amount (i.e., about 1 cc) of an unprocessed blood sample drawn from a person suspected to have cancer.
[0053]
[0054] In an exemplary embodiment, sensor 102 may include an exemplary electrochemical sensor. In an exemplary embodiment, sensor 102 may include substrate 103 and three electrodes 107, 108, and 109 formed on substrate 103. In an exemplary embodiment, three electrodes 107, 108, and 109 may include a set of electrochemical electrodes configured to be utilized for electrochemical measurements. In an exemplary embodiment, three electrodes 107, 108, and 109 may include a working electrode 107, a counter electrode 108, and a reference electrode 109.
[0055] In an exemplary embodiment, each respective electrode of three electrodes 107, 108, and 109 may include an electrically conductive layer deposited on substrate 103. In an exemplary embodiment, an exemplary electrically conductive layer may have a thickness in a range of about 5 nm to about 20 nm. In an exemplary embodiment, an exemplary electrically conductive layer may include a catalyst material for a process of growing carbon nanotubes (CNTs), for example, vertically aligned multi-walled CNTs (VAMWCNTs), thereon. In an exemplary embodiment, an exemplary electrically conductive layer may include a layer of at least one of nickel, gold, and combinations thereof. In an exemplary embodiment, an exemplary electrically conductive layer may include a circular distal end and a proximal end. In an exemplary embodiment, an exemplary circular distal end may form an exemplary circular sensing part similar to circular sensing parts 107a, 108a, and 109a and an exemplary proximal end may form an exemplary connecting pad similar to connecting pads 107b, 108b, and 109b. In an exemplary embodiment, each respective circular sensing part of circular sensing parts 107a, 108a, and 109a may have a diameter in a range of about 0.5 mm to about 3 mm. In an exemplary embodiment, each respective circular sensing part of circular sensing parts 107a, 108a, and 109a may have a diameter of about 2 mm. In an exemplary embodiment, circular sensing parts 107a, 108a, and 109a of three respective electrodes 107, 108, and 109 may be placed on substrate 103 apart from each other by a distance between about 1 mm and about 5 mm. In an exemplary embodiment, circular sensing parts 107a, 108a, and 109a of three respective electrodes 107, 108, and 109 may be placed on substrate 103 apart from each other by a triangular distance 110 (illustrated in
[0056] Referring back to
[0057] In an exemplary embodiment, sensor 102 may further include a second electrically passivating layer deposited on surface of sensor 102 except surface of circular sensing parts 107a, 108a, and 109a and surface of connecting pads 107b, 108b, and 109b of three respective electrodes 107, 108, and 109.
[0058] In an exemplary embodiment, each respective circular sensing part of circular sensing parts 107a, 108a, and 109a may be coated with a layer of carbon nanotubes (CNTs) deposited on the respective circular sensing part.
[0059] In an exemplary embodiment, system 100 may further include a holder and a cap.
[0060] In an exemplary embodiment, an exemplary cap 142 may include a circular element with a diameter equal or more than an outer diameter of holder 140. In an exemplary embodiment, cap 142 may be placed on a top side of holder 140 above sensor 102 and unprocessed blood sample 150. In an exemplary embodiment, cap 142 may be configured to protect sensor 102 and unprocessed blood sample 150 being in contact with each other away from pollutants and disturbing factors.
[0061] In an exemplary embodiment, electrical stimulator-analyzer device 104 may be electrically connected to sensor 102 at respective proximal ends of each respective electrode of three electrodes 107, 108, and 109 of sensor 102. In an exemplary embodiment, electrical stimulator-analyzer device 104 may be electrically connected to sensor 102 at connecting pads 107b, 108b, and 109b of three electrodes 107, 108, and 109 utilizing three respective electrically conductive lines 116, 118, and 120. In an exemplary embodiment, electrical stimulator-analyzer device 104 may include a potentiostat device. In an exemplary embodiment, electrical stimulator-analyzer device 104 may be configured to apply a set of voltages in a sweeping range from about −3 V to about +3 V, for example, a range of −0.8 V to +0.8 V, to sensor 102 with unprocessed blood sample 150 placed thereon and measure a produced set of electrical currents of sensor 102 responsive to the applied set of voltages. In an exemplary embodiment, electrical stimulator-analyzer device 104 may be electrically connected to processing unit 106 utilizing at least one of an electrically conductive line 122, a wireless connection (not illustrated), and combinations thereof. In an exemplary embodiment, the wireless connection may include Bluetooth devices or Bluetooth modules, which may be embedded in electrical stimulator-analyzer device 104 and processing unit 106. In an exemplary embodiment, electrical stimulator-analyzer device 104 may be further configured to send the measured set of electrical currents to processing unit 106.
[0062] In an exemplary embodiment, processing unit 106 may include a memory having processor-readable instructions stored therein and a processor. In an exemplary embodiment, the processor may be configured to access the memory and execute the processor-readable instructions. In an exemplary embodiment, executing the processor-readable instructions by the processor may configures the processor to perform a method. In an exemplary embodiment, the method may include an exemplary method for real-time detecting cancer described herein below.
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[0065] With further detail in regards to step 202,
[0066] With further detail in regards to step 301, an exemplary semi-conductive substrate may be cleaned. In an exemplary embodiment, an exemplary semi-conductive substrate may include semi-conductive layer 310 shown in part 310 of
[0067] In further detail with respect to step 302, step 302 may include forming a first electrically passivating layer on surface of an exemplary cleaned semi-conductive substrate. In an exemplary embodiment, an exemplary first electrically passivating layer 322 (similar to first electrically passivating layer 103b of
[0068] In further detail with respect to step 303, step 303 may include depositing a catalyst layer on an exemplary first electrically passivating layer as schematically shown in part 313 of
[0069] In further detail with respect to step 304, step 304 may include forming three exemplary electrodes 326, 328, and 330 (similar to exemplary three exemplary electrodes 107, 108, and 109 of
[0070] In an exemplary embodiment, each exemplary electrode 328 of three exemplary electrodes 326, 328, and 330 may include a respective circular head 328a (similar to each of circular sensing parts 107a, 108a, and 109a illustrated in
[0071] In further detail with respect to step 305, step 305 may include electrically passivating a surface of an exemplary sensor except three respective circular distal ends of exemplary three electrodes by coating a second electrically passivating layer on surface of an exemplary sensor except three respective circular distal ends of three exemplary electrodes. In an exemplary embodiment, step 305 may include electrically passivating a surface of exemplary sensor 102 except three respective circular heads 326a, 328a, and 330a) of electrodes 326, 328, and 330 as schematically shown in part 315 of
[0072] In further detail with respect to step 306, step 306 may include forming three arrays of CNTs on three respective circular distal ends of three exemplary electrodes of an exemplary sensor as schematically shown in part 316 of
[0073] In further detail with respect to step 307, step 307 may include removing parts of an exemplary second electrically passivating layer coated on three respective proximal ends of exemplary three electrodes of an exemplary sensor as schematically shown in part 317 of
[0074] Referring back to
[0075] In further detail with respect to step 206, step 206 may include applying a set of voltages to sensor 102. In an exemplary embodiment, applying the set of voltages to sensor 102 may include applying the set of voltages between exemplary reference electrode 109 and exemplary working electrode 107 of electrodes 107, 108, and 109 of sensor 102. In an exemplary embodiment, applying the set of voltages to sensor 102 may be done utilizing electrical stimulator-analyzer device 104. In an exemplary embodiment, applying the set of voltages to sensor 102 may include applying the set of voltages in a sweeping range from about −3 V to about +3 V between exemplary reference electrode 109 and exemplary working electrode 107 of electrodes 107, 108, and 109 of sensor 102. In an exemplary embodiment, applying the set of voltages to sensor 102 may include applying the set of voltages in the sweeping range from about −0.8 V to about +0.8 V between exemplary reference electrode 109 and exemplary working electrode 107 of electrodes 107, 108, and 109 of sensor 102.
[0076] In further detail with respect to step 208, step 208 may include measuring a produced set of electrical currents respective to the applied set of voltages to sensor 102. In an exemplary embodiment, a set of electrical currents may be generated between an exemplary counter electrode 108 and an exemplary working electrode 107 of the three electrodes 107, 108, and 109 of sensor 102 responsive to the applied set of voltages. In an exemplary embodiment, step 208 may include measuring the produced set of electrical currents between counter electrode 108 and working electrode 107 of three electrodes 107, 108, and 109 of sensor 102 respective to the applied set of voltages. In an exemplary embodiment, measuring the produced set of electrical currents may be carried out utilizing electrical stimulator-analyzer device 104. In an exemplary embodiment, measuring the produced set of electrical currents may further include sending the measured produced set of electrical currents to processing unit 106 by electrical stimulator-analyzer device 104.
[0077] In further detail with respect to step 210, step 210 may include measuring ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 by measuring a maximum level of the set of electrical currents. In an exemplary embodiment, measuring the maximum level of the set of electrical currents may be carried out by one or more processors similar, for example, processing unit 106. In an exemplary embodiment, measuring the ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 may include measuring a level of reactive oxygen species (ROS) in the unprocessed blood sample 150 by measuring a maximum electrical current of the measured set of electrical currents. In an exemplary embodiment, measuring the ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 may include receiving the measured produced set of electrical currents, plotting a cyclic voltammetry (CV) diagram by plotting the measured produced set of electrical currents versus the applied set of voltages, and measuring a peak value of electrical currents of the CV diagram.
[0078] In an exemplary embodiment, measuring the ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 may further include determining a level of ratio of LDNs to HDNs in the unprocessed blood sample using the measured maximum electrical current of the measured set of electrical currents. In an exemplary embodiment, determining the level of ratio of LDNs to HDNs in the unprocessed blood sample may include detecting the level of ratio of LDNs to HDNs in the unprocessed blood sample is more than 1 if the measured maximum electrical current of the measured set of electrical currents is less than a first threshold electrical current value and detecting the level of ratio of LDNs to HDNs in the unprocessed blood sample is less than 1 if the measured maximum electrical current of the measured set of electrical currents is more than a second threshold electrical current value.
[0079] In an exemplary embodiment, exemplary method 200 may further include generating the first threshold electrical current value and the second threshold electrical current value.
[0080] In further detail with respect to step 232, step 232 of generating a first dataset of a plurality of unprocessed blood samples associated with a plurality of cancer patients may include measuring a first set of electrical current peaks of the plurality of unprocessed blood samples associated with the plurality of cancer patients, measuring a first set of ratio of LDNs to HDNs in the plurality of unprocessed blood samples associated with the plurality of cancer patients utilizing a cell counter, and assigning each measured ratio of LDNs to HDNs of the first set of ratio of LDNs to HDNs to the respective measured electrical current peak of the first set of electrical current peaks.
[0081] In further detail with respect to step 234, step 234 of generating a second dataset of a plurality of unprocessed blood samples associated with a plurality of healthy persons may include measuring a second set of electrical current peaks of the plurality of unprocessed blood samples associated with the plurality of healthy persons, measuring a second set of ratio of LDNs to HDNs in the unprocessed blood samples associated with the plurality of healthy persons utilizing a cell counter, and assigning each measured ratio of LDNs to HDNs of the second set of ratio of LDNs to HDNs to the respective measured electrical current peak of the second set of electrical current peaks.
[0082] In further detail with respect to step 236, step 236 of determining the first threshold electrical current value and the second threshold electrical current value based on the first dataset and the second dataset may include determining the first threshold electrical current value equal to a maximum electrical current peak among the first set of electrical current peaks and determining the second threshold electrical current value equal to a minimum electrical current peak among the second set of electrical current peaks. In an exemplary embodiment, the first threshold electrical current value and the second threshold electrical current value may be equal to each other.
[0083] In an exemplary embodiment, the first threshold electrical current value and the second threshold electrical current value may depend on age of an exemplary suspected person. In an exemplary embodiment, the first threshold electrical current value may be equal to an electrical current peak of about 300 μA for an adult person. In an exemplary embodiment, the first threshold electrical current value may be equal to an electrical current peak of about 100 μA for a child. In an exemplary embodiment, the second threshold electrical current value may be equal to an electrical current peak of about 450 μA for an adult person. In an exemplary embodiment, the second threshold electrical current value may be equal to an electrical current peak of about 300 μA for a child.
[0084] Referring back to
[0085] In an exemplary embodiment, the first threshold electrical current value corresponding to a threshold LDNs/HDNs value of 1 may equal to 300 μA for adults and 100 μA for children. In an exemplary embodiment, “adults” as used herein may refer to people who are 18 years old or older. In an exemplary embodiment, “children or pediatrics cohort” as used herein may refer to people who are younger than 18 years, for example, children from new born to 13 years old. In an exemplary embodiment, detecting the cancer disease in the suspected person's body may include detecting the cancer disease in an adult suspected person's body if the measured maximum electrical current of the measured set of electrical currents is less than about 300 μA corresponding to a ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 being more than 1. In another exemplary embodiment, detecting the cancer disease in the suspected person's body may include detecting the cancer disease in a suspected child's body if the measured maximum electrical current of the measured set of electrical currents is less than about 100 μA corresponding to a ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 being more than 1.
[0086] In another exemplary embodiment, detecting cancer status of an exemplary suspected person may include detecting no cancer disease (a normal or healthy status) in the suspected person's body if the measured ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 is less than the threshold LDNs/HDNs value. In an exemplary embodiment, no cancer may be detected for an exemplary suspected person if a level of LDNs is less than HDNs in exemplary unprocessed blood sample 150. In an exemplary embodiment, no cancer may be detected for an exemplary suspected person if a level of ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 is less than about 1 if the measured maximum electrical current of the measured set of electrical currents is more than the second threshold electrical current value. In an exemplary embodiment, detecting no cancer disease (a normal or healthy status) in the suspected person's body may include detecting no cancer disease in the suspected person's body if the suspected person is an adult and if the measured maximum electrical current of the measured set of electrical currents is more than 450 μA corresponding to a ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 being less than 1. In another exemplary embodiment, detecting no cancer disease (a normal or healthy status) in the suspected person's body may include detecting no cancer disease in the suspected person's body if the suspected person is a child and if the measured maximum electrical current of the measured set of electrical currents is more than 300 μA corresponding to a ratio of LDNs to HDNs in exemplary unprocessed blood sample 150 being less than 1.
[0087]
[0088] If programmable logic is used, such logic may execute on a commercially available processing platform or a special purpose device. One ordinary skill in the art may appreciate that an embodiment of the disclosed subject matter can be practiced with various computer system configurations, including multi-core multiprocessor systems, minicomputers, mainframe computers, computers linked or clustered with distributed functions, as well as pervasive or miniature computers that may be embedded into virtually any device.
[0089] For instance, a computing device having at least one processor device and a memory may be used to implement the above-described embodiments. A processor device may be a single processor, a plurality of processors, or combinations thereof. Processor devices may have one or more processor “cores.”
[0090] An embodiment of the present disclosure is described in terms of this example computer system 500. After reading this description, it will become apparent to a person skilled in the relevant art how to implement the invention using other computer systems and/or computer architectures. Although operations may be described as a sequential process, some of the operations may in fact be performed in parallel, concurrently, and/or in a distributed environment, and with program code stored locally or remotely for access by single or multiprocessor machines. In addition, in some embodiments the order of operations may be rearranged without departing from the spirit of the disclosed subject matter.
[0091] Processor device 504 may be a special purpose or a general-purpose processor device. As will be appreciated by persons skilled in the relevant art, processor device 504 may also be a single processor in a multi-core/multiprocessor system, such system operating alone, or in a cluster of computing devices operating in a cluster or server farm. Processor device 504 may be connected to a communication infrastructure 506, for example, a bus, message queue, network, or multi-core message-passing scheme.
[0092] In an exemplary embodiment, computer system 500 may include a display interface 502, for example a video connector, to transfer data to a display unit 530, for example, a monitor. Computer system 500 may also include a main memory 508, for example, random access memory (RAM), and may also include a secondary memory 510. Secondary memory 510 may include, for example, a hard disk drive 512, and a removable storage drive 514. Removable storage drive 514 may include a floppy disk drive, a magnetic tape drive, an optical disk drive, a flash memory, or the like. Removable storage drive 514 may read from and/or write to a removable storage unit 518 in a well-known manner. Removable storage unit 518 may include a floppy disk, a magnetic tape, an optical disk, etc., which may be read by and written to by removable storage drive 514. As will be appreciated by persons skilled in the relevant art, removable storage unit 518 may include a computer usable storage medium having stored therein computer software and/or data.
[0093] In alternative embodiments, secondary memory 510 may include other similar means for allowing computer programs or other instructions to be loaded into computer system 500. Such means may include, for example, a removable storage unit 522 and an interface 520. Examples of such means may include a program cartridge and cartridge interface (such as that found in video game devices), a removable memory chip (such as an EPROM, or PROM) and associated socket, and other removable storage units 522 and interfaces 520 which allow software and data to be transferred from removable storage unit 522 to computer system 500.
[0094] Computer system 500 may also include a communications interface 524. Communications interface 524 allows software and data to be transferred between computer system 500 and external devices. Communications interface 524 may include a modem, a network interface (such as an Ethernet card), a communications port, a PCMCIA slot and card, or the like. Software and data transferred via communications interface 524 may be in the form of signals, which may be electronic, electromagnetic, optical, or other signals capable of being received by communications interface 524. These signals may be provided to communications interface 524 via a communications path 526. Communications path 526 carries signals and may be implemented using wire or cable, fiber optics, a phone line, a cellular phone link, an RF link or other communications channels.
[0095] In this document, the terms “computer program medium” and “computer usable medium” are used to generally refer to media such as removable storage unit 518, removable storage unit 522, and a hard disk installed in hard disk drive 512. Computer program medium and computer usable medium may also refer to memories, such as main memory 508 and secondary memory 510, which may be memory semiconductors (e.g. DRAMs, etc.).
[0096] Computer programs (also called computer control logic) are stored in main memory 508 and/or secondary memory 510. Computer programs may also be received via communications interface 524. Such computer programs, when executed, enable computer system 500 to implement different embodiments of the present disclosure as discussed herein. In particular, the computer programs, when executed, enable processor device 504 to implement the processes of the present disclosure, such as the operations in method 200 illustrated by
[0097] Embodiments of the present disclosure also may be directed to computer program products including software stored on any computer useable medium. Such software, when executed in one or more data processing device, causes a data processing device to operate as described herein. An embodiment of the present disclosure may employ any computer useable or readable medium. Examples of computer useable mediums include, but are not limited to, primary storage devices (e.g., any type of random access memory), secondary storage devices (e.g., hard drives, floppy disks, CD ROMS, ZIP disks, tapes, magnetic storage devices, and optical storage devices, MEMS, nanotechnological storage device, etc.).
Example 1: Sensor Fabrication Process
[0098] In this example, an exemplary sensor similar to sensor 102 was fabricated via a method similar to exemplary method 300 illustrated in
[0099] Repeatability of the fabricated sensor was examined by testing three blood samples from normal and cancer patients with a 3-times of test repeat on each of them.
Example 2: Detecting Cancer by Electrochemical Measuring of Reactive Oxygen Species (ROS) Levels in Blood
[0100] In this example, one hundred sixty patients of kids (2 months-13 years of age) and adults (20-73 years old) in two groups of newborn cancer with histologically confirmed cancer (n=60) and normal candidates (n=100) were tested here to investigate a correlation between ROS electrochemical peak results and abundancy ratio of HDNs to LDNs in their unprocessed blood samples. All cancer patients had advanced diseases (stages III-IV) in accordance with the American Joint Committee on Cancer criteria. Additionally, some treated patients with chemo/radiotherapy were investigated (n=20). Clinical, laboratory, and radiological characteristics and diagnosis and outcomes data were collected from all patients. Fresh blood samples prepared from 180 candidates were recorded. Also, patients' symptoms and therapeutic steps of cancer patients were recorded. For each patient, about 1 cc blood was used to measure released ROS/H.sub.2O.sub.2 from HDNs and LDNs in peripheral blood utilizing an exemplary system similar to system 100 including an exemplary sensor fabricated in EXAMPLE 1 hereinabove via an exemplary method similar to method 200. Furthermore, LDNs and HDNs were isolated from whole blood with magnetic isolation and Percoll gradient. Then, LDN and HDN cells were counted by cell counter. Also, LDN and HDN cells were stained with Giemsa following thin-layer cell preparation and ROS fluorescent assay of LDNs and HDNs was carried out using a fluorescent microscopy system.
[0101] Neutrophil Isolation from Whole Blood and Quantification:
[0102] Total circulating neutrophils were isolated from 5 ml cancer patients' blood by negative selection using a Whole Blood Neutrophil Isolation Kit for human. Then, LDN and HDNs were isolated by discontinuous Percoll density gradient method. 9 ml Percoll was added to 1 ml 10×PBS (Stem cell) to create a 100% solution. Then, 2.5 ml of 78% Percoll diluted in 1×PBS was added carefully to a 15 ml falcon tube followed by a 2 ml layer of 66% Percoll and, so total neutrophils resuspended in 2 ml 54% Percoll. Centrifugation at 1545×g without brake was performed for 30 minutes at room temperature. HDNs were recovered from the 78%/66% interface and LDNs from 66%/54% interface. Cell counts were performed using a cell counter. Afterward, HDNs and LDNs cells were fixed by dipping the glass slides in 70% ethanol for 2 min and allowed to dry at room temperature before staining. HDNs and LDNs cells were stained 50 min in Giemsa solution, and washed in Tap water. Finally, glass slides were air-dried shortly and the dried slides were inspected under a light microscope.
[0103] Ros Assay:
[0104] ROS generation was analyzed with a 5-chloromethyl-2′-7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) assay. This probe is changed to 2′-7′-dichlorofluorescein (DCF) with a green fluorescent property by esterase enzymes in the cells' cytosol. After culturing fibroblast and MCF-7 cells overnight, the cells were washed with PBS, and then 500 μL of CM-H2DCFDA solution was added with a concentration of 20 μM. After 30 min incubation at room temperature in dark ambient, cells were again washed and then imaged with a fluorescent microscope. The cells were incubated with 10 mM of NAC as a ROS scavenger and then treated with CM-H2DCFDA. For positive control, cells were incubated with 100 μM H.sub.2O.sub.2 and treated by CM-H2DCFDA. Afterwards, samples were imaged with a fluorescent microscopy system.
[0105] An exemplary sensor fabricated in EXAMPLE 1 hereinabove was used to detect ROS levels in about 1 cc of an unprocessed blood sample from cancer patients and normal cases. The unprocessed blood sample was a residue of conventionally received blood from volunteers due to their checkup by informing them and their satisfaction. Intensity of released ROS levels in peripheral blood was recorded by a cyclic voltammetry procedure via a method similar to method 200 described hereinabove. Coherent results were achieved after categorizing the measured ROS current peaks and comparing them with candidates' clinical diagnostic results and abundancy of LDNs versus HDNs in their blood samples. Table 1 shows baseline characteristics of 160 patients investigated herein, including group 1 (G1) of normal candidate/patients with non-cancer diseases and group 2 (G2) of patients with newborns cancer.
TABLE-US-00001 TABLE 1 Baseline characteristics of 160 investigated patients (G1: Normal candidate/patients with non-cancer diseases, G2: Patients with newborns cancer). State of patients Normal/Non Characteristic cancer diseases Malignant tumor Candidates Age Sex Race (n = 80) (n = 80) Children 2 months-13 Female Male White Appendicitis Ewing sarcoma/neck years old (n = 21) (n = 39) (n = 5) tumor mass (n = 3) Abdominal Rhabdomyosarcoma edema (n = 6) pelvis (n = 6) Abdominal hernia Ewing (n = 2) sarcoma/abdomen Left kidney stone tumor mass (n = 1) (n = 4) Liver Hydatid Brain tumor in the cyst (n = 3) left ventricle (n = 2) Hypertrophic Lung cancer (n = 3) pyloric stenosis Wilms tumor (n = 7) (n = 2) Hepatoblastoma Osteocyte foot (n = 2) bone (n = 2) Neuroblastoma Vesicostomy (n = 2) (n = 3) Osteosarcoma (n = 1) Normal (n = 6) Adults 20-73 Female Male White Fibrocystic Phyllodes (n = 100) (n = 0) changes/ (cancerous) (n = 5) Microcalcification IDC nuclear grade 2 (n = 20) (n = 60) Fibroadenoma/ IDC nuclear grade 3 Cystic mass (n = 25) (n = 10) high grade DCIS Florid ductal (n = 7) hyperplasia Low/inter mediate (n = 20) grade DCIS (n = 2) Normal (n = 50) ILC (n = 1)
[0106] Also, a calibration pattern was provided between responses measured and obtained by exemplary fabricated sensor and results of pathological state of the patients with tumors.
[0107] Furthermore, receiver operating characteristic (ROC) curve analysis was done to compare between peak level responses associated with neutrophil ROS production in blood measured by exemplary fabricated sensor and results of clinical diagnostics (gold standard). The clinical diagnostics, including pathological and radiological evaluation, was considered as the gold standard test for diagnosis cancerous specimens.
[0108] Moreover, specificity, sensitivity, accuracy, precision, and selectivity was calculated for exemplary method and fabricated sensor as presented in Table 3. Also, true and false positive and negative data are shown in detail in Table 3. Accordingly, exemplary method, system, and sensor described herein can be used as an accurate diagnostic tool for detecting cancer in suspicious cases by real-time testing of peripheral blood. Exemplary fabricated sensor showed selectivity of about 98% and specificity of about 96% for the presence of cancer in patients with a breast mass and no signs of other inflammatory diseases (which may activate cytokine storm). In conclusion, exemplary fabricated sensor has proper sensitivity and specificity and can be used as a diagnostic tool for detecting suspicious cases by real-time testing of peripheral blood.
TABLE-US-00002 TABLE 2 AUC for exemplary fabricated sensor and method versus clinical diagnostics results for blood samples from 160 patients Area Under the Curve Test Result Variable(s) for exemplary fabricated sensor Asymptotic Asymptotic 99% Confidence Interval Area Std. Error.sup.a Sig..sup.b Lower Bound Upper Bound 0.981 0.012 0.000 0.949 1.000 The test result variable(s): Exemplary fabricated sensor has at least one tie between the positive actual state group and the negative actual state group. Statistics may be biased. .sup.aUnder the nonparametric assumption .sup.bNull hypothesis: true area = 0.5
TABLE-US-00003 TABLE 3 Cross tabulation results for exemplary fabricated sensor, method, and system versus clinical diagnostics as a gold standard for blood samples from 160 patients Exemplary system/method versus Clinical diagnostics Crosstabulation Clinical diagnostics Nega- Posi- tive tive Total Exemplary Negative Count 77 1 78 system/ % within exemplary 98.7% 1.3% 100.0% method system/method % within Clinical 97.5% 1.2% 48.8% diagnostics Positive Count 2 80 82 % within exemplary 2.4% 97.6% 100.0% system/method % within Clinical 2.5% 98.8% 51.2% diagnostics Total Count 79 81 160 % within exemplary 49.4% 50.6% 100.0% system/method % within Clinical 100.0% 100.0% 100.0% diagnostics Sensitivity = 98.80%, Specificity = 97.50%, Accuracy = 98.70%, Precision = 97.60%, Selectivity = 96.33%
[0109] In addition, isolation and characterization of high-density and low-density Neutrophils in comparison with ROS florescent assay was done to check validity of exemplary method described and conducted herein based on electrochemical measurement of ROS levels in blood samples utilizing exemplary fabricated probe hereinabove. Isolation and counting a ratio of LDNs versus HDNs in patients' blood may reveal better a relation between sensor scores and pathological states of blood volunteers. Mature neutrophils are characterized by a polymorphonuclear nucleus, which is in general large with 2-5 lobes ‘segmented neutrophils’, whereas immature neutrophils are shown by a one-lobed curved or ring-shaped nucleus. LDNs (immature and some mature neutrophils) are more in blood of cancer patients.
[0110]
[0111] Fluorescent ROS assay may reveal a correlation between ROS/Hypoxic functions of cultured LDNs and HDNs and their recorded current peaks. As shown in
TABLE-US-00004 TABLE 4 Detailed information of HDN and LDN counts and measured current peaks in 20 normal cases (N = 10) and cancer patients (N = 10) HDN cell LDN cell current counts in counts in peak (μA) Patient 5 ml of 5 ml of LDN/HDN for 1 ml of ID State of patients blood blood ratio blood 1 Normal 2*10.sup.6 0.5*10.sup.6 0.25 1000 2 Normal 1.5*10.sup.6 0.5*10.sup.6 0.33 730 3 Normal 2*10.sup.6 0.5*10.sup.6 0.25 810 4 Normal 2*10.sup.6 .sup. 1*10.sup.6 0.5 531 5 Cancer (IDC nuclear grade 2) 2*10.sup.6 3.5*10.sup.6 1.75 255 6 Cancer (IDC nuclear grade 2) 1.5*10.sup.6 .sup. 4*10.sup.6 2.67 92 7 Cancer (IDC nuclear grade 2) 1*10.sup.6 .sup. 2*10.sup.6 2 149 8 Cancer (IDC nuclear grade 3) 0.5*10.sup.6 1.5*10.sup.6 3 61 9 Cancer (IDC nuclear grade 3) 0.5*10.sup.6 1.5*10.sup.6 3 71 10 Normal 1.5*10.sup.6 0.5*10.sup.6 0.33 609 11 Normal 1*10.sup.6 0.5*10.sup.6 0.5 489 12 Normal 6.5*10.sup.6 0.5*10.sup.6 0.08 1085 13 Cancer (IDC nuclear grade 3) 2*10.sup.6 .sup. 2*10.sup.6 1 298 14 Cancer (IDC nuclear grade 2) 1.5*10.sup.6 3.5*10.sup.6 2.33 129 15 Cancer (IDC nuclear grade 2) 2*10.sup.6 .sup. 4*10.sup.6 2 202 16 Normal 2*10.sup.6 0.5*10.sup.6 0.25 997 17 Cancer (IDC nuclear grade 3) 1*10.sup.6 .sup. 3*10.sup.6 3 65 18 Normal 3*10.sup.6 .sup. 1*10.sup.6 0.33 654 19 Normal 1*10.sup.6 0.5*10.sup.6 0.5 515 20 Cancer (IDC nuclear grade 3) 0.5*10.sup.6 .sup. 2*10.sup.6 4 41
[0112]
[0113] Percentage Changes of ROS Level and LDNs/HDNs Ratio:
[0114] Herein, percentage of changes of ROS level and LDNs/HDNs ratio for cases investigated hereinabove (reported in Table 4) was calculated according to Equations (1) and (2) herein below.
[0115] ROS level changes in cancer patient' blood (%) according to data summarized in Table 4 using Equation (1) is calculated as follow:
[0116] LDNs/HDNs ratio changes in cancer patients' blood (%) according to data summarized in Table 4 using Equation (2) is calculated as follow:
[0117] A significantly reduced ROS (82%) level in peripheral blood of cancer patients measured by an exemplary fabricated electrochemical sensor, which showed a great correlation with an increased ratio of LDNs/HDNs (65%) in such patients was observed.
Example 3: Effect of Chemo/Radiotherapy on Patients' Blood and Sensor Response
[0118] Table 5 shows detailed information of 20 patients with chemo/radiotherapy and measured electrochemical current peaks utilizing exemplary system, method, and fabricated sensor described hereinabove. As may be observed in Table 5, blood samples of non-treated patients with invasive tumors showed significantly lower levels of CV peaks than their blood after complete response (due to an associated oncologist's opinion) to chemo/radiotherapy. It revealed that exemplary system, method, and fabricated sensor has a capability of monitoring therapeutic effects on cancer patients by testing a 1 cc blood sample drawn from a patient. As COVID-19 may also increase Neutrophils' levels in blood due to cytokine storm, which may perturb cancer-associated response of exemplary sensor and method here, RT-PCR test and clinical evaluation was done on patients to be ensured from non-COVID involvement in them.
TABLE-US-00005 TABLE 5 Detailed information of 20 patients with chemo/radiotherapy and associated measured electrochemical current peaks. Therapy current peak Patient (Surgery, Chemo/ (μA) for 1 ml ID Cancer type radiotherapy (session)) of blood 161 IDC nuclear grade 3 Chemo (8) and Surgery (mastectomy) 1090 162 IDC nuclear grade 2 Chemo (5) 550 163 IDC nuclear grade 2/3 Chemo (6)/Radio (30) and surgery 939 (lumpectomy) 164 IDC nuclear grade 3 Chemo (6)/Radio (28) 731 165 IDC nuclear grade Chemo (8) 469 2/DCIS component (20%) 166 IDC nuclear grade 3 Chemo (6)/Radio (0) 393 167 IDC nuclear grade 2/3 Chemo (4)/Radio (0) 321 168 IDC nuclear grade 2 Chemo (5) and Surgery (mastectomy) 498 169 IDC nuclear grade 2 Chemo (8) 681 170 IDC nuclear grade 2/3 Chemo (6)/Radio (20) 537 171 IDC nuclear grade 3 Chemo (6) and Surgery (mastectomy) 786 172 IDC nuclear grade Chemo (6) 606 2/DCIS 173 IDC nuclear grade 3 Chemo (8) 815 174 IDC nuclear grade 2 Surgery (lumpectomy) and radio (0) 495 175 IDC nuclear grade 2 Surgery (lumpectomy) 562 176 IDC nuclear grade 2 Surgery (lumpectomy) 510 177 IDC nuclear grade 3 Chemo (8) 840 178 IDC nuclear grade 3 Chemo (4) 325 179 IDC nuclear grade 2 Chemo (6) and Surgery (mastectomy) 595 180 IDC nuclear grade 2/3 Chemo (8) and Surgery (mastectomy) 700
Example 4: Effect of Other Diseases on Sensor Response
[0119] Different kinds of pediatrics sickness with non-cancer diseases and normal candidates were studied as presented in Table 1. For adult investigation, electrochemical ROS detection was also performed on non-breast cancer diseases. Moreover, blood ROS levels were recorded in some patients with hypertension, heart disease, dyslipidemia, osteoporosis, and diabetes who had no tumoral diseases.
[0120] LDNs may be increased in blood due to cancer, infection, and inflammation. One of the inflammatory diseases is COVID-19 as a dangerous viral pandemic. It is often more severe in people over 60 years old or with health conditions like lung or heart disease, diabetes, or conditions that affect their immune system. In COVID-19 patients, a significant increase of immature neutrophil numbers may be observed in acute patients compared to healthy donors/recovered patients, while the number of mature neutrophils decreased. Also, a meaningful shift in a ratio between mature and immature neutrophils may associate with a severity of viral involvement. The presence of “low density inflammatory neutrophils” may be strongly correlated with disease severity, and IL-6 levels. In addition, immature neutrophil numbers may strongly associate with IL-6 and IP-10. IL-6 and IP-10 may be consistently upregulated during cytokine storms and may be correlated with severe acute respiratory distress syndrome (ARDS). In addition to inflammatory monocytes as a source of IL-6, immature neutrophils may also be a non-negligible source of IL-6 through COVID-19 induced cytokine storm. So, an increase in number of LDNs may occur in cytokine storm stage of COVID disease. To evaluate an effect of COVID-19 as the most frequent infectious disease currently on peripheral blood ROS, exemplary system, method, and fabricated sensor was tested on blood sample of 14 patients with COVID-19 who were hospitalized in the intensive care unit (ICU).
[0121] Table 6 shows electrochemical ROS peak currents measured for blood samples of COVID-19 infected patients with any comorbidity. ROS/H.sub.2O.sub.2 assisted current peaks for 87% of adults with COVID-19 infection were lower than about 300 μA, same as patients with breast cancer tumors. But it is worth noting that when a patient comes to a medical center for evaluation of her/his breast tumor, COVID-19 involvement is prechecked for her/him, and global vaccination drastically reduces a probability of COVID-19 involvement with cytokine storm in the people. Hence, herein candidates suspicious to have high-risk breast tumors with no trace of inflammatory diseases or pregnancy were evaluated.
TABLE-US-00006 TABLE 6 Electrochemical ROS peak currents in COVID-19 infected patients with any comorbidity. Patient Past disease RT- CT- I ID Age Gender history Symptoms ESR CRP PCR Scan (μA) 1 61 Male Lung Dyspnoea, 42 24.7 Pos. Pos. 125 cancer/DM Unconscious 2 72 Male Colon Myalgia or 16 20.3 Pos. Pos. 228 cancer fatigue, Dyspnoea, Lack of appetite 3 63 Male IHD, CVA, loss of 29 31.3 Pos. Pos. 355 DVT consciousness, hypoxemia 4 79 Male IHD, DM, Fever, 24 29.5 Pos. Pos. 149 Alzheimer's Myalgia or fatigue, loss of consciousness 5 73 Female PI, HLP, Fever, 27 21.3 Pos. Pos. 194 DM, HTN, Myalgia or DLP fatigue, loss of consciousness 6 48 Male HLP, Fever, 34 28.6 Pos. Pos. 202 PMH, IHD, Myalgia or DM fatigue, Dyspnoea, Chest pain 7 67 Male DM, Fever, 35 20.5 Pos. Pos. 125 Parkinson Myalgia or fatigue, Vomit, loss of consciousness 8 73 Male HTN, Fever, Cough, 10 14 Pos. Pos. 157 Pneumonia Dyspnoea, Diarrhoea 9 58 Female DM, HTN Fever, Cough, 20 25.8 Pos. Pos. 348 loss of consciousness 10 67 Male DM, HTN, Fever, Cough, 26 22.5 Pos. Pos. 125 Chest pain 11 62 Male DM Fever, 34 26.5 Pos. Pos. 149 Dyspnoea, fatigue, loss of consciousness 12 59 Female IHD, HTN Fever, fatigue, 17 20 Pos. Pos. 251 chest pain 13 70 Male DM, Fever, 15 19.3 Pos. Pos. 205 Parkinson Dyspnoea, Chest pain 14 73 Female IHD, DM, Fever, 32 27.3 Pos. Pos. 188 Alzheimer's Dyspnoea, loss of consciousness IHD: ischemic heart disease, CVA: Cerebrovascular accident, DVT: Deep vein thrombosis, PI: primary immunodeficiency, HLP: Hyperkeratosis lenticularis perstans, DLP: Dyslipidemia, PMH: Progressive macular hypomelanosis, DM: Diabetes mellitus, and HTN: Hypertension.
[0122] Furthermore, ESR and CRP (blood tests for detecting inflammation) were investigated in breast cancer patients. ROS peak levels recorded by exemplary system described here, ESR and CRP of these patients are shown in Table 7.
TABLE-US-00007 TABLE 7 Electrochemical ROS peak currents in patients with breast cancer tumor Patient ESR* CRP** I ID Age Gender Tumor type (mm/hr.) (mg/L) (μA) 1 39 Female ILC 22 7 118 2 38 Female Papillary with 40 1.9 106 Atypia 3 38 Female IDC 10 2.2 134 4 38 Female DCIS 20 3 102 5 54 Female IDC 18 4 96 6 40 Female IDC 34 10 89 7 55 Female IDC 7.3 2 97 8 46 Female IDC 0.5 1 104 9 41 Female IDC 5.4 5 124 10 43 Female IDC 11 3 128 11 39 Female IDC 14 0.5 171 12 72 Female IDC 19 2 202 13 39 Female IDC 21 3 123 14 50 Female IDC 41 6 178 15 42 Female IDC 45 8 147 *The normal range of ESR is 0-29 mm/hr for women **The normal range of CRP is less than 10 mg/L
[0123] Hence, patients with observable mass in their breast but asymptomatic signature about other inflammatory diseases would-be a candidate for exemplary system and method described here to detect their LDNs associated ROS levels in blood in favor of their tumor disease stage. In addition, a cytokine storm occurs when a patient has symptoms. However, there is a tumor in cancer and the suspected person who is referred for diagnosis has no symptoms in favor of inflammatory disease.
[0124] While the foregoing has described what are considered to be the best mode and/or other examples, it is understood that various modifications may be made therein and that the subject matter disclosed herein may be implemented in various forms and examples, and that the teachings may be applied in numerous applications, only some of which have been described herein. It is intended by the following claims to claim any and all applications, modifications and variations that fall within the true scope of the present teachings.
[0125] Unless otherwise stated, all measurements, values, ratings, positions, magnitudes, sizes, and other specifications that are set forth in this specification, including in the claims that follow, are approximate, not exact. They are intended to have a reasonable range that is consistent with the functions to which they relate and with what is customary in the art to which they pertain.
[0126] The scope of protection is limited solely by the claims that now follow. That scope is intended and should be interpreted to be as broad as is consistent with the ordinary meaning of the language that is used in the claims when interpreted in light of this specification and the prosecution history that follows and to encompass all structural and functional equivalents. Notwithstanding, none of the claims are intended to embrace subject matter that fails to satisfy the requirement of Sections 101, 102, or 103 of the Patent Act, nor should they be interpreted in such a way. Any unintended embracement of such subject matter is hereby disclaimed.
[0127] Except as stated immediately above, nothing that has been stated or illustrated is intended or should be interpreted to cause a dedication of any component, step, feature, object, benefit, advantage, or equivalent to the public, regardless of whether it is or is not recited in the claims.
[0128] It will be understood that the terms and expressions used herein have the ordinary meaning as is accorded to such terms and expressions with respect to their corresponding respective areas of inquiry and study except where specific meanings have otherwise been set forth herein. Relational terms such as first and second and the like may be used solely to distinguish one entity or action from another without necessarily requiring or implying any actual such relationship or order between such entities or actions. The terms “comprises,” “comprising,” or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. An element proceeded by “a” or “an” does not, without further constraints, preclude the existence of additional identical elements in the process, method, article, or apparatus that comprises the element.
[0129] The Abstract of the Disclosure is provided to allow the reader to quickly ascertain the nature of the technical disclosure. It is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims. In addition, in the foregoing Detailed Description, it can be seen that various features are grouped together in various embodiments. This is for purposes of streamlining the disclosure, and is not to be interpreted as reflecting an intention that the claimed embodiments require more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive subject matter lies in less than all features of a single disclosed embodiment. Thus, the following claims are hereby incorporated into the Detailed Description, with each claim standing on its own as a separately claimed subject matter.
[0130] While various embodiments have been described, the description is intended to be exemplary, rather than limiting and it will be apparent to those of ordinary skill in the art that many more embodiments and embodiments are possible that are within the scope of the embodiments. Although many possible combinations of features are shown in the accompanying figures and discussed in this detailed description, many other combinations of the disclosed features are possible. Any feature of any embodiment may be used in combination with or substituted for any other feature or element in any other embodiment unless specifically restricted. Therefore, it will be understood that any of the features shown and/or discussed in the present disclosure may be implemented together in any suitable combination. Accordingly, the embodiments are not to be restricted except in light of the attached claims and their equivalents. Also, various modifications and changes may be made within the scope of the attached claims.