Patent Description:
The term "stress" describes mental or physical reactions due to day to day life and/or clinical associated conditions including chronic disease conditions and/or not limited to what people feel when they are under mental, physical, social or emotional pressure. Based on the conditions creating stress, it may be categorized mainly as psychological or physiological stress, in a subject. Although it is normal to experience some type of stress from time to time, people or an animal who experience a high level of physiological or psychological stress or who experience it repeatedly over a long period of time may develop mental and/or physical health problems. Such stress can be caused both by daily responsibilities and routine events, as well as by more unusual events, such as a trauma or illness in a mammal including but not limited to him/her/oneself or a close family member. It can sometimes help to motivate people to finish a task, or perform well. But stress can also be harmful if one becomes over-stressed as the stress interferes with the ability to get on with normal life for too long. One may feel tired, unable to concentrate or may easily become irritated. Stress can also damage an individual's physical health. When people feel that they are unable to manage or control changes caused by any illness or daily life activities, they experience physiological as well as psychological stress. This has become increasingly recognized as a factor that can reduce the quality of life of patients suffering from chronic diseases such as cancer. There is even some evidence that extreme stress is associated with poorer clinical outcomes. Clinical guidelines are available to help doctors and nurses assess levels of stress and help patients manage it. Some studies have indicated a link between various psychological factors and an increased risk of developing cancer.

If one faces a stressful event, the body responds by activating the nervous system and releasing hormones such as adrenalin and cortisol. These hormones cause physiological changes in the body which help one to react quickly and effectively to get through the stressful situation. This is sometimes called the 'fight or flight' response. The hormones increase one's heart rate, breathing, blood pressure, metabolism, and muscle tension. One's pupils dilate and our perspiration rate increases. While these physical changes help one try to meet the challenges of the stressful situation, they can cause other physical or psychological symptoms. It can be linked to headaches, an upset stomach, back pain, and trouble sleeping. It can weaken a person's immune system, making it harder to fight off diseases. If a person already has a health problem, stress may make it worse. It can make a person moody, tense, or depressed. The relationships with others may suffer, and a person would not do well at work. Hence it is very important to manage all types of stress for leading a healthy life.

Stress management encompasses techniques intended to equip a subject with effective coping mechanisms for dealing with physiological and psychological stress. Techniques of stress management include self-understanding, self-management (e.g. becoming better-organized), conflict resolution, adopting a more positive attitude, breathing exercises, meditation, exercises, altering the diet, medical treatment (e.g. anti-anxiety drugs) and taking more rest regularly and effectively. A good diet with a well- balanced nutrition is very important for achieving a stress resistant body. (Ref: Combating Stress with a Balanced Nutritional Diet: By Jayne Ritchie-Stress Management Society and Bodychef). Diets rich in vitamins, minerals and essential amino acids can be helpful in this context.

Macular carotenoids, especially xanthophylls, such as lutein, zeaxanthin isomers (RR-Zeaxanthin/RS-meso-zeaxanthin), meso-zeaxanthin and the like and their enantiomers, metabolites, esters, salts, derivatives either alone or in combination are considered to be important nutritional elements for a human body. These carotenoids effectively protect biological tissue in a mammal such as retinas of eyes by acting as an antioxidant, where the carotenoids accumulate in very high densities in the macula. This leads to a significant reduction of a risk of developing diseases, including age-related macular degeneration (AMD). Macular carotenoids' antioxidant function also appears to benefit cardiovascular health, skin health, and brain health. Indeed, higher levels of carotenoids such as lutein and zeaxanthin, presumably through anti-inflammatory action (Choi et al. <NUM>) have been associated with reduced atherosclerosis (Kailora, <NUM>), protection from UV-induced erythema (Heinrich et al. <NUM>), and significantly better cognitive performance in the elderly (Johnson et al. <NUM>; Feeney et al. <NUM>; Vishwanathan et al.

<NPL>) relates to the relationship between a macular pigment, such as lutein (L), zeaxanthin (Z), and meso-zeaxanthin (MZ), and cognitive function in adults aged ≥<NUM> years. Xanthophyll carotenoids were found to have an impact on a cognitive function.

Epub <NUM> Oct <NUM>) relates to a relation between cognition and lutein and zeaxanthin concentrations in the brain tissue of decedents from a population-based study. It was found that zeaxanthin concentrations in brain tissue were significantly related to levels of a global cognitive function, memory retention, verbal fluency, and dementia severity after adjustment for age, sex, education, hypertension, and diabetes.

<NPL>) relates to lutein or zeaxanthin supplementation to protect lens protein, lipid, and DNA from oxidative damage and to improve the intracellular redox status upon oxidative stress. The data implies that sufficient intake of lutein and zeaxanthin may reduce the risk for senile cataract via protecting the lens from oxidative damage.

<CIT> relates to a composition to promote ocular health. The composition includes amounts of vitamin A; vitamin C; vitamin D; vitamin E; zinc; copper; selenium; non-vitamin A carotenoids, which include lutein and zeaxanthin; omega-<NUM> fatty acids, which include eicosapentaenoic acid and docosahexaenoic acid; taurine; alpha-lipoic acid; pine bark extract; astaxanthin; and Piper spp. The reference describes a method of a treatment for a subject experiencing oxidative or visual stress related degradation to the eye.

<CIT> relates to a method of enhancing an aspect of memory in a healthy individual, wherein the aspect of memory is selected from the group consisting of: associative memory, spatial memory and memory under stress comprising: administering a composition consisting of: a) an effective amount of either lutein or the combination of lutein and zeaxanthin; and b) an appropriate carrier; and observing the enhanced associative memory, spatial memory or memory under stress.

<CIT> relates to a total modular system of multivitamin and mineral supplementation for improving public health. One of the modular systems is a stress module oral dosage composition containing: about <NUM> IU of Beta Carotene, about <NUM> IU of Alpha Carotene, about <NUM> mcg of Lutein, about <NUM> mcg of Lycopene, about <NUM> mcg of Zeaxanthin, about <NUM>,<NUM> IU of Vitamin A, about <NUM> of Vitamin B<NUM>, about <NUM> of Vitamin B<NUM>, about <NUM> of Niacinamide, about <NUM> of Pantothenic Acid, about <NUM> of Vitamin B<NUM>, about <NUM> mcg of Biotin, about <NUM> mcg of Vitamin B<NUM>, about <NUM> of Vitamin C, about <NUM> IU of Vitamin E, about <NUM> of Calcium, about <NUM> mcg of Chromium, about <NUM> of Copper, about <NUM> of Iron, about <NUM> of Magnesium, about <NUM> mcg of Selenium, and about <NUM> of Zinc. The modular system is directed towards supplying the needs to persons during periods of physical and emotional stress, and for support for the immune system during and recovering from illness by using a combination of multivitamins and essential nutrients. The study focuses on the support of the immune system using a combination of multivitamins and minerals. The focus is not on any effect of any particular individual ingredient on physical and emotional stress.

<CIT> discloses the preparation and use of compositions containing epi-lutein and carotenoid compounds such as lutein and zeaxanthin for treating various ocular diseases.

<CIT> discloses a method for the prevention of age related macular degeneration by determining the susceptibility of an individual to age-related macular degeneration and, on the basis of that determination, selecting or identifying (and administering) a substance to the individual. <CIT> discloses a xanthophyll composition containing macular pigments consisting of trans-lutein and zeaxanthin isomers, the composition containing at least <NUM>% by weight of total xanthophylls, of which at least <NUM>% by weight being trans-lutein, at least <NUM>% by weight being (R,R)-zeaxanthin and at least <NUM>% by weight being (R,S)-zeaxanthin. <CIT> describes the effect of a supplement containing meso-zeaxanthin, zeaxanthin and lutein on macular responses.

Although many health benefits of the macular carotenoid rich diet are known, none of the references focus on their role in managing physiological and psychological stress. Stress is a health problem faced by the population at large due to a busy life style and it affects many vital organs of the body, such as the brain and nerves, heart, muscles and joints, digestive system and reproductive system, thus increasing burden on health system. Even though different exercises and counseling techniques are available for managing the stress, none of these is proven to be very effective for improving overall health status of a subject experiencing, suffering from, or likely to develop physiological or psychological stress. Thus there is a need of having an effective solution for managing physiological, psychological and chronic stress, particularly through dietary supplementation, which is convenient for the person in need of the managing the stress.

Taking into account several health benefits and high tissue densities achieved by carotenoids in different body tissues, it would be highly desirable to evaluate effects of carotenoids on improving overall health status by managing physiological and psychological variables causing the stress, in order to provide alternate, effective, and safe solutions to address problems of managing stressful conditions. Chronic psychological stress is fairly common in modern society, and may lead to many undesirable physical and psychological health outcomes. Stress in the form of anxiety or depression can affect behavior of an individual, thus affecting the performance and sometimes can turn to fatal conditions. Stress can lead to changes in the serum level of many hormones including, for example but not limited to glucocorticoids, insulin, vasopressin, catecholamines, growth hormone and/or prolactin. It can also alter the clinical status of many preexisting endocrine disorders, such as but not limited to the precipitation of adrenal crisis and thyroid storm. Stress can enhance secretion of a number of hormones including glucocorticoids, insulin, vasopressin, catecholamines, growth hormone and prolactin. The increase of the hormone levels in subjects such for example a human and/or other mammals can improve mobilization of energy sources and allows them to adapt to their new circumstances. Evidence of the protective role of macular carotenoids in maintaining eye health through the effect of the macular carotenoids as an antioxidant and anti-inflammatory has been presented. Embodiments herein are directed to their applications for management of stress by establishing co-relation(s) between improvement of distribution and density of a macular pigment in eyes and their macular carotenoids effect on relieving stress in a subject in need thereof, thus resulting into improving overall health status.

In one aspect, the present disclosure relates to a macular carotenoid composition comprising lutein including trans-lutein, (R,R)-zeaxanthin and (R,S)-zeaxanthin, along with one or more food grade excipients, for use in increasing macular pigment optical density (MPOD) levels and treating psychological stress in a subject, wherein the macular carotenoid composition is derived from a plant extract/oleoresin containing xanthophylls/xanthophylls esters and comprises at least <NUM>% by weight of total xanthophylls, of which at least <NUM>% by weight being trans-lutein, at least <NUM>% by weight being (R,R)-zeaxanthin and at least <NUM>% by weight being (R,S)-zeaxanthin, wherein the ratio of trans-lutein to (R,R)-zeaxanthin and (R,S)-zeaxanthin is in the range of <NUM>:<NUM> to <NUM>:<NUM> and the ratio of (R,R)-zeaxanthin and (R,S)-zeaxanthin is in the range of <NUM>:<NUM> to <NUM>:<NUM>, and wherein the composition is administered daily and comprises at least <NUM>/kg body weight of lutein and at least <NUM>/kg body weight of (R,R)-zeaxanthin and (R,S)-zeaxanthin.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the lutein of <NUM>/kg body weight.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the zeaxanthin isomer(s) of <NUM>/kg body weight.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the combination of <NUM>/kg of the lutein, and <NUM>/kg body weight of the zeaxanthin isomer(s) to the subject.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the zeaxanthin isomer(s) of at least <NUM>/kg body weight.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the combination of the lutein of at least <NUM>/kg body weight, and the zeaxanthin isomer(s) of at least <NUM>/kg body weight.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the lutein of at least <NUM>/kg body weight.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the combination of the lutein of at least <NUM>/kg body weight and the zeaxanthin isomer(s) of at least <NUM>/kg body weight.

In an embodiment, the macular carotenoid composition is administered at a daily dose of the combination of the lutein of <NUM>/kg body weight and the zeaxanthin isomer(s) of <NUM>/kg body weight.

In an embodiment, the macular carotenoid composition is administered at a daily dose at least <NUM>/kg body weight of the lutein or lutein isomer such as t-lutein, and/or at least <NUM>/kg body weight of the zeaxanthin isomer(s), such as meso-zeaxanthin and R,R zeaxanthin, either alone or in combination.

It will be appreciated that any of the mg/kg values stated above can represent a lower limit in a range of the mg/kg amount of the respective carotenoid (i.e. the lutein and/or zeaxanthin isomer(s)) present in the dose. It will also be appreciated that any of the mg/kg values above can represent an upper limit of the mg/kg amount of the respective carotenoid, either alone, or where relatively higher mg/kg values can be used as an upper limit in a range together with one of the relatively lower mg/kg values described above as the lower limit.

The macular carotenoid composition includes along with the active ingredient(s), e.g. carotenoid(s), at least one food grade excipient as described herein.

In an embodiment, the methods herein include evaluating overall health status after the administration of the macular carotenoid composition.

An embodiment herein is directed to a method of identifying a subject in need thereof to increase MPOD, by administering a macular carotenoid composition to such a subject in an effective daily dose and evaluating an improvement of optical density and/or of distribution of macular pigments.

The higher the level of MPOD, the better protection one has against retinal damage from blue light and oxidative stress. The MPOD is associated with brain function, executive function, and processing speed. The layer of macular pigment is located in the inner retina, and it is remote from the metabolic activities involved in the pathogenesis of age-related macular degeneration (AMD) which occur in the outer retina. The generation of radical oxygen species (ROS) in retinal pigment epithelium (RPE) cells and the resulting increase in oxidative retinal stress in the outer retina. When ROS levels exceed the RPE cell's antioxidative capacity, the result is increased levels of retinal oxidative stress and increased inducement of the inflammatory response. The antioxidant enzymes superoxide dismutase (SOD), catalase, glutathione peroxidase and glutathione reductase, for example, display reduced activities in the brains. A decline in the normal antioxidant defense mechanisms can increase the vulnerability of the brain to the deleterious effects of oxidative damage. ROS can cause severe damage to mitochondrial and cellular proteins, lipids, and nucleic acids and increase psychological and physiological stress.

In an embodiment, a macular carotenoid composition may be administered on a daily basis in an effective amount, such as for example at a mg/kg body weight dose of each lutein and/or meso-zeaxanthin, and at a mg/kg body weight dose of zeaxanthin (e.g. R,R-zeaxanthin), and then evaluating reduction of psychological stress and associated hormones and conditions by co-relating the administered amount(s) of the carotenoid(s) with an improvement in optical density or distribution of macular pigments.

In an embodiment, a macular carotenoid composition may be administered on a daily basis in an effective amount, such as for example at levels of <NUM> to <NUM>/day of the active ingredient(s), for a period of <NUM> to <NUM> months or longer, and assessing the parameters for evaluation of reduction in psychological and physiological stress at least at baseline, around <NUM> months, and around <NUM> months, and annually thereafter.

In an embodiment, <NUM> to <NUM> daily dose of trans-lutein corresponds to administration of a macular carotenoid composition at a dose of at least <NUM>/kg body weight of lutein. It will also be appreciated that a daily dose of at least <NUM>/kg body weight of the zeaxanthin isomer(s) (e.g. meso- and/or R,R-) can also be included. See reported example below.

In an embodiment, the daily dose may be in the range of <NUM> to <NUM> of lutein (e.g. trans-lutein). The daily dose also includes an amount of zeaxanthin isomer(s). In an embodiment, the amount of zeaxanthin isomer(s) can be about <NUM>. In an embodiment, the amount of zeaxanthin isomer(s) can be about <NUM> of meso-zeaxanthin and <NUM> of R,R-zeaxanthin. In an embodiment, other doses of lutein can be <NUM> and <NUM>, where the amount of zeaxanthin isomer(s) can be adjusted accordingly.

In an embodiment, the period of administration is at least one month. It will be appreciated that the period of administration can be continued for a period till the stress management is evident through MPOD improvement, cortisol reduction and other parameters. It will also be appreciated that administration can be taken for extended time periods, without any side effects or harmful reactions in the body. It will also be appreciated that the evaluation can be done at <NUM> time, which is baseline, while monthly and/or quarterly evaluations can be continued for a year or longer until the suitable stress management is achieved and/or risk factors are managed.

An embodiment includes administering to a subject in need thereof, an effective amount of a daily dose of lutein and/or zeaxanthin isomer(s) and evaluating reduction in physiological stress by co-relating the administered amount(s) of the carotenoid(s) with a reduction in cortisol levels. For example, the daily dose includes lutein, meso-zeaxanthin, and R,R-zeaxanthin.

An embodiment also includes administering to a subject in need thereof, an effective amount of a daily dose of at least <NUM> of the active ingredient(s) of the macular carotenoid composition and assessing the levels of serum lutein and zeaxanthin isomers, blood cortisol levels and macular pigment optical density (MPOD) via relevant techniques. Behavioral data are also obtained via questionnaire to assess psychological stress, anxiety and depression in the subjects administered with the macular carotenoid composition.

The term 'co-relating' herein includes a response observed in subjects after administration of the specific doses of a composition that is proportional, e.g. directly proportional, to the improvement in optical density or distribution and/or reduction in cortisol levels, thus dose and effect can be co-related to each other. Co-relation between the dose and effect (response) can be observed through effect of various doses on MPOD, e.g. at baseline, subjects' estimation of their level of psychological stress was significantly correlated to their retinal lutein status. To check effect of carotenoid supplementation on stress reduction, subjects' psychological stress measures were evaluated at the final study visit, for example at about <NUM> weeks and compared to their baseline levels. Psychological stress was found to be significantly reduced in the groups administered with the compositions corresponding to <NUM> and <NUM> of t-lutein, but not in group which received <NUM> dose or placebo group. Hence there is a co-relation between levels of carotenoid administered and the effect observed. It may be also that at some level beyond a <NUM> daily dose, retinal response to carotenoid supplementation levels off.

The macular carotenoid compositions and uses described herein impact on three physiological systems that are involved in the stress response, which include the nervous system, the endocrine system, and the immune system.

Macular carotenoid compositions herein can provide uses for managing stress of a subject by administering a daily dose of lutein either alone or in combination with effective doses of zeaxanthin isomer(s). These carotenoids are administered in combination with each other and in combination with at least one food grade excipient, for example a fat, fatty acid, oil, antioxidant and the like, and combinations thereof, to a subject.

In an embodiment, a macular carotenoid composition may be administered on a daily basis in an effective amount, such as for example at levels of the active ingredient(s) of at least <NUM> to <NUM>/day for a period of <NUM> to <NUM> months and assessing the parameters for evaluation of reduction in psychological and physiological stress at baseline, <NUM> months and <NUM> months.

Uses herein include administering an effective amount of a macular carotenoid composition comprising at least or about <NUM>% by weight of total xanthophylls to an individual in need thereof suffering from stress. The xanthophylls contain at least <NUM>% by weight trans-lutein, at least <NUM>% by weight (R,R)-zeaxanthin, and at least <NUM>% by weight (R,S)-zeaxanthin , and the remaining concentration may be other carotenoids. In an embodiment, the amount of trans-lutein can be <NUM>% by weight or higher than <NUM>% by weight.

In an embodiment herein an effective daily dose of a macular carotenoid composition is used for management and treatment of a subject, experiencing, suffering or likely to be affected by psychological stress, or conditions of the like hampering routine activities and overall well-being.

The term 'treatment' refers to the condition wherein the subject suffering from stress, in particular psychological stress, is identified by relevant techniques and the macular carotenoid compositions as described herein, are administered in effective doses to such individual over a period of time, in order to treat his/her condition and to also take care of risk factors associated with the stress condition. The subject who is in need of such treatment can be also identified by asking a set of questionnaire, which give indication about overall health status and thus sets ahead need for treatment of such subject.

The term 'prevention' or 'prophylaxis' refers to administering the macular carotenoid compositions described herein, in an effective amount to subjects who do not show any current symptoms of stress, but are prone to psychological stress due to hectic routine, work pressure, family conditions, health problems, social status. As the macular carotenoid compositions can be used as nutritional supplements over a prolonged time without any side effects, these can be administered to such subjects as a prophylactic or preventive measure.

The term "manage, managing, or management of stress" as used herein, unless otherwise indicated, describes controlling and improving health conditions of a subject in need thereof, in terms of improving the associated risk factors based on such stress, which affect routine daily activities and additional work load, thus lowering the output expected from an individual. The stress may be physiological, psychological or any other related type of stress. Physiological stress may arise because of chronic or acute diseases, thus affecting specific body organs. This may be experienced by subjects suffering from ailments like diabetes, hypertension, and/or cancer, thus affecting other body systems. The psychological stress may affect number of physiological functions and activities of an individual under stress. One of the risk factors may be hampering visual performance or overall well-being of an individual, which is due to lowering of density of macular pigments (MPOD) or disturbing the distribution of macular pigments in a subject's eye. Another risk factor may be increase in blood cortisol levels or oxidative or inflammatory markers, which are indicative of psychological stress. Improving or reducing one or more of these risk factors by administering an effective amount of a the macular carotenoid composition herein can help in reducing stress and to improve overall health status of a subject, who is in need of such treatment. Macular carotenoid compositions and methods as used herein, encompasses both a prophylactic and therapeutic regimen and also includes self-treatment (e.g. a subject without the assistance of any intermediary ingests or applies a carotenoid to himself or herself).

The macular carotenoid compositions herein improve density and/or distribution of macular pigments in an eye, thus relieving stress in a subject in need thereof. Macular carotenoid compositions herein can also reduce stress by reducing cortisol levels in blood.

Embodiments herein can improve overall health status of a subject after administration of macular carotenoid composition(s) herein, and the improvement may be evaluated through the questionnaire and recording the feedback of the subjects.

Macular carotenoid compositions herein are safe for consumption and can be employed for management of relieving psychological stress as well as associated risk factors when administered in an effective amount to a subject in need thereof. The reduction in stress is shown by improvement in density and distribution of macular pigment as well as reduction in blood cortisol levels, which ultimately results in improvement in overall health status and well-being of a subject.

Applicant has carried out rigorous experimentation for the evaluation of effect(s) of macular carotenoid composition on healthy volunteers as well as subjects suffering from stress and found that it is effective in increasing MPOD levels, reducing cortisol levels, reducing anxiety and depression, thus managing stress and improving overall health status, when administered in an effective daily dose.

Stress in a subject can be co-related with reduced or disturbed density and/or distribution of macular pigments in retina of a subject, where administration of an effective amount of macular carotenoid composition(s) herein improves density and/or distribution of macular pigments, thus managing or relieving psychological stress and improving overall health status.

In one embodiment herein, stress in a subject is also presented in the form of anxiety or depression, or disturbed overall health status, which is seen to be taken care of by administering an effective amount of the macular carotenoid composition over a period of at least <NUM> to <NUM> months, and evaluating behavioral data by using relevant questionnaire to assess psychological stress such as anxiety and depression, in healthy subject, and/or in a subject in need thereof. The macular carotenoid compositions herein can be in the form of delivery systems, which are convenient for administration and ensure dose accuracy and efficacy, such as powder, granules or beadlets for reconstitution in a suitable vehicle or beverage medium, tablets, capsules, oil suspensions, soft gel capsules, meant for oral administration, as well as semisolid and liquid formulations for other than enteral routes of administration. The delivery of the macular carotenoid compositions or preparations can be packaged and used in, for example, flavoring mixtures that also contain salt or other spices for use in flavoring foods such as for example but not limited to soups, salads, popcorn, casseroles, and the like, baking mixes for use in preparing cakes, cookies, brownies, and the like, dried soup mixes, additives for milk such as chocolate-flavored powders, and other granular formulations, such as but not limited to functional foods and beverages. The delivery system may include an enteric coating, a sustained release coating, and/or a release control coating. The delivery system may release lutein and/or meso-zeaxanthin, and zeaxanthin about <NUM> minutes to <NUM> hours from the time of administering the composition to the subject.

Macular carotenoid compositions described herein relieve or manage psychological stress and also reduce anxiety and depression, thus improving overall health status of healthy individual or a subject suffering from stress.

Macular carotenoid compositions herein also improve distribution and density of macular pigments in the eyes, thus improving visual performance of a subject in need thereof, thus relieving or managing psychological stress.

Macular carotenoid compositions herein can be administered to a subject for periods of <NUM> to <NUM> months and maintain higher levels of serum carotenoids, thus resulting into reduction in oxidative stress and inflammation, beneficial for reducing stress, cortisol levels and anxiety. Macular carotenoid compositions herein are safe for consumption and can be employed for management, treatment or prevention of stress as well as the associated risk factors of stress, and to improve overall health status of a subject, when administered in effective amounts over prescribed time period.

The following terms, among others, are used to describe the macular carotenoid compositions and uses herein. It is to be understood that a term which is not specifically defined is to be given a meaning consistent with the use of that term within the context of the macular carotenoid compositions and methods herein and as understood by those of ordinary skill.

The term "subject" is used throughout the specification within a context to describe a mammal, an animal or a human being, to which management, treatment, and/or prevention including prophylactic treatment (and prophylaxis), with macular carotenoid compositions herein is provided. For management, treatment and/or prophylaxis of those conditions, specific parameters or disease states which are specific for human beings to generate stress are considered. Human beings may be subjected to various conditions such as changes in routine, extra work load, disturbed family conditions and relationships and the like conditions which affect normal functioning of such subject. Chronic and disease conditions such as diabetes, hypertension, systemic and local infections, cancer, physical injury, and the medical treatment thereof may also cause stress in individuals. Aside from applications for humans, the macular carotenoid compositions herein have additional uses in the veterinary world. Conditions under which animals would benefit due to administration of macular carotenoid compositions herein are particularly those which are subjected to stress due to training procedures and education for specific purposes, e.g. for hunting dogs, guide dogs, police dogs, etc., or animals used in the movie industry. Animals which can benefit from macular carotenoid compositions herein include those animals which are subject to stressful conditions such as, for example, after capture or transport or may be due to housing conditions such as change of domicile or owner. Animals which are subject to stress would also include those which are racing animals (e.g. dogs, horses, camels), or used in various sports, performing animals (such as circus animals and those appearing on stage, television or in the movies) and horses which perform dressage and other highly disciplined routines. Preferred "animals" may include but are not limited to pets or companion animals and farm animals. Examples of pets are dogs, cats, birds, aquarium fish, guinea pigs, (jack) rabbits, hares, and ferrets. Examples of farm animals are aquaculture fish, pigs, horses, ruminants (cattle, sheep and goats), and poultry.

The term "effective amount" is used herein, unless otherwise indicated, to describe an amount of a compound or composition which, in context, is used to produce or affect an intended result, whether that result relates to the management of a subject's stress by improving overall health status. These amounts may reflect specific concentrations of active ingredient(s) which are present as part of a total composition, when the doses are measured as an amount(s) corresponding to individual nutrient(s) such as lutein, lutein isomers, zeaxanthin, meso-zeaxanthin, and/or other zeaxanthin isomers, as present in the composition The doses administered per kg body weight corresponding to specific nutrient of the composition may vary, but the effect produced by specific doses is measurable qualitatively and/or quantitatively for a particular health application under test. This term subsumes all other effective amount or effective concentration terms (including the term "amount sufficient") which are otherwise described in the present application.

The term "overall health status" refers to the health status of a subject which is a multidimensional concept that is usually measured in terms of emotional well-being, physical well-being and satisfactory social functioning. There is no single "standard" measurement of overall health status for individuals or population groups of animals. Subject's overall health status may be measured by an observer (e.g., a physician), who performs an examination and rates the individual along any of several dimensions, including parameters reflecting overall well-being of an individual for sound performance during daily routine activities. The questions that can be asked or effects observed in the subjects under test can be related to assessing nervousness or shakiness inside the body, faintness or dizziness, feeling annoyed or irritated, a troublesome feeling, poor appetite, loneliness, and/or fear of a crowd or situations, and the like. The overall health status is evaluated by asking such questions which are called as Sub-Health Status Questionnaire (SHSQ) and rating the individual responses on a scale and the data are aggregated to assess those tested and for a population.

The term "zeaxanthin" or "zeaxanthin isomers" is used throughout this specification within a context to describe the compositions, and can include RR-zeaxanthin and/or RS-meso-zeaxanthin (meso-zeaxanthin).

The term "macular carotenoids" is used throughout the specification within a context to describe the compositions, and include lutein and/or zeaxanthin isomers such as R,R-zeaxanthin and/or R,S-meso-zeaxanthin herein.

Therefore the term "subject in need thereof" as used herein, unless otherwise mentioned, means an individual who is suffering from one or more risk factors or symptoms indicating psychological stress, which may affect normal physiological functions and work conditions. Subjects in need of stress management can be identified by measuring density or distribution of macular pigments in retina of a subject and selecting the subjects having lowered macular pigment optical density or distribution.

The term "macular carotenoid composition" as used herein, unless described otherwise, describes a xanthophyll composition containing high amounts of lutein including trans-lutein, (R,R)-zeaxanthin and (R,S)-zeaxanthin, in combination, and including one or more food grade excipients, such as fatty acid, fatty acid ester, triglyceride, oil, antioxidant, stabilizer and the like. The macular carotenoid compositions are derived from the plant extract and/or oleoresin containing xanthophylls and/or xanthophylls esters, which is safe for consumption and useful for nutrition and health applications. The macular carotenoid composition may be administered in the form which is convenient for administration and ensures dose accuracy, selected from, but not limited to, powders, granules, beadlets for reconstitution, filled as such in sachets, or formulated into tablets, capsules, soft gel capsules, oil suspensions, or formulated as semisolids or liquids for enteric and other routes of administration.

The macular carotenoid compositions are formulated by using one or more nutrients or food grade excipients selected from a group of, but not limited to fats, fatty acid, oils, medium chain triglycerides, antioxidant, surfactants, vitamins, diluents, taste masking agents, stabilizers, and the like, and combinations thereof.

The subjects are evaluated for improvement of overall health status using a questionnaire (SHSQ) and recording the responses to the questionnaire to assess management of psychological stress.

In an embodiment, the subject in need thereof is administered with an effective amount of a macular carotenoid composition herein, comprised of lutein including trans-lutein, (R,R)-zeaxanthin and (R,S)-zeaxanthin, as a daily dose, either over a period of prescribed time period or as a nutritional supplement (e.g. in combination with multivitamins and/or mineral supplementation) for a life-long time period, based on a need of a subject. The macular carotenoids are used along with one or more nutrients and/or food grade excipients.

As per another embodiment, the subject in need thereof is administered with an effective amount of a macular carotenoid composition as described herein, comprised of at least <NUM> to <NUM>/day of active ingredient(s) in the macular carotenoid composition and parameters for stress evaluation are monitored at baseline level, at <NUM> months as well as at the end of <NUM> months of administration.

As mentioned above, the xanthophyll composition is comprised of at least <NUM>% by weight of total xanthophylls. The xanthophylls include at least <NUM>% by weight of trans-lutein, at least <NUM>% by weight of (R, R)-zeaxanthin, and at least <NUM>% by weight of (R,S)-zeaxanthin. The ratio of trans-lutein and zeaxanthin isomers is in the range of <NUM>:<NUM> to <NUM>:<NUM>. The ratio of the isomers of zeaxanthin is in the range of <NUM>: <NUM> to <NUM>:<NUM>.

The macular carotenoid compositions herein can satisfy the safety and regulatory considerations, because of the use of generally recognized as safe (GRAS) reagents and hence, is generally safe for consumption and useful for nutrition and health care.

Macular carotenoid compositions herein are administered to a healthy subject and/or a subject in need thereof, to manage, treat and/or prevent psychological stress, when administered in daily effective doses. Evaluation of the subjects is carried out by measurement of optical density or distribution of macular pigments in the retina, recording the responses to a questionnaire to assess overall health status of a subject. The levels of serum macular carotenoids are also monitored during and after the period of administering the macular carotenoid composition to the subject. Behavioral pattern of subjects under test and anxiety and/or depression states are evaluated using relevant questionnaire. Macular carotenoid compositions herein may be administered over a prescribed time period or over a prolonged time, such as life-long administration of the macular carotenoid composition as per need of a subject.

In an embodiment, the macular carotenoid compositions described herein are in an oral form, for example, a tablet, a capsule, a soft capsule, granules, liquid, effervescent tablet, etc., a suppository form, an eye preparation form, a nasal preparation form, a topical form, for example ointment, gel, cream, lotion, a patch, a sublingual tablet or patch, etc..

Phlebotomy may be performed and macular pigment optical density (MPOD) is measured via heterochromatic flicker photometry. An eccentric area of fixation in human retina commonly referred to as a preferred retinal locus/loci (PRL) is considered as the "standard" measure and the <NUM>' retinal locus (preferred retinal loci) is used for analysis, wherein visual performance is evaluated after administering an effective amount of a macular carotenoid composition herein. Increased MPOD levels are correlated to reduction in stress and improvement in overall health status of a subject, who is administered with an effective amount of a macular carotenoid composition herein. Serum lutein and zeaxanthin isomers levels are monitored and measured by HPLC technique. Overall health status of a subject is also evaluated by asking questions to subjects and recording the answers and/or recording observations in certain situations to which the subject is exposed and recording the observations, and assessing the anxiety, depression condition and overall stress management.

The details of the methods and macular carotenoid compositions herein, its objects, and advantages are explained hereunder in greater detail in relation to non-limiting exemplary illustrations.

The purpose of this study using a macular carotenoid composition described herein was to characterize the response dynamics in both blood and retina, for three different daily levels of macular carotenoid composition corresponding to t-lutein in amounts of <NUM>, <NUM>, and <NUM> dose of the active ingredient (i.e. t-lutein), versus a placebo. Data regarding subjects' stress and overall health status were also obtained. The macular carotenoid composition was in a formulation of a soft gelatin capsule and was administered orally. The <NUM>, <NUM>, and <NUM> doses relate to a daily dose in the macular carotenoid composition administered having at least <NUM>/kg body weight of lutein and <NUM>/kg body weight of zeaxanthin isomers (small amount of zeaxanthin is included, e.g. meso-zeaxanthin and R,R zeaxanthin).

This study was a double-blind, placebo-controlled <NUM> week trial. Informed consent was obtained from each subject, and the study adhered to the tenets of the Declaration of Helsinki. Twenty-eight healthy subjects, aged <NUM>-<NUM> years, participated in this study. Subjects were randomly assigned to study groups: placebo (n = <NUM>), a macular carotenoid composition corresponding to <NUM> lutein (n = <NUM>), a macular carotenoid composition corresponding to <NUM> lutein (n = <NUM>), and a macular carotenoid composition corresponding to <NUM> lutein (n = <NUM>). Subjects were randomized to different treatments. Each subject had one capsule per day orally for a period of <NUM> weeks. After being familiarized with the study, subjects were instructed to visit the laboratory every <NUM> weeks, in order to participate in vision testing and phlebotomy. Phlebotomy was conducted after fasting for about <NUM> hours, and subjects were given some food (e.g. a bagel or a breakfast bar) and water immediately after the blood sampling. A Macular pigment measurement was conducted shortly thereafter. The spatial profile of MPOD was assessed via heterochromatic flicker photometry, using a device described by Wooten et al.

Effect of a macular carotenoid composition herein was also assessed by measuring levels of cortisol in blood by administering the doses corresponding to <NUM>- to <NUM>-mg lutein, for <NUM> weeks to the subjects under test. The baseline and final (<NUM> week) visits also included extensive questionnaires designed to acquire information regarding subjects' diet, health, and psychological stress level. For the health assessment, the Sub-Health Status Questionnaire (SHSQ-<NUM>; Wang and Yan, <NUM>) was utilized to determine overall health status. To ensure that the subjects met the inclusion criteria for participation of this study, biometric data (e.g. height, weight, body fat percentage), as well as health habits data (e.g. whether or not a smoker) of the subjects were obtained at each visit.

In some embodiments, the above <NUM> to <NUM> doses, for example, can also be lutein from <NUM>/kg to <NUM>/kg for macular carotenoid compositions corresponding to <NUM> of t-lutein, (e.g. corresponding to body weights ranging from <NUM> to <NUM>, infant to obese).

In some embodiments, lutein can be at <NUM>/kg to <NUM>/kg for macular carotenoid compositions corresponding to <NUM> of t- lutein (calculated for body weights <NUM> to <NUM>); while calculating for zeaxanthin and isomers-can also correspond to <NUM>/kg to <NUM>/kg corresponding to composition containing <NUM> of t-lutein, which contains not less than <NUM> of zeaxanthin isomers.

In some embodiments, lutein can further be at <NUM>/kg to <NUM>/kg for macular carotenoid compositions containing <NUM> t-lutein and not less than <NUM> zeaxanthin (for weights range <NUM> to <NUM>);
In an embodiment, lutein may range from at or about <NUM> to at or about <NUM>/kg, and zeaxanthin isomers may range from at or about <NUM>/kg to at or about <NUM>/kg of zeaxanthin isomers. In an embodiment, such relative amounts are contained in macular carotenoid composition containing <NUM> to <NUM> of t-lutein.

In an embodiment, such macular carotenoid compositions herein can be administered as a daily dose in a range from at or about <NUM> to at or about <NUM>/kg body weight for lutein, and/or in a range from at or about <NUM> to at or about <NUM>/kg body weight for zeaxanthin isomers.

The overall results for the MPOD response to the various dose levels of the carotenoid, as a function of time in the study, are shown in <FIG>. Although data were obtained at week <NUM>, a response in the retina (albeit minor) was not detectable until the second study visit (i.e. week <NUM>), and so the week <NUM> is the first data point presented for each group. Significant increases from the baseline were determined for the <NUM> group at the <NUM>-week visit (p = <NUM>); for the <NUM> group at weeks <NUM>, <NUM>, and <NUM> (p < <NUM>), and for the <NUM> group at weeks <NUM>, <NUM>, and <NUM> (p < <NUM>). Because it is considered to be the "standard" measure, the <NUM>' retinal locus (Preferred retinal loci) was used for this analysis. Although the <NUM>' measure explained the most variance in the data, similar results were found for all retinal loci tested, especially those even closer to the center of the fovea (e.g. <NUM>' and <NUM>' loci). From <FIG> it can be seen that the response to each level of composition was roughly linear as a function of time, and the response was shown to be greater as time passed as a function of the level of the carotenoid. The data suggest a significant increase in response with increased doses. The increase with the <NUM> group is closer in magnitude to the increase with the <NUM> group, compared with the increase with the <NUM> group versus the increase with the <NUM> group.

Subjects' level of psychological stress was significantly correlated to their MPOD (r = -<NUM>; p = <NUM>; see <FIG>). In order to determine the potential of macular carotenoid compositions which include for example to <NUM> to <NUM> of trans-lutein, and which may include in some embodiments combinations with meso-zeaxanthin and/or zeaxanthin (e.g. R,R-zeaxanthin), one example composition tested herein is a Lutemax <NUM> composition by Applicant, which is disclosed for example in co-pending <CIT>. Supplementations of such macular carotenoid compositions were evaluated to determine stress reduction, and the subjects' psychological stress were measured at the final study visit (week <NUM>) and compared to their baseline levels.

In other embodiments, macular carotenoid compositions can also include those from Applicant's <CIT>.

<FIG> shows the subjects' level of psychological stress at baseline, as a function of MPOD. Red line is least-squares fit to the data (Y = -<NUM>. 012X +<NUM>). r = -<NUM>; p = <NUM>. Paired-sample t-tests were used for this purpose. Psychological stress was found to be significantly reduced in the <NUM> (t = <NUM>; p = <NUM>) and <NUM> (t = <NUM>; p = <NUM>) groups, but not <NUM> or placebo groups (see Table <NUM>). It appears therefore that the macular carotenoid compositions corresponding to <NUM> and <NUM> of t-lutein account for the significant reduction in the psychological stress.

A significant correlation between the retinal lutein status and overall health status (as assessed by the SHSQ-<NUM> - see Appendix I) was obtained(r = -<NUM>; p = <NUM>; see <FIG>). As with the psychological stress data, effects of the macular carotenoid composition on overall health status of subjects (as assessed via the SHSQ-<NUM>) were determined via comparing their baseline scores to their final visit scores with paired-samples t-tests. At all dose levels of the carotenoid (<NUM>, <NUM>, <NUM> of t-lutein (lutein)) in the macular carotenoid compositions, it was found that overall health status improved significantly over the <NUM> week study period (see Table <NUM>).

Referring to the SHSQ, "SHSQ-<NUM>" scores means scores calculated for each respondent by summing the ratings for <NUM> questions. Although annexure shows more than <NUM> questions, these are reduced to <NUM>, by combining two questions to form one sentence. Thus total questions asked to the subjects were <NUM>. Each subject was asked to rate a specific statement, based on how often they suffered various specific complaints in the preceding <NUM> months. The raw scores of <NUM> to <NUM> on the questionnaire were recoded as <NUM> to <NUM>. SHS scores were calculated for each respondent by summing the ratings for the <NUM> items. The SHSQ is considered as indicative of stress conditions in the subjects, and is used to understand behavioral patterns of a group of subjects, and to understand overall health status of the subjects administered the compositions as described herein.

<FIG> shows the subjects' overall health status at baseline, as a function of MPOD.

The line is a least-squares fit to the data (Y = -<NUM>. 90X +<NUM>). r = -<NUM>; p = <NUM>.

In order to visualize the change in sub-optimal health scores over the <NUM>-week study period, <FIG> presents a box-and-whisker plot of the baseline and the final visit data for all <NUM> lutein supplementation subjects. A paired-samples t-test determined a statistically significant improvement in the overall health status over the study period (t = <NUM>; p < <NUM>). Questionnaires evaluating psychological stress levels, overall health status (suboptimal health questionnaire [SHSQ]), and diet were completed at baseline and final visits.

In addition to a significant decrease in sub-optimal health issues, the subjects also exhibit a markedly reduced score range and its variability in their responses for the <NUM> week measure versus their baselines (see <FIG>).

The term sub optimal health issues is related to Sub optimal health questionnaire (SHSQ) provided in Annexure I. A set of questions is prepared which is asked to the study subjects to assess what is the health status before administering the macular carotenoid composition and what is at the end of study, to understand improvement in overall health status of a subject. (Wang W, Yan Y (<NUM>). Suboptimal health: a new health dimension for translational medicine. ClinTransl Med. <NUM>;<NUM>(<NUM>):<NUM>.

In <FIG>, baseline and final visit SHSQ-<NUM> scores are compared with box-and-whisker plots. Asterisks indicate lower and upper range of values, bars are +/- <NUM> standard deviation from the mean.

<FIG> shows subjects' overall health status scores, as a function of psychological stress level, at baseline. The line is a least-squares fit to data (Y = <NUM>. 35X + <NUM>). r = <NUM>; p = <NUM>.

MPOD increased significantly in subjects administered with the macular carotenoid compositions herein, compared to a placebo (which exhibited little to no change throughout the trial). Also, it was found that a significant relationship existed between MPOD and psychological stress: r = -<NUM>; p = <NUM> (more MPOD = lower stress). The retinal response was somewhat linear, with greater increases seen in those taking the higher (e.g. <NUM>) dose of lutein. A repeated-measures analysis of variance (ANOVA) revealed significant increases from the baseline, which were determined with the <NUM> group at the <NUM>-week visit (p = <NUM>); with the <NUM> group at visits at weeks <NUM>, <NUM>, and <NUM> (p < <NUM>), and with the <NUM> group at visits at weeks <NUM>, <NUM>, and <NUM> (p < <NUM>). The cross-sectional evaluation of MPOD versus psychological stress revealed a significant relationship between the two, such that the subjects having higher levels of MPOD tended to maintain a lower psychological stress profile (r = -<NUM>; p = <NUM>). After <NUM> weeks of lutein supplementation, psychological stress levels were found to be reduced significantly in the <NUM> (p = <NUM>) and <NUM> (p = <NUM>) groups. The placebo group did not change in this regard. In terms of overall health, the subjects' scores on the SHSQ were found to correlate significantly at the baseline with MPOD, such that those having higher MPOD tended to have fewer health-related problems (e.g. they were sick less often, suffered less from allergies; r = -<NUM>; p = <NUM>). After <NUM> weeks of macular carotenoid composition administration, each group, other than the placebo group, exhibited a significant reduction in health-related problems (<NUM>: p = <NUM>; <NUM>: p = <NUM>; <NUM>: p = <NUM>).

In the group administered with the macular carotenoid composition described herein, stress and cortisol were significantly lowered after <NUM>-<NUM> weeks.

Based on the results of this study, ingestion of a macular carotenoid composition herein can produce a significant positive response in a retina. The shapes of the response curves suggest that a response is somewhat linear - higher doses result in a greater response. In terms of systemic/psychological effects, this is the first study to show a relationship between a macular carotenoid composition and psychological stress, and that between a macular carotenoid composition and overall health. These findings have very important implications as an effective, safe, and alternate technique for managing psychological stress by improving an overall health status.

Macular carotenoid compositions herein have been readily bioavailable, judging from the robust retinal response in subjects. Compared to a placebo, a significant positive response was seen after <NUM> weeks in the group even with the lowest (<NUM>) dose. Moreover, a significant retinal response was seen in the <NUM>- and <NUM>-mg groups after only <NUM> weeks of administration with a macular carotenoid composition herein. Second, daily administering of a macular carotenoid composition herein corresponding to <NUM>- or <NUM>-mg of lutein significantly reduces psychological stress after <NUM> weeks of administering the macular carotenoid composition. In our study this effect was seen with the macular carotenoid composition corresponding to two higher doses (<NUM> and <NUM>) of lutein, which indicates that relatively high levels of daily intake can provide effects on psychological stress reduction.

In the group administered with a macular carotenoid composition herein, stress and cortisol was significantly lowered after <NUM>-<NUM> weeks. Scores of overall health status demonstrate significantly improvement over the <NUM> week study period, for all levels of macular carotenoid compositions (see Table <NUM>), but not for the placebo group. Lastly the results of the study and consideration of relationships of the macular carotenoid composition with both stress and health as shown in <FIG> indicate interrelationships among bodily systems (such as the nervous, endocrine, and immune systems), and that the status of these systems can ultimately impact management of physiological and psychological stress.

This <NUM>-month, double-blind, placebo-controlled trial was conducted to evaluate the effects of administering carotenoid on blood cortisol levels, subjects' psychological stress ratings, behavioral measures of mood, and symptoms of sub-optimal health.

Method of evaluation: <NUM> young (mean age = <NUM> yrs. ; <NUM> males), healthy subjects were evaluated in this study. Subjects were randomly assigned to three groups: Group <NUM> (placebo; n = <NUM>), Group <NUM> (n = <NUM>; <NUM> / day total carotenoids, and Group <NUM> (n = <NUM>; <NUM> / day total carotenoids (n = <NUM>). All parameters in the study were assessed at baseline, <NUM> months, and <NUM> months. Some of the blood markers such as Interleukins, TNF alpha and Brain-derived neurotrophic factor (BDNF) were also evaluated during this supplementation. Total study period was <NUM> months, the evaluation was done at both <NUM> and <NUM> months to check effects at definite intervals.

BDNF, which is a protein in humans, is encoded by the BDNF gene. BDNF is a member of the neurotrophin family of growth factors, such as the canonical Nerve Growth Factor. Exposure to stress and the stress hormone corticosterone has been shown to decrease the expression of BDNF, and, if exposure is persistent, this leads to an eventual atrophy of the hippocampus. Supporting this, voluntary exercise, caloric restriction, intellectual stimulation and various treatments for depression increase expression of BDNF in the brain.

Serum lutein and zeaxanthin isomers were measured by high performance liquid chromatography (HPLC), blood cortisol via (ELISA), and macular pigment optical density (MPOD) via customized heterochromatic flicker photometry. Behavioral data were obtained via questionnaire. Psychological stress was determined using the Psychological Stress Measure (PSM-<NUM>), scores of anxiety and depression were obtained via the Beck inventories, overall psychological health via the Brief Symptom Inventory (BSI), and symptoms of suboptimal health were evaluated with the Suboptimal Health Status Questionnaire (SHSQ-<NUM>).

<FIG> shows BDNF and MPOD at baseline (time = <NUM>).

<FIG> shows the change in BDNF as a result of <NUM> months administering carotenoids.

At baseline, BDNF and MPOD expressions were significantly limited and these were increased linearly with the <NUM> months administering of the composition as shown in <FIG>.

At baseline, significant correlations were obtained between MPOD and Beck Anxiety scores (r = -<NUM>; p = <NUM>), MPOD and BSI scores (r = -. <NUM>; p = <NUM>), and between blood cortisol levels and PSM-<NUM> scores (r = <NUM>; p < <NUM>). Marginally significant correlations were obtained for MPOD and blood cortisol levels (r = -<NUM>; p = <NUM>), MPOD and psychological stress (r = - <NUM>; p = <NUM>), and MPOD and symptoms of suboptimal health (r = -<NUM>; p = <NUM>). After <NUM> months of administering macular carotenoid, a repeated-measures ANOVA revealed that psychological stress, blood cortisol levels, anxiety scores, and symptoms of suboptimal health were significantly lower for Groups <NUM> and <NUM> than those for Group <NUM> (p < <NUM> for all). These outcomes either were maintained at their <NUM> month levels or decreased further (but not significantly) at <NUM> months. Serum levels of lutein and zeaxanthin isomers also increased significantly after <NUM> months (p < <NUM>) in Groups <NUM> and <NUM>, and appeared to a plateau thereafter, as indicated by similar values at <NUM> months to those at <NUM> months. Group <NUM> (placebo) did not improve in any of these respects (p > <NUM> for all). In Groups <NUM> and <NUM>, MPOD was found to increase linearly and significantly at both <NUM> and <NUM> months (p < <NUM>). Group <NUM> exhibited no increase in MPOD across the study period (p = <NUM>).

Observations: It appears that the administration of the macular carotenoids lutein, zeaxanthin, and meso-zeaxanthin, at levels of at least <NUM> / daily, leads to significant psychological and physiological benefit. Reduction of stress, cortisol, and anxiety is observed after <NUM> month administering of the carotenoids, and is maintained at <NUM> months. Trial carried out is for <NUM> months, means administration of composition for <NUM> months. During the trials, after <NUM> months administration, positive effects are observed and the stress reduction continued to be evident after administration for <NUM> months. This indicates that the effects are not transient, or short-lived. The mechanism for these findings could involve either deposition of carotenoids in neural tissue, the increase in serum levels with administering carotenoids, or both. The effects, however, leveled off after <NUM> months whereas MPOD continued to increase. It is therefore possible that the reduction of stress, cortisol, and anxiety is dependent upon systemic reduction of oxidative stress and inflammation afforded by relatively high circulating levels of lutein, zeaxanthin, and meso-zeaxanthin. According to this study, the macular carotenoids have been shown to promote wide-ranging, beneficial effects on a number of biological and physiological systems in the body that are themselves impacted by stress (e.g. neural and cardiovascular systems).

Below is one example of a SHSQ (Suboptimal Health Status Questionnaire).

In order to judge overall health status and to identify the subjects with sub-optimal health (showing symptoms of stress), following questions were asked to the subjects.

Read each of the following items carefully and circle that which best describes how much that problem has distressed or bothered you during the past <NUM> days including today. Circle only one number for each problem. Do not skip any items. Read the example below before beginning. If you have any questions, please ask the doctor now.

Claim 1:
A macular carotenoid composition comprising lutein including trans-lutein, (R,R)-zeaxanthin and (R,S)-zeaxanthin, along with one or more food grade excipients, for use in increasing macular pigment optical density (MPOD) levels and treating psychological stress in a subject, wherein the macular carotenoid composition is derived from a plant extract/oleoresin containing xanthophylls/xanthophylls esters and comprises at least <NUM>% by weight of total xanthophylls, of which at least <NUM>% by weight being trans-lutein, at least <NUM>% by weight being (R,R)-zeaxanthin and at least <NUM>% by weight being (R,S)-zeaxanthin, wherein the ratio of trans-lutein to (R,R)-zeaxanthin and (R,S)-zeaxanthin is in the range of <NUM>:<NUM> to <NUM>:<NUM> and the ratio of (R,R)-zeaxanthin and (R,S)-zeaxanthin is in the range of <NUM>:<NUM> to <NUM>:<NUM>, and wherein the composition is administered daily and comprises at least <NUM>/kg body weight of lutein and at least <NUM>/kg body weight of (R,R)-zeaxanthin and (R,S)-zeaxanthin.