Methods and kits for determining impaired neuro-metabolism associated with mental condition and dietary treatment thereof

Abstract

The disclosure provides methods and kits, including ex vivo and computer- implemented methods, for diagnosing and classifying mental health condition, and managing thereof using neuro nutrition.

Description

METHODS AND KITS FOR DETERMINING IMPAIRED NEURO-METABOLISM ASSOCIATED WITH MENTAL CONDITION AND DIETARY TREATMENT THEREOFFIELD OF INVENTION[1] The present disclosure generally relates to diagnosis of mental health conditions and management thereof using neuro nutrition.BACKGROUND OF INVENTION[2] Mental health encompasses emotional, psychological, and social well-being, influencing how individuals think, feel, and act. Mental Conditions (MC) involve disruptions in these aspects, impacting daily functioning and quality of life. The main contributing factors to a MC of an individual can be divided in 3 groups: genetic, external (e.g. social stressors, challenging life events) and biological, e.g. hormonal, microbiota, etc.[3] In 2019, 1 in every 8 people (970 million) worldwide were living with a mental condition (MC). In 2022, mental health was considered a higher health concern than cancer for the first time ever, only being surpassed by COVID- 19.[4] Despite the gravity of the matter, only 1 in 10 people with MC symptoms improve as a result of treatment. The root causes for the rather poor improvement rates in MC are lack of reliable and objective diagnostic tools (e.g., biomarkers) or misdiagnosis due to inaccuracy of current diagnostic methods, and ineffective treatments.[5] Most MC are assessed and diagnosed with subjective, manual, and inaccurate symptom data revealed by the patients to their physicians in the form of conversations, interviews, or written questionnaires followed by physicians’ subjective interpretations. This subjectivity can lead to misdiagnosis or underdiagnosis, as symptoms may overlap across different disorders, making it hard to distinguish between different MC.[6] Despite many candidate methods relying on biomarkers for diagnosis on MC, none of those methods have yet provided proven improvements in MC diagnostics, let alone treatment efficiency. This leads to reduced chances of being accurately diagnosed (only 10% of patientsare correctly diagnosed) and increases the risk of receiving inadequate or ineffective treatments.[7] After an assessment, psychotherapy and pharmacotherapy are recommended as the main first-line treatments, often combined. However, in 85-90% no improvement is observed despite pharmacological therapies and over 50% do not improve despite psychotherapy. Alternative “mindfulness” methods such as mental health apps, yoga, or the so-called brain food (i.e., neuro nutrition) suggested for use in subjects with MC, while being able to improve an overall well-being of subjects, have no clinical relevance or reliability for neither improved diagnostics nor improvement of mental conditions.[8] Moreover, due to the ineffectiveness of treatments, a high drop out of patients (over 50% drops off the treatment early).[9] This is also manifested by a decline in the usage of digital apps among customers. Of over 10,000 apps only 2% are downloaded, and less than 4 percent of users were still using the app 15 days after the download.

[0010] Accordingly, there is an unmet need for diagnostic and therapeutic approaches for managing MC, in particular an objective and accurate diagnosis. However, poor understanding of the individual patients’ neuro metabolism barricade success of such quantitative approach.

[0011] Neuro metabolism refers to the biochemical processes (from consuming nutrients to extracting rest products in urine) organized in metabolic pathways affecting metabolism and function of the central and peripheral nervous system (including initial processes regulating metabolism of newly consumed nutrients and through the final excretion of unwanted side products).

[0012] Neuro metabolic processes also determine the way nutrition affects the body and the mind, and neuro-nutrition is an emerging field focusing on dietary interventions to support the function of the nervous system.

[0013] When these processes do not function optimally, it can lead to a metabolic impairment of the nervous system, and consequently to a neurological dysfunction, resulting in development and progression of MC and / or associated symptoms. For instance, a dysfunctionalmetabolism of serotonin can result in an impairment of a serotonergic pathway, and contribute to MC, such as anxiety or depression.

[0014] There is therefore an unmet need for quantitative diagnostic approaches for managing MC, capable of considering patients’ neuro metabolic status, and distinguishing between often comorbid MC (e.g. depression, anxiety and stress disorders), even when manifested at different levels of severities, as well as for a personalized therapeutic approach, tailored according to the diagnosis.

[0015] There thus remains a need for a method and system that can enable precise identification of impaired steps of various neurometabolic pathways of a subject, generating a neurometabolic fingerprint of the subject based thereon and deriving underlying neurometabolic impairments and secondary impairments that in turn can aide in a more accurate mental health diagnosis of the subject. There further remains a need for nutritional interventions tailored to address specific aspects of an individual's mental health based on the neurometabolic fingerprint and root cause analysis.SUMMARY OF INVENTION

[0016] According to some aspects, the present disclosure provides methods and kits, including ex vivo and computer-implemented methods, for diagnosing and managing a mental condition (MC) of a subject in need thereof.

[0017] In some embodiments thereof, by detecting / determining a level of a plurality of neurobiomarkers, such as neuro substances (NS) precursors, metabolites and / or co-factors, participating at various steps of each of a plurality of metabolic pathways, a neuro-metabolic fingerprint is computed and outputted using pre-programmed algorithms.

[0018] Advantageously, by mapping of a neurometabolic pathway at various steps thereof, the step of the pathway actually impaired may be identified rather than just the level of a single molecule / metabolite (often an end-product of the pathway). This is particularly beneficial since pathway balance is crucial for proper pathway function. For example, taking selective serotonin reuptake (SRS) inhibitors when tryptophan (a serotonin precursor) is low may have a contradictive effect in that the high (often ultra-high) serotonin levels may cause the body toslow down the metabolism of tryptophan or increase serotonin metabolism, thereby effectively decreasing treatment efficacy. Instead, if tryptophan consumption is increased when low tryptophan levels are identified (while leaving all other steps essentially unaltered), the balance of the serotonergic pathway may naturally be restored.

[0019] Restoring pathway imbalance is further important since many neuro-metabolic pathways are intercorrelated. For example, if dopamine levels are low, it typically leads to low levels of norepinephrine and thus restoring one pathway imbalance may restore the imbalance of a second neuro-metabolic pathway imbalance, while pharmaceutical treatment may lead to pathway imbalances in more than one pathway.

[0020] The intercorrelation between various neurometabolic pathways further results in the often-observed bundle of mental health issues experienced by mental health patients, which in turn makes an exact diagnosis challenging. Advantageously, by mapping of the various neuro- metabolic pathways of the patients into a fingerprint enables identification and differentiation between root causes and secondary causes of a patients’ mental health issues, which in turn make diagnosis more and consequently treatment more effective. In fact, according to some embodiments, the diagnosis of the patient may be solely based on the neurometabolic fingerprint identified, thus obviating the need for a prior diagnosis by a healthcare professional and even the use of the complex questionnaires typically utilized for diagnosing mental health problems.

[0021] Accordingly, embodiments of the disclosure further provides an advantageous method for classifying biological samples of a subject into one or more categories of mental health conditions (MC) by identifying the one or more impaired neuro-metabolic step, thereby distinguishing between at least two mental health conditions, such as but not limited to depression and anxiety, which surprisingly were both found to be related to impaired norepinephrine, phenylethylamine (PEA), serotonin and / or GABA pathways.

[0022] As a further advantage the hereindisclosed algorithmic platform can output neuro nutritional solutions that positively affect the MC and / or MC symptoms by providing a personalized recommendation of a neuro nutrition treatment regimen including functional food, nutrients and supplements, and / or nutraceutical compositions, rich with precursors, metabolites, and / or co-factors, which are predicted by the algorithm to modulate the impairedneuro metabolic step, thereby restoring the neuro metabolism to its proper / normal levels, and promoting healthy brain and nerve system function and mental health.

[0023] In this regard, since pathway balance is critical, specific personalized formulations of precursors, metabolites, and / or co-factors, directed to repair / restore a specific step(s) identified as impaired, may be formulated by the platform. As a result, the method may further include prescribing a neurometabolic treatment regimen. The method may also include treating patients in need thereof, according to the prescribed neurometabolic treatment regimen.

[0024] According to some embodiments, there is provided an ex vivo method for managing a mental health condition of a subject in need thereof, the method comprising: (a) detecting / determining a level of a plurality of neuro -biomarkers in a previously obtained biological urine sample of the subject, wherein the plurality of neuro-biomarkers comprises a neuro substance (NS) and at least one precursor, metabolite and / or co-factor; (b) applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing between the detected / determined levels of at least two neurobiomarkers belonging to a same metabolic pathway; (c) identify one or more impaired neurometabolic steps in any of the plurality of neuro metabolic pathways by applying a second algorithm on the fingerprint; and (d) providing a recommendation of a neuro-nutritional treatment regimen for the subject, based on the identified impairment in the one or more neurometabolic step, wherein the neuro-nutritional treatment regimen is designed to alter the metabolism of the impaired step of a neuro metabolic pathway.

[0025] In some embodiments, the method further comprising obtaining a questionnaire result, wherein the questionnaire result comprises the subject’s answers as to whether or not and / or to what level of severity he / she is experiencing one or more of a plurality of mental health associated symptoms. In some related embodiments, identifying the one or more impaired neuro-metabolic steps further comprises incorporating / analyzing obtained questionnaire result. According to some embodiments, the questionnaire is solely used for building the framework (as further elaborated herein).

[0026] According to some embodiments, generating the metabolic fingerprint further comprises comparing the detected / determined levels to a baseline physiological level / range.

[0027] In some embodiments, the precursor is an initial precursor of a first metabolic reaction / step in the pathway and / or the metabolite is a final metabolite of a last metabolic reaction / step in the pathway

[0028] In some embodiments, detecting / determining a level of the plurality of neuro - biomarkers further comprise measuring a level of a mineral, a vitamin, an enzyme, or any combination thereof.

[0029] In some embodiments, the neuro-nutritional treatment comprises a precursor and one or more of a vitamin, a mineral, an enzyme, or any combination thereof.

[0030] In some embodiments, the neuro-nutritional treatment comprises a food, a functional food, and / or a supplement, or any combination thereof.

[0031] In some embodiments, the treatment regimen comprises administration / consumption by the subject of the provided neuro-nutrition for a period of at least 8 weeks.

[0032] In some embodiments, the biological urine sample comprises a dry urine card.

[0033] According to some embodiments, the plurality of biomarkers comprises at least one biomarker selected from the neuro substances listed in Table 2. According to some embodiments, the plurality of mental health associated symptoms comprises at least one symptom selected from Table 1. According to some embodiments, the plurality of neuro metabolic pathways comprises at least one metabolic pathway selected from Table 2.

[0034] According to some embodiments, the method further comprises treating the subject with a nutraceutical composition based on the treatment recommendation.

[0035] According to another aspect, there is provided an ex vivo method for classifying a mental health condition of a subject in need thereof, the method comprising: (a) detecting / determining a level of a plurality of neuro -biomarkers in a previously obtained biological urine sample of the subject, wherein the plurality of neuro-biomarkers comprises a neuro substance (NS) and at least one precursor, metabolite and / or co-factor; (b) applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing between the detected / determined levels of at least two neurobiomarkers belonging to a same metabolic pathway; (c) identify one or more impaired neuro- metabolic steps in any of the plurality of neuro metabolic pathways by applying a second algorithm on the fingerprint; and (d) classifying the subject into one or more classes of mentalhealth conditions / categories based on the impaired step of the neuro -metabolic pathway, wherein the classifying comprises distinguishing between at least two mental health conditions.

[0036] In some embodiments, distinguishing between at least two mental health conditions comprises distinguishing between depression and anxiety.

[0037] According to another aspect, there is provided a diagnostic kit for ex vivo detection of neuro-biomarkers associated with mental health, the kit comprising a plurality of urine cards impregnated with agents for detecting a plurality of neuro -biomarkers in a urine sample. According to some embodiments, the kit comprises at least two, at least three or at least four urine cards. According to some embodiments, the kit comprises 7*2-6 urine cards for a week of testing. According to some embodiments, the kit further comprises a questionnaire or link thereto. A non-limiting example of a suitable urine card can be found at https: / / www.zrtlab.com / sample-types / dried-urine / .

[0038] In some embodiments, the plurality of biomarkers comprises at least one biomarker selected from the neuro substances listed in Table 2.

[0039] According to another aspect, there is provided a method for distinguishing between depression and anxiety, the method comprises: (a) detecting / determining a level of a plurality of neuro -biomarkers in a previously obtained biological urine sample of the subject, wherein the plurality of neuro-biomarkers comprises a neuro substance (NS) and at least one precursor, metabolite and / or co-factor; (b) applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing between the detected / determined levels of at least two neuro-biomarkers belonging to a same metabolic pathway; (c) identify one or more impaired neuro -metabolic steps in any of the plurality of neuro metabolic pathways by applying a second algorithm on the fingerprint; and (d) classifying the subject into one or more classes of mental health conditions / categories selected from Anxiety or fear-related disorders (6B00-6B0Z) and Depression and Mood disorders (6A70-7Z) based on the impaired step of the neuro-metabolic pathway.

[0040] In some embodiments, the neuro substance (NS) includes norepinephrine and at least one additional NS selected from the neuro substances listed in Table 2. In some embodiments, the neuro substance (NS) includes norepinephrine and at least one additional NS selected from, PAE, serotonin and GABA, or any combination thereof.

[0041] According to some embodiments, there provided method for managing a mental health condition of a subject in need thereof, the method comprising:(a) obtaining at least one urine sample of the subject;(b) detecting / determining a level of a plurality of neuro -biomarkers of each of a plurality of neuro -metabolic pathways in the urine sample, wherein the plurality of neuro-biomarkers is selected from a neuro substance (NS) and at least one precursor, metabolite and / or co-factor;(c) applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing between the detected / determined levels of at least two neurobiomarkers belonging to a same metabolic pathway (intra-pathway ratio) and comparing a level of the plurality of neuro -biomarkers to a respective level range identified as healthy;(d) applying a second algorithm on the metabolic fingerprint, wherein the algorithm is configured to identify one or more impaired neuro -metabolic steps in any of the plurality of neuro metabolic pathways and to classify the identified one or more impaired neuro-metabolic steps into primary and secondary impairments; and(e) outputting a neuro-nutritional treatment regimen for the subject, based on the identified primary impairment in the one or more neuro-metabolic step, wherein the neuro-nutritional treatment regimen is designed to improve metabolism of the impaired step of a neuro metabolic pathway.

[0042] According to some embodiments, the method further includes an initial step of exposing a dry urine card to a urine sample of the subject.

[0043] According to some embodiments, the first algorithm is a clustering algorithm.

[0044] According to some embodiments, the second algorithm is an anomaly detection algorithm and / or an inference algorithm.

[0045] According to some embodiments, the plurality of biomarkers comprises at least 10 different biomarkers.

[0046] According to some embodiments, the plurality of neurometabolic pathways comprise at least 3 different neurometabolic pathways.

[0047] According to some embodiments, the precursor is an initial precursor of a first metabolic reaction / step in the pathway and / or wherein the metabolite is a final metabolite of a last metabolic reaction / step in the pathway.

[0048] According to some embodiments, detecting / determining a level of the plurality of neuro-biomarkers further comprise measuring a level of a mineral, a vitamin, an enzyme, or any combination thereof.

[0049] According to some embodiments, the neuro-nutritional treatment comprises a precursor and one or more of a vitamin, a mineral, an enzyme, or any combination thereof.

[0050] According to some embodiments, the neuro-nutritional treatment comprises a food, a functional food, and / or a supplement, or any combination thereof.

[0051] According to some embodiments, the treatment regimen comprises administration / consumption by the subject of the provided neuro-nutrition for a period of at least 8 weeks.

[0052] According to some embodiments, the at least one urine sample is a dry urine sample impregnated on a urine card. According to some embodiments, the at least one urine sample comprises at least 2 dry urine samples obtained at different timepoints of a day. According to some embodiments, the at least one urine sample comprises at least 4 dry urine samples obtained at different timepoints of a day. According to some embodiments, the detecting / determining the level of the plurality of neuro -biomarkers comprises averaging the levels detected in each of the dry urine samples obtained at different timepoints of a day.

[0053] According to some embodiments, the plurality of biomarkers comprises at least one biomarker selected from the neuro substances listed in Table 2. According to some embodiments, the plurality of mental health associated symptoms comprises at least one symptom selected from Table 1. According to some embodiments, the plurality of neuro metabolic pathways comprises at least one metabolic pathway selected from Table 2.

[0054] According to some embodiments, the method further comprises treating the subject with a nutraceutical composition based on the treatment recommendation.

[0055] Certain embodiments of the present disclosure may include some, all, or none of the above advantages. One or more technical advantages may be readily apparent to those skilledin the art from the figures, descriptions and claims included herein. Moreover, while specific advantages have been enumerated above, various embodiments may include all, some or none of the enumerated advantages.BRIEF DESCRIPTION OF THE FIGURES

[0056] The invention will now be described in relation to certain examples and embodiments with reference to the following illustrative figures.

[0057] FIG. 1A is an illustrative flowchart of the herein disclosed method managing a mental health condition of a subject in need thereof.

[0058] FIG. IB illustrates assessment of the biochemical steps of a neuro metabolic pathway, here the dopaminergic pathway from the initial nutrient / precursor to the last metabolite, presenting 5 neuro-biomarkers in total, that can be measured in a biological sample, such as urine sample.

[0059] FIG. 1C illustrates the evaluation of concentration of the plurality of biomarkers, according to the present invention. The levels of the biomarkers belonging to biochemical steps of a neuro metabolic pathway are compared between themselves (ascending / descending steps of the solid line), and also to their level range in biological samples derived from healthy individuals (dashed lines), thereby generating a unique fingerprint including the relations between the biomarkers, and reflecting the structure and the functionality of the neuro metabolic pathway (represented by the shape of the solid line itself).

[0060] FIG. ID illustrates an example of an analysis of the metabolic fingerprint and determination of the impaired step(s) of several pathways.

[0061] FIG. IE illustrates an example of an analysis of the metabolic fingerprint based on the analysis of several pathways, determining and distinguishing between categories of MC having common symptoms (e.g. depression vs. anxiety) and further determining the underlying impairment e.g. serotonergic based anxiety or phenylethylamine based anxiety). The fingerprint data may be analyzed in association with the symptom data and its level of severity derived from patient’s questionnaires.

[0062] FIG. IF illustrates an example of a recommended personalized treatment, based on the determination of the MC as anxiety and its underlying neuro-metabolic impairment as Phenylethylamine-based anxiety, but not serotonin-based anxiety.

[0063] FIG. 2A-FIG. 2E show an exemplary outline of the algorithmic flow of the hereindisclosed method managing a mental health condition of a subject in need thereof, such as method 100 of FIG. 1A).

[0064] FIGs. 3A-3J are exemplary graphs presenting correlations between levels of neurobiomarkers associated with neuro-metabolic pathways as measured in dry urine and a degree of self-assessed MC associated symptoms.

[0065] FIG. 3A presents a correlation between reduction in levels of the neuro -biomarker serotonin indicative of impaired serotonergic pathway (responsible for anabolism of serotonin) and increment in severity of an anxiety symptom indicative of impaired mental health. Degree / severity of symptoms is presented as non-existing, mild, moderate, or severe.

[0066] FIG. 3B presents a correlation between reduction in levels of the neuro -biomarker tryptophan and increment in severity of an anxiety symptom indicative of impaired mental health. Degree / severity of symptoms is presented as non-existing, mild, moderate, or severe.

[0067] FIG. 3C schematically illustrates steps in the biosynthesis / anabolism of serotonin and the precursors (tryptophan and 5 -hydroxy tryptophan (5-HTP)) and co-factors (tetrahydrobiopterin (BH4), Fe, tryptophan hydroxylase (TPH), vitamin B6, and aromatic L- amino acid decarboxylase (AADC)) that participate in each step, as well as a step in the biodegradation / catabolism of serotonin to its metabolite 5 -hydroxy indole 3-acetic acid (5- HIAA) using Cu, vitamin B6, monoamine oxidase (MAO) and aldehyde reductase (AR) as cofactors.

[0068] FIG. 3D presents a correlation between reduction in levels of the neuro -biomarker tyrosine and increment in severity of fatigue (lack of energy) symptom indicative of impaired mental health. Degree / severity of symptoms is presented as non-existing, mild, moderate, or severe.

[0069] FIG. 3E presents a correlation between reduction in levels of the neuro -biomarker dopamine and increment in severity of fatigue (lack of energy) symptom indicative of impairedmental health. Degree / severity of symptoms is presented as non-existing, mild, moderate, or severe.

[0070] FIG. 3F presents a correlation between reduction in levels of the neuro -biomarker norepinephrine and increment in severity of fatigue (lack of energy) symptom indicative of impaired mental health. Degree / severity of symptoms is presented as non-existing, mild, moderate, or severe.

[0071] FIG. 3G presents a correlation between reduction in levels of the neuro -biomarker norepinephrine and increment in severity of depression (low mood) symptom indicative of impaired mental health. Degree / severity of symptoms is presented as non-existing, mild, or moderate.

[0072] FIG. 3H presents a correlation between reduction in levels of the neuro -biomarker serotonin and increment in severity of depression (low mood) symptom indicative of impaired mental health. Degree / severity of symptoms is presented as non-existing, mild, or moderate.

[0073] FIG. 31 presents a correlation between reduction in levels of the neuro -biomarker tryptophan and increment in severity of depression (low mood) symptom indicative of impaired mental health. Degree / severity of symptoms is presented as non-existing, mild, or moderate.

[0074] FIG. 3J presents a correlation between reduction in levels of the neuro -biomarker dopamine and increment in severity of addictive behavior. Degree / severity of symptoms is presented as non-existing, mild, or moderate.

[0075] FIG. 4A shows the levels of neuro -biomarkers measured for a patient, and respective normal ranges.

[0076] FIG. 4B shows the metabolic fingerprint of the dopaminergic pathway of the patient.

[0077] FIG. 4C shows the metabolic fingerprint of the serotonergic pathway of the patient.

[0078] FIG. 4D shows the metabolic fingerprint of the dopaminergic pathway of the patient in comparison to healthy ranges (shaded in grey).

[0079] FIG. 4E shows the metabolic fingerprint of the serotonergic pathway of the patient in comparison to healthy ranges (shaded in grey).

[0080] FIG. 4F shows the dopaminergic pathway points of imbalance / impairment.

[0081] FIG. 4G shows the serotonergic pathway points of imbalance / impairment.

[0082] FIG. 4H shows the primary and secondary causes of dopaminergic and serotonergic pathway imbalances.

[0083] FIG. 41 shows the metabolic fingerprint of the dopaminergic pathway of the patient before and after treatment.

[0084] FIG. 4J shows the metabolic fingerprint of the serotonergic pathway of the patient before and after treatment.

[0085] FIG. 4K shows the mental health issues of the patient before and after treatment.DETAILED DESCRIPTION

[0086] In the following description, various aspects of the disclosure will be described. For the purpose of explanation, specific configurations and details are set forth in order to provide a thorough understanding of the different aspects of the disclosure. However, it will also be apparent to one skilled in the art that the disclosure may be practiced without specific details being presented herein. Furthermore, well-known features may be omitted or simplified in order not to obscure the disclosure.Definitions

[0087] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In case of conflict, the patent specification, including definitions, governs.

[0088] 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, “a” and “an” are used herein to refer to one or more than one (i.e., to at least one) of the stated object, unless the context clearly dictates otherwise. By way of example, “a biomarker” means one or more biomarker(s).

[0089] The term “may” refer to an optional / possible approach / possibility, but not a requirement. The term “can” refer to a permissible / plausible approach / possibility, but not a requirement.

[0090] As used herein, "optional" or "optionally" means that the subsequently described event or circumstance does or does not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0091] As used herein, the term "about" when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass deviations / variations of ±20% or in some embodiments ±10%, or in some embodiments ±5%, or in some embodiments ±1%, or in some embodiments ±0.1% from the specified value, as such deviations are appropriate to perform the disclosed methods.

[0092] As used herein, the terms “essentially” and “substantially” are synonymous and when referring to a stated material such as a composition, a substance, and the like, is meant to encompass variations of in some embodiments, ±0.1%, or in some embodiments, ±1%, or in some embodiments, ±2%, or in some embodiments, ±5% from a stated amount, as such variations / deviations are appropriate to perform the disclosed methods. According to some embodiments, the term “essentially devoid of’ may refer to a stated material as either entirely absent or present in a residual amount, such as less than 5%, or less than 2%, or less than 1%, or less than 0.1% are present. Each possibility is a separate embodiment. According to some embodiments, the term “substantially made of’ may refer to a stated material as either entirely present or absent in a neglectable amount, such as more than 95%, or more than 98%, or more than 99%, or more than 99.9% are present. Each possibility is a separate embodiment.

[0093] As used herein, the term “comprising” is synonymous with the terms "including" "containing," or "characterized by," and is inclusive or open-ended i.e. does not exclude additional, unrecited elements. According to some embodiments, the term comprising may be replaced with the term with the term “consisting of’ which excludes any element, step, or ingredient not specified in the claim. According to some embodiments, the term comprising may be replaced with the term “consisting essentially of’ which limits the scope of a claim to the specified materials or steps "and those that do not materially affect the basic and novel characteristics" of the claimed invention.

[0094] As used herein, the terms “prevent”, “reduce”, “attenuate”, “ameliorate”, “inhibit” are used interchangeably.

[0095] As used herein, the terms “enhanced”, “increased”, “elevated” are used interchangeably.

[0096] As used herein, the term “plurality” may refer to at least two (two or more). According to some embodiments, the plurality includes at least two, or at least three, or at least five, at least about ten, at least about twenty, or at least about thirty, or at least about forty, or at least about fifty, or at least about sixty, or at least about seventy, or at least about eighty, or at least about ninety, or at least about one hundred, or at least about two hundreds, or more. Each possibility is a separate embodiment.

[0097] As used herein, the terms "subject", "patient" or "individual" may be used interchangeably and refer to a mammal, preferably a human. According to some embodiments, the subject is a subject suffering from a mental condition. According to some embodiments, the subject is a subject suffering from a plurality of mental condition associated categories, such as at least 2 at least 3, at least 4 or at least 5 mental condition categories. Each possibility is a separate embodiment. As a non-limiting example, the subject may suffer from both anxiety and depression, or both ADHD and depression etc. According to some embodiments, the subject is a subject suffering from a plurality of mental condition associated symptoms, such as at least 2 at least 3, at least 4 or at least 5 or at least 10 mental condition associated symptoms. Each possibility is a separate embodiment. Non-limiting examples of symptoms are provided in table 1 herein. As used herein, the term “mental condition” (”MC”), “mental health condition” or “mental health associated condition” may be interchangeably used to refer to physiological or psychological symptom(s) experienced by a subject, affecting its emotional, psychological, and social well-being.

[0098] According to some embodiments, the present invention is also directed towards prevention. Since it is known in the art that the concentration of neuro biomarkers may change and deviate from normal / typical / optimal physiological baseline levels or range, even before the full scope / extent / severity of the symptom(s) of a condition / disorder are experienced by the subject, the neuro metabolic profiles / fingerprints may be useful for early diagnosis and prevention of development or progression of mental conditions. In other words, such advantageous metabolic fingerprints may serve as a base not only for diagnosis and treatment,but also for early diagnosis and prophylactic treatment, i.e., prior to manifestation of any obvious mental health related symptom or prior to its progression with respect to severity of symptoms.

[0099] According to some embodiments, the mental condition manifests at least one symptom and / or condition associated with mental health. Each embodiment is a separate possibility.

[0100] According to some embodiments, non-limiting examples of mental health associated symptom and / or condition may include any of the symptoms and / or conditions listed in hereinbelow Table 1.

[0101] According to related embodiments, mental health associated symptom and / or condition may include at least one symptom and / or condition selected from: anxious, dizzy spells, OCD, panic attacks, urinary urge increased, premenstrual dysphoric disorder, ringing in ears, aches and pains, acne, allergies, breasts, tender, joint pain, mania (extreme shifts in mood and energy), rapid heartbeat, neck or back pain, eating disorders, sugar cravings, weight gain - breasts or hips, weight gain - hips, weight gain - waist, erection decreased, fatigue - evening, fatigue mental, fatigue - morning, libido decreased, triglycerides elevated, cholesterol high, hoarseness, hot flashes, infertility, rapid aging, uterine fibroids, bone loss, muscle size decreased, numbness - feet or hands, apathy, autism spectrum disorder, burned out feeling, constipation, depressed, flexibility decreased, mood swings, nails breaking or brittle, tearful, ADD / ADHD (attention / focus difficulties), foggy thinking, forgetfulness, mental sharpness decreased, memory lapse, stamina decreased, body temperature cold, bleeding changes, goiter, hair - dry or brittle, heart palpitations, skin thinning, sweating decreased, swelling or puffy eyes / face, pulse rate slow, urine flow decreased, blood pressure high, blood pressure low, blood sugar low, breast cancer, breasts - fibrocystic, chemical sensitivity, developmental delays, fibromyalgia, hair or skin oily, hair - increased facial or body, hair - scalp loss, hearing loss, incontinence, prostate cancer, prostate problems, aggressive behavior, irritable, sleep disturbed, sleeping difficulty, headaches, muscle soreness, nervous, night sweats, stress, vaginal dryness, water retention, and addictive behaviors; or any combination thereof. Each possibility is a separate embodiment.

[0102] According to related embodiments, the symptoms are classified into categories of associated mental health effects / mental health conditions. According to some embodiments, non-limiting examples of categories of associated mental health effects / mental healthconditions include the categories listed in hereinbelow Table 1. According to some embodiments, categories of mental health effects / mental health conditions includes at least one category selected from: (1) Sleep quality, Sleep wake disorders; (2) Stress and burnout, PTSD; (3) Memory, focus and attention; (4) Anxiety, excessive worry and trauma, fear-related disorders (5) Mood disorders and depression; (6) Energy and libido, fatigue and decreased energy; (7) Appetite balance; (8) Susceptibility to addictions; (9) Self-regulation, impulsive control disorders; and (10) Immune system, neuroinflammation and immune defense; or any combination thereof.

[0103] In some embodiments, categories of mental conditions includes 10 ICD 11: (1) Sleep wake disorders (7A00-7A8Z); (2) Stress, burn out, PSTD (6B40-6B4Z, QE00-9Z, QD00-9Z); (3) Memory, focus, attention (incl. autism spectrum disorder 6A02; ADHD 6A05, mild neurocognitive disorder 6D71, dementia 6D80-8Z); (4) Anxiety or fear-related disorders (6B00-6B0Z); (5) Depression and Mood disorders (6A70-7Z); (6) Fatigue and decreased energy (MG22) important; (7) Appetite balance (Eating Disorders 6B80-6B0Z, e.g. Anorexia Nervosa, 6B81 Bulimia Nervosa, 6B82 Binge Eating Disorder, 6B83 Avoidant-Restrictive Food Intake Disorder) life threatening; (8) Susceptibility to addictions (Disorders Due to Substance use or Addictive Behaviors 6C40-6C0Z); (9) Self-regulation (impulse control disorders 6C70-73, 6C45.73-6C47.73, 6C50-51, 6E66, 6B25); and (10) Neuroinflammation.

[0104] Table 1: physical and psychological symptoms and / or conditions and their related mental health category according to ICD 11.

[0105] As used herein, the term "treating" or “managing” may be used interchangeably and refers to an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptom(s) or condition(s) associated with mental health, diminishment of the extent of disease, stabilization of the state of disease, prevention of deterioration of the disease or condition, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total). In some embodiments, the managing, or the treating, includes initial analysis of impairments in neuro metabolic pathways associated with mental health.

[0106] The term “treatment” as used herein may refer to both therapeutic treatment and prophylactic or preventative measures which are based on neuro nutrition, that alter metabolism of an impaired metabolic pathway. In some embodiments, those in need of treatment include those already having a disorder or a symptom as well as those in which the disorder or the symptom is to be prevented. In some embodiments, the treatment includes nutraceutical compositions. In some embodiments, the nutraceutical compositions include precursors, metabolites, and / or co-factors, directed to repair / restore a specific step(s) of a neurometabolic pathway(s) identified as impaired. In some embodiments, the treatment is not a pharmaceutical treatment, and the nutraceutical compositions do not include pharmaceuticals.

[0107] The expression “neuro nutrition” generally refers to nutrients including food and drinks and supplements needed for a subject to maintain a healthy nervous system and neurocognitive functionality and resilience. According to some embodiments, neuro nutrition includes neuro substances (NS) precursor, metabolites, co-factors or any combination thereof. Each possibility is a separate embodiment.

[0108] As used herein, the term “neuro biomarkers” may refer to any of a neuro-substances (NS), metabolites, precursors / nutrients, and / or co-factors belonging to / participating in a neuro metabolic pathway (i.e., pathways affecting the nervous system). Measuring and determining the level / concentration of a neuro biomarker(s) and further comparing between the determined levels of neuro biomarker(s) belonging to a same step and / or pathway may provide information about the integrity of the pathway that biosynthesizes or biodegrades a neuro substance (NS), by a series of step-by-step anabolic or catabolic biochemical reactions, whereas imbalanced concentrations may be indicative of specific impairment in a specific metabolic step of the pathway.

[0109] As used herein, the terms “neuro substance precursor” and “precursor” may be used interchangeably and refer to molecules being metabolized as part of a neuro metabolic pathway, but may also more specifically refer to the end product of the anabolic steps (biosynthesis steps) of a neuro metabolic pathway, and therefore may refer to the molecule (e.g., a neurotransmitter, a hormone) that directly exerts the biological effect of the pathway on the nervous system, possibly through binding to a pre- or post-synaptic receptor or through any other biological mechanism (thereby distinguishing most NS from most precursors or most metabolites that serve mainly as building block for biosynthesis or as side products of biodegradation, or from co-factors that promote neuro metabolic reaction). Nevertheless, the NS of a certain pathway may serve as a precursor or metabolites of a different metabolic reaction.

[0110] A neuro metabolic pathway is usually named according to the main NS that is being biosynthesized (anabolism) or biodegraded (catabolism) by the pathway, reference is made to the 18 non-limiting neuro metabolic pathways and their corresponding NS listed in Table 2 as well as to the specific NS marked in bold which were used in the example as neuro biomarkers (i.e., NS that their concentration was measured). According to related embodiments, NS includes at least one NS selected from the list in Table 2, or any combination hereof.

[0111] A non-limiting example of a precursor is tryptophan (which is metabolized into serotonin). Other non-limiting examples of NS precursors are disclosed in Table 2.

[0112] Table 2: neuro substances and related neuro metabolic pathways. Marked in bold are at least 26 biomarkers measured in dry urine samples collected from the subjects.

[0113] As used herein, the terms “metabolite” and “precursor metabolite” refer to substances, typically small molecules that are an intermediate or an end product of metabolism, i.e. derived from a precursor.

[0114] As used herein the term “co-factor” may refer to a mineral, a vitamin, or an enzyme taking part / aiding in the metabolism of a neuro-metabolic pathway. In some embodiments, a co-factor includes a mineral, a vitamin, an enzyme, or any combination thereof. Each possibility is a separate embodiment.

[0115] As used herein, the term “imbalanced concentrations” may refer to a deviation in the ratio between the levels / amounts of several neuro biomarkers, preferably biomarkers participating in a same metabolic pathway, but possibly also between pathways. The term may also refer to a deviation in the level / amount of a neuro biomarker from the normal physiological levels. According to some embodiments, a level of a neuro -biomarker is determined as being imbalanced if the levels detected deviate by at least 5%, at least 10% or at least 15% from that of a normal levels or level range. Each possibility is a separate embodiment.

[0116] The term “baseline physiological level” as used herein refers to a range of standard physiological concentrations of biomarkers measured in biological samples provided by healthy, non- symptomatic individuals at least with respect to symptoms and / or conditions associated with mental health. The terms “baseline physiological level”, “normal / standard physiological range” may be interchangeably used when referring to the range of healthy physiological concentrations. According to some embodiments, the normal level may be levels previously defined in literature. According to some embodiments, the normal level may be levels determined while excluding subject’s suffering from mental health conditions.

[0117] As used herein, the term “neuro metabolic fingerprint” refers to the determining of the neuro metabolic status of a neuro metabolic pathway or a plurality of neuro metabolic pathways, in a biological sample. This is performed by assessing whether the metabolic steps, preferably each one of the steps, of the chain of biochemical reactions that constitute a certain pathway function properly, or whether one or more of them is impaired, and to what extent, based on the determined levels / concentrations of the neuro biomarkers (preferably each one,but in any case more than two, of the biomarkers, including NS, precursors, metabolites and co-factors) that takes part in the steps / biochemical reactions of the chain of a certain pathway. The assessment of any impairments in the chain of neuro metabolic steps / biochemical reactions is performed by comparing the relative amounts of the neuro biomarkers of a pathway (also referred to herein as “intra-pathway ratio). The assessment of any impairments in the chain of neuro metabolic steps / biochemical reactions further includes comparing the levels of the neuro biomarkers in biological samples of symptomatic subjects with the normal physiological level / range determined in biological samples of non-symptomatic, healthy subjects. The ‘mapping’ of the network of neuro metabolic chains / pathways, in order to generate a metabolic signature of the network based on the determined / characterized levels of the neuro substances (NS), precursors, metabolites, and co-factors, and the relative expression thereof (i.e., the ratio between them) also includes the context of their metabolic / biochemical hierarchy that takes under consideration the sequential order of the biochemical reactions that constitute the steps of the chain of a certain pathway (i.e., intra-pathway) and may also take under consideration the sequential and / or parallel order of the biochemical reactions that are shared by pathways or influence other pathways (i.e., the inter-pathway connection). Therefore, a metabolic signature / fingerprint / map provides a comprehensive understanding / characterization of the neuro metabolic status of symptomatic subjects and can be linked to the subject’s mental health status, by considering the mental health related symptoms and / or conditions and level of severity, provided in the questionnaire. The terms “mapping”, “signature”, and “fingerprint” may be interchangeably used.

[0118] In some embodiments, imbalanced concentrations include deviation in the ratio between the levels / amounts of at least 2 neuro biomarkers participating in a same metabolic pathway. In some embodiments, imbalanced concentrations include deviation in the ratio between the levels / amounts of at least 2 neuro biomarkers participating in a same or different metabolic pathway(s). Each possibility is a separate embodiment.

[0119] As used herein, the term “neuro nutritional treatment” refers to an advantageous personalized recommendation for treatment, or treatment per se, provided to a subject based on consumption / administration of neuro nutrition (i.e., a dietary regimen) including: food and drinks, functional food and drinks, supplements, and / or nutraceutical compositions, naturally rich or artificially enriched with precursors, metabolites, and / or co-factors predicted to restore proper function of specific neuro metabolic pathway, thereby promoting healthy brain andnerve system function and mental health. According to some embodiments, the neuro nutritional treatment is a nutraceutical composition (also referred to a formulation which specifically includes the precursor(s), metabolite(s) and / or co-factor(s) of an impaired metabolic step.

[0120] As used herein, the term “nutraceuticals” or “nutraceutical compositions” refers to nutritional products (i.e., food) serving medical purposes usually made from whole food with extra benefits due to concentrations and combinations of nutrients / substances that participate in metabolism, including precursors, metabolites, and / or co-factors, or any combination thereof that contributes.

[0121] As used herein, the term “functional food” refers to food and beverages relatively (to other food sources) rich or enriched with certain precursors, metabolites and / or co-factors that makes the functional food potentially more beneficial for preserving / restoring healthy neuro metabolism. Accordingly, consumption of suitable food, suitable functional food, suitable food supplements, or suitable nutraceutical compositions, may be predicted by the algorithm as to positively modulate / influence an impaired neuro metabolic step, thereby restoring the neuro metabolic pathway to its optimal / normal functioning.

[0122] In some embodiments, the neuro nutritional treatment or the recommendation for a neuro nutritional treatment of the present disclosure refers to a personalized treatment. In some embodiments, the treatment or the recommendation for treatment is devoid of pharmaceuticals. In some embodiments, the treatment or the recommendation for treatment may include a combination of neuro nutritional treatment with pharmaceutical treatment and / or psychotherapy.

[0123] As used herein, the term “administering” includes routes of administration which allow the food, supplements, and nutraceutical compositions of the invention to perform their intended function. In some embodiments, routes of administration include, but is not necessarily limited to administration via the digestive tract. In some embodiments, the preferred way of administration is consumption by eating or drinking.

[0124] According to an aspect of the disclosure, there is provided an ex vivo method for managing a mental condition (MC) of a subject in need thereof, the method comprising: (a) detecting / determining a level of a plurality of neuro -biomarkers of each of a plurality of neuro- metabolic pathways in a previously obtained biological urine sample of the subject, whereinthe plurality of neuro -biomarkers include a neuro substance (NS) and at least one precursor, metabolite and / or co-factor; (b) computing, preferably by applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers, a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing between the detected / determined levels of the at least two neuro-biomarkers belonging to a same metabolic pathway (intra-pathway ratio) and comparing between the detected / determined levels of the at least two neuro-biomarkers to that of a healthy individual (healthy / MC ratio); (c) identifying one or more impaired neuro-metabolic steps in any of the plurality of neuro metabolic pathways, and classifying the identified one or more impaired neuro-metabolic steps into primary and secondary impairments preferably by applying a second algorithm on the fingerprint, the second algorithm configured to compare the levels of the; (d) computing, preferably by applying a trained machine learning model on the detected / determined levels of the at least two neuro -biomarkers and the impaired neuro-metabolic steps identified to classify the impaired steps as primary or secondary causes of the MC symptoms of a subject and (e) providing a personalized recommendation of a neuro -nutritional treatment regimen for the subject, based on the identified impairment in the one or more neuro-metabolic step, wherein the neuro-nutritional treatment regimen is designed to alter the metabolism of the impaired step of the impaired neuro metabolic pathway identified as being a primary cause.

[0125] According to some embodiments, step b further comprises comparing levels of the neuro-biomarkers between pathways (inter-pathway comparison).

[0126] In some embodiments, the metabolic fingerprint comprises comparing between the detected / determined levels of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, or more, neuro-biomarkers belonging to a same metabolic pathway. Each possibility is a separate embodiment.

[0127] In some embodiments, generating the metabolic fingerprint further comprises comparing the detected / determined levels of the plurality of neuro-biomarkers to a baseline physiological level / range.

[0128] In some embodiments, the neuro-nutritional treatment is for replacing pharmaceutical treatment. In some optional embodiments, the personalized recommendation for treatment further comprises recommendation for pharmaceutical treatment in addition to the neuro- nutritional treatment regimen.

[0129] In some embodiments of the ex vivo method for managing a mental health condition of a subject, at least steps (b) and (c), and optionally step (d) and (e) are performed by the computer.

[0130] Reference is now made to FIG 1A, which is an illustrative flowchart of a method 100 for managing a mental health condition, according to some embodiments. It is understood that while some steps are necessarily sequential and must be conducted in the indicated order, other steps may be conducted simultaneously or even in a different order. One of the ordinary skills in art will readily understand the order of which steps are rigid, and which can be changed.

[0131] In step 110 of method 100, the levels of neuro -biomarkers, in a urine sample of a subject, are measured. According to some embodiments, some of the biomarkers are nutrients (e.g. tyrosine), some of them signaling molecules (e.g. DOPA or dopamine), and / or some of them are metabolites (e.g. vanillylmandelic acid VMA). According to some embodiments, the biomarker may have multiple roles, e.g. being a metabolite and possessing certain in / direct signaling capabilities. A non-limiting example of such dual-function biomarker is DOPAC.

[0132] In some embodiments, detecting / determining a level of a plurality of neuro-biomarkers includes measuring / determining the amount / concentration of at least one neuro-substances (NS) and at least one of a metabolite, precursor, and / or a co-factor, or any combination thereof. Each possibility is a separate embodiment.

[0133] Non-limiting examples of methods for detecting / determining a level of a plurality of neuro-biomarkers includes, according to some embodiments, chromatography, an immunoassay such as ELISA-based assay, nucleic acid probe-based assay such as aptamerbased assay, PCR, next generation sequencing, an electrochemistry-based assay, a lateral-flow assay, a nanobead-based assay, a microfluidics-based assay, and an oligonucleotide-templated reaction.

[0134] In some embodiments, the biological sample is a previously obtained biological sample. In some embodiments, the biological sample is a urine sample and / or a blood sample. Each possibility is a separate embodiment. In some embodiments, the biological sample is a urine sample. In some embodiments, the urine sample is a dry urine sample. In some embodiments, the blood sample is a dry blood sample. In some embodiments, the urine sample is impregnated on a urine card. In some embodiments, the urine sample includes at least two urine samples taken at different timepoints of the day (e.g. morning and evening). In some embodiments, theurine sample includes at least three urine samples taken at different timepoints of the day. In some embodiments, the urine sample includes at least four urine samples taken at different timepoints of the day.

[0135] In some embodiments, neuro-biomarkers are measured more than one time a day, such as at least twice or at least 4 times during the day. In some embodiments, neuro -biomarkers are measured in the morning at awakening time, two hours after waking up, in the evening before dinner (e.g., 5p.m-7p.m), and before going to sleep.

[0136] In some embodiments, detecting / determining a level of a plurality of neuro-biomarkers includes determining the levels per gram of creatinine.

[0137] In some embodiments, the plurality of neuro -biomarkers includes at least a neuro substance (NS), at least 2 NS, at least 3 NS, at least 4 NS, at least 5 NS, at least 6 NS, at least 7 NS, at least 8 NS, at least 9 NS, at least 10 NS, at least 11 NS, at least 12 NS, at least 13 NS, at least 14 NS, at least 15 NS, at least 16 NS, at least 17 NS, at least 18 NS, at least 19 NS, at least 20 NS, at least 21 NS, at least 22 NS, at least 23 NS, at least 24 NS, at least 25 NS, at least 26 NS, at least 27 NS, at least 28 NS, at least 29 NS, at least 30 NS, or more. Each possibility is a separate embodiment.

[0138] In some embodiments, determining the level of a plurality of neuro metabolic biomarkers includes determining the level of at least one precursor, metabolite, or co -factor, at least 2 precursors, metabolites, and / or co-factors, at least 3, at least 4, at least about 5, at least about 8, at least about 10, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45 or more, precursors, metabolites, and / or co-factors. Each possibility is a separate embodiment.

[0139] In some embodiments, the plurality of neuro -biomarkers includes at least one, metabolite, precursor, or co-factor, at least 2 metabolites, precursors, or co-factors, or any combination thereof; at least 3 metabolites, precursors, or co-factors, or any combination thereof; at least 4 metabolites, precursors, or co-factors, or any combination thereof; at least 5 metabolites, precursors, or co-factors, or any combination thereof; at least 6 metabolites, precursors, or co-factors, or any combination thereof; at least 7 metabolites, precursors, or cofactors, or any combination thereof; at least 8 metabolites, precursors, or co-factors, or any combination thereof; at least 9 metabolites, precursors, or co-factors, or any combination thereof; at least 10 metabolites, precursors, or co-factors, or any combination thereof; at least11 metabolites, precursors, or co-factors, or any combination thereof; at least 12 metabolites, precursors, or co-factors, or any combination thereof; at least 13 metabolites, precursors, or cofactors, or any combination thereof; at least 14 metabolites, precursors, or co-factors, or any combination thereof; at least 15 metabolites, precursors, or co-factors, or any combination thereof; at least 16 metabolites, precursors, or co-factors, or any combination thereof; at least 17 metabolites, precursors, or co-factors, or any combination thereof; at least 18 metabolites, precursors, or co-factors, or any combination thereof; at least 19 metabolites, precursors, or cofactors, or any combination thereof; at least 20 metabolites, precursors, or co-factors, or any combination thereof; at least 25 metabolites, precursors, or co-factors, or any combination thereof; at least 30 metabolites, precursors, or co-factors, at least 35 metabolites, precursors, or co-factors, at least 40 metabolites, precursors, or co-factors, at least 45 metabolites, precursors, or co-factors, at least 50 metabolites, precursors, or co-factors, or any combination thereof; or more. Each possibility is a separate embodiment.

[0140] According to related embodiments, neuro biomarkers may include at least one neurosubstances (NS), and at least one metabolites, precursors, and / or co-factors selected from: 3- Hydroxy kynurenine; 5-Hydroxyindoleacetic acid (5-HIAA); Creatinine; 3,4-Dihydroxyphenylacetic acid (DOPAC); Dopamine; Epinephrine; Gamma-aminobutyric acid (GABA); Glutamate; Glutamine (non-essential amino acid); Glycine (non-essential amino acid); Histamine; Histidine; Homovanillic acid (HVA); Kynurenic acid; Kynurenine; N- Methylhistamine; Norepinephrine; Normetanephrine; Phenethylamine (PEA); Serotonin; Taurine; Tryptophan; Tyramine; Tyrosine (non-essential amino acid); Vanillylmandelic acid (VMA); Xanthurenic acid; Acetylcholine; other non-essential amino acids selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, proline and serine; other essential amino acids selected from BCAA (branched-chain amino acids pathways, such as leucine, isoleucine, and valine), phenylalanine and methionine; cofactors selected from hormones, amino acids, their metabolites and derivates, vitamins, minerals and acids such as Carnitine, L-lysine and L-methionine, , Citrulline, Copper, Cortisol, DHA, EPA, Ferritin, Linoleic Acid, Lauric Acid, Lysine, Magnesium, Molybdenum, Manganese, NAD (nicotinamide adenine dinucleotide), Oleic Acid, Omega 3 Index, Ornithine, Proline, Selenium, Vitamin A, Vitamin A Retinol, Vitamin B2, B3, B5, B6, B9, B12, Vitamin C, Vitamin D, Vitamin E Alpha Tocopherol and Zinc; cofactors as enzymes enabling steps of metabolic pathways selected from BH4 tetrahydrobiopterin, MTHF methyl tetrahydrofolate, SAMe S- adenosyl methionine, AADC aromatic L-amino acid decarboxylase, AANMT arylalkylamineN-methyltransferase, ALDH aldehyde dehydrogenase, AR aldehyde reductase, CDO cysteine dioxygenase, COMT catechol-O-methyltransferase, CSAD cysteinesulfinic acid decarboxylase, DBH dopamine beta hydroxylase, FKF N-Formyl kynurenine formamidase, GAD glutamate decarboxylase, GLS glutaminase, GS glutamine synthetase, HD hypotaurine dehydrogenase, HDC histidine decarboxylase HIOMT hydroxyindole-O-methyltransferase, HNMT histamine N-methyltransferase, IDO indoleamine 2,3-dioxygenase, KAT kynurenine aminotransferase, KMO kynurenine hydroxylase / monooxygenase, MAO monoamine oxidase, M6H melatonin 6 hydroxylase, M6ST melatonin 6 sulfotransferase, PAH phenylalanine hydroxylase, PNMT phenylethanolamine N-methyltransferase, SHMT serine hydroxymethyltransferase, TD tyrosine decarboxylase, TDO tryptophan 2,3-dioxygenase, TH tyrosine hydroxylase, ThrA threonine aldolase and TPH tryptophan hydroxylase; or any combination thereof. Each possibility is a separate embodiment.

[0141] According to related embodiments, NS includes at least one NS selected from: tryptophan, serotonin, 5-HIAA, GABA, Glycine, Taurine, Glutamate, Glutamine, Histidine, Histamine, N-Methylhistamine, PEA, Tyrosine, Tyramine, Dopamine, DOPAC, HVA, Norepinephrine, Normetanephrine, Epinephrine, Kynurenine, Kynurenic acid, 3- Hydroxykynurenine, Xanthurenic acid, and Acetylcholine, or any combination thereof. Each possibility is a separate embodiment.

[0142] According to related embodiments, NS includes at least one NS selected from: 3- Hydroxy kynurenine; 5-Hydroxyindoleacetic acid (5-HIAA); Creatinine; 3,4- Dihydroxyphenylacetic acid (DOPAC); Dopamine; Epinephrine; Gamma-aminobutyric acid (GABA); Glutamate; Glutamine (non-essential amino acid); Glycine (non-essential amino acid); Histamine; Histidine; Homovanillic acid (HVA); Kynurenic acid; Kynurenine; N- Methylhistamine; Norepinephrine; Normetanephrine; Phenethylamine (PEA); Serotonin; Taurine; Tryptophan; Tyramine; Tyrosine (non-essential amino acid); Vanillylmandelic acid (VMA); Xanthurenic acid; Acetylcholine and any combination thereof. Each possibility is a seperate embodiment.

[0143] The disclosure exemplifies correlations between neuro biomarkers concentrations detected in urine samples and health related symptoms at different levels of perceived severity. Non-limiting examples of such correlation include, according to some embodiments, an association between GABA and anxiety, association between serotonin and anxiety, association between phenylethylamine and anxiety, association between GABA andfatigue / low energy, association between tyrosine and fatigue / low energy, association between GABA and memory degeneration, association between norepinephrine depression / low mood, and association between norepinephrine and self-regulation. Each possibility is a separate embodiment.

[0144] It was found by the inventors of the present application that urine is an excellent specimen that enables identifying statistically relevant correlations between the levels of neurobiomarkers and mental conditions (e.g., low mood and depression, anxiety, low motivation and energy, sleep difficulties, appetite balance etc.). The reasons are multiple:

[0145] Ultimately, all metabolic products are excreted in the urine. In some embodiments, the biomarker obtained found in the urine sample is a metabolized urine marker.

[0146] While some biomarkers may be measured in blood, urine testing is often advantageous since biomarker levels blood can change quickly and may also be affected by the stress of testing.

[0147] Neuro-biomarkers are present in small amounts in the urine of all people, sick and healthy. They allow determination of average intervals in healthy individuals and low / high intervals associated with symptoms and deterioration of health. An example of previously reported normal urine levels of certain neuro -biomarkers (tryptophan, taurine, glutamate, glycine, histidine, etc.) can be found here: https: / / www Jabcorp.com / resource / urine-amino- aci d-reference-intery als . However, importantly, the previously reported intervals have not been associated with neuro-metabolic pathways or mental health conditions. According to some embodiments, the normal or non- symptomatic intervals have been redefined in context of mental health. For example, it was found that non- symptomatic populations often have biomarker levels outside the previously reported interval indicating that mental health symptomatic populations may have been included in the previously reported “normal” intervals.

[0148] In step 120 of method 100, the biomarkers are grouped into pathway steps and the levels of a biomarker obtained for a patient are compared to a healthy reference interval. In this step, the intra-pathway ratio levels of neuro-biomarkers in symptomatic persons is also determined in order to identify inefficient pathway steps. According to some embodiments, the ratios are likewise compared to those of healthy individuals.

[0149] In step 130 of method 100 a neuro-metabolic fingerprint of the patient is generated, meaning the patient’s own pattern of neuro-metabolism, is broken down for each metabolic pathway. According to some embodiments, generating the fingerprint includes computing the intra-pathway ratios, health / MC ratios for each step as well as links to other of the patient’s neurometabolic pathways, to map metabolic pathway imbalances.

[0150] In some embodiments, neuro metabolic fingerprint includes a neuro metabolic pathway or a plurality of neuro metabolic pathways. Each possibility is a separate embodiment. In some embodiments, neuro metabolic fingerprint includes at least a neuro metabolic pathway. In some embodiments, neuro metabolic fingerprint includes at least 1 neuro metabolic pathway, at least 2 neuro metabolic pathways, at least 3 neuro metabolic pathways, at least 4 neuro metabolic pathways, at least 5 neuro metabolic pathways, at least 6 neuro metabolic pathways, at least 7 neuro metabolic pathways, at least 8 neuro metabolic pathways, at least 9 neuro metabolic pathways, at least 10 neuro metabolic pathways, or more neuro metabolic pathways. Each possibility is a separate embodiment.

[0151] It is to be understood that even if a symptomatic individual has a level of a biomarker, for example serotonin, in the same range as other healthy individuals, by analyzing the steps of the serotonergic pathway, and comparing the levels / ratio of neuro biomarkers of each step (or at least some steps) with the previous and the following one, an impaired metabolism in that pathway can be discovered. In accordance, the importance of the herein disclosed method lies not solely in the measuring of neuro biomarkers of the symptomatic subject and comparing them with the previously defined levels of healthy individuals, but in the analysis of the levels of neuro biomarkers of the symptomatic subject in relation to each other (also referred to as “intra-pathway ratio”), organized in neuro-metabolic pathways.

[0152] In some embodiments, generating a neuro metabolic fingerprint includes comparing between the detected / determined levels of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, or more, neuro metabolic biomarkers belonging to a same metabolic pathway. Each possibility is a separate embodiment.

[0153] In some embodiments, generating a neuro metabolic fingerprint includes determining the level of neuro metabolic biomarkers belonging to a plurality of neuro metabolic pathways (i.e., 2 or more neuro metabolic pathways). Each possibility is a separate embodiment.

[0154] In some embodiments, generating a neuro metabolic fingerprint includes determining the level of neuro metabolic biomarkers belonging to 2 or more neuro metabolic pathways, 3 or more neuro metabolic pathways, 4 or more neuro metabolic pathways, about 5 or more neuro metabolic pathways, about 10 or more neuro metabolic pathways, about 15 or more neuro metabolic pathways, about 18 or more neuro metabolic pathways, about 20 or more neuro metabolic pathways, or about 25 or more neuro metabolic pathways. Each possibility is a separate embodiment.

[0155] In some embodiments, generating a neuro metabolic fingerprint includes determining the level of neuro metabolic biomarkers belonging to one or more steps in the chain of a neuro metabolic pathway, 2 or more steps in the chain of a neuro metabolic pathway, 3 or more steps in the chain of a neuro metabolic pathway, preferably generating a neuro metabolic fingerprint includes determining the level of neuro metabolic biomarkers belonging to all the steps in the chain of the neuro metabolic pathway.

[0156] In some embodiments, generating a neuro metabolic fingerprint includes complete mapping / characterization of the steps of a metabolic chain of a neuro metabolic pathway, wherein the complete characterization includes determining the levels of the neuro substances (NS), precursors, metabolites, and co-factors and the sequential order of the biochemical reactions that constitute the steps of the chain of the pathway (i.e., intra-pathway).

[0157] In some embodiments, generating a neuro metabolic fingerprint further includes the sequential and / or parallel order of the biochemical reactions that are shared by pathways or influence other pathways (i.e., the inter-pathway connection). Each possibility is a separate embodiment.

[0158] In some embodiments, generating a neuro metabolic fingerprint includes characterizing the metabolic chains starting from the initial precursor / nutrient.

[0159] For example, the tryptophan / serotonergic pathway consists of the following chain of metabolic steps: (1) Tryptophan; (2) 5HTP; (3) serotonin; (4) 5-HIAA Tryptophan; (5) Kynurenine; (6) Kynurenic acid; (7) 3 -OH Kynurenine; (8) Xanthurenic acid. Measuring only the main NS (serotonin) can lead to false conclusions, but measuring also the initial nutrient obtained from food (tryptophan) promotes understanding whether the impairment in the pathway relates to insufficient consumption / intake of the nutrient tryptophan in the food (i.e., the initial precursor), or to the or an impaired metabolism of the subject. In this non-limitingexample, the subject shows a low intake of tryptophan and a high level of serotonin, due to a too high synthesis of serotonin. If only serotonin is measured, and the level is too high, lowering the intake of tryptophan even further is not helpful and could affect other vital tryptophandependent processes. Instead, lowering the precursors and / or cofactors of the synthesis of serotonin (vitamins, minerals, enzymes responsible for this step) rather than lowering serotonin itself or tryptophan - could be much more effective.

[0160] In some embodiments, generating a neuro metabolic fingerprint includes determining the level of a plurality of neuro metabolic biomarkers (i.e., 2 or more neuro metabolic biomarkers). Each possibility is a separate embodiment.

[0161] In some embodiments, generating a neuro metabolic fingerprint includes determining the level of 2 or more neuro metabolic biomarkers, 3 or more neuro metabolic biomarkers, 4 or more neuro metabolic biomarkers, about 5 or more neuro metabolic biomarkers, about 8 or more neuro metabolic biomarkers, about 10 or more neuro metabolic biomarkers, about 12 or more neuro metabolic biomarkers, about 15 or more neuro metabolic biomarkers, about 20 or more neuro metabolic biomarkers, about 25 or more neuro metabolic biomarkers, about 30 or more neuro metabolic biomarkers, or about 35 or more neuro metabolic biomarkers. Each possibility is a separate embodiment.

[0162] In some embodiments, generating a neuro metabolic fingerprint includes analyzing a deviation in the level / amount of a neuro biomarker from the normal physiological levels, or deviation in the ratio between the levels / amounts of several neuro biomarkers). Each possibility is a separate embodiment.

[0163] In step 140 of method 100, the metabolic fingerprint and the identified impairments are analyzed. According to some embodiments, the knowledge of deviations per se does not provide the answers regarding the reasons of the impairments, which is imperative in identification of the root cause and dedicated treatment thereof.

[0164] A low level of a certain step of the pathway can depend on many different factors. One of the common factors is an insufficient synthesis, depending on the low levels of either precursors, or cofactors, or both. Another reason for a low level of a molecule could be an elevated level of metabolism. Other factors could originate from enzymology, e.g. being caused by genetic inefficiencies in the production of metabolizing enzymes.

[0165] Therefore, in step 150, the fingerprint is further analyzed to identify root causes of the identified point of imbalance. According to some embodiments, identifying a root cause comprises classifying the points of imbalances as primary and secondary. According to some embodiments, subtypes of mental condition(s) are identified, along with their neuro-metabolic causes and / or contributing factors.

[0166] In step 160, a treatment recommendation in the form of neuro-nutritional treatment regimen, is provided, based on the identified impairment in the one or more neuro-metabolic step. According to some embodiments, the neuro-nutritional treatment regimen is designed to alter the metabolism of the impaired neuro metabolic pathway upstream or downstream to the neuro substance (NS).

[0167] It is to be understood that the neuro-nutritional treatment does often not aim to increase / decrease the concentration levels or combinations of measured biomarkers, metabolites etc. Instead, the precursors and cofactors of an impaired step (nutrients, vitamins, minerals, enzymes) that modulate up or down such step may be provided, thereby indirectly increasing / decreasing the synthesis or metabolism of the metabolite.

[0168] According to some embodiments, method 100 further includes step 170 of treating the subject with a nutraceutical composition, based on the treatment recommendation.

[0169] In some embodiments, treatment regimen includes at least 8 weeks of treatment. In some embodiments, the treatment regimen comprises administration / consumption by the subject of the provided neuro-nutrition for a period of at least about 8 weeks, at least about 12 weeks, at least about 16 weeks, at least about 20 weeks, at least about 30 weeks, at least about 40 weeks, or more. Each possibility is a separate embodiment.

[0170] Advantageously, in some embodiment, the recommendation for a neuro nutritional treatment includes a personalized recommendation regarding consumption / administration of food including drinks, functional food and drinks, supplements, and / or nutraceutical and / or pharmaceutical compositions, naturally rich or artificially enriched with precursors, metabolites, and / or co-factors, or any combination thereof. Each possibility is a separate embodiment.

[0171] Advantageously, in some embodiment, the personalized neuro nutritional treatment includes consumption / administration of food including drinks, functional food and drinks,supplements, and / or nutraceutical compositions, naturally rich or artificially enriched with precursors, metabolites, and / or co-factors, or any combination thereof. Each possibility is a separate embodiment.

[0172] Advantageously, in some embodiments, the neuro nutritional treatment is predicted to restore proper function of specific neuro metabolic pathway / step, thereby promoting healthy brain function and mental health.

[0173] In some embodiments, the neuro nutritional treatment includes food, functional food, supplements, and / or nutraceutical and / or pharmaceutical compositions, or any combination thereof. Each possibility is a separate embodiment.

[0174] In some embodiments, the neuro nutrition includes naturally rich or artificially enriched food including drinks, functional food / drinks, supplements, and nutraceutical compositions, or any combination thereof. Each possibility is a separate embodiment.

[0175] In some embodiments, the neuro nutrition includes food, functional food, supplements, and nutraceutical compositions, naturally rich or artificially enriched with precursors, metabolites, and / or co-factors, or any combination thereof. Each possibility is a separate embodiment.

[0176] In some embodiments, the treatment of the present invention includes pharmaceuticals. In some embodiments, the neuro nutritional treatment further includes pharmaceuticals.

[0177] According to some embodiments, non-limiting examples of functional foods include, fruits (e.g. berries, kiwi, pears, peaches, apples, oranges, bananas), vegetables (e.g. broccoli, cauliflower, kale, spinach, zucchini), nuts (e.g. almonds, cashews, pistachios, macadamia nuts, Brazil nuts), seeds (e.g. chia, flax, hemp, pumpkin), legumes (e.g. black beans, chickpeas, navy beans, lentils), whole 10 grains (e.g. oats, barley, buckwheat, brown rice, couscous), seafood (e.g. salmon, sardines, anchovies, mackerel, cod), fermented foods (e.g. tempeh, kombucha, kimchi, kefir, sauerkraut), herbs and spices (e.g. turmeric, cinnamon, ginger, cayenne pepper), and beverages (e.g. coffee, green tea, black tea).

[0178] According to some embodiments, non-limiting examples of functional food rich in vitamin B2 is selected from dried spirulina, dried parsley, dried coriander, dried shitake mushrooms, paprika, had goat cheese, feta cheese, dry roasted, almonds, tea, sun-dried tomatoes, sesame paste (tahini), cooked wild salmon, safflower seeds, sunflower seeds,caraway seeds, portobellos, Greek yoghurt, and fava beans, or any combination thereof. Each possibility is a separate embodiment.

[0179] According to some embodiments, non-limiting examples of functional food rich in vitamin B 12 is selected from cooked wild salmon, cooked, cod, Swiss cheese, mozzarella, feta cheese, whey protein powder, Greek yoghurt, and raw sauerkraut, or any combination thereof. Each possibility is a separate embodiment.

[0180] According to some embodiments, non-limiting examples of functional food rich in vitamin B3 is selected from dry roasted or raw peanuts, dried gingko nuts, dried shitake mushrooms, peanut butter, dried spirulina seaweed, dried coriander, dried parsley, ground ginger, paprika, wild cooked salmon, hemp seeds, sun-dried tomatoes, and chia seeds, or any combination thereof. Each possibility is a separate embodiment.

[0181] According to some embodiments, non-limiting examples of functional food rich in vitamin B6 is selected from ground sage, dried spearmint, dried chives, garlic powder, dried rosemary, paprika, pistachio nuts, hazelnuts, sunflower seeds, safflower seeds, hemp seeds, garlic, dried shitake mushrooms, wild cooked salmon, low moisture prunes, dried coriander, ground ginger, dried gingko nuts, peanut butter, fennel seeds, and low moisture dried apricots, or any combination thereof. Each possibility is a separate embodiment.

[0182] According to some embodiments, non-limiting examples of functional food rich in vitamin B9 is selected from dried spearmint, edamame, dried basil, rosemary, dried coriander, ground sage, soy-based protein powder, raw peanuts, sunflower seeds, spinach, turnip greens, cooked lentils, cooked pinto beans, kelp seaweed, soy protein powder, cooked chickpeas, dried shitake mushrooms, ground mustard seeds, safflower seeds, cooked mung beans, cooked black beans, and cooked asparagus, or any combination thereof. Each possibility is a separate embodiment.

[0183] According to some embodiments, non-limiting examples of functional food rich in vitamin C is selected from dried chives, dried coriander, raw guava, black currants, sauteed green bell peppers, sauteed red bell peppers, raw kale, kiwi, raw broccoli, raw green cauliflower, cooked broccoli, raw papaya, raw turnip greens, strawberries, raw red cabbage, cooked kohlrabi, oranges, lemons, orange juice, clementines, pineapple, cooked cauliflower, raw green peas, sun-dried tomatoes, or any combination thereof. Each possibility is a separate embodiment.

[0184] According to some embodiments, non-limiting examples of functional food rich in cupper (Cu) is selected from dried spirulina seaweed, dried shitake mushrooms, cocoa powder, soy protein powder, sesame seeds, poppy seeds, hemp seeds, dried sunflower seeds, safflower seeds, dried pumpkin seeds, raw or dry roasted cashews, Brazil nuts, walnuts, pine nuts, hazelnuts, raw or roasted pistachio nuts, raw peanuts, raw or dry roasted almonds, dried basil, dried coriander, ground turmeric, sesame paste (tahini), dried spearmint, sun-dried tomatoes, chia seeds, caraway seeds, dried chives, paprika, goat cheese, peanut butter, low-moisture dried apricots or prunes, dried gingko nuts, turnip greens, portobellos, raw tofu, and cooked chickpeas, or any combination thereof. Each possibility is a separate embodiment.

[0185] According to some embodiments, non-limiting examples of functional food rich in cysteine or cystine is selected from soy protein powder, hemp seeds, dried spirulina seaweed, dried sunflower seeds, sesame seeds, pumpkin seeds, safflower seeds, poppy seeds, chia seeds, raw cashews, raw or roasted peanuts, Brazil nuts, roasted pistachio nuts, pine nuts, hazelnuts, almonds, walnuts, sesame paste (Tahini), dried parsley, Gruyere cheese, Swiss cheese, gouda cheese, parmesan, boiled edamame, cooled wild salmon, cocoa powder, paprika powder, peanut butter, dried spearmint, dried basil, dried shiitake mushrooms, boiled lupin beans, sun- dried tomatoes, falafel, and rye bread, or any combination thereof. Each possibility is a separate embodiment.

[0186] According to some embodiments, non-limiting examples of functional food rich in DOPA or L-DOPA is selected from mununa pruriens fresh beans, other types of Mucuna family, Holtanii, Andreana, Aterrino and Gigantean, Tamarindus indica seeds, Sesbania bispinosa seeds, Entada scandens seeds, Acacia leucophlea seeds, Bauhinia variegate seeds, Canavalia gladiate seeds, Vigna aconifolia, and unguiculata, or any combination thereof. Each possibility is a separate embodiment.

[0187] According to some embodiments, non-limiting examples of functional food rich in Fe is selected from dried basil, dried spearmint, dried marjoram, dried coriander, dried rosemary, dried parsley, dried chives, cumin seeds, sesame seeds, dried pumpkin seeds, hemp seeds, ground turmeric, ground sage, ground paprika, ground ginger, curry powder, dried spirulina seaweed, soy protein powder, cocoa powder, morel mushrooms, chantarelle mushrooms raw, sun-dried tomatoes, sesame paste (Tahini), lemon grass, chia seeds, dried sunflower seeds, safflower seeds, Goji berries, cashew nuts, dried pine nuts, hazelnuts, raw peanuts, dry pistachio nuts, almonds, olives, low moisture dried apricots, low moisture prunes, boilededamame, oat (uncooked), cooked large white beans, cooked spinach, and falafel, or any combination thereof. Each possibility is a separate embodiment.

[0188] According to some embodiments, non-limiting examples of functional food rich in GABA is selected from GABA tea, flowers of Chinese Windmill Palm (Trachycarnus fortune) and Astragalus manaholicus, or any combination thereof. Each possibility is a separate embodiment.

[0189] According to some embodiments, non-limiting examples of functional food rich in glutamic acid or glutamine is selected from soy protein powder, parmesan, provolone, Gruyere cheese, Swiss cheese, hard goat cheese, mozzarella, cheddar, camembert, cottage cheese, feta cheese, dried spirulina seaweed, hemp seeds, dried pumpkin seeds, dried sunflower seeds, ground mustard seeds, poppy seeds, sesame seeds, safflower seeds, caraway seeds, fennel seeds, almonds, raw peanuts, roasted peanuts, raw cashews, dry roasted or raw pistachio nuts, roasted cashews, hazelnuts, Brazil nuts, pine nuts, sun-dried tomatoes, peanut butter, dried chives, dried parsley, dried basil, sesame paste (Tahini), cooked wild salmon, cooked cod, cooked lupin beans, chia seeds, firm tofu, edamame, cocoa powder, rye whole grain bread, dried shiitake mushrooms, falafel, curry powder, paprika powder, dried spearmint, onion powder, and dried gingko nuts, or any combination thereof. Each possibility is a separate embodiment.

[0190] According to some embodiments, non-limiting examples of functional food rich in glycine is selected from soy protein powder, dried spirulina seaweed, dried pumpkin seeds, ground mustard seeds, caraway seeds, safflower seeds, pistachio nuts, dried parsley, dried basil, dried chives, paprika, dried oregano, ground ginger, ground turmeric, hemp seeds, dried sunflower seeds, sesame seeds, dried spearmint, poppy seeds, chia seeds, raw or roasted peanuts, almonds, pistachio nuts, cashew, walnuts, Brazil nuts, hazelnuts, pine nuts, pecans, peanut butter, wild cooked salmon, cooked cod, sesame paste (Tahini), chia seeds, cocoa powder, boiled edamame, tofu, parmesan, Gruyere cheese, mozzarella, boiled lupin beans, falafel, dried gingko nuts, edamame, and dried shiitake mushrooms, or any combination thereof. Each possibility is a separate embodiment.

[0191] According to some embodiments, non-limiting examples of functional food rich in histidine is selected from soy protein powder, parmesan, Gruyere cheese, Edam cheese, gouda cheese, mozzarella, hard goat cheese, camembert, feta cheese, dried spirulina seaweed,hemp seeds, dried pumpkin seeds, caraway seeds, dried sunflower seeds, poppy seeds, fennel seeds, dried parsley, dried basil, dried spearmint, wild cooked salmon, cooked cod, raw peanuts, roasted peanuts, almonds, roasted pistachio nuts, cashews, hazelnuts, Brazil nuts, walnuts, pine nuts, dried pine nuts, roasted peans, peanut butter, chia seeds, sesame paste (Tahini), boiled edamame, boiled lupin beans, firm tofu, falafel, cocoa powder, cottage cheese, curry powder, cooked large white beans, edamame, boiled red kidney beans, or any combination thereof. Each possibility is a separate embodiment.

[0192] According to some embodiments, non-limiting examples of functional food rich in Mn (Mangan) is selected from ground chives, ground ginger, cardamon, ground turmeric, cinnamon, curry powder, ground nutmeg, paprika, dried spearmint, dried basil, dried parsley, dried coriander, dried oregano, dried rosemary, dried chives, dried mango, hemp seeds, celery seeds, poppy seeds, fennel seeds, dried pumpkin seeds, cumin seeds, sesame seeds, safflower seeds, dried sunflower seeds, coriander seeds, caraway seeds, hazelnuts, pecans, pine nuts, almonds, raw peanuts, roasted peanuts, raw cashews, Brazil nuts, pistachio nuts, lemon grass, uncooked oats, cocoa powder, wild frozen blueberries, chia seeds, dried spirulina seaweed, sun-dried tomatoes, peanut butter, sesame paste (Tahini), cooked lima beans, firm tofu, hummus, dried shiitake mushrooms, mashed sweet potatoes, cooked chickpeas, and boiled edamame, or any combination thereof. Each possibility is a separate embodiment.

[0193] According to some embodiments, non-limiting examples of functional food rich in Mg (Magnesium) is selected from dried basil, dried coriander, dried chives, dried spearmint, dried parsley, dried oregano, dried rosemary, hemp seeds, dried pumpkin seeds, celery seeds, cumin seeds, sesame seeds, safflower seeds, coriander seeds, dried sunflower seeds, pumpkin seeds, caraway seeds, cocoa powder, Brazil nuts, raw cashews, almonds, pine nuts, peanuts, hazelnuts, pecans, pistachio nuts, chia seeds, curry powder, cardamon, paprika, ground ginger, ground turmeric, ground nutmeg, dried spirulina seaweed, sun-dried tomatoes, uncooked oats, dried shiitake mushrooms, sesame paste (Tahini), cooked spinach, boiled edamame, falafel, hummus, cooked lima beans, cooked black beans, cooked white beans, and cooked quinoa, or any combination thereof. Each possibility is a separate embodiment.

[0194] According to some embodiments, non-limiting examples of functional food rich in Mo is selected from soy protein powder, dried spirulina seaweed, parmesan, Gruyere cheese, Mozzarella, hard goat cheese, Camembert, feta cheese, dried pumpkin seeds, ground mustard seeds, dried sunflower seeds, caraway seeds, safflower seeds, poppy seeds, fennel seeds, driedparsley, dried basil, dried spearmint, dried chives, hemp seeds, raw peanuts, almonds, pistachio nuts, raw cashews, hazelnuts, Brazil nuts, pine nuts, peanut butter, chia seeds, wild cooked salmon, cooked cod, sesame paste (Tahini), cocoa powder, uncooked oats, boiled edamame, firm tofu, falafel, boiled lupin beans, cooked pinto peans, cooked large white beans, boiled red kidney beans, black beans, paprika, curry powder, ground turmeric, cottage cheese, and dried shiitake mushrooms, or any combination thereof. Each possibility is a separate embodiment.

[0195] According to some embodiments, non-limiting examples of functional food rich in in Cu and Fe stimulating the production of kynurenine (a metabolite of tryptophan) is selected from dried basil, dried spearmint, dried marjoram, dried coriander, dried rosemary, dried parsley, dried chives, cumin seeds, sesame seeds, dried pumpkin seeds, hemp seeds, ground turmeric, ground sage, ground paprika, ground ginger, curry powder, dried spirulina seaweed, soy protein powder, cocoa powder, morel mushrooms, chantarelle mushrooms raw, sun-dried tomatoes, sesame paste (Tahini), lemon grass, chia seeds, dried sunflower seeds, safflower seeds, Goji berries, cashew nuts, dried pine nuts, hazelnuts, raw peanuts, dry pistachio nuts, almonds, olives, low moisture dried apricots, low moisture prunes, boiled edamame, oat (uncooked), cooked large white beans, cooked spinach, falafel, and dried shitake mushrooms, poppy seeds, Brazil nuts, walnuts, caraway seeds, goat cheese, peanut butter, dried gingko nuts, turnip greens, portobellos, raw tofu, and cooked chickpeas, or any combination thereof. Each possibility is a separate embodiment.

[0196] According to some embodiments, non-limiting examples of functional food stimulating the production of kynurenic acid (a neuroactive metabolite of kynurenine) is selected from ground sage, dried spearmint, dried chives, garlic powder, dried rosemary, paprika, pistachio nuts, hazelnuts, sunflower seeds, safflower seeds, hemp seeds, garlic, dried shitake mushrooms, wild cooked salmon, low moisture prunes, dried coriander, ground ginger, dried gingko nuts, peanut butter, fennel seeds, and low moisture dried apricots, or any combination thereof. Each possibility is a separate embodiment.

[0197] According to some embodiments, non-limiting examples of functional food stimulating the production of 3 -Hydroxy kynurenine (a neuroactive metabolite of kynurenine) is selected from dried spirulina, dried parsley, dried coriander, dried shitake mushrooms, paprika, had goat cheese, feta cheese, dry roasted almonds, tea, sun-dried tomatoes, sesame paste (tahini), cooked wild salmon, safflower seeds, sunflower seeds, caraway seeds, portobellos, Greek yoghurt, and fava beans, or any combination thereof. Each possibility is a separate embodiment.

[0198] According to some embodiments, non-limiting examples of functional food stimulating the production of xanthurenic acid (an indirect metabolite of the kynurenine and a direct metabolite of 3 -Hydroxy kynurenine) is selected from ground sage, dried spearmint, dried chives, garlic powder, dried rosemary, paprika, pistachio nuts, hazelnuts, sunflower seeds, safflower seeds, hemp seeds, garlic, dried shitake mushrooms, wild cooked salmon, low moisture prunes, dried coriander, ground ginger, dried gingko nuts, peanut butter, fennel seeds, and low moisture dried apricots, or any combination thereof. Each possibility is a separate embodiment.

[0199] In some embodiments, a co-factor includes a mineral, a vitamin, an enzyme, or any combination thereof. Each possibility is a separate embodiment.

[0200] In some embodiments, the neuro-nutritional treatment comprises a food (including beverages), a functional food (including beverages), and / or a supplement, or any combination thereof.

[0201] In some embodiments, the treatment regimen comprises administration / consumption by the subject of the provided neuro -nutrition for a period of at least 8 weeks. In some embodiments, the treatment regimen comprises administration / consumption by the subject of the provided neuro-nutrition for a period of at least about 8 weeks, at least about 12 weeks, at least about 16 weeks, at least about 20 weeks, at least about 30 weeks, at least about 40 weeks, or more. Each possibility is a separate embodiment.

[0202] According to some embodiments, method 100 is executed using various algorithms and / or models, as exemplified with reference to FIG. 2A-FIG. 2E.

[0203] Advantageously, using algorithms, including supervised or unsupervised Al-algorithms for learning and classification, the convolution of the data, including the abovementioned correlations, is studied in an unprecedented depth. This serves as a basis for computing / generating more complex and non-obvious layers, relationships, and patterns distinctively reflecting the neuro metabolic status of the subject, connecting them to the subject’s mental health status, and yielding / producing an advantageous and non-obvious neuro biochemical signatures / fingerprints that can predict a mental health status of a subject and point towards specific impaired metabolic steps just by association to a biochemical status of a plurality of neuro metabolic biomarkers, and pathways belonging thereto.

[0204] Further advantageous, is the algorithm providing a personalized recommendation, for a neuro nutritional / diet regimen, based on consumption / administration of specific nutrients rich / enriched with those metabolites, precursors, and co-factors (but not the NS itself) that are related to the impairment (the impaired step) and are predicted to modulate the level of a neurosubstance (NS) of interest. By consumption / administration of food, functional food, supplements, and / or nutraceutical compositions, naturally or artificially enriched with those metabolites, precursors, and co-factors in order to alter the metabolism of the impaired metabolic step and restore the integrity of the pathway.

[0205] In some embodiments, the method includes applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing between the detected / determined levels of at least 2 neuro-biomarkers belonging to a same metabolic pathway.

[0206] In some embodiments, the method includes applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing the detected / determined levels to a baseline physiological level / range.

[0207] In some embodiments, the method includes applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways.

[0208] In some embodiments, generating the metabolic fingerprint comprises comparing the detected / determined levels to a baseline physiological level / range.

[0209] In some embodiments, the algorithm is an Al-algorithm. In some embodiments, the first algorithm and the second algorithm are the same or different. Each possibility is a separate embodiment.

[0210] In some embodiments, the Al algorithm is selected from one or more of supervised learning, unsupervised learning, semi-supervised learning, reinforced learning, self-supervised learning, transfer learning, meta-learning, evolutionary algorithms, or any combination thereof. Each possibility is a separate embodiment.

[0211] In some embodiments, the Al algorithm is a supervised machine learning algorithm capable of regression and / or classification selected from one or more of Support- vector machines, Linear regression, Logistic regression, Random Forest, Naive Bayes, Linear discriminant analysis, Decision trees, K-nearest neighbor algorithm, Deep Neural networks, Neural networks (Multilayer perceptron), Gradient Boosting Algorithms, Linear Discriminant Analysis, Ridge Regression and Lasso Regression, Elastic Net, Bayesian Regression, Multiclass Classification Algorithms, Similarity learning, or any combination thereof. Each possibility is a separate embodiment.

[0212] In some embodiments, the algorithm includes experts’ knowledge.

[0213] In some embodiments, the method further comprising obtaining a questionnaire result, wherein the questionnaire result comprises the subject’s answers as to whether or not and / or to what level of severity he / she is experiencing one or more of a plurality of mental health associated symptoms.

[0214] In some embodiments, the level of severity to which the subject is experiencing one or more of a plurality of mental health associated symptoms and / or conditions is perceived by the subject as moderate and / or severe, ranked 3 or 4 out of 4.

[0215] In some embodiments, the method includes identifying one or more impaired neuro metabolic steps further comprises incorporating / analyzing obtained questionnaire result. In some embodiments, an impaired neuro metabolic step includes imbalanced concentrations of at least one neuro-biomarker (i.e., imbalanced with respect to the normal physiological range).

[0216] According to yet another aspect of the disclosure, there is provided a diagnostic kit for ex vivo detection of neuro-biomarkers associated with mental health, the kit comprising a urine card impregnated with a plurality of agents for detecting a plurality of neuro-biomarkers in a urine sample and instructions for use. According to some embodiments, the kit includes at least 2 urine cards, at least 3 urine cards or at least 4 urine cards. According to some embodiments the kit also comprises a questionnaire for evaluation of mental health or a link to same.

[0217] In some embodiments, the plurality of biomarkers comprises at least one biomarker selected from the neuro substances listed in Table 2. In some embodiments, the plurality of biomarkers comprises at least 2 biomarkers, at least 3 biomarkers, at least 4 biomarkers, at least 5 biomarkers, at least 6 biomarkers, at least 7 biomarkers, at least 8 biomarkers, at least 9biomarkers, at least 10 biomarkers, at least 15 biomarkers, at least 20 biomarkers, or more selected from the neuro substances listed in Table 2

[0218] In some embodiments, the agents for detecting a plurality of neuro -biomarkers include any of chromatography, an antibody, an immunoassay such as ELISA-based assay or similar, nucleic acid probe-based assay such as aptamer-based assay, primers for PCR, next generation sequencing, an electrochemistry-based assay, a lateral-flow assay, a nanobead-based assay, a microfluidics-based assay, and an oligonucleotide-templated reaction, or any combination thereof. Each possibility is a separate embodiment.

[0219] According to yet another aspect, there is provided a method for distinguishing between depression and anxiety, the method comprises: (a) detecting / determining a level of a plurality of neuro -biomarkers in a previously obtained biological urine sample of the subject, wherein the plurality of neuro-biomarkers comprises a neuro substance (NS) and at least one precursor, metabolite and / or co-factor; (b) applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing the detected / determined levels to a baseline physiological level / range; (c) identify one or more impaired neuro -metabolic steps in any of the plurality of neuro metabolic pathways by applying a second algorithm on the fingerprint; and (d) classifying the subject into one or more classes of mental health conditions / categories selected from the 4thcategory (6B00-6B0Z) and / or 5thcategory (6A70-7Z) in Table 1 based on the impaired step of the neuro-metabolic pathway.

[0220] In some embodiment, the neuro substance (NS) includes norepinephrine and at least one NS selected from, PEA, serotonin and GABA, or any combination thereof.

[0221] According to another embodiment, the status of the mental health of the subject is related to Sleep wake disorders, Stress, burn out, PSTD, neurocognitive impairment, disorders and functioning incl Alzheimer’s and Parkinson’s, Anxiety or fear-related disorders, Mood disorders and depression, Fatigue and decreased energy, Appetite balance (Eating Disorders, e.g. Anorexia Nervosa, Bulimia Nervosa, Binge Eating Disorder, Avoidant-Restrictive Food Intake Disorder), Susceptibility to addictions (Disorders Due to Substance use or Addictive Behaviors), Self-regulation (impulse control disorders), Neuroinflammation and immune defense.

[0222] The following examples are presented in order to more fully illustrate some embodiments of the invention. They should in no way be construed, however, as limiting thebroad scope of the invention. One skilled in the art can readily devise many variations and modifications of the principles disclosed herein without departing from the scope of the invention.

[0223] Reference is now made to FIG. 1B-FIG. IF, which demonstrate the flow of method 100 according to a real-world example, namely a patient diagnosed with depression, prescribed SSRIs, yet still suffering from severe fatigue, severe anxiety, and moderate depression symptoms.

[0224] In the first step (corresponding to step 110 of method 100) the level of several groups of biomarkers were measured in a urine sample of the patient. FIG IB illustrates the neurobiomarkers of the dopaminergic pathway, from the initial nutrient (tyrosine) to the last metabolite (HVA). Biomarkers of other pathways, such as the serotonergic, dopaminergic pathways etc. were also measured.

[0225] Next, as shown in FIG 1C an analysis of the levels of the patient’s neuro-biomarkers (here the dopaminergic pathway biomarkers) were assessed and compared to a pre-defined healthy interval. As seen in this case, the nutritional intake of the main precursors was measured and found to be within a healthy interval, suggesting there is no nutritional issue and that the intake of the initial nutrient and precursor is sufficient. Similarly, the synthesis of DOPA from the tyrosine was also measured and found to be within a normal range. However, the levels of dopamine measured were below normal followed by a normal metabolism of dopamine into DOPAC and HVA (intra-pathway ratio). At this stage, the reasons for the insufficient levels of dopamine are unknown.

[0226] Following this step and as seen from FIG. ID, a neuro-metabolic fingerprint was then generated for the patient. As seen, the norepinephrine pathway and serotonergic pathways were also found to be out of balance at all steps thereof.

[0227] The fingerprint was then analyzed to identify points of imbalance in the various pathways. As shown in FIG. ID, each point of imbalance was identified by a number:

[0228] 1. Synthesis of dopamine: shows insufficient levels of cofactor (Vitamin B6).

[0229] 2. Metabolism of dopamine: no deviations

[0230] 3. Production of Norepinephrine: correlated with low levels of dopamine in catecholamine synthesis

[0231] 4. Low levels of normetanephrine: correlated with low levels of precursors in catecholamine synthesis

[0232] 5. Low levels of VMA: correlated with low levels of catecholamine synthesis

[0233] 6. Low levels of tryptophan: a nutritional issue (too low intake of this important amino acid)

[0234] 7. Insufficient synthesis of serotonin: correlated with a deficiency in the precursor tryptophan

[0235] 8. Low level of metabolite 5HIAA: correlated with a deficiency in serotonergic precursors and nutrients.

[0236] Based on the numerous points of imbalance identified, two primary causes were identified (see encircled numbers 1 and 2 in FIG. IF), namely

[0237] a) Insufficient synthesis of dopamine (subtype: dopaminergic pathway based) leading to severe fatigue (as reported by the subject).

[0238] b) Insufficient intake of amino acid tryptophan associated with the severe anxiety (subtype: serotonergic pathway based) reported by the subject, insufficient synthesis of serotonin and.

[0239] In addition, two secondary causes were also identified (see the numbers 3 and 4 encircled with stippled line in FIG. IF), namely:

[0240] i) Low levels of norepinephrine (as a consequence of the insufficient levels of dopamine and thus catecholamine synthesis - inter-pathway effect) associated with the moderate depression reported by the subject, and;

[0241] ii) low levels of serotonin (as a result of the insufficient intake of tryptophan), also responsible for the severe anxiety.

[0242] Based on the identified primary and secondary causes and the complementary diagnosis, the following recommendations were provided:

[0243] 1. Increasing intake of vitamin B6. Vitamin B6 is an important cofactor in the dopaminergic pathway, specifically it is required for efficient synthesis of dopamine. Accordingly, by increasing vitamin B6 levels, the patient’s own metabolism of DOPA into dopamine will be improved and the symptoms of fatigue addressed. Moreover, as a result of increased levels of dopamine, which is a precursor to norepinephrine, the norepinephrine pathway is also expected to rise, addressing the symptoms of depression and anxiety

[0244] 2. Increasing intake of tryptophan, which, using the patient’s own metabolism, will naturally cause a rise in serotonin and restore the balance of the serotonergic pathway, thereby likewise, also addressing the symptoms of depression and anxiety.

[0245] Taking serotonin reuptake inhibitors, dopamine reuptake inhibitors or norepinephrine reuptake inhibitors is on the other hand not recommended in that it may further exacerbate the pathway imbalances.

[0246] Reference is now made to FIG. 2A-FIG. 2E which provide an illustrative example of the algorithmic flow of method 100. While some of the steps / data relies of known expert knowledge (indicated as “pre-defined”) other steps / data is generated by the inventors of the present invention (indicated as “HL”)

[0247] As shown in FIG. 2A, the first step is generating the framework based on which the neuro-biomarkers are selected and their normal level intervals determined. This step is also referred to as a set-up step in that it need not be repeated for each patient, although one of ordinal skills in the art will recognize that continuous or periodic updates may be made to the framework.

[0248] Generating the framework relies on input data in the form of prior knowledge. This includes a list of 10 mental condition categories, the 68 symptoms found to characterize these mental condition categories, and their severity. It further relies on the levels of the biomarkers obtained for a plurality of symptomatic and non- symptomatic individuals (objective data).

[0249] In addition to the prior knowledge data, 25 neuro-metabolic biomarkers (found in urine samples) were identified by the inventors of the present invention (indicated as HL) as being associated with symptomatic patients. The inventors of the present invention further computed the normal intervals of the 25 neurometabolic markers using a bounded optimization algorithm configured to verify that an optimal range covers the normal interval.

[0250] By applying an adaptive cross-validation algorithm on the totality of input data, correlations between symptoms, symptom severity and urine neuro-biomarker values and their interdependencies were identified and the framework formed, as exemplified in FIGs 3A-FIG. 3J.

[0251] Once the framework is completed, the first step of the method can be carried out, in the form of measurement of neuro -biomarkers in urine samples of unknown subjects. These measured levels can then serve as an input to a threshold algorithm configured to determine whether and / or to which extent the measured values lie within the determined normal interval.

[0252] Next, a neurometabolic fingerprint is generated for each patient, as illustrated in FIG. 2B. This step is based on pre-defined neurometabolic pathways, their steps and the precursors and co-factors taking part as well as on neuro-metabolic fingerprints generated for non- symptomatic patients, when building the framework. According to some embodiments, this step may include grouping the measured biomarkers and their levels into pathways and comparison of the fingerprint generated to the non-symptomatic fingerprint, for example by using a clustering algorithm.

[0253] Once a fingerprint has been generated, anomaly detection and causal inference algorithms can be applied to identify deviations and contributing factors. Moreover, causal analysis and differential diagnosis algorithms may be applied to establish primary and secondary conditions - also referred to herein as root cause analysis (see FIG. 2C).

[0254] Once root causes have been identified a treatment recommendation may be outputted by the algorithm (FIG. 2D). The treatment recommendation is designed to increase or decrease synthesis and / or metabolism of a particular step of the relevant pathway by 1. increasing / decreasing the availability of a precursors and / or a cofactor and / or an enzyme responsible for the synthesis / metabolism of the relevant step in the impaired pathway 2. Increasing or decreasing activity of another, competing or non-competing, not necessarily impaired, but correlated pathway.

[0255] Advantageously, based on the hereindisclosed algorithm the outputted treatment recommendation may be personalized based on which pathway and which step were identified as impaired, the type of impairment (e.g. insufficient synthesis or too high metabolism etc.), and identified primary and secondary conditions.EXAMPLESExperimental set-up - building the framework:

[0256] 49 symptomatic, non-medicated, subjects and 24 non- symptomatic subjects agreed to participate in this study aimed to find correlations between mental health related condition and / or symptoms and neuro biomarkers, in particular neurotransmitters, in biological samples. The subjects provided medical information and biological samples that were used for establishing a data resource including medical data and biochemical data.

[0257] Medical pre-clinical data was derived from self-assessed questionnaire completed by the subjects in order to evaluate their mental health status. In the questionnaire, the subjects were asked to self-rate about 87 types of physical and psychological symptoms related to mental health conditions, and encompassing 10 types of MC categories, according to the subject’s perceived level of severity, on a scale of 1-4, wherein 1 represents non-exitance of a symptom, and 2-4 represent mild, moderate and severe, respectively.

[0258] The pre-clinical data of the subjects’ mental health status included lists of about 87 types of physical and psychological symptoms that are associated with mental health, and which are grouped into 10 categories of mental health effect / MC (as set forth in Table 1).

[0259] Biochemical data was derived from biological samples provided by the subjects for evaluation of their neuro metabolic status / pathways. Each subject provided 4 dry urine samples (dry urine cards) which were analyzed for the amount / concentration of 26 neuro biomarkers encompassing 18 neuro metabolic pathways. The 4 urine samples were collected in the morning at awakening time, two hours after waking up, in the evening before dinner, and before going to sleep.

[0260] The biochemical data included measurements of the 26 neuro biomarkers belonging to the 18 different neuro metabolic pathways (see Table 2), some of the pathways, such as, but not limited to: the kynurenine / kynurenic acid pathway, the kynurenine / xanthurenic acid pathway, the glutaminergic pathway - were fully characterized in terms of metabolites / precursors levels, neurotransmitters levels, participating in the biosynthesis or biodegradation pathway- while other pathways, such as, but not limited to: the serotonergic pathway, the serotonergic / melatonin pathway, the histaminergic pathway, the dopaminergic pathway, and the catecholamine pathway 1 (norepinephrine) - were almost completely characterized leaving only one or two metabolites / precursors or neuro substances unmeasured.

[0261] Next, the medical and biochemical data were compared, by applying on the data Adaptive Cross-Validation Algorithms to find correlations between the symptoms and / or conditions, their level of severity and the category of related mental health effect, as selfreported by the subjects (i.e., the mental health status), and the levels of the plurality of neuro biomarkers, including for example, but not limited to: precursors and metabolites acting as substrates, neurotransmitters, amino acids and hormones related to neuro metabolic pathways as measured in the dry urine samples provided by the non-medicated symptomatic subjects, in comparison to the non-symptomatic, healthy control subjects. This analysis results in identification of one or more impaired / defective / flawed neuro-metabolic pathway.

[0262] More specifically, the algorithm was applied on biochemical data derived from biological samples which were biochemically characterized for the amount / concentration of a plurality of neuro biomarkers (at least 26) - constituting a neuro biomarkers profile / signature - and which were labelled according to the corresponding medical pre-clinical data they derived from, for example, the self-reported questionnaires of the symptomatic subject and / or non-symptomatic healthy control subjects, who self-evaluated including self-rating of perceived severity of the symptoms and / or conditions they suffer from on a scale of, for example, a scale of 1-4. The algorithm then finds the correlation between the biochemical profile of the labelled biological samples including at least the levels of one or more precursor or metabolite, and the severity of the symptoms and / or condition (mental health status as selfevaluated), and identifies the sample(s) as belonging to subject(s) predicted (at some level of probability) to suffer from a neuro-metabolic impairment in one or more specific neuro metabolic pathway(s).

[0263] As each subject may suffer from one or more symptoms and / or conditions associated with unique neuro biomarkers profile / signature, the clinical and biochemical data repository / resource used for training and validation may include comorbidities associated neuro biomarkers profiles that continues to grow as more samples being characterized.

[0264] By dissecting each step of the pathway in view of the abovementioned data analysis, the specific step(s) of the metabolic pathway exhibiting a flawed / impaired functionality is revealed, and a personalized treatment aimed to compensate / complement / restore the defective functionality of the neuro metabolic pathway is facilitated, namely by advising on administration / consumption of neuro-nutrients from food, functional foods, and natural supplements, and optionally on nutraceutical composition.

[0265] Statistical analyses - in the statistical assessment of the correlations between the specified neuro-metabolic biomarkers and symptoms of the mental conditions MC, different predictive models are used with the purpose of, based on the biomarkers, predicting whether a patient has a MC or not. As a start, single neuro -biomarker is associated with a risk factor in a simplistic model. The basis for such associations between each biomarker and a risk factor are correlations between them. An aggregated risk across biomarkers guides the outcome prediction. After performing the analysis of single biomarkers, the analysis focuses on pairs and groups of them. In more complex models, a correlation between pairs and groups of biomarkers and the corresponding risk factors are established. After that part is completed, the single neuro-biomarkers or their groups are organized in pathways, and the analysis of the neuro-metabolic pathways begins. The model compares the outcomes of the questionnaires gathering the symptom information and the severity and determination of MC based on the biomarkers. By comparing the predictions of the model against the outcomes of the subjective patient data from the questionnaires, the sensitivity and specificity of the approach is recomputed and improved. The specificity is measured using an earlier validation based on a split of the sample in diagnosed and healthy intervals. On the basis of subjective symptom data and their correlation with the levels / concentrations of neuro-biomarkers. The correlated levels of neuro-biomarkers were analyzed, and intervals: "diagnosed" (with corresponding severity), and "healthy" were created.

[0266] Chemical / biochemical analyses - To differentiate the molecules of the measured neuro biomarkers, a mass spectrometry and high-performance liquid chromatography (HPLC) systems were employed. Some of the molecules can be read using mass spectrometry and the identification of chemical profile of neuro substances based on the separation of gaseous ions in electric and magnetic fields according to their mass-to-charge ratios. Other molecules required HPLC and a separation and detection of them based on their distinct velocities through the chromatographic column. The process involved two phases: the mobile phase and the stationary phase.

[0267] Identifying statistically significant correlations

[0268] Then statistically significant correlations between metabolites found in the dry urine samples and the medical health conditions / symptoms were identified.

[0269] Anxiety

[0270] A statistically significant correlation was found between the self-reported level of severity of anxiety - a symptom and / or a condition belonging to a category of mental health of anxiety, excessive worry and trauma (referring to the 4thcategory in Table 1) - and the urine levels of the neuro biomarkers and neurotransmitter serotonin which is related to the serotonergic pathway and to the serotonergic / melatonin pathway (referring to the 1stand 2ndneuro metabolic pathway in Table 2).

[0271] For example, as can be seen in FIG. 3A, the level of serotonin in urine samples was found to be correlated with anxiety in that the level was significantly higher in non- symptomatic subjects as compared to subjects who self-rated their anxiety as mild, moderate or severe. Interestingly, the level found in non- symptomatic subjects (143 pg / g Cr in average of 21 subjects) was found to be significantly higher than the previously reported normal range (61.0-103.2 pg / g Cr). However, in patients suffering from even mild levels of anxiety the levels of metabolized serotonin were about 30% lower.

[0272] Similarly, as can be seen in FIG. 3B, the levels of tryptophan (the serotonergic pathway precursor) were likewise correlated with anxiety in that in non-symptomatic subjects' tryptophan levels were found to be significantly lower (7,552 pg / g Cr), than that observed for subjects suffering from severe anxiety (13,056 pg / g Cr).

[0273] It is understood that, while these results indicate that imbalances in the serotonergic pathway are correlated with anxiety, further analysis must be done in order to understand the underlying cause. As can be seen in FIG. 3C in order to dissect the impaired / defective step(s) of the serotonergic pathway data about the amount / concentration of precursors and metabolites such as tryptophan and 5-HTP is considered, as well as co-factors including for example, but not limited to: BH4, Fe, TPH, vitamin B6, AADC, Cu, MAO and AR must be assessed.

[0274] In a first non-limiting scenario for the underlying cause of the low levels of serotonin, it may be found that the levels of precursors / metabolites in the serotonergic pathway such as tryptophan and 5-HTP are normal indicating an effective functionality of step 1 (see FIG. 3C), thus indicating that the low levels of serotonin and its metabolite 5-HIAA (steps 2 and 3) may be indicative a flawed anabolic step 2 (5-HTP -> serotonin).

[0275] Therefore, for the subject under evaluation in this first non-limiting scenario a possible suggested neuro-metabolic intervention should promote / stimulate / induce serotonin biosynthesis from 5-HTP, and may include increasing administration / intake of the one or more precursors and / or co-factors that participates in execution of the anabolism of step 2 includingfor example one or more of 5-HTP, vitamin B6 and / or AADC in the form of a nutritional supplement.

[0276] In a second non-limiting scenario for the underlying cause of the low levels of serotonin: normal levels of precursors / metabolites in the serotonergic pathway such as tryptophan and 5-HTP suggesting an effective functionality of step 1 (referring to FIG. 3D and Table 2), and normal levels of serotonin suggests an effective functionality of step 2 may be found. However, reduced levels of the metabolite 5-HIAA (steps 3) may be indicative of a flawed catabolic step 3 (serotonin -> 5-HIAA).

[0277] Therefore, for the subject under evaluation in this second non-limiting scenario a possible suggested neuro-metabolic intervention should promote / stimulate / induce 5-HIAA biosynthesis from serotonin, and may include increasing administration / intake of the one or more precursors and / or co-factors that participates in execution of the catabolism of step 3 including for example one or more of Cu, vitamin B6 MAO and / or AR in the form of a nutritional supplement.

[0278] Fatigue

[0279] As can be seen in FIG. 3D, the levels of tyrosine in urine samples were found to be correlated with fatigue in that the levels of tyrosine in non-symptomatic subjects was found to be significantly higher than the levels in subjects suffering from fatigue and the levels were found to decrease with an increase in the severity of the condition. As tyrosine is a neurobiomarker of several neurometabolic pathways, the result may indicate that one or more of the tyraminergic pathway, the dopaminergic pathway, the catecholamine pathway 1 (norepinephrine), and / or the catecholamine pathway 2 (epinephrine) are correlated with the mental health condition of fatigue / low energy.

[0280] As can be seen in FIG. 2E, the levels of dopamine the levels of tyrosine in urine samples were found to be correlated with fatigue in that the level in non-symptomatic subjects was significantly higher than that found in subjects suffering from severe levels of fatigue. This demonstrates a correlation between reduced dopamine levels, the glutaminergic pathway and mental health symptoms of fatigue and further demonstrates that a same symptom may have a different root cause and thus require a different treatment.

[0281] As can be seen in FIG. 3F, the levels of norepinephrine in urine samples were also found to be correlated with fatigue in that its level in non-symptomatic subjects was significantly higher than the levels in subjects suffering from severe levels of fatigue, thereby yet again demonstrating that a same symptom may have a different root cause and thus require a different treatment.

[0282] Depression

[0283] As can be seen in FIG. 3G, the levels of norepinephrine the levels of norepinephrine in urine samples were also found to be correlated with depression in that the level in non- symptomatic subjects was significantly higher than in subjects who self-reported that they suffer from depression. In fact, the levels of norepinephrine were found to be continuously lower in correlation with the severity of the condition. This result demonstrates that a pathway imbalance may negatively impact more than one mental health condition (e.g. both fatigue and depression).

[0284] As can be seen in FIG. 3G and FIG. 3H, the levels of serotonin and tryptophan in urine samples were also found to be correlated with depression in that their level in non-symptomatic subjects were significantly higher than that observed in symptomatic subjects who self-reported that they suffer from various degree of depression.

[0285] Addictive behavior

[0286] As can be seen in FIG. 31, the levels of metabolized dopamine (dopamine found in urine) were found to correlate with addictive behavior in that in non-symptomatic subjects the level was significantly higher compared to subjects who self-reported that they suffer from addictive behaviors, even at mild levels.

[0287] Additional correlations between neuro -biomarkers found in urine and various mental health conditions and symptoms were found (results not shown). These correlations were used to build the framework enabling the algorithm to output a root cause of a subject’s mental health condition(s), to classify the conditions and causes into primary and secondary and to provide a personalized treatment regimen.Example 1 - levels of neuro biomarkers and related neuro metabolic pathways in dry urine samples.

[0288] The levels of neuro-biomarkers of a subject (subject A) diagnosed with depression and suffering from attention deficits and had difficulties focusing in addition to the depressive symptoms) were measured. The subject had previously been prescribed SSRI (selective serotonin reuptake inhibitors). After 6 months of medication the subject did not experience an improvement and discontinued pharmacological therapy. The subject was not taking any medication 6 months in advance to the neuro-metabolic analysis.

[0289] The tables below show the levels of 26 different biomarkers, and the 13 associated neuro metabolic pathways they encompass / belong to. Values are in pg per g Creatinine (pg / g Cr) unless indicated they are in in mg per g Creatinine (mg / g Cr) or in pmol per g Creatinine (pmol / g Cr). Off range values are underlined. Related neuro metabolic pathway numbering corresponds to the pathway number in hereinabove Table 2.1. Serotonergic pathway2. Serotoner ic / melatonin pathway3. Kynurenine / kynurenic acid pathwayKynurenine / xanthurenic acid pathwayGlutaminergic pathwayGlycinergic pathwayHistaminergic pathwayCysteine and trans- sulfuration pathwayTyraminergic pathwayEA pathwayopaminergic pathwayatecholamine pathway 1 (norepinephrine) pathwayatecholamine pathway 2 (epinephrine) pathway

[0290] The neuro-metabolic profile of subject A did not show any metabolic impairments of the serotonergic pathway. Instead, impairments of PEA pathway were present, especially in the synthesis and metabolism of phenylethylamine, impacting the subjects’ ability to maintain attention and focus.

[0291] Also, an impairment of GABA pathway was determined, which corresponded to a neuro-metabolic profile of anxiety, and more specifically a subtype of anxiety based on the metabolic impairment of the synthesis of Gamma-aminobutyric acid.

[0292] As a result, the diagnosis was changed. A new medical assessment of the subject was provided as the subject was now diagnosed with anxiety, and ADHD (and not, as earlier, depression).

[0293] The subject could be supported with the precursors and co-factors for the synthesis and metabolism of PEA, to restore the attention and focus functionality of PEA pathway. Also, precursors and cofactors of the synthesis of GABA helped to restore the functionality of the major inhibitory (calming) NS.

[0294] In order to test if a personalized dietary regimen including neuro-nutrients based on food, functional foods, and natural supplements can alleviate symptoms and / or conditions related to mental health, the symptomatic subject of Example 1, who was re-diagnosed with PAE-based anxiety and impairments in the GABA pathway, instead of depression, was asked to follow a set of customized recommendations including consumption of personalized neuro nutrition for a minimum period of 8 weeks.

[0295] The neuro nutrition regimen was aimed to support the synthesis and metabolism of phenylethylamine , the subject was ordered to increase the intake of phenylalanine (an addition of e.g. 150 g cooked soy beans per day, alternatively 100 g dried pumpkin seeds), vitamin B6 (e.g. 100 g whole grain cereals added to daily nutrition), Cu (e.g. 14 grams=2 table spoons spirulina added per day) and vitamin B2 (e.g. 100 g dried almonds added to daily nutrition). To stimulate the synthesis of GABA, the glutaminergic pathway was supported with Mg, Mn and vitamin B6.

[0296] At the end of the 8 weeks, the subject was asked to provide additional 4 biological samples of dry urine cards for biochemical analysis of several relevant biomarkers, as well as to re-evaluate his symptoms and / or conditions using the same 1-4 scale of perceived level of severity, where 1 represents non-exitance of a symptom, and 2-4 represent mild, moderate and severe, respectively.

[0297] The amounts / concentrations of the measured biomarkers before and after treatment are presented in Table 3 below, and the self-reported ranking of his symptoms and / or conditions before and after treatment are presented in Table 4 below.Table 3: Biomarkers levels of the symptomatic subjectTable 4: Ranking of the perceived severity of symptoms and / or conditions, as selfreported by the subject

[0298] The data in Table 3 and Table 4 suggests that the subject suffered from neuro metabolic impairments manifested by low GABA levels, high histidine levels (allergies and sensitivities), low glycine levels, and low PEA levels, and that the treatment with personalized neuronutrition based dietary regimen alleviated his symptoms related to mental condition, as indicated by -30% change in total scoring after treatment.

[0299] In conclusion, the analysis enabled a revised / changed diagnosis, where a GABA pathway-based anxiety was determined (an earlier diagnosis of depression was no longer considered adequate), and the subject was evaluated for and diagnosed with ADHD. Frequent low mood symptoms were a result of difficulties experienced in study and professional context, where subjects faced obvious difficulties due to untreated ADHD challenges. Non adequateand not effective medications were no longer prescribed, and severe side effects of not adequate pharmaceuticals (increased suicidal risk due to the usage of anti-depressants among patients 18-25) could be avoided. The support of PEA pathway and addressing its impairments, together with ADHD medication, allowed the subject to successfully study and work. The low mood symptoms were reduced.Example 2 - clinical example.

[0300] The hereindisclosed method was executed on a 49-year-old male patient suffering from severe sleep difficulties, severe fatigue as well as moderate decrease in mental sharpness, forgetfulness, depression, decreased flexibility, addictive behavior and anxiety. The patient also suffered from mild degree of irritation, and morning fatigue.

[0301] Along the line of method 100, initially the level of the biomarkers listed in FIG. 4A was measured.

[0302] The levels were then input into an algorithm and were applied to group the neurobiomarkers into pathways and to draw a neuro-metabolic fingerprint. Results for the dopaminergic and the serotonergic pathways are shown in FIG. 4B and FIG. 4C, respectively. In total, 18 neuro-metabolic pathways were drawn (data not shown).

[0303] Next, the levels measured of each pathway were compared to a healthy reference interval as shown in FIG. 4D (reference interval shaded in grey) and in Table 5 below for the dopaminergic pathway and as shown in FIG. 4E and Table 6, for the serotonergic pathway. As seen from FIG. 4D, tyrosine level, a precursor in the dopaminergic pathway, was pronouncedly lower than the healthy interval (shaded in grey) and dopamine levels were also below health, whereas DOPAC and HVA levels were within normal range. As seen from FIG. 4E, tryptophan levels, a precursor in the serotonergic pathway, were pronouncedly lower than the healthy interval (shaded in grey), while serotonin and 5-HIAA levels were within normal range. High levels of PEA were also found (see FIG. 4A).

[0304] Table 5 - Dopaminergic pathway level comparison

[0305] Table 6 - Serotonergic pathway level comparison

[0306] Next, the fingerprint of each of the pathways were inputted for analysis and impairments (also referred to as “points of imbalance”) identified.

[0307] As seen from FIG. 4F, which shows the analysis for the dopaminergic pathway, 4 points of imbalance were identified:

[0308] 1. Levels of tyrosine: insufficient. Tyrosine is a non-essential amino acid, which can be obtained from nutrition or is produced by the cells from an essential amino acid phenylalanine which must be obtained from nutrition.

[0309] 2. Synthesis of dopamine: insufficient, resulting in insufficient levels of dopamine.

[0310] 3. Levels of dopamine: insufficient.

[0311] The rest of the pathway is within the reference interval typical for non-symptomatic patients.

[0312] As seen from FIG. 4G, which shows the analysis for the serotonergic pathway, 2 points of imbalance were identified:

[0313] 1. Levels of tryptophan: insufficient. Tryptophan is an essential amino acid which needs to be obtained from nutrition.

[0314] 2. The rest of the pathway is within the optimal interval typical for non-symptomatic patients

[0315] Next a (complementary) diagnosis was output based on the fingerprint and the framework built for the method (as described hereinabove). In this case, the following was inputted:

[0316] As seen from FIG. 4H, the patient was identified as having 2 primary causes (also referred to as ’’root cause”) and 1 secondary cause for his mental health issues:

[0317] A. Primary - insufficient levels of tyrosine and insufficient synthesis of dopamine result in low levels of norpinephrine. As seen from FIG. 3D, FIG. 3E and FIG. 3F all three biomarkers are corelated with fatigue. Norpinephrine is also correlated with depression.

[0318] B. Primary - insufficient levels (intake) of the amino acid tryptophan. As seen from FIG. 3H, low levels of tryptophan is corelated with depression. Low levels of tryptophan has also been correlated with anxiety (not shown).

[0319] C. Secondary - high levels of phenethylamine (PEA) are correlated with sleep difficulties and racing thoughts. PEA is produced from phenylalanine, an essential amino acid which must be obtained from the diet. PEA prolongs the signaling of dopamine, norepinephrine and serotonin.

[0320] Next a treatment regimen was outputted based on the fingerprints and the complementary diagnosis:

[0321] 1. Increase intake of tyrosine and iron, an important precursor and cofator enabling the synthesis of dopamine, respectively. The increased intake of tyrosine and iron was prescribed to adress the symptoms of fatigue and depression. By elevating the levels of precursors and cofactors (instead of increasing dopamine directly), the body increases its natural production of dopamine (restores pathway imbalance), which is far better than synthetically increasing dopamine levels (e.g. via adminstration of dopamine reuptake inhibitors).

[0322] 2. Increase intake of the essential amino acid tryptophan and vitamine B6 both in order to increase the levels of tryptophan and also to indirectly elevate the synthesis of serotonine, thereby addressing the symptoms of anxiety. Of note, it was found that while tryptophan is a precursor of serotonin it also appears to have a serotonin-independent correlation to anxiety. Accordingly, in addition to the fact that naturally improving serotonin levels via increased intake of tryptophan has a better effect than synthtic elevation (e.g. via serotonin reuptake inhibitors), the elevated level of tryptophan itself improves anxiety, an effect that cannot be obtained via consumption of serotonin reuptake inhibitors.

[0323] 3. No direct treatment for the high PEA levels is required. This since, as a result of increased dopamine and serotonin levels, the high levels of PEA are expected to decrease, reducing the symptoms of sleeep difficulties and racing thoughts.

[0324] After 8 weeks of treatment with tyrosine, iron, tryptophan and vitamine B6, the pateints status was reevaluated by remeasuring the level of biomarkers, generating fingerprint and analysis of the fingerprint for impariments

[0325] The results for the dopaminergic and serotonergic pathways can be seen in FIG. 41 and FIG. 4J, respectively, as well as in Table 7 and Table 8, respectively.

[0326] Table 7 - dopaminergic pathway before / after treatment.

[0327] Table 8 - serotonergic pathway before / after treatment.

[0328] As seen from FIG. 41, while tyrosine levels were still below the reference interval, the levels were significantly higher after treatment. This in turn also resulted in an increase in the level of dopamine which was now found to be within the normal range solely as a result of the increase intake of tyrosine and the improved synthesis of dopamine due to the increase in the co-factor iron.

[0329] As seen from FIG. 4J, as a result of the treatment, tryptophan levels were significantly elevated and were now found to be in the normal range. A slight increase in the levels of serotonin was also observed.

[0330] Importantly, as seen from FIG. 4K, the symptoms were drastically reduced and oftentimes even eliminated, as a result of the treatment (without taking any medicament).While certain embodiments of the invention have been illustrated and described, it will be clear that the invention is not limited to the embodiments described herein. Numerous modifications, changes, variations, substitutions and equivalents will be apparent to those skilled in the art without departing from the spirit and scope of the present invention as described by the claims which follow.

Claims

CLAIMS1. An ex vivo method for managing a mental health condition of a subject in need thereof, the method comprising:(a) obtaining at least one urine sample of the subject(b) detecting / determining a level of a plurality of neuro -biomarkers of each of a plurality of neuro-metabolic pathways in the urine sample, wherein the plurality of neuro-biomarkers is selected from a neuro substance (NS) and at least one precursor, metabolite and / or co-factor;(c) applying a first algorithm on each of the detected / determined levels of the plurality of neuro biomarkers to generate a metabolic fingerprint of a plurality of pathways, wherein generating the metabolic fingerprint comprises comparing between the detected / determined levels of at least two neurobiomarkers belonging to a same metabolic pathway (intra-pathway ratio) and comparing a level of the plurality of neuro -biomarkers to a respective level range identified as healthy;(d) applying a second algorithm on the metabolic fingerprint, wherein the algorithm is configured to identify one or more impaired neuro-metabolic steps in any of the plurality of neuro metabolic pathways and to classify the identified one or more impaired neuro-metabolic steps into primary and secondary impairments; and(e) outputting a neuro-nutritional treatment regimen for the subject, based on the identified primary impairment in the one or more neuro-metabolic step, wherein the neuro-nutritional treatment regimen is designed to improve metabolism of the impaired step of a neuro metabolic pathway.

2. The method of claim 1, wherein the first algorithm is a clustering algorithm.

3. The method of claim 1 or 2, wherein the second algorithm is an anomaly detection algorithm and / or am inference algorithm.

4. The method of any one of claims 1-3, wherein the plurality of biomarkers comprises at least 10 different biomarkers.

5. The method of any one of claims 1-4, wherein the plurality of neurometabolic pathways comprise at least 3 different neurometabolic pathways.

6. The method of any one of claim 1-5, wherein the precursor is an initial precursor of a first metabolic reaction / step in the pathway and / or wherein the metabolite is a final metabolite of a last metabolic reaction / step in the pathway.

7. The method of any one of claims 1-6, wherein detecting / determining a level of the plurality of neuro-biomarkers further comprise measuring a level of a mineral, a vitamin, an enzyme, or any combination thereof.

8. The method of any one of claims 1-7, wherein the neuro-nutritional treatment comprises a precursor and one or more of a vitamin, a mineral, an enzyme, or any combination thereof.

9. The method of any one of claims 1-8, wherein the neuro-nutritional treatment comprises a food, a functional food, and / or a supplement, or any combination thereof.

10. The method of any one of claims 1-9, wherein the treatment regimen comprises administration / consumption by the subject of the provided neuro -nutrition for a period of at least 8 weeks.

11. The method of any one of claims 1-10, wherein the at least one urine sample is a dry urine sample impregnated on a urine card.

12. The method of claim 11, wherein the at least one urine sample comprises at least 2 dry urine samples obtained at different timepoints of a day.

13. The method of claim 12, wherein the at least one urine sample comprises at least 4 dry urine samples obtained at different timepoints of a day.

14. The method of any one of claims 11-13, wherein the detecting / determining the level of the plurality of neuro-biomarkers comprises averaging the levels detected in each of the dry urine samples obtained at different timepoints of a day.

15. The method of any one of claims 1-14, wherein the plurality of biomarkers comprises at least one biomarker selected from the neuro substances listed in Table 2.

16. The method of any one of claims 1-15, wherein the plurality of mental health associated symptoms comprises at least one symptom selected from Table 1.

17. The method of any one of claims 1-15, wherein the plurality of neuro metabolic pathways comprises at least one metabolic pathway selected from Table 2.

18. The method of any one of claims 1-16, further comprising treating the subject with a nutraceutical composition based on the treatment recommendation.

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