Patent Application
20200305849
The present invention is directed generally to systems and methods for detecting and tracking gut microbiome volatile organic compounds.
The thought of being “gassed” often invokes images of gas masks being frantically donned to protect from inhalation injury, but the fact is, we are gassed daily from within our bodies through the byproducts of microbial colonization, metabolism and fermentation as well as various organ and enzymatic processes. Microbes residing in the gastrointestinal (GI) tract not only play a major role in producing gases, but also in consuming them. Gases are the smallest bioactive molecules utilized for sustaining life and can easily and rapidly traverse cell membranes without the need of binding to specific cell membrane receptors. Once gases enter a cell they can interact with intracellular enzymes and ion channels and can cause post-translational modifications to proteins creating genetic changes.
Among the most abundant intraluminal gases are carbon dioxide (CO2), hydrogen (H2), methane (CH4), hydrogen sulfide (H2S), nitrogen (N2), oxygen (O2) nitric oxide (NO), ammonia (NH3), carbon monoxide (CO) and short-chain fatty acids (SCFA). These “gasotransmitters” have been shown to act as mediators and modulators along with other trace volatile organic compounds (VOCs) in the bidirectional communication between the gut, immune system, and brain, which is known as the “microbiota-gut-brain axis.” These gases also serve as nutrients for various tissues, such as nerve cells. Thus, alterations in the gut microbiome (GM) balance, whether through diet, stress, radiation exposure, antibiotic or probiotic usage, disease, infection, or confinement can alter the production of gasotransmitters and cause direct functional consequences to host health, creating a very insidious type of biological threat. Some diseases and disorders reporting alterations in metabolic gas production include GI cancers, mental illness, diabetes, and infertility to name a few.
Optimizing gasotransmitter pools through diet and/or symbiotic microbes may prove beneficial for protecting the microbiome, brain and immune function and preventing opportunistic infections. Thus, accurately and quickly measuring the GM gasotransmitters (GM VOCs) may provide benefits to overall human health. However, there may be a need for improving how GM VOC analysis is performed in various settings. Improvements may be needed in the context of confined spaces, such as on cruise ships, space crafts, and space stations. Improvements are also needed in order to produce a commercially viable GM VOC analyzer that is affordable for hospitals, clinics, and the average consumer in the home-setting.
For spaceflight, gas analyzers must be low weight, low power consuming, easy to operate, take little storage space, and use limited consumables, which is also beneficial for commercial product development on Earth. However, many of the rapid trace VOC detection methods and devices are large devices with long analysis times that require multiple sample preparation steps. Other faster and smaller trace gas analyzers have low detection accuracy and reliability due to the decreased resolution. These devices also do not offer real-time sample collection methods needed for GM VOC detection. Accurate calibration is also a concern when environmental changes occur or when the environment is unknown, such as with temperature, humidity, gas volume, flow rates, and/or distance from the gas source. Many gas analyzers are also not easily calibrated or operated by laypersons and untrained users making trace chemical analysis expensive and difficult for an at-home user, increasing errors in results and taking extended periods of time to operate. In the professional setting, time is extremely valuable not only for the delivery of results, but also for the personnel involved including astronauts, military and ship personnel and within hospitals, clinics, and labs. Some gas sensor systems are also known to be affected by small variations in CO2 and water in the environment which would not be feasible in the high CO2 environment of a spacecraft or in humid environments on cruise ships or in lavatory facilities where GM VOC analysis may take place. Other systems may only be capable of detecting a few VOCs and/or unable to detect trace levels of VOCs.
Embodiments of the invention are directed to methods, systems and devices for (quickly) detecting and tracking GM VOCs. The present invention represents a gas analyzer for use in the vicinity of a toilet, toilet accessory, or device that captures waste from organisms. There is no need to directly analyze waste material as the sensitivity of the instrument ranges from part per billion to part per thousand.
An embodiment of the present invention is directed to a gas collection and analysis system for detecting microbiomes. The system may include a waste disposal site, a sorbent cartridge coupled adjacent to the waste disposal site, wherein the sorbent cartridge includes a sorbent media configured to absorb a gas sample from within the waste disposal site, a sorbent heating block configured to heat the sorbent cartridge and release the gas sample absorbed by the sorbent media, and a pumping system coupling the sorbent cartridge to an ionized gas analyzer mass spectrometer, the pumping system configured to direct the flow of the released gas sample from the sorbent media to the gas analyzer mass spectrometer for chemical analysis.
In some embodiments, the system may further include one or more sensors coupled to the waste disposal site configured to control the pumping system and heating of the sorbent heating block.
In some embodiments, the ionized gas analyzer mass spectrometer includes a quadrupole, an ion trap, a time of flight mass spectrometer, an ion mobility, a differential mobility, or a field asymmetric ion mobility spectrometer.
In some embodiments, the system may further include a controller configured to process data of the ionized gas analyzer mass spectrometer and/or concentration of absorbed and releases gases in real-time and over period of time.
In some embodiments, the ion mobility measurement may be done by IMS, DMS, FAIMS, or hyphenated techniques of ion mobility and mass spectrometry with gas chromatography.
In some embodiments, the ionization mechanism includes nickel-63 foil, electron ionization, corona discharge, chemical ionization, photoionization, electro-spray, atmospheric pressure chemical ionization, thermal ionization, DART, desorption electro spray, or metal spray.
In some embodiments, the sorbent cartridge is reusable, thereby allowing the system to obtain gas samples of multiple, different users.
In some embodiments, the sorbent cartridge may include a carboxen sorbent, a liquid sorbent, or one or more membranes.
In some embodiments, the sorbent cartridge may include a single or multiple sorbent media for analysis.
In some embodiments, the waste disposal site is a toilet.
In some embodiments, the waste disposal site is a waste collection bag.
In some embodiments, the sorbent cartridge may be configured to be attached to a disposable mouthpiece to obtain the gas sample from a breath analysis.
In some embodiments, the gas sample may be utilized for online analysis via gas chromatography.
In some embodiments, the ionized gas analyzer mass spectrometer utilizes gas chromatography.
In some embodiments, the system may be configured to monitor and track the gas samples of a user for evaluation.
Another embodiment of the present invention is directed to a method for detecting microbiomes of a user. The method may include providing a gas collection and analysis system comprising a waste disposal site, wherein the waste disposal site, a sorbent cartridge coupled adjacent to the waste disposal site, wherein the sorbent cartridge includes a sorbent media configured to absorb a gas sample from within the waste disposal site, a sorbent heating block configured to heat the sorbent cartridge and release the gas sample absorbed by the sorbent media, and a pumping system coupling the sorbent cartridge to an ionized gas analyzer mass spectrometer, the pumping system configured to direct the flow of the released gas sample from the sorbent media to the gas analyzer mass spectrometer for chemical analysis; having the user sit on the waste disposal site; absorbing a gas sample of the user from the waste disposal site into the sorbent cartridge; having the user stand up from the waste disposal site; heating the sorbet cartridge to release a gas sample from the sorbent cartridge; pumping the released gas sample from the sorbent cartridge to the gas analyzer mass spectrometer; determining the chemical compounds in the gas sample based on a chemical analysis from the gas analyzer; and providing the user health data based on the chemical analysis.
In some embodiments, the method may include removing the sorbent cartridge from the waste disposal site for analysis at a remote location.
In some embodiments, the gas collection and analysis system may further include one or more sensors coupled to the waste disposal site, the sensors being configured to control the pumping system and heating of the sorbent heating bloc when the user stands up from the waste disposal site.
In some embodiments, the gas collection and analysis system may further include a controller configured to process data of the ionized gas analyzer mass spectrometer and/or concentration of absorbed and releases gases to the user in real-time and over period of time.
In some embodiments, the sorbent cartridge is reusable, thereby allowing the system to obtain gas samples of multiple, different users.
Some embodiments of the present invention are directed to Earth-based systems, methods, and devices.
Some embodiments of the present invention are directed to space-based systems, methods, and devices.
Some embodiments include a modular inlet system that can direct, optionally vacuum, air from the headspace of a toilet, preconcentrate the GM VOCs on a sorbent media housed within a sorbent cartridge, and release the gases into the detection chamber of either a local or remote gas analyzer for analysis. An embodiment for spaceflight purposes may also include a waste collection bag with membrane port to avoid particulate contamination and a sampling line directing airflow to the sorbent cartridge inlet system. Whilst another embodiment allows for the sorbent cartridge (using a sorbent tube configuration) to be used alone as a gas collection device for capturing breath samples as an indirect measure of the GM VOCs and then deposited into the heated block for thermal desorption and analysis.
Machine learning methods (artificial intelligence or AI) can be applied to identify patterns from the data in real-time that correspond to health diagnosis information. The invention may be applied as a health screening and health tracking device (i.e., early detection of infections, cancer, ingested toxins); to monitor fertility (i.e., ovulation and menstrual changes); and to monitor/study the health of humans living in confined spaces, extreme environments, or within vulnerable populations where infections can spread rapidly (i.e., ships, space stations, military bases, hospitals, long-term care facilities).
It is noted that aspects of the invention described with respect to one embodiment, may be incorporated in a different embodiment although not specifically described relative thereto. That is, all embodiments and/or features of any embodiment can be combined in any way and/or combination. Applicant reserves the right to change any originally filed claim and/or file any new claim accordingly, including the right to be able to amend any originally filed claim to depend from and/or incorporate any feature of any other claim or claims although not originally claimed in that manner. These and other objects and/or aspects of the present invention are explained in detail in the specification set forth below. Further features, advantages and details of the present invention will be appreciated by those of ordinary skill in the art from a reading of the figures and the detailed description of the preferred embodiments that follow, such description being merely illustrative of the present invention.
The present invention now is described more fully hereinafter with reference to the accompanying drawings, in which embodiments of the invention are shown. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
The present invention now will be described more fully hereinafter with reference to the accompanying drawings, in which illustrative embodiments of the invention are shown.
In the figures, certain layers, components or features may be exaggerated for clarity, and broken lines illustrate optional features or operations unless specified otherwise. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
It will be understood that, although the terms first, second, etc. may be used herein to describe various elements, components, regions, layers and/or sections, these elements, components, regions, layers and/or sections should not be limited by these terms. These terms are only used to distinguish one element, component, region, layer or section from another region, layer or section. Thus, a first element, component, region, layer or section discussed below could be termed a second element, component, region, layer or section without departing from the teachings of the present invention. The sequence of operations (or steps) is not limited to the order presented in the claims or figures unless specifically indicated otherwise.
Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. Well-known functions or constructions may not be described in detail for brevity and/or clarity.
The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “comprises” and/or “comprising”, when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items.
As used herein, phrases such as “between X and Y” and “between about X and Y” should be interpreted to include X and Y. As used herein, phrases such as “between about X and Y” mean “between about X and about Y.” As used herein, phrases such as “from about X to Y” mean “from about X to about Y.”
The following descriptions are example embodiments of the present invention. The system of the present invention may be generally referred to herein as “Egest™.”
Embodiments of the invention may provide a basis to monitor and track an individual's health via gases and log irregularities due to changes in their gaseous profile due to illness or other complications in their biology. Target gases include, carbon dioxide (CO2), hydrogen (H2), methane (CH4), hydrogen sulfide (H2S), nitrogen (N2), oxygen (O2) nitric oxide (NO), ammonia (NH3), carbon monoxide (CO) and short-chain fatty acids (SCFA) as well as trace VOCs from various compartments of the human volatolome (e.g., feces, urine, blood, saliva, perspiration, milk, emesis, flatus, breath).
Pursuant to embodiments of the present invention, a gas collection and analysis system for use in or near the vicinity of a waste collection system for the gas analysis and detection of microbiomes is provided. The gas collection and analysis system may allow the detection of numerous trace GM VOCs in various environmental conditions. Methods of using a gas collection and analysis system are also provided.
In some embodiments, the present invention may use a conventional sample collection procedure known as thermal desorption for volatile and semi volatile compounds. Collection of these volatile and semi-volatile compounds into sorbent cartridges (see, e.g.,
GM VOCs produced during the waste elimination process concentrate on a preconcentration medium (sorbent media) housed within a sorbent preconcentration container (sorbent cartridge) that is integrated into or near a toilet seat. Note that the present invention does not limit the sorbent cartridge to strictly toilets. For example, in some embodiments, the present invention may be used with waste collection bags and breath tubes through a valve assembly and pump or disposable mouthpiece, respectively (see, e.g.,
Preconcentration sorbent cartridges allow for large volumes of air to pass over or through a sorbent media to collect and concentrate the emitted VOCs for analysis. This may be beneficial for the present invention because the length of time spent using the lavatory facility varies and the GM VOCs may become diluted by air if sampled directly. Therefore, concentrating the GM VOCs may result in acquiring more sample to be delivered into the detection chamber of the gas analyzer at once, using a conventional thermal desorption process which can provide greater sensitivity and specificity during analysis and avoids losing VOCs that may be of low concentration.
In some embodiments, the sorbent material may comprise of one or more sorbents which may allow it to capture a variety of VOCs with varying chemical characteristics (i.e., size, charge, boiling point, etc.). For example, in some embodiments, the sorbent media may be comprised of C2-C26, Tenax, graphitized carbon, Carbopack, Carbosieves, Carboxen, and/or liquid polymer. In some embodiments, the sorbents may be suitable for collecting and/or analyzing biosamples of an organism, human, or animal including urine, feces, saliva, breath, sputum and the like, that will not irreversibly bind the VOCs of interest and will release the VOCs collected for analysis by a gas analyzer.
Referring to
In some embodiments, the sorbent cartridge may enclose the sorbent material within an interior surface of the cartridge such that the sorbent material cannot be touched, capture oil or other contaminants from users' fingers or other surfaces. In some embodiments, the sorbent cartridge may be made of stainless steel, aluminum, glass or other material that can sustain high temperatures (up to 300° C.) and that does not out-gas or introduce contaminates into the sample collection when the cartridge is heated. The sorbent cartridges of the present invention are a special device that is different than a typical carbon filter that is only used to purify the air. Conventional filters have exposed carbon which also allows the user to touch the carbon during installation, which may leave oil residues on the filter from the skin—giving false results. Conventional filters also cannot typically be heated to a desired level (i.e., about 150° C. or higher) as the sorbent cartridge is formed of plastic material that, even if capable of heating, can off-gas its own VOCs once heated which can skew data. The carbon types typically used in conventional filters will most likely irreversibly bind chemicals given the purpose of prior filters is commonly to absorb odors, thus if it were capable of heating it would not release the captured chemicals into the system for analysis.
In order to decrease sample collection and analysis time to achieve a rapid analysis of a collected sample, in some embodiments, the size of the sorbent cartridge orifice is configured such that the cartridge may capture the greatest quantity of VOCs possible and also allow the VOCs to be released quickly and efficiently for analysis using a conventional thermal desorption process.
In some embodiments, the sorbent cartridge may be configured to be removed and analyzed by a gas analyzer at a different location. In some embodiments, the sorbent cartridge may be configured such that the cartridge is directly connected to the heated inlet of the gas analyzer.
Referring to
In some embodiments, the system can be turned on manually or automatically. For example, in some embodiments, the system may comprise a pressure sensor in the toilet seat (e.g., in the rubber grips of the seat), or coupled to the toilet and/or the toilet seat which can detect when a user is seated. In some embodiments, when the system senses a user on the toilet, the pump may be activated to automatically begin evacuating the air from the headspace of the waste collector (e.g., toilet or other) and through the sorbent cartridge. As air is evacuated from the waste collector, waste VOCs may be collected or trapped within the sorbent media and the air vented. In some embodiments, when the user stands, the (pressure) sensors (in the seat) may direct the system to end the sample preconcentration collection process (optionally closing/opening any valves). In some embodiments, the system may use a light sensor, body heat sensor, opto-electronic sensor, camera or proximity sensor or combinations of the same.
In some embodiments of the present invention, the sorbent cartridge may be configured to be removed from an ambient (not-heated) inlet compartment, and placed in a biohazard package, for example, if the cartridge is transported to an alternate site for analysis. An ambient (not-heated) gas preconcentration inlet system with removable sorbent cartridge connected to the back of a standard toilet seat according to embodiments of the present invention is illustrated in
When transferred to an alternate site for analysis, the sorbent cartridge may be inserted into a thermal desorption (heated) inlet which is coupled to a gas analyzer. The thermal desorption inlet allows the gas molecules to be released from the sorbent medium for sample analysis. Benefits of this embodiment may include reduced cost to the user related to not needing to purchase a chemical analyzer. This may also allow for batch analysis of sorbent cartridges to be performed for multiple users, multiple sorbent cartridges with only one chemical analyzer that is located at a central site such as a core laboratory within a hospital, academic institution, military base, ship, space station, etc.
In some embodiments, the system of the present invention may house the sorbent cartridge within a heater mantle (e.g., heater block) located at the waste disposal site where the inlet system directly interfaces with the chemical analyzer of choice. For example, a preconcentration thermal (heated) desorption gas inlet system connected to a standard toilet seat according to embodiments of the present invention is illustrated in
As shown in
In some embodiments, the sorbent cartridge is not coupled to a toilet/toilet seat, but rather, is connected at one end to a disposable mouthpiece to capture VOCs from the breath during a forced expiration. As shown in
Referring to
Exemplary methods of using the gas collection and analysis system according to embodiments of the present invention are also provided. For example, as shown in
In some embodiments, analysis of the collected sample begins by heating the sorbent medium to release the gas molecules. In some embodiments, operations of the system may be controlled (e.g., programmatically) such as operating a heater and controlling actuation of any valves or vacuum pumps, as well as modifications to a user through an interface for selectable settings. The temperature profile may control heating of the sorbent, and molecules desorb according to their affinity with the sorbent. This affinity is similar to a molecule's boiling point, with low boiling small molecules desorbing first, then higher boiling molecules desorbing as the temperature rises. In some embodiments, resistive heating may be used with thermocouples to measure the temperature of the heater. The heating of the sample may be induction through a heater cartridge. In some embodiments, a small metal (e.g., aluminum) heat block/sleeve may be used. The heat block/sleeve may extend over at least a majority, typically at least 80%, or all the length of the sorbent cartridge. In some embodiments, the toilet seat may be insulated/heat shielded to protect the user for burn injury. In some embodiments, heating of the sample may be controlled to initiate only after the user stands. In some embodiments, the heat block may use a lever/latch push and lock mechanism to load the sorbent cartridge, for example, similar to the unheated ambient version described herein. It may be heated from ambient to about 150° C. to release the volatiles collected in the sorbent material.
In some embodiments, the released gaseous molecules are pulled or pushed into an ionization region for analysis. The ionization region is a region where the gaseous molecules become ions. The gaseous molecules become ions through ionization and are charged molecules with a positive or negative charge. A multitude of sources may be used to form ions and can vary from instrument to instrument, some of the most common types applicable in this analysis. In some embodiments, the ion sources useful in the formation of ions for analysis may include radioactive nickel-63 foil, electron ionization, corona discharge, chemical ionization, photoionization, electrospray, atmospheric pressure chemical ionization, thermal ionization, DART, desorption electrospray, or metal spray.
The gas analysis for the present invention occurs via mass spectrometry (quadrupole, ion trap, time of flight). If a mass spectrometer is not available, another type of analyzer that may be used with the present invention includes ion mobility and analysis occurs based on the mobility of an ion through a buffer gas. In some embodiments, the ion mobility measurement may be done via IMS, DMS, FAIMS, or hyphenated techniques of ion mobility and mass spectrometry. In some embodiments, hyphenated techniques may include IMS-MS, GC-MS, GC-IMS to analyze directly or from sorbent materials where the gas is collected at waste collection point (i.e., toilet). Alternatively, in some embodiments, the analyzer may be separate from the waste collection, and a sorbent cartridge used to collect and tag sampling for analysis at the instrument at a different location.
In some embodiments, following analysis of a sample, the sorbent cartridges may be reusable. This may allow for reducing consumables cost. In some embodiments, to clean and sterilize the sorbent cartridge, a bake-out program may b used to heat the cartridge while in the thermal desorption inlet (heater block) to a high temperature for a predetermined amount of time (e.g., about 250° C. to about 300° C. for about 10 minutes). Thus, a user may not be required to change the sorbent cartridge out each time after a user eliminates waste at the waste collection site where a thermal desorption inlet is incorporated, thereby allowing the sorbent cartridge to be multi-use rather than single use-disposable.
Referring to
In some embodiments, the data provided by the chemical analyzer can be used in a variety of ways. In some embodiments, AI (machine learning) may be used to determine correlations with diet/eating habits, environmental factors, behavioral data, and the VOC patterns discovered by the invention. Independent of how the correlation data is generated, the information generated may be used to track and/or adjust diet/eating habits, for tracking historical obesity data, geographical mapping of disease incidence, food imports, domestic crops, pesticides, etc. for health assessments and screenings.
In some embodiments, the system of the present invention may be installed on a toilet seat at a user's home, hospital, clinic, nursing home or other patient facility, as well as on ships, military bases or other location where health monitoring may be needed for screening and prevention of infectious disease or other illness. In some embodiments, the system may be used for academic research purposes and clinical studies.
In some embodiments, the system of the present invention may be used at the International Space Station (ISS) or kept at a home base for real-time testing with astronauts before, during, and after spaceflights. The system may be used to compare collected data with metabolomics data from the Astronaut Microbiome Project. Further embodiments may include developing unique software for the system of the present invention using algorithms to perform artificial intelligence (AI) machine learning of VOC patterns and determine correlations with diet/eating habits, environmental factors, and behavioral data onboard the ISS. These studies may help determine how VOC levels correlate with space metabolomics data; specific health anomalies experienced in microgravity; and determining how increasing metabolic activity is indicative of a causal effect between the existence of a microbial colony and/or a disease process.
Designing an efficient life support system can be important to maintain the minimum requirements for sustaining human life in space. This includes reliable rapid turnaround tools for detecting disease during spaceflight. Spaceflight is more complicated because the alterations in the gut microbiome are not well understood. Additional data may be collected by the system of the present invention to help determine issues that arise during spaceflight so that they can be avoided or alleviated. The system may track the microbiome of astronauts before, during, and after ISS missions. The Astronaut Microbiome project is evaluating culture-independent methods for studying the microbiome of astronauts, surfaces, and water on the ISS, largely using RNA and DNA metagenomic sequencing methods, which is timely and expensive.
Special benefits for astronauts may be realized by embodiments of the invention. High priority risks for deep space travel include increased radiation exposure, cognitive decline and behavioral conditions, inadequate food supply, unknown medication stability, and inflight medical conditions. Radiation can cause substantial damage to the microbiome. Cognitive and behavioral effects may also occur during spaceflight due to aberrations in the microbiome. Sterile meals and heavily filtered air reduce microbial diversity which can increase opportunistic infections. Currently, the recommended countermeasure to deal with bacterial infections in the ISS is the administration of antibiotics which may not be effective due to the increased virulence of bacteria in microgravity and may also induce dysbiosis with use. Additionally, probiotic treatment may result in unintended symptoms due to overuse creating imbalanced microbiomes. A rapid turnaround detection system, such as the system described herein, may offer a novel tool for studying the microbiome in space and mitigating these risks during long-duration spaceflight missions.
The foregoing is illustrative of the present invention and is not to be construed as limiting thereof. Although exemplary embodiments of this invention have been described, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. Accordingly, all such modifications are intended to be included within the scope of this invention as defined in the claims. The invention is defined by the following claims, with equivalents of the claims to be included therein.
The present application claims priority from and the benefit of U.S. Provisional Application Ser. No. 62/826,485, filed Mar. 29, 2019, the disclosure of which is hereby incorporated herein in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| 62826485 | Mar 2019 | US |
