FIELD OF THE INVENTION
This invention relates to CognoBar, a nutraceutical composition for
amelioration of cognitive decline in acute stress.
BACKGROUND OF THE INVENTION;
Climatic conditions above the thermally neutral, comfortable zone may
produce unwanted effects on humans of which reduction in mental
performance is one accompanied by unfavorable changes in behavior and
mood. High levels of environmental stress or long hours of exposure to extreme
environmental conditions have significant impairment on performance tasks of
animals. In humans, significant decrements on several types of cognitive
functions like alertness, reaction time, logic skills, and short-term memory in
cold and hypoxia, have been observed. But studies with human volunteers
have not consistently shown positive performance effect with tyrosine
supplementation which might be due to differences in the severity of stress
produced by the experimental paradigm or individual responses to acute
stress.
Among the various environmental factors, high heat and humidity in
particular have devastating effect on human performance. Individual's ability to
accomplish specified tasks is affected by heat stress, high level of which leads
to reduction in both physical and psychological performance. Its impact on
cognition can complicate assignments at higher levels. Command, control
and communications can impair rational decision making and the ability to
analyze complex situations may also be affected adversely. While the
physiological responses to environmental heat have been well established, its
impact on cognition is less understood. In hot* and humid conditions, the
cognitive functions most vulnerable are probably the maintenance of vigilance
and short-term working memory.
2
Stressful events cause an increase in transmission of noradrenergic
neurons in the frontal cortex, because these neurons are activated by stress.
There are ample evidences relating catecholamine (CA), stress and behavior.
Exposure to stress activates central and peripheral nervous system and the
ability of these neurons to continue to release their neurotransmitter is
important in enabling the individual to cope with the stress. Norepinephrine
(NE) depletion is associated with behavioral depression and decreased
exploration and motor behavior in animals and lowered attention span in
humans. Catecholamines are important in animals and lowered attention span
in humans. Catecholamines are important for learning and memory processes.
Since stress results in increased catecholamine synthesis and depletion, it is
likely that the impact of tyrosine supplementation will become more
pronounced in this state.
L-Tyrosine is the precursor amino acid for synthesis of CA
neurotransmitters i.e. dopamine, norepinephrine and epinephrine. It is
reported that its administration specifically accelerates brain CA concentration,
turnover and release from physiologically active CA neurons. Being a normal
constituent of food, it is rapidly metabolized and therefore, unlikely to have
long-term toxicity or side-effects.
The ability of tyrosine to enhance synthesis of CAs in and their release
from rapidly firing neurons has been demonstrated by using a variety of
experimental manipulations. In rats receiving a tyrosine rich diet, neither NE
depletion nor behaviour impairments were found after stress induction. In
humans, similar results were obtained. Supplementation of 100 mg/kg bw
tyrosine caused a reduction in the number of psychomotor impairments and
stress symptoms during exposure to cold hypoxia, compared to a placebo.
Administration of tyrosine, either systemically just prior to initiation of the
stress or as a dietary supplement has been shown to protect animals from both
neurochemical and behavioural consequences of stress. Immobilisation, cold,
3
tail shock was used as stressors in several studies. A significant linear decline
has been shown in NE levels as a function of stress duration in several brain
areas associated with 60-70% decline in activity. A single intra peritoneal dose
of tyrosine (200 mg/kg bw) prior to tail shock protected animals from acute
behavioral depression by stress. Further it is reported that performance on
Morris water maze in saline injected rats was impaired due to stress which
significantly improved in post stress performance in Morris water maze in
animals receiving tyrosine. Stress induced increase in CA metabolites i.e.
homovanillic acid (HVA) and vanillylmandelic acid (VMA) in brain was
prevented by treatment with tyrosine.
Little is known about stress resistance by tyrosine. Some beneficial
effects of tyrosine on mood state of depressed patients were observed. In one
study, beneficial effect of tyrosine on human behaviour was examined where
soldiers were subjected to hypobaric hypoxia (4200 and 4700m) and cold (15°C)
and tested by standardized behavioral tests of mood and mental performance.
A significantly better cognitive performance and reaction time and a
significantly increased plasma tyrosine level was observed in subjects
administered with oral dose of tyrosine (100 mg/kg bw) as compared to placebo
and stress treated subjects.
Defensive measures such as minimizing the risk factors, psychotherapy
and chemotherapy are available but they are likely to affect decision making
and impair performance. Therefore, a non-pharmacological substance such as
a normal food constituent with the potential to reduce or reverse the stress
response would not only be useful to military combatants but also to other
individuals experiencing cognitive decline due to acute stressful conditions.
Further, till date no study using objective tools to measure information
processing and its impairment like event related potentials (ERPs) in evaluating
the effect of tyrosine containing supplement under stressful condition like
acute heat stress has been reported.
4
The present study appears to be the first one to evaluate the impact of
tyrosine as a countermeasure on the heat exposed cognitive decline in terms of
its electrophysiological component P300 and CNV. Event related potentials
(ERPs) are the objective tools to measure information processing and its
impairment. ERP P300 and contingent negative variation (CNV) provides a
neurophysiological means of investigating higher cerebral function that is
related to attention, cognition, information processing and working memory.
OBJECTIVES OF THE INVENTION
The primary objective of this invention is to formulate a nutraceutical
composition for amelioration of acute heat and cold stress induced cognitive
decline.
Another objective of the invention is to develop a nutraceutical
composition which is non-pharmaceutical and non-toxic in nature with no side
effects and contraindications.
Yet another objective of the invention is that the composition helps in
amelioration of cognitive decline during acute stress in actual field conditions
(environmental and psychological) and also in simulated stress conditions.
Still further objective of the invention is to prepare a composition which
helps to retain learning, attention, immediate memory and reaction time.
An additional object of the invention is that the preparation should be
easily chewable and palatable and is prepared with ingredients from natural
resources.
The foregoing has outlined some of the pertinent objectives of the
invention. These objectives should not be construed to be merely illustrative of
some of the more prominent features and applications of the intended
invention. Many other beneficial results can be obtained by applying the
5
disclosed invention in a different manner or modifying the invention within the
scope of disclosure.
Accordingly, other objectives and a full understanding of the invention
and the detailed description of the preferred embodiment in addition to the
scope of invention are to be defined by the claims undertaken.
These and other objectives and advantages of the invention will be
apparent from the ensuing description.
BRIEF DESCRIPTION OF ACCOMPANYING DRAWINGS
Further objectives and advantages of this invention will be more
apparent from the ensuing description when read in conjunction with the
accompanying drawings wherein:
Fig. 1 : P300 Latencies: Bar Diagram shows an increase in P300 Latency
during heat exposure with placebo. After CognoBar, it decreases
near to normal level. Values are mean ± SEM. **p<0.01 when
compared to baseline; ##p<0.01 when compared to placebo.
Fig.2 : CNV Latencies: During heat exposure there was an increment in
Ml00 latency after placebo administration but after CognoBar it
reduces. Values are mean ± SEM. *p<0.05 as compared to
compared to baseline; #p<0.05 when compared to placebo.
Fig.3 : CNV amplitude: M100 amplitude decreased after placebo
treatment but increased after CognoBar treatment compared to
placebo. Values are mean ± SEM. **p<0.01 as compared to
baseline; #p<0.05 when compared to placebo.
Fig.4 : CNV Reaction Time: Bar diagram shows that during heat exposure
reaction time increased after placebo treatment but it reduces with
CognoBar supplementation. Values are mean ± SEM. **p<0.01
when compared to baseline; ##p<0.01 as compared to placebo.
6
Fig.5 : Aromatic amino acids: Bar diagram show a significant decrease in
plasma tyrosine level in placebo group after stress exposure. With
CognoBar supplementation the plasma tyrosine levels significantly
increase. Values are Mean ± SEM. * p< 0.05, ***p<0.001 as
compared to baseline. ### p< 0.001 as compared to placebo post.
Fig.6 : Plasma Monoamines: Plasma nor-epinephrine (NE) is decreased
post exposure in placebo but increases significantly with CognoBar
e supplementation. Values are Mean ± SEM. # p < 0.05 as
compared to placebo post. EPI: epinephrine, DA: dopamine.
Fig.7 : Plasma Serotonin: Bar diagram shows a decrease in plasma 5-HT
levels in placebo post exposure, whereas, an increase in CognoBar
group was observed post exposure. Values are Mean ± SEM.
##p<0.02 as compared to placebo post.
Fig.8 : A marginal increase in vanillylmandelic acid (VMA) was observed in
tyrosine supplemented group compared to baseline and placebo.
Homovanillic acid (HVA) levels were significantly higher in tyrosine
compared to baseline and placebo. A significant increase 5-
Hydroxyindole acetic acid (5-HIAA) levels was observed in tyrosine
group than baseline. Values are Mean ± SEM. ** p< 0.01 when
compared to baseline. # p< 0.05 when compared to placebo post.
Fig.9 : Bar diagram shows as increment in P300 latency during cold
exposure with placebo. After CognoBar it decreases. Values are
mean ± SEM. *p<0.05 as compared to placebo.
Fig. 10 : Bar diagram shows that in cold stress CognoBar supplementation
reduces the reaction time compared to placebo treatment. Values
are mean ± SEM. *p<0.05 as compared to placebo.
7
Fig. 11 : More number of correct responses was observed with CognoBar
supplementation in cold stress compared to placebo. Values are
mean ± SEM. *p<0.05 as compared to placebo.
Fig. 12 : Less error were observed with Cognobar supplementation
compared to placebo treatment under cold stress. Values are mean
± SEM. *p<0.05 compared to placebo.
Fig. 13 : Bar diagram shows a fewer errors with Cognobar supplementation
compared to placebo treatment under cold stress Values are mean
± SEM. *p<0.05 as compared to placebo.
Fig. 14 : Bar diagram shows lower latency during cold exposure with
CognoBar. Values are mean ± SEM. *p<0.05 as compared to
placebo.
While the invention is described in conjunction with the illustrated
embodiment, it is understood that it is not intended to limit the invention to
such embodiment. On the contrary, it is intended to cover all alternatives,
modifications and equivalents as may be included within the spirit and scope
of the invention disclosure as defined by the claims.
STATEMENT OF THE INVENTION
According to the invention, there is provided CognoBar, A nutraceutical,
for amelioration of cognitive decline in acute stress, comprising of the following
constituents present in the composition by weight: Carbohydrates 67-71%;
Protein 15-18% (including 12-14 % L-Tyrosine); Fat 8-10%; Moisture 3- 5%
and Dietary Fibre 1.5 -4%.
8
DETAILED DESCRIPTION OF THE INVENTION
At the outset of the description, which follows, it is to be understood that
the ensuing description only illustrate a particular form of the invention.
However, such a particular form is only an exemplary embodiment and the
teachings of the invention are not intended to be taken restrictively.
For the purpose of promoting and understanding of the principles of the
invention, reference is now to be made to the embodiments illustrates and the
specific language would be used to describe the same. It is nevertheless to be
understood that no limitations of the scope of the invention is hereby intended,
such alterations and further modifications in the illustrated bag and such
further applications of the principles of the invention as illustrated therein
being contemplated as would normally occur to one skilled in the art to which
the invention relates.
The beneficial effects of CognoBar supplementation on cognitive
performance reported in the present work are consistent with the hypothesis
that uncontrollable stress causes an increase in the firing rate of
catecholaminergic neurons. This accelerated firing activates the enzyme
tyrosine-hydroxylase, making it more tyrosine sensitive. Hence, the enzyme
becomes more responsive to increased intraneural tyrosine, through oral/
parenteral administration of tyrosine. Tyrosine supplementation prior to stress
increases synthesis of catecholamines required for optimization of cognitive
functions. The hypothesis is supported by studies demonstrating that brain
CAs, particularly NE and DA, are depleted in certain regions of the brain by
exposure to a variety of stressors and that this depletion is associated with
decrement of cognitive performance.
9
In one embodiment, the CongoBar, the nutraceutical leads to a
significant increase in plasma tyrosine levels under acute stressful conditions
and Improves overall plasma catecholamine status after supplementation prior
to stress exposure.
In another embodiment, the CognoBar, the nutraceutical enhances
selective, divided and sustained attention under acute stressful conditions and
Improves ability to discriminate between target and non-target stimuli and
overall concentration and focus and proper classification and recognition of
stimuli.
Yet in another embodiment, the CognoBar, the nutraceutical improves
orientation towards a signal and ability to get warned and also Improves
general mood and behavior making an individual perseverant and calm thereby
preparing an individual for multitasking under acutely stressful conditions.
In another embodiment, CognoBar, the nutraceutical Reduces anger
like behavior arising due to acute stressful condition.
According to the invention, there is provided a nutraceutical
composition namely CongoBar for amelioration of cognitive decline in acute
stress. The composition comprises of following ingredients:
Constituents
Carbohydrates
Proteins
Fats
Moisture
Dietary fibre
% Composition (W/W)
67-71
15-18 (including 12-14 grams L-tyrosine)
8-10
3-5
1.5-4
10
Preferably the aforesaid constituents are present in the composition in
the following percentage:
Constituents
Carbohydrates
Proteins
Fats
Moisture
Dietary fibre
% Composition (W/W)
69-70
16-17 (including 13-14 grams L-tyrosine)
8-9
2.5-3.5
2-3
The ingredients used to prepare this bar are from natural sources: White
oats, Puffed Rice, White Sesame seeds, Milk pow^der, Isabgol (Psyllium husk),
Sugar, Raisins, Dates syrup and Tyrosine.
The constituents raw materials used for the supplementation of
carbohydrate, fat and protein are White oats -20%; Puffed Rice - 20%; White
Sesame seeds - 8%; Milk powder -5%; Isabgol (Psyllium husk) -2%; Sugar -
20%; Raisins -8%; Dates syrup - 4% and Tyrosine -13%.
Example I : Nutraceutical bar composition
Constituents
Carbohydrates
Proteins
Fats
Moisture
Dietary fibre
g per 100 g
70.00
17.00 (including 13 grams L-tyrosine)
09.00
03.00
02.70
11
Example II :Nutraceutical bar composition
Constituents
Carbohydrates
Proteins
Fat
Moisture
Dietary Fiber
CognoBar
(Per 100 Grams)
68.50 gm. approx.
17.00 gm. approx. (including 13 grams
L-Tyrosine)
10.00 gm. approx.
2.50 gm. approx.
2.00 gm. approx.
Study Protocol
The study was carried out in two phases. Phase I: simulated (hot)
environment and phase II: field (cold) condition.
Participants
Phase I [simulated (hot) environment], Human volunteer (n=10) and
phase II [field (cold) condition] human volunteers (n=30), soldiers from Indian
Army in the age group of 20-30 years, weight ranging 65+2 kg participated in
this study. The volunteers were screened for their general health and had
thorough ENT check-ups including audiometry to rule out any ear pathology
affecting hearing. They were free from any neurological disorder and cardiac
problems. They were not taking any psycho-stimulant drugs during the course
of the study. The protocol of the experiment was approved by the Ethics
Committee of the Institute. All the guidelines of Helsinki protocol were followed.
Study design and procedure
Both phase I and phase II studies were double blind, random, cross over
and placebo controlled.
Phase I
12
On the test day, the fasting blood sample and urine of each volunteer
was collected prior to measurement of their baseline physiological parameters
followed by battery of cognitive tasks and questionnaires. The volunteers were
then administered with either CognoBar or placebo bar and asked to wait for
90 minutes followed by stress exposure of 90 minutes duration.
Phase II
The soldiers who were posted for more than six months in cold
environment participated in the study. On the test day, all the parameters were
recorded 90 min after supplementation with either CognoBar or placebo bar.
Stress exposure:
Phase I : Each volunteer was exposed to simulated heat (45°C + 30% RH) in
human climatic chamber with capability of temperature control within 0.5 °C of
the desired temperature and humidity within 1% of the desired value. Prerecorded
gun noise (90-95 dB) was used simultaneously as another stressor
and the noise levels were well within the permitted limits.
Phase II: The study was carried out in Reasi district, Jammu. The location is a
remote, cold (-5 °C to + 5°C) and LIC area.
Administration of CognoBar
Volunteers in the supplemented group were administered with a high
energy CognoBar of 50g. Similarly, the placebo group received placebo bar of
50g which did not contain 1-tyrosine. The bars were matched for taste and
texture to ensure volunteers could not distinguish CognoBar from placebo
bars.
Sample Collection
Samples (blood and urine) were collected prior to bar supplementation
and immediately post stress exposure. Blood sample was collected in
13
heparinised vacutainers. Plasma was separated by centrifuging the blood
samples at 3000g for 10 minutes at 4°C and stored at-80° till further assay.
Urine samples were stored in acidic medium (Porteer and Brodie, 1969) till
further analysis.
Event related potentials
Preparation of participants
All the participants reported to the Neurophysiology Laboratory with a
clean, oil-free scalp. Scalp was gently rubbed with NuPrep™ skin prepping gel
and 10-20 TM conductive EEG paste was applied to different locations on the
scalp according to the internationally accepted 10-20 electrode placement
system. Ag-AgCI disc electrodes were used for recording. Auditory stimuli were
presented to the left ear using a headphone. The responses were recorded.
Artifact rejection was set at 90 as a default setting by the software and notch
filter was set on the 'on' button to filter out the SOiHz interference.
P300 recording
The response was recorded while the volunteers rested in supine
position. Electrodes were placed at Al and A2 (reference), FPz (common
ground), Cz and Pz (active) locations.
Two different tone sounds, one non-target and the other target, were
presented using the standard auditory odd-ball paradigm. The volunteers were
asked to count the target stimuli; the number of tones counted at the end of
each session was noted. Three-hundred sweeps were considered for the
generation of waveforms, with a target: non-target stimuli ratio of 20:80; the
actual count was determined by the software.
14
After completion of the recordings, the positive and negative peaks were
marked, based on which latencies (ms) and amplitudes (fiV) of peaks were
derived. (Ray et.al.,2012)
CNV recording
The recording was standardized (Loveless and Sanford, 1974; Poon et.al.,
1974). Electrodes were placed at El and with E2 (eye movement), Al
(reference), Fpz (common ground), C3, C4, Cz, and Fz (active). Participants
were explained the protocol of the recording. A pair of LED-goggles was used to
deliver visual stimuli and a headphone was used to deliver auditory stimuli.
Two stimuli were delivered: the first Varning' stimulus (SI) was a single tone
sound. After duration of a second, the second 'imperative' stimulus (S2) was
presented in the form of trains of flashes, and the subject was asked to press a
button to terminate the flash. The time which elapsed between S2 and the
motor response was recorded as the reaction time (Roy et.al., 2012).
HPLC of plasma and urine
High performance liquid chromatography using reverse phase C18
column was used to estimate the concentration of aromatic amino acids viz.
tyrosine, phenylalanine and tryptophane, catecholamines viz. norepinephrine,
epinephrine, and dopamine and serotonin in plasma samples.
Neurotransmitter metabolites, Vanillyl mandelic acid (VMA), Homovanillinic
Acid (HVA) and 5-Hydroxyindole acetic acid (5-HIAA) in urine using standard
kit protocol were assayed. Waters HPLC system with an autosampler, HPLC
pump and electrochemical detector was used to analyze the samples.
Cognitive tests and Questionnaries
Subjects were evaluated using multiple, dependent measures of
symptoms, mood, and cognitive performance, Cognitive performance was
measured using a computerized performance assessment battery. Personality
15
and mood questionnaire was administered by paper and pencil. Subjects
performed all these testes before and during the simulated environmental
stressors except the personality test which was administered once before stress
exposure.
Profile of Mood States Questionnaire
POMS Questionnaire is an inventory of self-reported mood states that
describe emotional sentiments of people (McNair et.al., 1971). The volunteer
has to read each word and mark his response, indicating to what extend (such
as Not at all, A little, Moderately, Quite a bit, extremely) did these words
describe his feelings during the past two or three days, including the day of
questionnaire administration.
Simple Reaction Time (SRT)
This is a test of selective attention. It measures the ability of an
individual to respond to a visual stimulus. In this test dots of different colors
are shown on computer screen. Each dot appears one after another on
computer screen. Volunteers are asked to focus on the red colored dot(s) and
press space bar of the keyboard whenever they encounter the same.
Choice Reaction Time (CRT)
This is a test of divided attention. It measures the ability of an individual
to respond to three different visual stimuli. In this test, dots of different colors
appear one after the other on computer screen. The volunteers are asked to
focus on red, blue and green colored dots and press left, down, right arrow key
for red, green, blue dot respectively, whenever they encounter the same.
Vigilance Test (VT)
This is a test of sustained attention (Mackworth, 1948) and measures the
alertness and concentration of individual. In this test clock like structure is
16
shown on the computer screen and a large black needle (pointer) in circular
background. The needle makes short jumps approximately after every second
and sometimes it makes double jump. The volunteers had to detect and
respond to the same (double jump) by pressing left arrow key on keyboard.
CANTAB Neuropsychological tests
For the evaluation of cognitive function Cambridge Automated
Neuropsychological Test Battery (CANTAB) (Torgersen et al. 2012) was used
which is a semi-automated neuropsychological test battery applied on a lap-top
PC, developed at the University of Cambridge. CANTAB contains different
neuropsychological tests in five cognitive domains: Visual memory, semantic/
verbal memory, decision making and response control, executive function and
attention.
Paired Associate Learning (PAL)
PAL is a test of episodic and visual memory but also depends on the
ability of spatial planning. The performance on PAL depends on input mainly
from the temporal lobe, but also from the frontal lobe. Volunteers had to
remember the location of different patterns appearing on the screen and then
point out where on the screen the pattern initially was shown. Increasing
difficulty level ranging from two to eight patterns to be remembered (Sahakian
etal. 1988).
Pattern recognition memory (PRM)
Subjects were required to learn a series of 12 simple, but abstract,
coloured visual patterns located in the centre of the screen. Each pattern was
presented for 3 sec. In the recognition phase, V2 pairs of coloured patterns
were then presented on the screen one at a time and the subject had to identify
the target pattern from a 'distractor' pattern. This procedure was repeated with
12 new patterns, and the subject was given a total score (maximum=24)
17
expressed as a percentage (Mehta et al. 1999). This test is sensitive to
dysfunction in medial temporal areas of the brain and relatively insensitive to
dysfunction in the frontal lobe.
Spatial Working Memory (SWM)
SWM is a test of the subject's ability to retain spatial information and to
manipulate remembered items in working memory. It is a self-ordered task,
which also assesses heuristic strategy. This test is a sensitive measure of
frontal lobe and 'executive' dysfunction.T he test begins with a number of
coloured squares (boxes) being shown on the screen. The aim of this test is
that, by process of elimination, the subject should find one blue 'token' in each
of a number of boxes and use them to fill up an empty column on the right
hand side of the screen. The number of boxes is gradually increased from three
to eight boxes (or to ten in the second mode). The colour and position of the
boxes used are changed from trial to trial to discourage the use of stereotyped
search strategies.
The subject must touch each box in turn 'until one opens with a blue
token inside. When a blue token has been found, the subject has to place it in
the right column by touching the right-hand side of the screen. The subject
must then begin a new search for the next blue token. It may be in any of the
boxes that so far have been empty. This is repeated, until a blue token has
been found in every box on the current screen. Touching any box in which a
blue token has already been found is an error. The subject decides the order in
which the boxes are searched. The computer determines the number of empty
boxes that must be visited (discounting errors). Performance at the harder
levels of this task is enhanced by the use of a heuristic search strategy (Owen
etal. 1990).
Choice Reaction Time (CRT)
18
CRT is a 2-choice reaction time test. An arrow-shaped stimulus is
displayed on either the left or the right side of the screen. The subject must
press the left hand button on the press pad if the stimulus is displayed on the
left hand side of the screen, and right hand button on the press pad if the
stimulus is displayed on the right hand side of the screen.
Statistical analysis
The data presented are mean + SEM. One way analysis of variance
(ANOVA) followed by Student's Newman-Keuls post hoc test was performed to
analyze the data. Subjective data was analyzed by Dunn's multiple comparison
test. Significance level was set at p<0.05.
5. Results
Event Related Potentials
P300
No significant effect on N100 latency of ERP was found following
placebo/CognoBar supplementation and after stress exposure. P300 latency
was significantly increased (p<0.01) after stress' exposure with placebo than
baseline. When CognoBar condition was compared with placebo there was a
significant reduction (p<0.01) in P300 latency. No significant change was
observed in P300 latency of CognoBar when compared with baseline. After
CognoBar administration P300 latencies returned close to pre placebo or
baseline value. (Figure 1)
Contingent Negative Variation
There was a significant increase (p<0.05) in MlOO latency of placebo as
compared to baseline. MlOO latency was significantly decreased (p<0.05) after
CognoBar supplementation than placebo. There was no significant change in
M200 latency in both placebo and CognoBar supplemented groups. Amplitude
of MlOO was significantly degreased (p<0.01) in placebo than baseline. After
19
CognoBar administration amplitude of Ml00 was significantly increased
(p<0.05) than placebo and return close to baseline value. M200 amplitude was
decreased in placebo than baseline but not significantly. No significant change
was observed in M200 amplitude between placebo and CognoBar. (Figure 2 863)
Reaction time
Reaction time was significantly increased (p<0.01) in placebo than
baseline and reduced significantly (p<0.01) after GognoBar administration than
placebo. No significant change was observed in CognoBar when compared with
baseline. (Fig.4)
Amino acid precursors
Analysis of HPLC data showed a significant decrease in plasma tyrosine
concentration in placebo group post exposure (p<0.05). Whereas, a significant
(p<0.01) increase in plasma tyrosine was in observed CognoBar supplemented
group post exposure. Further, comparison of plasma tyrosine levels in placebo
and CognoBar supplemented group. No significant change in plasma levels of
other aromatic amino acids (phenylalanine and tryptophan) in both the groups
was observed although a slight increase in CognoBar group and decrease in
placebo group post exposure was seen. (Figure 5)
Plasma monoamines
Prominent effect of CognoBar supplementation was found in plasma NE
levels in both the groups post stress exposure. Results indicate a significant
(p<0.05) increase in NE level in CognoBar group compared to placebo post
exposure. Also an increase in NE levels post CognoBar supplementation
compared to baseline values was observed. Whereas, decreased NE levels post
exposure in placebo group compared to their baseline level was seen, though
the difference was not statistically significant. A slight increase in levels of
catecholamine neurotransmittes epinephrine (EPI) and dopamine (DA) in post
20
CognoBar group was observed compared to baseline and post placebo, but the
difference was statistically not significant. (Figure 6)
With respect to serotonin (5-HT) concentration, a marginal decrease in
placebo group compared to its baseline value was observed following stress
exposure. Whereas, a reverse trend in CognoBar supplemented group was seen
i.e. the 5-HT level was significantly (p<0.02) higher in post CognoBar group
compared to post placebo. (Figure 7)
Neurotransmitter metabolites
Analysis of results of urinary metabolites revealed a slight increment in
VMA levels in CognoBar group compared to baseline and placebo. Significantly
higher levels of HVA in CognoBar group compared to baseline (p<0.01) and
placebo (p<0.05) was observed. 5-HIAA was significantly more (p<0.01) in
CognoBar group compared to baseline. (Figure 8)
Cognitive performance and mood profile
Analysis of psychological tasks revealed a significant (p<0.001)
improvement in selective attention (SRT) and sustained attention (VT) (p<0.02)
in CognoBar supplemented group compared to the placebo group post stress
exposure. Divided attention (CRT) also showed a marginal improvement,
though not significant. A significantly higher (p<0.001) reaction time (RT) was
observed in placebo group post exposure compared to baseline value. In case of
CognoBar supplemented group, RT tends to normalize towards the baseline
value although it differed significantly (p<0.02). A highly significant (p<0.001)
difference in RT was observed between post placebo and post CognoBar groups.
One of the mood profile component, anger, showed a significant
(p<0.001) increase in placebo group post exposure compared to baseline. Also,
a significantly (p<0.01) higher anger was observed post exposure in CognoBar
21
group compared to baseline value but it was significantly lower (p<0.001) than
post exposure value of placebo group.
Table 1. Psychological tasks
Baseline
Placebo
CognoBar
Selective
Attention
Divided
Attention
Sustained
Attention
AngerfMood
Profile)
(Scores)
45.0 ± 0.42
40.5 ± 0.40
***
44.7 ± 0.67 »»«
66.3 ± 16.7
44.0 ± 11.6
75.8 ± 19.0
5.8 ± 1.70
5.5 ± 0.60
7.4 ± 0.40 ##
2.29 ± 0.27
8.28 ± 0.35
***
5.85 ± 0.32
*
Reaction Time
(milli second)
433.57 ± 5.64
515.01 ±5.59
***
451.88 ±3.91
###
*p<0.05; *** p < 0.001 ascompared to baseline ; ##p<0.02; ###p<0.001 as compared to placebo
Phase II
P300
Latency of P300 (Fig. 9) was significantly (p<0.05; 3,39%) reduced after
CognoBar supplementation than placebo. There was no significant change in
P200 and N200 latencies. (Figure 9)
CNV
Significant decrement (p<0.05; 20.8%) in reaction time was observed with
CognoBar supplementation as compared to placebo. (Figure 10)
CANTAB
Pattern Recognition Memory
Significant improvement (p<0.05; 13.20%) in percent correct response (Fig. 11)
was observed with CognoBar supplementation.
Paired Associates Learning
Significant decrement (p<0.05; 40.34%) in total 'error (Fig. 12) was observed
with CognoBar supplementation.
22
Spatial Working Memory
Supplementation with CognoBar significantly decreased (p<0.05; 32.3%) the
total error (Fig. 13) compared to placebo in SWM task.
Choice Reaction Time
CRT Correct Latency (Fig. 14) was significantly reduced (p<0.05; 9.92%) after
CognoBar supplementation.
It can be concluded that pretreatment with CognoBar containing
aromatic amino acid I-tyrosine ameliorates 'the acute stress induced
decrements in signal processing and cognitive performance. The composition
serves as an effective non-pharmacological countermeasure to optimize
cognitive performance during acute stressful environmental conditions.
ADVANTAGES
a) The composition is non- pharmaceutical in nature: The constituents raw
materials used for the supplementation of carbohydrate, fat and protein are
White oats -20%; Puffed Rice - 20%; White Sesame seeds - 8%; Milk powder -
5%; Isabgol (Psyllium husk) -2%; Sugar - 20%; Raisins -8%; Dates syrup - 4%
and Tyrosine -13%. *
b) Helps in amelioration of cognitive decline during acute hot and cold stress
(environmental and psychological).
c) Helps in amelioration of cognitive decline in simulated stressful conditions
(hot).
P300 latency:
• Hot environment: Increased 10.20%
• CognoBar: Reduced 11.37%
Ml00 latency of CNV:
23
• Hot environment: Increased 3.78%
• CognoBar: Reduced 4.34%
Ml00 amplitude of CNV:
• Hot environment: Decreased 46.30%
• CognoBar: Increased 41.70%
CNV Reaction Time:
• Hot environment: Increased 25.62%
• CognoBar: Reduced 24.40%
d) Helps in amelioration of cognitive decline under cold field conditions.
P300 latency:
• CognoBar in cold field environment: 3.39% lower than placebo
CNV Reaction Time:
• CognoBar in cold field environment: 20.80% lesser than placebo
e) Helps to retain learning, attention, immediate memory and reaction time.
Heat Stress
Selective attention:
• CognoBar: Improved by 10.37% compared to placebo
Sustained attention: .
• CognoBar: Improved by 34.54% compared to placebo
Divided attention:
• CognoBar: Improved by 72.27% compared to placebo
3-Choice Reaction Time:
• CognoBar: Decreased by 12.26% than placebo
Cold Stress
Pattern Recognition Memory:
i
24
• CognoBar in cold field environment: Improved by 13.20% than placebo
Total Error of Paired Associates Learning:
• CognoBar in cold field environment: Decreased by 40.34% than placebo
Total Error of Spatial Working Memory:
• CognoBar in cold field environment: Decreased by 32.30% than placebo
Correct Latency of Choice Reaction Time:
• CognoBar in cold field environment: Decreased by 9.92% than placebo
f) The composition is pharmaceutically inert except the ameliorating action of
the active principal L-tyrosine (a non-pharmaceutical adjuvant).
g) The preparation is a square cuboid, easily chewable and highly palatable.
h) The ingredients used to prepare this bar are from natural sources: White
oats, Puffed Rice, White Sesame seeds, Milk powder, Isabgol (Psyllium husk),
Sugar, Raisins, Dates syrup and Tyrosine.
i) The composition is non-toxic in nature.
j) The composition has no side effects.
References
Loveless, N.E., 8B Sanford, A.J. (1974). Slow potential correlates of preparatory
set. Biological Psychology, 1, 303-314.
Mackworth, N. H. (1948). The breakdown of vigilance during prolonged visual
search. Quarterly Journal of Experimental Psychology, 1, 6-21.
McNair, D. M., Lorr, M., 8& Droppleman, L.F. (1971). Profile of mood states
manual. San Diego, CA: Educational and industrial testing services.
25 *
Mehta MA, Sahakian BJ, McKenna PJ, Robbins TW. Systemic sulpiride in
young adult volunteers simulates the profile of cognitive deficits in
Parkinson's disease. Psychopharmacology 1999; 146: 162-74.
Owen AM, Downes J J , Sahakian BJ, Polkey CE, Robbins TW. Planning and
spatial working memory following frontal lobe lesions in man.
Neuropsychologia 1990; 28: 1021-34.
Poon, L.W., Thompson, L.W., Williams, Jr. R.B., & Marsh, G.R. (1974).
Changes of antero-posterior distribution *of CNV and late positive
component as a function of information processing demands.
Psychophysiology, 11, 660-673.
Porter, I. A., & Brodie, J. (1969). Boric Acid Preservation of Urine Samples.
British Medical Journal, 2, 353-355.
Ray, K., Chatterjee, A., Panjwani, U., Kumar, S., Sahu, S., Ghosh, S., Thakur,
L., & Anand, J. P. (2012). Modafinil improves event related potentials P300
and contingent negative variation after 24 hours sleep deprivation. Life
Sciences, 91, 94-99,
Sahakian BJ , Morris RG , Evenden JL , Heald A , Levy R , Philpot M , Robbins
TW . A comparative study of visuospatial memory and learning in
alzheimer-type dementia and parkinson's disease. Brain 1988; 111: 695-
718.
Torgersen J, Flaatten H, Engelsen BA, Gramstad A. Clinical Validation of
Cambridge Neuropsychological Test Automated Battery in a Norwegian
Epilepsy Population. Journal of Behavioral and Brain Science 2012; 2:
108-16.
26 t
All documents cited in the- description are incorporated herein by
reference. The present invention is not to be limited in scope by the specific
embodiments and examples which are intended as illustration of a number of
aspects of the scope of this invention. Those skilled in the art will know or to be
able to ascertain using no more than routine experimentation many
equivalents to the specific embodiments of the invention described herein.
It is to be further noted that present invention is susceptible to
modifications adoptions and changes by those skilled in the art. Such variant
embodiments employing the concepts and features of this invention are
intended to be within the scope of the present invention which is further set
forth under the claims:
27
*

WE CLAIM
1. CognoBar, A nutraceutical, for amelioration of cognitive decline in acute
stress, comprising of the following constituents present in the
composition by weight:
Carbohydrates .... 67-71%
Protein 15-18% (including
12-14 %L-Tyrosine)
Fat .... 8-10%
Moisture .... 3- 5%
Dietary Fibre ... 1.5-4%
2. CognoBar, A nutraceutical, as claimed in claim 1 wherein the following
constituents are present preferably in the composition by weight:
Carbohydrates .... 69-70%
Protein ..... 16-17% (including
13-14 % L-Tyrosine)
Fat .... 8-9%
Moisture .... 2.5-3.5%
Dietary Fibre ... 2-3% t
3. CognoBar, A nutraceutical, as claimed in claim 1 wherein the
constituents for the said CongoBar is preferably from natural sources
like White Oats, Puffed Rice, White Sesame seeds, Milk Powder, Isabgol
(Psyllium husk), Sugar, raisins, Dates Syrup and Tyrosine.
4. CognoBar : A nutraceutical, as claimed in claim 1 wherein the
constituents raw materials used for the supplementation of
carbohydrates, fats and protein are White Oats: 20%, Puffed Rice: 20%,
28
White Sesame seeds: 8%, Milk Powder: 5%, Isabgol (Psyllium husk): 2%,
i
Sugar: 20%, raisins: 8%, Dates Syrup: 4% and Tyrosine: 13%.
5. CognoBar : A nutraceutical, as claimed in claim 1 wherein CognoBar
supplementation reduces the increase in P300 latency due to hot
environment (10.20%) by 11.37%.
6. CognoBar, A nutraceutical, as claimed in claim 1 wherein CognoBar
supplementation offsets the increase in Ml00 latency of CNV and CNV
reaction time due to hot environment.
7. CognoBar, A nutraceutical, as claimed in claim 1 which helps in
amelioration of cognitive decline under cold field conditions.
8. CognoBar, A nutraceutical, as claimed in claim 1 which helps to retain
learning, attention, immediate memory and reaction time.
9. CognoBar, A nutraceutical, as claimed in claim 1 for amelioration of
cognitive decline in acute stress, as herein described substantially with
reference to the accompanying drawings.