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Gender Dimorphism in Aspartame-Induced Impairment of Spatial Cognition and Insulin Sensitivity

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Gender Dimorphism in Aspartame-Induced Impairment
of Spatial Cognition and Insulin Sensitivity
Kate S.Collison
,Nadine J.Makhoul
,Marya Z.Zaidi
,Soad M.Saleh
,Bernard Andres
,Angela Inglis
Rana Al-Rabiah
,Futwan A.Al-Mohanna
Cell Biology and Diabetes Research Unit,Department of Cell Biology,King Faisal Specialist Hospital and Research Centre,Riyadh,Saudi Arabia,
Al-Faisal University
Medical School,Riyadh,Saudi Arabia
Previous studies have linked aspartame consumption to impaired retention of learned behavior in rodents.Prenatal
exposure to aspartame has also been shown to impair odor-associative learning in guinea pigs;and recently,aspartame-fed
hyperlipidemic zebrafish exhibited weight gain,hyperglycemia and acute swimming defects.We therefore investigated the
effects of chronic lifetime exposure to aspartame,commencing
in utero
,on changes in blood glucose parameters,spatial
learning and memory in C57BL/6J mice.Morris Water Maze (MWM) testing was used to assess learning and memory,and a
random-fed insulin tolerance test was performed to assess glucose homeostasis.Pearson correlation analysis was used to
investigate the associations between body characteristics and MWM performance outcome variables.At 17 weeks of age,
male aspartame-fed mice exhibited weight gain,elevated fasting glucose levels and decreased insulin sensitivity compared
to controls (P
0.05).Females were less affected,but had significantly raised fasting glucose levels.During spatial learning
trials in the MWM (acquisition training),the escape latencies of male aspartame-fed mice were consistently higher than
controls,indicative of learning impairment.Thigmotactic behavior and time spent floating directionless was increased in
aspartame mice,who also spent less time searching in the target quadrant of the maze (P
0.05).Spatial learning of female
aspartame-fed mice was not significantly different from controls.Reference memory during a probe test was affected in
both genders,with the aspartame-fed mice spending significantly less time searching for the former location of the
platform.Interestingly,the extent of visceral fat deposition correlated positively with non-spatial search strategies such as
floating and thigmotaxis,and negatively with time spent in the target quadrant and swimming across the location of the
escape platform.These data suggest that lifetime exposure to aspartame,commencing
in utero,
may affect spatial cognition
and glucose homeostasis in C57BL/6J mice,particularly in males.
Collison KS,Makhoul NJ,Zaidi MZ,Saleh SM,Andres B,et al.(2012) Gender Dimorphism in Aspartame-Induced Impairment of Spatial Cognition and
Insulin Sensitivity.PLoS ONE 7(4):e31570.doi:10.1371/journal.pone.0031570
Christopher Morrison,Pennington Biomedical Research Center,United States of America
October 15,2011;
January 11,2012;
April 3,2012
2012 Collison et al.This is an open-access article distributed under the terms of the Creative Commons Attribution License,which permits
unrestricted use,distribution,and reproduction in any medium,provided the original author and source are credited.
The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.
Competing Interests:
The authors have declared that no competing interests exist.
Previous studies have shown that chronic consumption of the
dipeptide artificial sweetener aspartame may affect the T-maze
cognitive performance of male rats,promoting impairment of
retention of learned behavior when compared to the performance
of controls [1].The Acceptable Daily Intake for aspartame
currently stands at 50 mg/Kg body weight in the United States,
and 40 mg/Kg in Europe.Once ingested,aspartame (L-aspartyl-
phenylalanine methyl ester) is rapidly metabolized to its metabolic
components phenylalanine,aspartate,and methanol in the ratio of
50:40:10 w/w/w [2].Whilst the neurological effects of methanol
have been well documented,[3] aspartate,like glutamate,has
been shown to cause brain lesions [4],obesity [5] and impaired
memory retention [6] in rodents exposed to these excitatory amino
acids (EAA).
The aspartame metabolite phenylalanine is an essential amino
acid which occurs naturally in the breast milk of mammals;
however high levels of phenylalanine are a health hazard to those
born with phenylketonuria (PKU),a metabolic disorder caused by
an inherited mutation in the phenylalanine hydroxylase (PAH)
gene which prevents phenylalanine from being metabolized
correctly.This results in a detrimental accumulation of the amino
acid,leading to developmental defects,seizures and mental
retardation [7].Normal mammalian plasma levels of phenylala-
nine are approx.30–50
M (0.5–0.8 mg/dL),however 1 in 50
individuals are heterozygous for the mutation in the phenylalanine
hydroxylase gene [8],resulting in significantly higher levels of
fasting plasma phenylalanine compared to non-carriers [9],
together with a reduced phenylalanine clearance rate after
intravenous loading [10].Repeated ingestion of 8 servings of
aspartame-sweetened beverages by PAH heterozygous individuals
incurred plasma phenylalanine levels of up to 165
M [11],
although this was still well below the levels reported to cause
neurotoxicity during acute administration in primates.Addition-
ally,genetically mutated PAH-deficient homozygous
BTBR mice have six times the level of brain phenylalanine
compared to their heterozygous counterparts [12],resulting in
abnormal CNS synapses and dendritic spines [13] together with
pathological cognitive impairment [14].
In vitro
,phenylalanine has
been demonstrated to specifically and reversibly attenuate
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glutamatergic synaptic transmission by competing with the glycine
binding site of N-methyl D-aspartate (NMDA) receptor [15,16].
Additionally,the ratio of GluN2A/GluN2B NMDA receptor
subunit expression is significantly increased in the hyperphenyl-
alaninemic Pah
PKU mouse model,suggesting a potential
mechanism whereby elevated levels of phenylalanine may impair
brain development and function [12,17].Since their discovery in
the early 1950s,NMDA receptors have been implicated in many
crucial functions of central importance,including learning and
memory,neuronal plasticity and neurotoxicity [18,19];and they
are the only known receptor that is regulated both by a ligand
(usually glutamate) and also by voltage [20].There are at least five
binding sites on the NMDA receptor which regulate its activity
including glutamate,glycine,magnesium,zinc and a fifth site that
binds to the hallucinogenic substance phencyclidine [21].The
central role of NMDA receptors in the process of learning and
memory has been confirmed by the extensive use of NMDA
receptor agonists and antagonists to study long term potentiation
in memory acquisition and maintenance [22].
Whereas it is generally agreed that aspartate crosses the
placenta only to a limited degree [23],phenylalanine is actively
transported across the placenta [24],resulting in an increase in this
aromatic amino acid at the expense of the maternal concentration
[25].During pregnancy,aspartame administration by gavage
resulted in impaired performance of the offspring in an odor-
aversion test administered to guinea-pigs within the first month of
life [26].This insightful study confirms previous reports in a
second species,that aspartame administered pre- and postnatally
to rats can result in impaired cognitive performance of the
offspring [27].Furthermore,aspartame has been shown to cause
brain inflammation,hyperglycemia and fatalities in a third species:
the hyperlipidemic zebrafish model [28].Aspartame-fed zebrafish
also exhibited swimming defects which were interpreted as
possibly due to damage in the brain and neurons (28).
The timing,dosage and route of EAA administration in vivo
appears to be of critical importance,since acute exposure to high
doses of dipeptide aspartame during adulthood has no effect on
cognitive ability in either humans [29] or rodents [30].However a
single i.p.injection 500 mg/Kg aspartate was sufficient to cause
memory impairment and neuronal damage in adult mice
undergoing passive avoidance testing [6];and intracranial
injections of phenylalanine caused permanent amnesia in 1 day
old chicks [31].Similarly,perinatal exposure to glutamate results
in delayed onset neuroendocrine dysfunction together with
cognitive deficiencies [32–38],whereas exposure to considerable
amounts of dietary glutamate in adulthood is apparently without
effect [39].Interestingly,we [40] and others [41] have noted
gender-specific differences in behavior in response to Monosodium
Glutamate (MSG).Gender dimorphism in MSG-induced impair-
ment of the growth hormone/IGF-1 axis has also been
investigated [42],and it would be of interest to ascertain whether
gender dimorphism in glucose homeostasis exists in response to
aspartame consumption.
The aims of the present study were therefore to examine the
effect of lifetime exposure to aspartame,commencing in utero,on
weight gain,spatial cognition,insulin sensitivity and glucose
parameters of male and female C57BL/6J mice.We used a
dosage of aspartame which approximated the ADI for aspartame
in the US (approx.50 mg/kg body weight).Insulin sensitivity was
assessed by a randomfed insulin tolerance test (ITT),together with
measurements of fasting glucose and insulin levels;and cognitive
performance was assessed in the Morris Water Maze (MWM).The
relationship between visceral fat deposition and cognitive function
was determined by analyzing the correlation between body
characteristics,glucose and insulin parameters and performance
targets in the MWM test,using Pearson correlation analysis.
Materials and Methods
Animals and Diets
C57BL/6J mice of both sexes were obtained from the Jackson
Laboratory and housed/caged in a controlled environment (3 to a
cage in pathogen-free conditions of 12 h light/dark cycle,
2262uC) and fed a standard chow diet with water ad libitum until
6 weeks of age.The two diet groups used in this study were (1) ad
libitum Standard Chow (Control diet) with ad libitum drinking
water.(2) Ad libitum Standard Chow,with ad libitum drinking water
containing 0.25 g/L aspartame as the only source of drinking
water (Asp-Phe methyl ester,catalog A5139 Sigma Aldrich).After
a 3-week period of adjustment,male and female mice were bred,
weaned and maintained on these respective diets for the times
stated.Mean body weight was assessed at 6 and 17 weeks of age.
Food and water intake was assessed at 7 weeks and again at 15
weeks of age,in pre-weighed mice over a four day period,by
weighing the food pellets and water bottles to the nearest 0.1 g.
Mean food/water consumption was calculated by subtraction,and
expressed as g/20 g body weight (bw) and mls/20 g bw
respectively.Mean aspartame consumption was calculated from
the amount of aspartame-water consumed,and expressed in mg
per Kg bw.Body length to the nearest mm was assessed at 6 and
17 weeks of age using a woven tape measure.The breeding and
care of the animals were in accordance with the protocols
approved by the Animal Care and Use Committee of the King
Faisal Specialist Hospital & Research Centre.At the conclusion of
the study,animals were humanely euthanized with a mixture of
xylazine and ketamine (10 mg/kg and 100 mg/kg respectively);
and the liver and visceral fat were carefully dissected out and
removed,rinsed twice in PBS,blotted dry and weighted to the
nearest 0.01 g.These tissues were rapidly snap-frozen for use in
further studies.
Measurement of Fasting SerumGlucose,Insulin and Lipid
Overnight fasting blood glucose was measured using the
Ascensia Contour glucometer (Bayer HealthCare,IN,USA).
Fasting serum insulin was measured using the mouse insulin
ELISA kit from Millipore/Linco (Uppsala,Sweden).Homeostatic
Model Assessment Index (HOMA-IR) values,a measure of insulin
resistance,were calculated according to the established formula:
(fasting serum insulin
IU/ml) * (fasting serum glucose mM)/22.5
[43].Serum Triglyceride (Tg),total cholesterol (T-CHOL),and
HDL-C concentrations were measured in overnight fasted mice
using the Reflovet Plus instrument (Roche,F.Hoffmann-La
Roche Ltd,Basel,Switzerland) as previously described [40].
Morris Water Maze (MWM) Testing Apparatus
The spatial learning abilities of the C57BL/6J mice were
assessed at 16 weeks of age (mature adulthood) in a MWM task
[44],since our previous studies indicate that this is the optimal
time for assessing the effect of dietary interventions on cognitive
behavior [40].The apparatus consisted of a white circular pool of
150 cm diameter and 50 cm height,filled with water made
opaque by the addition of a small amount of non-toxic white paint
(30 cm deep) and maintained at 21–22uC.A circular escape
platform (11 cm in diameter) was placed in a fixed South-West
location hidden 0.5 cm below the surface of the water,and 3
stationary geometric visual cues were kept in the room throughout
the period of testing as previously described [40].A digital camera
Aspartame-Induced Impairment of Spatial Cognition
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was positioned above the centre of the tank and linked to a
tracking system in order to record the performance of the
experimental subjects (HVS Image Analysis VP-200,HVS Image,
MWM Procedure:Assessment of Learning abilities
Mice were given four consecutive days of acquisition training
sessions that consisted of four trials per day with an inter-trial
interval of 20 minutes.In order to investigate the effect of diet on
allocentric spatial reference memory,the position of the hidden
platform remained fixed however the entry point was pseudo-
randomly selected from one of the 4 compass locations each
day,and the same sequence of starting points was used for all the
mice tested.Mice were given 120 seconds to find the platformand
if the subject failed to locate the platform within this period,it was
guided onto it.All mice were allowed to rest on the platform for a
30 second interval after each trial.At the end of the training block,
mice were put in a pre-warmed drying cage and allowed to dry
prior to being returned to their experimental cages.
MWM Escape Strategies Analysis
During the acquisition phases of learning,mice adopt a number
of increasingly sophisticated escape strategies as part of allocentric
spatial memory acquisition [45].Initial strategies such as
thigmotaxis and random searching which do not involve any
spatial or directional preference progress to strategies usually
described as scanning (directionless searching of the interior
portion of the pool),chaining (circular swimming at an
approximately fixed distance greater than 16 cm from the wall),
followed by directed searching for the escape platform.
MWM Probe Test for Spatial Memory Assessment
A Probe Test was run on day 5,after 4 days of acquisition.
Before the probe test,the hidden platform was removed and the
mouse was introduced from the north quadrant,where it was left
to search for the platform for 60 seconds.Time spent in the target
quadrant,number of platform crossings,and annulus crossing
index (ACI:defined as the number of crosses over the platform
position in the target quadrant,relative to crosses over the
corresponding platforms in the remaining three quadrants) were
Random-fed Insulin Tolerance Test (ITT)
The effect of aspartame on glucose parameters was determined
using a random-fed insulin tolerance test (ITT) administered to 19-
week old mice.Because swimming exercise has been shown to
alter insulin sensitivity and glucose homeostasis [46],we
performed the ITT on additional mice who had not been
subjected to the water maze protocol (n=18 per diet/gender
group).For the ITT,an intraperitoneal injection of insulin (Sigma,
IL) at a dose of 0.75 U/kg body weight was administered,and
whole blood glucose levels were measured from the tail vein at 0,
15,30,45 and 60 minutes after injection.
Data Analysis
Data were presented as mean 6SEM for body-weight,serum
lipid profile,glucose and insulin variables separately in male and
female mice.For statistical comparisons between the two diet
groups,unpaired Student’s t test was applied using Graph Pad
Instat software (version 3,California,USA).For the MWM
experiments,the following variables characterizing the perfor-
mance of mice in the MWM were chosen for analysis:Latency,
defined as time taken (in seconds) for a mouse to reach and climb
the platform,and the length of each Swim Path (in meters).
Locomotor activity was analyzed using the average Swim Speed
(M/s),and the percent of time that the animals were relatively
stationary (with swim speed below the 0.06-m/sec threshold) was
recorded as Floating Time (%).Thigmotaxic swimming activity
Table 1.Effect of aspartame consumption on weight gain,adiposity,glucose homeostasis and lipid profile.
Male Female
Control Aspartame Control Aspartame
Body Weight (6 weeks) 17.6 6 0.13 17.22 6 0.35 14.31
6 0.17 14.34
6 0.22
Body Weight (17 weeks) 24.02 6 0.48 24.96 6 0.4 18.91
6 0.21 19.42
6 0.27
% Weight Gain 36.42 6 2.44 45.31* 6 2.71 32.3 6 1.45 35.63
6 1.5
Length (cm) 9.65 6 0.08 10.04** 6 0.08 9.21 6 0.07 9.52**
6 0.06
Visceral Fat (g) 0.09 6 0.02 0.24** 6 0.02 0.08 6 0.01 0.13**
6 0.01
RW of Visceral Fat (g/20g BW) 0.08 6 0.01 0.192** 6 0.02 0.09 6 0.01 0.13**
6 0.01
Liver Weight (g) 0.89 6 0.02 1.03* 6 0.06 0.76 6 0.03 0.71
6 0.02
RW of Liver (g/20g BW) 0.75 6 0.02 0.82 6 0.04 0.81 6 0.03 0.736* 6 0.02
Fasting Glucose (mM) 3.1 6 0.18 4.61** 6 0.43 2.93 6 0.13 3.65**
6 0.17
Fasting Insulin (uIU/ml) 17.32 6 2.54 16.67 6 1.86 10.37
6 0.8 8*
6 0.58
HOMA-IR 2.36 6 0.35 3.19 6 0.24 1.35
6 0.11 1.29
6 0.1
TG (mg/dL) 104.17 6 7.42 107.41 6 5.88 123.25 6 6.67 98.24* 6 6.32
T-CHOL (mg/dL) 121.5 6 1.08 121.08 6 1.4 121.92 6 0.91 122 6 0.46
HDL-C (mg/dL) 77.13 6 3.55 63.25** 6 3.04 54.47
6 2.73 41.45**
6 2.23
LDL (mg/dL) 23.54 6 3.10 36.34* 6 3.42 42.80 6 2.63 60.89**
6 2.41
BW,Body Weight;RW,Relative weight.
Data presented are means 6 SEM,n=12 per group.P-value,.05 and,0.01 based on t-test comparisons of diet groups within sexes are indicated by * and **;and
comparison of sexes by 1 and 11 respectively.
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was defined as percent of time swimming parallel to the pool’s wall
within a 16-cm distance from the wall.Spatial cognition of the
platform location during acquisition trials and during the probe
day was evaluated by the analysis of the dwelling time in each of
the pool’s quadrants.Overall treatment effects were examined
using a repeated measures generalized linear model using SPSS
13.0 statistical software (SPSS Inc.,Chicago,IL).For the analysis
of the escape strategies,the Swim Paths for each mouse was
plotted and categorized into one of the following search
strategies:(1) Direct swim towards the location of the platform;
(2) Directed search,in which the animal swims in a search
pathway directly towards the platform;(3) Focal search:searching
in the quadrant containing the hidden platform.(4) Scanning,
where the search path is restricted to a limited,often central,area
of the pool;(5) Randomsearching in which the animal swims over
the entire area of the pool.(6) Chaining,which is circular
swimming at a fixed distance from the wall;(7) Thigmotaxis (wall-
hugging swim):a persistent swim along the outer 16 cm of the
pool.(8) Floating:a state of inactivity without forward movement.
The use of each search strategy was presented as a percent of
incidences during each trial over the whole analyzed experimental
Effect of Aspartame on Weight Gain,Blood Glucose and
Insulin Parameters
Averaged aspartame ingestion in the drinking water was
55.1464.74 mg/Kg body weight.Water and food consumption
were unaffected by the addition of aspartame compared to
controls (data not shown).Weight gain was significantly higher in
the male aspartame diet group compared to controls (Table 1,
P=0.024),but female weight gain was unaffected by diet.Body
length and visceral fat deposition were both increased in
aspartame-fed mice of both sexes (Table 1,P,0.01).Females
weighed less than males,and had less fat deposition following
aspartame treatment,although there was no difference between
the absolute or relative weight of the visceral fat collected from
control animals.Liver weight was affected by diet as well as
gender,with livers from aspartame-fed males weighing more than
control males;and more than those of female aspartame-fed mice
(P,0.05 and P,0.01 respectively).Additionally,aspartame-fed
mice had elevated fasting blood glucose levels compared to non-
consumers of both sexes,although females had lower levels than
males (Table 1,P,0.01).Fasting insulin and HOMA-IR levels in
female mice were significantly lower than males,although all were
within the normal range (Table 1).Pearson correlation analysis
was used in order to examine associations between body weight,
fat deposition,indices of glucose homeostasis and lipid profile.As
expected,body weight at six weeks of age strongly correlated with
weight at 17 weeks (Table 2,r =0.888,P,0.001).We also found
correlations between weight gain and visceral fat,body length,
fasting glucose,insulin and HOMA-IR (Table 2,r =0.322,0.546,
0.440,0.371 and 0.548 respectively,P,0.01).
A Random-fed insulin tolerance test administered at 19 weeks
of age showed that glucose levels in male aspartame-fed mice
were 120.2%higher than control mice following insulin challenge
(Fig 1A,P=0.001);and remained significantly elevated above
controls for up to 30 minutes,suggesting impairment of glucose
and insulin regulation.The mean Area Under the Curve (AUC)
in male aspartame-fed mice was significantly higher than control,
suggesting deregulation of glucose homeostasis (Table 3:
U/L/60 min vs 4829.586131.96
60 min:P,0.001),Mean AUC in female mice showed a similar
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trend without reaching significance (5194.636208.04
60 min vs 4819.936190.36
U/L/60 min).The half-life of
glucose in the aspartame mice showed a non-significant trend
towards elevation compared to control (Table 3).Collectively
these data indicate that aspartame treatment affects C57BL/6J
weight gain (6–17 weeks of age),visceral fat deposition and
glucose homeostasis particularly in males,and to a lesser extent in
Figure 1.Aspartame consumption reduces insulin sensitivity of male C57BL/6J mice during a random-fed insulin tolerance test.
Values are mean6SEM of (A) male glucose levels and (B) female glucose levels,n=18 per group,*** P,0.001 compared to controls.
Table 3.Effect of aspartame consumption on blood glucose levels during a random–fed insulin tolerance test.
Control Aspartame
(mM x min) 4829.58 6 131.96 5829.79 6 197.07,.001
(%/min) 2.92 6 0.28 2.32 6 0.29 0.15
TK (min) 39.69 6 14.05 51.61 6 16.63 0.59
(mM x min) 4819.93 6 190.36 5194.63 6 208.04 0.19
(%/min) 2.71 6 0.37 2.24 6 0.26 0.31
TK (min) 34.19 6 4.4 23.28 6 9.59 0.31
AUC,area under the curve;K,clearance rate;TK,half-life.
Values are means6SEMs,n=18 per group.
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Gender Dimorphism in Aspartame-Induced Impairment
of Learning
Spatial learning strategies in the MWM.
During the
acquisition phase of the MWM testing (days 1 to 4),significant
differences in place learning performance of aspartame-fed mice
became apparent.Cued escape latencies across sessions of mice
from both diet groups are shown in Fig 2.All mice improved their
performance over the 4 days of training,however,latencies of
male aspartame-treated mice were higher than control latencies on
the last 2 days of the 4-day trial (Fig 2A,P,0.05),suggesting that
aspartame may have impaired learning (acquisition) skills in these
mice.Female aspartame-fed mice had similar latencies to non-
consumers (Fig 2B).The duration of time spent in the target
quadrant where the hidden platform was located is shown in
figure 2C (males) and 2D (females).As testing progressed,mice
learned to spend more time searching for the platform within the
target quadrant,but aspartame-fed males spent significantly less
time searching for the platform in the target quadrant (Fig 2C,
P,0.05).A comparison of mean distance (in meters) to reach the
goal platform showed that aspartame-fed mice were farther from
their goal than controls on all four days of acquisition training,
suggestive of impaired learning (Fig 2E,P,0.05).Swim speed was
unaffected by diet (data not shown).
Floating and thigmotaxis behavior.
To gain further insight
into the reduced spatial learning of aspartame-fed mice,we studied
the non-spatial behavioral parameters of these mice and that of
controls.Floating behavior,characterized by periods of immobility
and defined as percent of time with swimspeed below a 0.06m/sec
threshold,revealed a significant effect of diet during the
acquisition period,with male aspartame-fed mice exhibiting
increased floating behavior on days 2,3 and 4 compared to
controls (Fig 3A,P,0.001).Thigmotaxis,defined as percentage of
Figure 2.Effect of aspartame consumption on spatial learning in C57BL/6J mice:gender-specific differences.Acquisition curves of
escape latency in male (A) and female (B) aspartame-fed and control diet mice.Percentage time spent in target quad in male (C) and female (D) mice.
Mean Distance to goal in male (E) and female (F) mice.Each group consisted of 12 mice;* P,0.05,** P,0.01,*** P,0.001.
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time spent swimming within 16cm of the pool wall,was greatly
increased in aspartame-treated males throughout the whole
experiment (Fig 3C,P,0.01),whereas female thigmotactic
behavior decreased throughout the acquisition period,and
showed no diet effect.Taken together,our data suggests that
males exposed to chronic dietary aspartame had impaired spatial
learning abilities together with increased non-spatial strategies.
Effects of aspartame on qualitative aspects of
To assess the effect of diet on qualitative aspects of
learning which could account for the apparent differences in
latency,we analyzed the respective search strategies displayed by
the mice to locate the hidden platform on the four days of
acquisition training.As the training progressed,control male and
female mice spent increasing amounts of time in directed
searching and direct swimming towards the platform,in addition
to using other strategies which require spatial and directional
searching,including focal searching (Fig 4A,B).Non-spatial
behavior,such as thigmotaxis and floating,decreased during the
4 days of training such that by Day 4,male control mice spent no
time either swimming around the periphery of the maze,or
floating directionless.Conversely,compared to control,
aspartame-fed mice exhibited significantly less time acquiring
spatial and directional strategies to find the hidden platform
during the acquisition training (Fig 4B,P,0.05).
Effect of aspartame on spatial memory in the MWM
Probe Test.
Regardless of gender,diet-induced differences in
spatial retention were apparent during a probe test,the recognized
measure of spatial memory,performed after the 4 days of
acquisition training.Once the platform had been removed,
aspartame-fed mice of both genders spent significantly less time
swimming towards the former position of the hidden platform
compared to control,(Fig 5A & B,P,0.05).Additionally,male
aspartame-fed mice crossed the former location of the platform
significantly less frequently than controls (Fig 5 C,P,0.01),and
had a lower Annulus Crossing Index (Fig 5E,P,0.01),implying
impairment of spatial memory.Platform crossing and annulus
crossing indices of female mice showed a similar trend without
reaching statistical significance (Fig 5 D & F).Overlapping swim
paths (n=12) of aspartame-fed and control male and female mice
are shown in figure 5 G & H.The location of the hidden platform
can be visualized through the aspartame-mice swim path,
indicating that the animal did not frequently cross the location
of the platform during the trial.
To complete our analysis of the performance of the mice during
this water maze test,we next used repeated measures generalized
linear modeling analysis of MWM performance variables
throughout the 4 days of acquisition training and during the
probe test,in males and females consuming the aspartame diet
compared to controls (Table 4).In males,significant differences in
floating and thigmotactic behavior between controls and aspar-
tame-fed mice were apparent on four out of the five days of testing,
and the mean distance to goal (a measure of how close mice swam
towards the platform) was significantly different between the two
diet groups on all days of testing (P,0.05).The performance of
females however,was only significantly different on the last day of
acquisition training and during the probe test.
Figure 3.Effects of aspartame consumption on non-cognitive behavior.Percentage of time spent floating in male (A) and female (B)
aspartame-fed and control diet mice.Thigmotaxis in male (C) and female (D) mice,n=12 per group;* P,0.05,** P,0.01,*** P,0.001.
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Correlations between Water Maze Performance Output
Variables and Body Characteristics
In order to understand more about the possible mechanisms
behind the reduction in escape latencies in aspartame-fed mice,we
next examined correlations between body characteristics and
MWM performance indicators (Table 5).Surprisingly visceral fat
deposition correlated positively with floating time on all days of the
trials,and also with thigmotaxis on day 3,4 and the day of the
probe test (P,0.05).Fat deposition also correlated with escape
latency time on these days,and negatively with time spent in the
target quarter,platform crossings and ACI.Finally,in order to
validate our analysis,we performed correlation analysis on all
MWM outcome variables in all mice analyzed simultaneously.As
expected,we found a strong negative correlation between latency
and floating time and between latency and mean distance to goal
(Table S1).Escape latency also correlated negatively with time in
target quadrant,number platform crossings,and the ACI.Taken
together,these correlations suggest that our analysis is valid,and
lends credence to observations concerning adiposity and non-
spatial learning strategies.
Our results suggest that neonatal exposure to aspartame
consumed as part of the diet of pregnant mice,together with
continued chronic exposure of the offspring to dietary aspartame
throughout the first 20 weeks of life,may result in increased weight
gain compared to controls together with impairment of insulin
sensitivity and cognitive performance,most notably in males.Food
and water intake was not affected by aspartame administration
within the ADI.The results of our analysis support previous
observations that aspartame may cause impairment in learning
and memory particularly when administered chronically [1,47] or
neonatally [26].Additionally,damage to hypothalamic morphol-
Figure 4.Effects of diet and gender on spatial and non-spatial escape strategies during the MWMtest.(A) Orientation of entry point
and escape platformon trial days 1–4.Arrows indicate location of entry point.(B) Distribution of search strategies in aspartame-fed mice compared to
controls on trial days 1–4.As the acquisition training advanced,control C57BL/6J mice exhibited progressive behavioral changes from random
chaining,thigmotaxis and floating into predominantly spatial strategies such as direct swimand direct search.Mice in the aspartame group exhibited
less spatial strategies and more non-spatial behavior throughout the trials.
Aspartame-Induced Impairment of Spatial Cognition
PLoS ONE | 8 April 2012 | Volume 7 | Issue 4 | e31570
ogy has been reported in neonatal rodents ingesting high amounts
of aspartame [48].
Aspartame is metabolized rapidly into methanol,phenylalanine
and aspartate [2];and oral administration of aspartame (200 mg/
Kg) has been shown to increase levels of rat brain phenylalanine
and its metabolite tyrosine,whilst decreasing levels of leucine,
isoleucine and valine [49].Whereas it is generally accepted that
aspartate does not readily cross the placenta,phenylalanine and
tryosine are readily transported to the fetal tissues;resulting in an
increase in phenylalanine at the expense of the maternal
concentration [25].In rodents,phenylalanine is readily converted
into the neurotransmitter precursor tyrosine by the hepatic
enzyme PAH [50];however if the activity of this enzyme is
reduced or absent,the high levels of accumulated phenylalanine
Figure 5.Results froma probe trial intended to measure spatial memory.Percentage of time spent searching in the former location of the
platform(Target Quad) in (A) males and (B) females,compared to time spent in the adjacent quads.Aspartame-fed males showed a reduction in the
number of times they crossed over former location of the platform(C:PlatformCrossings and (E:Annulus Crossing Index),n=12 per group,* P,0.05,
** P,0.01,*** P,0.001.There was no significance in the reduction of platformcrossings and Annulus Crossing Index in females.Actual overlapping
swim paths of the control (G,I) and aspartame-fed mice (H,J).Swim paths illustrate less intense swimming in the location of the platform by
aspartame-fed mice,compared to controls.
Aspartame-Induced Impairment of Spatial Cognition
PLoS ONE | 9 April 2012 | Volume 7 | Issue 4 | e31570
may be converted into other metabolites such as phenylpyruvate,
phenylacetate and phenyllactate [50,51].Crucially,studies have
shown that in rodents,PAH activity is undetectable until the final
days of gestation and birth,whereupon the gene is activated by
glucocorticoids and cyclic AMP [52–54].Experimentally induced
hyperphenylalaninemia has been shown to result in spatial and
non-spatial deficits in cognition and learning that are not related to
impairment of locomotor skills [55,56].In humans,deficiency of
PAH due to genetic mutations in the gene results in phenylketon-
uria (PKU),which is characterized by neurotoxic hyperphenylal-
aninemia and microcephaly,together with visuo-spatial,executive
and attention deficits [57,58].The mechanisms responsible for the
hyperphenylalaninemia-induced brain damage are still largely
unknown;however hyperphenylalaninemia has recently been
shown to promote oxidative stress in rodent brains,which may
contribute to the neurotoxicity in phenylketonuria [59,60].
Oxidative stress is the result of the aberrant production of reactive
oxygen and/or nitrogen species,or a decrease in the capacity of
antioxidant defenses for example glutathione;and has been linked
to a number of neurodegenerative diseases and to the cognitive
decline associated with aging [61].Importantly,subcutaneous
injections of aspartame have recently been shown to increase rat
brain thiobarbituric acid-reactive substances (TBARS;markers of
lipid peroxidation) and decrease glutathione levels [62].Collec-
tively,these observations may provide clues as to a mechanism
whereby aspartame metabolites;phenylalanine in particular,may
contribute to the impairment in spatial learning and memory that
we observed in mice exposed to aspartame in utero and during the
first months of life.
During acquisition of the water maze task,young rodents
typically improve their performance as indicated by a progressive
reduction in escape latencies over successive training sessions.
Upon introduction to the maze,mice initially adopt non-spatial
behaviors including thigmotaxis,scanning and chaining [45,63].
As training progresses,this behavior gives way to spatial learning,
resulting in more cued swimming towards the hidden platform,
more time spent in the target quadrant,and shorter escape
latencies.Within the context of this paradigm,aspartame-fed mice
exhibited significant differences in learning strategies at four
months of age,which resulted in longer escape latencies compared
to controls,together with quantitative and qualitative differences
in behavioral strategies employed.Towards the end of acquisition
training,aspartame-fed mice spent significantly more time
swimming around the periphery of the pool and passively floating
compared to control mice,which may be indicative of an
ineffectual non-spatial swim strategy [64].Increased thigmotaxis
behavior linked to a deficit in responding to visual cues has
previously been noted after experimentally induced lesions to the
dorsal-striatum:a compound structure of the brain believed to be
involved in stimulus-response learning [65].Additionally,lesions
to the hippocampus [66] and NMDA receptor blockade using
specific antagonists have also shown to result in increased
thigmotactic behavior [67,68].
Impairment of spatial memory in the aspartame diet group was
suggested by a significant reduction in time spent swimming
towards the former platform location during the probe test.
Additionally,thigmotactic behavior and passive floating during the
probe test was increased in male aspartame-fed mice compared to
controls.Taken together this suggests that chronic exposure to
aspartame may impair rodent spatial memory.It has previously
been suggested that exposure to high doses of aspartame at a late
stage of pregnancy may result in a delay in visual placing response
in the offspring,which is a measure of sensorimotor activity [69].
However,a second study failed to duplicate these findings [70].
In addition to the effects of aspartame on rodent cognitive
performance in the water maze,aspartame appeared to raise
fasting blood glucose levels in both sexes.The relationship
between peripheral blood glucose levels and cognition has been
Table 4.Generalized linear modeling of performance variables in aspartame-fed C57BL/6J mice compared to controls.
Day 1 Day 2 Day 3 Day 4 Probe Test
Latency 0.862 0.061
Thigmotaxis time (%)
.0001 0.236
Floating time (%) 0.348
Mean distance to goal (m)
Time in target quadrant (%)
.01 0.076
.05 0.112 0.112
Time in non-target quadrants (%)
.01 0.076
.05 0.112 0.111
Platform Crosses 0.715 0.547
.001 0.800
Annulus Crossing Index 0.781 0.569
.01 0.553
Latency 0.817 0.359 0.644 0.090 0.474
Thigmotaxis (% time ) 0.623 0.957 0.452
.05 0.818
Floating (% time) 0.191 0.347 0.640
.01 0.175
Mean distance to goal (m) 0.626 0.822 0.291
Time in target quadrant (%) 0.321 0.897 0.605 0.176
Time in non-target quadrants (%) 0.321 0.895 0.602 0.175
Platform Crosses 0.225 0.339 0.650 0.153 0.277
Annulus Crossing Index 0.263 0.536 0.290 0.619 0.199
Significant differences in performance between the control and the diet groups are indicated in bold P-value#0.05,for trial days 1 to 4 and probe day,n=12 per
Aspartame-Induced Impairment of Spatial Cognition
PLoS ONE | 10 April 2012 | Volume 7 | Issue 4 | e31570
well-documented and suggests that there is a homeostatic
neuroglycemic range within which optimal cognitive function
occurs [71].Hyperglycemia may damage the microvasculature of
the blood-brain barrier and/or modify insulin availability in the
brain,disrupting normal brain function and cognition.Interest-
ingly hyperglycemia,increased body weight and swimming defects
were recently observed in hyperlipidic zebrafish exposed to
aspartame [28],which tends to support our present observations.
However,our data using doses of aspartame approximating the
current ADI contrasts with a previous report which concluded that
long term consumption of aspartame from six weeks onwards did
not increase weight gain [72].This apparent contradiction could
conceivably be due to the fact the aspartame in that toxicology
study was administered fromthe 6
week of life onwards at a dose
of 2–4 g/Kg body weight (40–80 times the current ADI),resulting
in a significant reduction in food intake for animals consuming the
higher quantity of the dipeptide sweetener.Interestingly human
studies have found a positive correlation between the consumption
of artificial sweeteners and weight gain [73,74];and surprisingly in
diabetic subjects,aspartame-containing meals elevated blood
glucose and insulin levels to the same extent as that of higher-
calorie sucrose-containing meals [75].
A third novel observation in our study relates to gender-specific
differences in insulin sensitivity and cognitive performance,with
males apparently exhibiting a greater adiposity,weight gain,
glucose dysregulation and cognitive impairment compared to
females.Gender-specific differences in behavior have also been
documented in response to Monosodium Glutamate (MSG) a
commonly consumed food additive.MSG-treated males appear to
be more adversely affected than females [41,76],and have a
greater increase in adipose tissue deposition [77] and insulin
resistance [34] than females.Therefore the possibility exists that
although both males and females showed equal increases in fasting
glucose levels in response to aspartame,the gender-specific
differences in cognitive performance may be due to differences
in the extent of adiposity and insulin resistance,both of which are
associated with cognitive performance [78].A final intriguing
outcome from our study was the correlation we found between
Table 5.Summary of correlation analysis between body characteristics and spatial memory variables in the MWM test.
% weight
Body Weight
Visceral Fat
(mg/dL) LDL (mg/dL)
Latency (s)
Day 2 0.027 20.092 0.005 0.095 0.050 0.132 0.242 20.054 0.094
Day 3 0.232 0.075 0.357* 0.023 0.246 0.148 0.191 20.065 0.130
Day 4 0.063 0.029 0.428** 20.076 0.124 0.010 0.195 20.070 0.139
Probe Test 0.211 0.076 0.431** 0.017 0.207 0.150 20.116 20.257 0.281*
Thigmotaxis time (%)
Day 2 0.290* 0.113 0.265 0.085 0.187 0.195 0.097 0.053 20.025
Day 3 0.327* 0.263 0.488*** 0.002 0.409** 0.255 0.206 0.169 20.108
Day 4 0.163 0.291* 0.553*** 0.046 0.396** 0.240 0.048 0.094 20.020
Probe Test 20.010 0.211 0.356* 0.001 0.259 0.133 0.061 0.123 20.097
Floating Time (%)
Day 1 20.157 0.046 0.402** 20.115 20.045 20.089 0.096 0.088 20.058
Day 2 20.103 20.052 0.340* 20.134 0.174 0.005 0.111 20.074 0.104
Day 3 0.086 0.062 0.487*** 20.224 0.369** 0.006 0.278* 20.045 0.158
Day 4 0.162 0.054 0.441** 20.128 0.349* 0.076 0.214 20.035 0.156
Probe Test 0.258 0.188 0.457** 20.077 0.433** 0.177 0.202 0.012 0.074
Time in Target Quadrants (%)
Day 1 20.164 0.075
0.284* 0.136 20.234 20.024 20.164 0.158 20.230
Day 2 20.245 20.224 20.188
0.361* 20.078
0.355* 20.188 20.102 0.086
Day 3 20.137 20.162
0.349* 20.135 20.021 20.121 20.056 0.037 20.041
Day 4 20.085 20.243
0.294* 20.193
0.330* 0.167 0.024 0.048
Probe Test 20.054 0.160 20.134 20.057 20.002 20.055 20.114 0.347* -0.368**
Platform Crosses
Day 2 20.028 0.130 0.071 20.020 20.010 20.034
0.387** 0.034 20.122
Day 4 20.122 0.002
0.369** 0.094 20.260 20.051 20.160 0.079 20.146
Probe Test 20.265 20.120
0.401** 0.023 20.157 20.080 20.005 0.223 20.221
Day 3 20.188 20.118
0.384** 20.051 20.182 20.131 20.030 0.042 20.073
Day 4 20.133 20.199
0.275* 20.173 0.060 20.125 20.161 20.118 0.079
Probe Test 20.213 20.027
0.359* 20.004 20.074 20.052 0.048 0.317*
Significant correlations are shown in bold with *,** and *** indicating a P-value of#.05,#.01 and#.001 respectively,n=12 per group
Aspartame-Induced Impairment of Spatial Cognition
PLoS ONE | 11 April 2012 | Volume 7 | Issue 4 | e31570
visceral adiposity and the adoption of non-spatial escape strategies
(thigmotaxis and floating behavior) during the MWM test.In
general,non-spatial behavior in the MWM test is associated with
an inability to adopt spatial cognitive abilities [79],and others
have found that high-fat diets which promote obesity also impair
spatial memory in rodents [80,81].
Our study terminated when the mice were 20 weeks of age
(mature adulthood);however it would be of interest to ascertain
whether the effects that we noted resulting from lifetime exposure
to aspartame will still be apparent in an aging mouse model.Our
unpublished observation,together with previous studies [81]
suggests that the water maze performance of older C57Bl/6J mice
decreases markedly with aging.Further studies are warranted to
assess the effects of aspartame on metabolism and cognition in
aging mice,and in mice from different strains.In conclusion,we
have demonstrated that compared to controls,neonatal exposure
of rodents to dietary aspartame,combined with chronic aspartame
consumption throughout early life,may result in impairment of
glucose and insulin homeostasis,together with a reduction in
cognitive performance.Several gender differences were observed,
with males exhibiting greater sensitivity to aspartame exposure.
Our data supports previous observations that chronic exposure to
aspartame may result in memory deficits in rodents.
Supporting Information
Table S1.
Correlations between body characteristics
and spatial memory variables in the MWM test.
We thank Jonathan Caigas,Rhea Mondreal,Rosario Ubungen,Razan
Bakheet and Qammar Al-Haffar for excellent technical assistance;and our
gratitude goes to Mr Hakim Al-Enazi for his unparalleled help in
coordinating research resources.
Author Contributions
Conceived and designed the experiments:KSC NJMFAA.Performed the
experiments:NJMMZZ SMS BA AI RAR.Analyzed the data:KSC NJM
MZZ.Wrote the paper:KSC.
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Oxidative Stress and Monoamine Neurotransmitter Levels in Lipopolysaccha-
ride-Treated Mice.Neurotox Res.Aug 6.[Epub ahead of print].
63.Wolff M,Savova M,Malleret G,Segu L,Buhot MC (2002) Differential learning
abilities of 129T2/Sv and C57BL/6J mice as assessed in three water maze
protocols.Behav Brain Res 136(2):463–474.
64.Wolfer DP,Stagljar-Bozicevic M,Errington ML,Lipp HP (1998) Spatial
memory and learning in transgenic mice:fact or artifact?News Physiol Sci 13:
65.Devan BD,White NM (1999) Parallel information processing in the dorsal
striatum:relation to hippocampal function.J Neurosci 19(7):2789–2798.
66.Hostetter G,Thomas GJ (1967) Evaluation of enhanced thigmotaxis as a
condition of impaired maze learning by rats with hippocampal lesions.J Comp
Physiol Psychol 63(1):105–110.
67.Cain DP,Saucier D,Hall J,Hargreaves EL,Boon F (1996) Detailed behavioral
analysis of water maze acquisition under APV or CNQX:contribution of
sensorimotor disturbances to drug-induced acquisition deficits.Behav Neurosci
68.Saucier D,Hargreaves EL,Boon F,Vanderwolf CH,Cain DP (1996) Detailed
behavioral analysis of water maze acquisition under systemic NMDA or
muscarinic antagonism:nonspatial pretraining eliminates spatial learning
deficits.Behav Neurosci 110(1):103–116.
69.Mahalik MP,Gautieri RF (1984) Reflex responsiveness of CF-1 mouse neonates
following maternal aspartame exposure.Res Commun Psychol Psychiatry Behav
70.McAnulty PA,Collier MJ,Enticott J,Tesh JM,Mayhew DA,et al.(1989)
Absence of developmental effects in CF-1 mice exposed to aspartame in utero.
Fundam Appl Toxicol 13(2):296–302.
71.Cox DJ,Kovatchev BP,Gonder-Frederick LA,Summers KH,McCall A,et al.
(2005) Relationship between hyperglycemia and cognitive performance among
adults with type 1 and type 2 diabetes.Diabetes Care 28 (1),71–77.
72.Ishii H,Koshimizu T,Usami S,Fujimoto T (1981) Toxicity of aspartame and its
diketopiperazine for Wistar rats by dietary administration for 104 weeks,
Toxicology 21(2):91–94.
73.Yang Q (2010) Gain weight by ‘‘going diet?’’ Artificial sweeteners and the
neurobiology of sugar cravings:Neuroscience,Yale J Biol Med 83(2):101–108.
74.Fowler SP,Williams K,Resendez RG,Hunt KJ,Hazuda HP,et al.(2008)
Fueling the obesity epidemic?Artificially sweetened beverage use and long-term
weight gain.Obesity (Silver Spring Md.) 16:1894–1900.
75.Ferland A,Brassard P,Poirier P (2007) Is aspartame really safer in reducing the
risk of hypoglycemia during exercise in patients with type 2 diabetes?.Diabetes
Care 30(7):e59.
76.Hlina´k Z,Gandalovicova´ D,Krejcı
I (2005) Behavioral deficits in adult rats
treated neonatally with glutamate.Neurotoxicol Teratol 27(3):465–473.
77.Nascimento Curi CM,Marmo MR,Egami M,Ribeiro EB,Andrade IS,et al.
(1991) Effect of monosodium glutamate treatment during neonatal development
on lipogenesis rate and lipoprotein lipase activity in adult rats.Biochem Int.
78.Yaffe K (2007) Metabolic syndrome and cognitive decline.Curr Alzheimer Res;
79.Janus C (2004) Search strategies used by APP transgenic mice during navigation
in the Morris water maze.Learn Mem;11(3):337–46.
80.Jurdak N,Lichtenstein AH,Kanarek RB (2008) Diet-induced obesity and spatial
cognition in young male rats.Nutr Neurosci.11(2):48–54.
81.Benice TS,Rizk A,Kohama S,Pfankuch T,Raber J (2006) Sex-differences in
age-related cognitive decline in C57BL/6J mice associated with increased brain
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science 137(2):413–23.
Aspartame-Induced Impairment of Spatial Cognition
PLoS ONE | 13 April 2012 | Volume 7 | Issue 4 | e31570


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The Official Blog for TeleVox Solutions

The Official Blog for TeleVox Solutions

West Corporation

Posted on September 30, 2013 by West Corporation

High-Tech Engagement Across Generations

patient engagement across generations

NOTE: The following is an excerpt from TeleVox’s Healthy World report, “Technology Beyond the Exam Room”, which illustrates that as digital technology becomes increasingly ubiquitous in American culture, a higher prevalence of its usage should be expected within our healthcare system. Download it HERE!

Consumers are currently living in an age revolutionized by digital communication. They are in a constant state of upgrading, as technology seems to update faster than most people can grasp how it’s used. Cell phones are old news within six months and computers are considered out-of-date within a year. Most everyone wants the latest and greatest version of whatever will allow them to stay plugged in, connected and ahead of the game.

Those ages 18–34, known as Generation Y, are typically the most savvy when it comes to being in-the-digital-know. They own and purchase more tablets and tech-advanced cell phones, have the most up-to-date social media pages, and are usually setting the standard for new digital and gaming verbiage in pop culture. Generation X, those ages 35–44, are racing right alongside their younger counterparts. However, instead of mastering the digital social realm, the Gen Xers are focused more on the practical and business functionality of these leading technologies.

Of equal importance is that in this lightning-speed race to the digital top those born in the Baby Boomer era, current senior citizens, can be left in the dust.

When taking into consideration that 85 percent of the U.S. population report wanting to communicate with their healthcare provider through emails, text messages and voicemails, rather than in-person visits, it’s important to remember what age group may need to communicate with their doctors most frequently.

Eighty-one percent of the Baby Boomer population surveyed said they visit a healthcare professional at least once a year, compared with the 78 percent of Gen Yers and 79 percent of Gen Xers reporting a yearly visit. As these Boomers start requiring a doctor’s care more frequently, younger generations must note the importance of educating them on the benefits of being plugged in.

Closing the Age Gap in Communication
Though the Boomer group would prefer a pen and paper over an instant message, and are not likely to follow MTV’s latest on Twitter no one should be quick to think that this generation doesn’t know how to text or log in into Facebook. Most Boomers’ usage of social media and other forms of digital communication is greatly increasing as it allows them the accessibility to connect with family, loved ones, and now their physicians.

The average adult over the age of 55 manages six to eight medications daily. As their pill boxes grow, it becomes difficult to remember when to take each pill, whether to take it with food, if the side effects they are experiencing are normal, and so on. Unfortunately, without the proper between-visit guidance, approximately 50 percent of the 2 billion prescriptions filled each year are not taken correctly. It’s estimated that 125,000 people with treatable diagnoses die each year simply because they do not take their medications as prescribed.

Ongoing follow-ups from physicians, via text message, email or voicemail can ease stress, both for the patient and the patient’s family. As more tech-savvy seniors emerge, they’re learning how to engage their doctors digitally.

TeleVox Solutions

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Shipping & Returns

Shipping Information

USPS Priority Mail Flat Rate Shipping.

Flat Rate Pricing. No weighing or calculating needed up to 70 lbs. Flat Rate offers one rate to any state. TEN Clothing ships only within the U.S. at this time.

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Please keep in mind that most of our accessories are made to order. We have very limited quantities in stock and can only guarantee delivery between 3 and 6 weeks after your order has been placed and depending on quantity requested. TEN believes in quality over quantity - slow fashion.

Returns & Exchanges

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Pacific Tennis Store

Pacific X Tack PRO 3 Pack - Black

Pacific X Tack PRO 3 Pack - Black
  • Grips Oberfläche: Feucht, Klebrig
  • Grips perforated: Synthetic grip
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X Tack PRO 3 Pack - Black Pacific
X Tack PRO 3 Pack - Black
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Pacific X Tack PRO 3 Pack - Black

Perforated overgrip for particularly good racket control

The Pacific X Tack Pro Perfo Overgrip convinces with its moist and adhesive surface and excellent sweat absorption.

With this synthetic overgrip you get a particularly grippy design with extra adhesive surface for ideal racket control. Thanks to the perforation, it absorbs sweat even better than the well-known Pacific X Tack version of the overgrip.

Convince yourself of the adhesiveness of the Pacific X Tack Pro Perfo overgrips!

  • sticky synthetic overgrip for a particularly good racket control
  • extremely handy and extra-sticky surface
  • with Tennis-Point logo
  • perforated version of the well-known Pacific X Tack overgrip with even better sweat absorption

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Teacher of English, West London

Closing date:

Kensington Aldridge Academy (KAA)West London

Date posted:
Contract type: Full Time
Salary: MPS (Inner London)
Contract term: Permanent
Suitable for NQTs: Yes

About KAA

KAA is a growing 11-18 academy. In our first three years we have built a school which has the very highest standards, both in terms of academic teaching and life beyond the classroom. We now have 720 students in Years 7, 8, 9, 10, and 200 in Y12 & 13. Our school is heavily oversubscribed at both Year 7 and Sixth Form.

At KAA we are building a school which has the very highest standards - both in terms of academic teaching and life beyond the classroom. In July 2014 the Department of Education reported we are “one of the best new academies they had ever seen” in terms of our planning and curriculum development. In November of that year we were inspected by Miriam Rosen, former HMCI, who wrote “the academy has made a remarkable start and is already securing highly positive outcomes for students.” We look forward to our first full Ofsted inspection in due course and aim to achieve ‘outstanding’ in all areas.

We are currently looking for a talented and committed teacher of English to join our growing English department.

The successful candidate will have:

  • Evidence of being, or having the clear potential to be, an outstanding teacher of their subject.
  • Significantly contributed to the work of a successful department.
  • Experience of leading the planning and implementation of a curriculum initiative.
  • Experience (can be limited) of delivering staff training and undertaking professional development of other teachers.
  • Contributed to intervention programmes at a department/year group/whole school level.
  • Good knowledge of data, tracking and monitoring of discrete groups of students.
  • NQTs may apply if they can demonstrate high-potential through the process.

Deadline: Apply by 9 am on Monday 30th October 2017

Please send your completed application form to Lavinia Baboi at recruitment@kaa.org.uk

Please also complete the Equality and Diversity Monitoring Form using the following link: https://goo.gl/forms/eh8N7EiLSyU6BIuE2

This post is subject to an enhanced Disclosure and Barring Service check. The post holder must be committed to safeguarding the welfare of children

Job location

More about Kensington Aldridge Academy (KAA)
Type: Mainstream School
Phase: Secondary with sixth form
Funding status: State - Academy
Gender: Mixed
Age Range: 11 - 18 years
Address: Temporary Address, Scrubs Lane, North Kensington, London, W12 0DE, United Kingdom
Telephone: +44 207 313 5800
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The Royal Borough of Kensington & Chelsea is a fantastic place to work. KAA is situated near Portobello Road, full of great shops, restaurants and bars. The school is part of the inspiring new £66 million pound Kensington Academy and Leisure Cent...


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Geodynamics enters the biogas energy market

17 July 2015

Thomson Geer’s Brisbane Corporate team advised ASX listed Geodynamics Limited in relation to its off market takeover scrip offer for all the shares in an unlisted public company, Quantum Power Limited (QPL).  The bid values QPL at approximately $8 million.

Geodynamics is transforming from a geothermal power developer to a participant in the broader cleantech sector. The acquisition will represent its entry into the biogas energy market.

QPL is a market leader in the Australian biogas market specialising in the engineering, procurement and installation of biogas production systems and biogas fuelled power generation for large agribusinesses.  With Geodynamics’ additional financial capacity, project management and engineering capability, the merger will enable QPL to accelerate delivery of its existing projects and pursue larger opportunities.


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T.I writes Another Open Letter

Musician T.I writes Another Open Letter to US

“As I write this letter, I can’t help but express my frustration and concern for US. I also can’t help but question my contribution to the status of where we are. I understand that I too have been guilty of doing the very things that put us here. Even still, right now the state of our community is more alarming than ever. And while I know this letter may be misunderstood by many, unduly dissected and misconstrued by most others, I still need to write and speak about these things that deeply concern me.
You see, all of US sometimes feel that one of the most difficult things in the world is to talk to US about US in an effort to help US. However, we cannot dismiss the fact that we can’t fix what we do not address. You see we’ve somehow allowed ourselves to think it’s okay to neglect and abandon our responsibility to ourselves and to our community. Everything cannot be blamed on THE SYSTEM.
We can no longer afford to be entertained beyond the point of accountability. We can’t keep selling each other out and be willing to do anything to be seen, to be famous or to be rich.
Our obsession with material things and lack of self-worth is evident in our need for an abundance of momentary luxuries and must-have amenities that have no true value for real, man. And I mean, we do it just to impress people that could care less if your children or your children’s children have anything left to show for your life after you gone. You see somewhere along the way, we’ve allowed the pursuit of fame and things to blind us to the issues that prevent us from moving upward. So, we cannot continue to get caught up in having material abundance and yet be fundamentally and spiritually broke.
Every one of us must do something to contribute to transforming our community for the better. We have been underserved, underprivileged and unfortunate for far too long. There are no more excuses. It’s not enough to have limited progress and allow our expectations and sense of purpose to evaporate. So, if that means we must sacrifice some nights at the club and give up buying the latest designer handbags and sneakers…well then damn, so be it. I can only hope that one day soon we’ll understand our true economic buying power by investing in our own communities and putting our money into businesses that keep our dollars in our community.
It is imperative for US to parent our children and educate them outside of the school systems, as our education system was not designed to lift US out of oppression. If we know that the pipelines to prisons are multiplying, well we must ask ourselves what can we do to end it? We must keep ourselves busy with finding ways to generate wealth for generations to come and work to pass down things to our children for them to pass down to their children. You see man, who wants to fight who on Instagram should not be as important as fighting for equality…and who’s f**king who should never be as important as creating a strategy to help break the cycle [of] oppression.
You see, if we don’t acknowledge and accept where we are failing, then we’ll never do the work that it takes to make it right for US. See, we are the benefactors of too much sacrifice and blood, sweat and tears to allow petty differences and trivial discrepancies to continue to separate and divide US, while we are being used, exploited and conquered. We should not live one more day ignoring the sh** that suffocates and strangles US keeping US imprisoned in our own communities. We cannot afford to not stick and work together for the betterment of our community.
As we look to the future for US, there could not be a better time to assess where we go from here. And while there’s currently great turmoil, there is even greater opportunity for US to work together to transform our community. Far too many of our children are fatherless, far too many of our mothers are standing in the prison waiting rooms and far too many of our young people feel hopeless.
We must work to reshape the need for our children to want to live so fast even if it means dying too young. Now, I am not here to demonize or vilify, but to pledge my commitment to help US do what needs to be done. You see if we continue to devalue ourselves we cannot realistically expect respect from others.
Now, how can I sit here in a place of privilege and say such things and make such suggestions you ask? Well, I answer simply because I know what it’s like. I know what it’s like to not know where you’re going or how to get there. I know what it’s like to have no one around you who can teach you how to be what you hope to become.
I know what it’s like to be so distracted by your surroundings and in the moment that it’s seemingly impossible to not get caught up in ‘em. I know what it’s like to feel so much smaller than the activities of your environment that you can’t see how not to succumb to ‘em. I know what it’s like to not be able to focus in class due to real life hunger pangs. I know what it’s like to be disruptive just to pass the time and take your mind off what’s lacking at home. I know what it’s like to be laughed at by your teacher when you tell them what you hope to be in life.
I also know what it’s like to be told by your teacher that you’ll never amount to anything.
I know what it’s like to feel hopeless and to feel like you’re not good enough just because of where you’re from. I know what it’s like to be profiled and to be abused by the police. I know what it’s like to be racially profiled, treated unjustly and abused by the police just because of how you look.
But even more importantly than knowing all these things, I know what it’s like to overcome ‘em. Now, I know it’s not easy…but all of US can do something.”

Respectfully yours,
Tip Harris

Read Also:See One Of The Biggest Guitar In The World

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THE1960POST: T.I writes Another Open Letter
T.I writes Another Open Letter
T.I writes Another Open Letter
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Blue Stud

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This silky soft, squishy jelly ribbed vibrator actually has the curve you see in the photos. It can straighten out completely if you pull the head upwards however making insertion super comfortable. The penis shaped head squishes inwards with your fingers, yet it's sturdy enough not to buckle under the pressure of anal insertion.

The ribbed shaft remains sturdy for added texture of you're in the mood for some thrusting. The motor rests underneath the penis shaped tip and quietly tosses vibrations around the entire top half of the shaft. This large prostate vibrator for men is a clever option for guys who already have everything!

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How to Retire in Your 30's

8 Lessons to Learn From Two Millennials Who Retired With $1 Million

young couple

When would you like to retire?

Sixty-five? Fifty?

You don’t have to wait that long, you know.

A California couple recently retired in their mid-30’s with $1 million in the bank, according to Forbes magazine.

OK, true, that remarkable feat of personal finance would be nearly impossible for most of us because we have, you know, lives. And kids. I could never pull off something like this with four boys to raise.

The couple —  Travis and Amanda — has no children. They earned good money as tech professionals and had already socked away $350,000 when they decided to chuck their jobs forever. They also have iron willpower as you’ll see. Still, there’s a lot to learn from these two Millennials about the fundamentals of building a retirement nest egg.

1. Know Your Objective

When Travis got laid-off from an IT job in 2012 he discovered a new passion — the freedom that came with not working. He and Amanda decided to retire as soon as possible. They figured they needed $1 million to make that happen. The couple planned to live on 3 percent — 4 percent of their portfolio’s value every year and expected a 7 percent annual growth rate.

How do you envision your retirement? Will you travel the world? Start a small business? Own a beach house? Knowing what you want post-career is the first step towards making those dreams come true.

2. Get It Together

Organizing your finances allows you to know where you are and how to move ahead. Travis and Amanda gathered all their money-related accounts into the free budgeting site and did a thorough review of their assets and expenses. This lead to spending cuts and the (easy) decision to combine several 401ks from former employers.

3. Savings First

The couple saved as much as 65 percent of their pay during the three years it took to amass $1 million. They lived in a rent controlled $2,200 Oakland house (a bargain by the Bay, for sure) and aggressively cut costs by doing stuff like driving less, and hanging the laundry out to dry in the free wind.

Here’s a tip for boosting your savings rate: Pay yourself first. Have your savings auto-deducted from your paycheck, whether going into the company 401k or another retirement fund. You can’t spend what you never see.

4. Beware of Fees and Taxes

Fees are the great return-killer. You should review and question every fee you pay, even on funds inside your 401k. Amanda and Travis put much of their retirement money in low-cost ETFs and index funds.  These paid off nicely, as the couple rode a more than 60 percent increase in the S&P 500 from 2012 until 2015.

The couple planned ahead and was able to avoid the 10 percent IRA early withdrawal penalty by using a Roth IRA conversion ladder. In this forward-looking strategy, they transferred a certain amount of money each year from their traditional IRA to their Roth. Once 5 years had passed from the initial IRA to Roth conversion, they were able to tap their Roth contributions in an annual laddered sequence and avoid the early withdraw penalty.

5. Your Job Is Your Real Moneymaker

 Easy to forget, your paycheck is the centerpiece of your investment plan. Anything you can do to hike your salary or add other income (a second job or rental property, for example), will dramatically advance your objectives.

Much as he hated work, Travis returned to a cubicle purely to make the couple’s retirement dream come true. He switched jobs three times in three years to obtain salary increases. Amanda stuck it out in her job as a chemical engineer. At their earnings peak, the couple was making a combined $200,000.

6. Move and Save

It’s considerably easier to save when your cost of living is low. When Amanda and Travis retired, they left the psycho-expensive Bay Area for a $270,000 house in Asheville, NC. The couple chose the mountain town because the cost of living is relatively low.

They also believe their house will be easy to rent to tourists while the couple continues to globe trot.

7. Simplify and Declutter

As they approached their retirement goal, Amanda and Travis sold much of the stuff in their two-story house. They headed into retirement with only what they used on a daily basis.

8. Plan Your Retirement Spending

We tend to focus on saving when we discuss retirement planning. But you need to think carefully about your post-work spending if you want your nest egg to see you through two or three decades. We’re living longer these days, remember the 1,000 Bucks A Month Rule from previous my previous AJC columns.

Travis and Amanda are very disciplined. They refuse to spend more than 4 percent of their portfolio’s current value per year. As a result, they sometimes have to cut spending when their portfolio value dips. They stuck to this rule even during their retirement dream driving trip from San Francisco to Costa Rica.

They made that journey on the cheap, driving (and sleeping in) an old Toyota 4runner. In Costa Rica, they leased a house for $1,000 a month — equal to $30 a night. They ate at home and skipped the touristy stuff.

Travis and Amanda told Forbes they will never work again. But they left the door open to having children. It’s tough to raise kids these days on a household income of just $30,000 or $40,000. But if anyone can do it, I’m betting on Travis and Amanda.

Their extreme example is inspiring. If these two thirty-somethings can save about $650,000 in three years, surely we can reach our retirement goals in 20 or 30 years.

All it takes is a goal, a plan, and commitment. Especially commitment.

Related: How To Retire At 50

The Balance does not provide tax, investment, or financial services and advice. The information is being presented without consideration of the investment objectives, risk tolerance or financial circumstances of any specific investor and might not be suitable for all investors. Past performance is not indicative of future results. Investing involves risk including the possible loss of principal.


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Qualifications Interview Questions

How to Answer Interview Questions About Your Qualifications For the Job

two men at table for job interview
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The purpose of a job interview is to demonstrate why you are the right person for the job. The best way to do that is to show how your skills, education, and experience have prepared you to do this job better than any other candidate under consideration. This means relating your qualifications to the job for which you’re being interviewed.

It's critical to show the hiring manager not just that you have the qualifications, but that you can apply them.

Go beyond simply reminding the interviewer of your relevant certifications, for example, or even the jobs you’ve held that have prepared you for this role. Be prepared to make a case for why your unique experience makes you the best choice.

The good news is that the hiring manager will most likely give you plenty of opportunities to demonstrate your aptitude for the role. Most job interviews involve a series of questions from the hiring manager, some trickier than others. Keep your ears open for questions about your qualifications, and be ready with responses that demonstrate why you’re a good fit.

Review typical job interview questions about your qualifications and practice your responses.

Interview Questions About Your Qualifications

    Are you a self-motivator? - Best Answers

    Do you prefer to work independently or on a team? - Best Answers

    How do you evaluate success? - Best Answers

    How do you handle stress and pressure? - Best Answers

    How do you plan to achieve those goals? - Best Answers

    How will your greatest strength help you perform? - Best Answers

    How would you describe yourself? - Best Answers

    If the people who know you were asked why you should be hired, what would they say?

Best Answers

    Is there anything else we should know about you? - Best Answers

    Tell me about yourself. - Best Answers

    What are you looking for in your next job? What is important to you? - Best Answers

    What are your goals for the next five years and 10 years? - Best Answers

    What can you contribute to this company? - Best Answers

    What do you find are the most difficult decisions to make? - Best Answers

    What experience do you have? - Best Answers

    What interests you about this job? - Best Answers

    What is your greatest strength? - Best Answers

    What motivates you? - Best Answers

    What type of work environment do you prefer? - Best Answers

    Why are you the best person for the job? - Best Answers

    Why do you want this job? - Best Answers

    Why do you want to work here? - Best Answers

    Why should we hire you? - Best Answers

Demonstrating Your Qualifications

In addition to describing your qualifications for the job, offer to demonstrate them. For example, if you're interviewing for a job that requires superior writing skills, demonstrate your qualifications by providing a writing sample. (However, if you provide a sample based on an assignment you completed for a previous employer, be sure you redact confidential or sensitive information.)

Another example of demonstrating your qualifications is to explain to the interviewer what you hope to accomplish during your first 30 days or 60 days on the job. Expressing your future accomplishments can give the interviewer and hiring manager insight into the type of work you're capable of performing.

Tips for Answering Tricky Job Interview Questions

No matter how much you practice, you’re bound to get a curveball job interview question or two during the process. To prepare, practice your responses and keep these other tips in mind:

1. Listen.

One of the most common job interview mistakes is talking too much. Another is not paying attention to what the interviewer is saying. Both are functions of not engaging with the hiring manager.

Remember that a job interview is a conversation, not a monologue.

You’re there to connect with the other person, and that means really listening to what they have to say.

2. Take your time.

If you don’t know the answer to a question, or can’t come up with a response off the top of your head, take a moment. It will feel unnatural to you, like 30 seconds of pondering is actually half an hour of silence, but it’s much better to take a beat than to rush. It’s OK to ask for more information, clarification about what the hiring manager wants, or for a moment to cogitate. In fact, doing so may make you seem more confident and trustworthy.

3. Avoid negativity.

Don’t badmouth your old boss, coworkers, or company. To do so is to tell the hiring manager that you might turn on your new employer at any moment. Not exactly a selling point.

Further, be kind to yourself. If you’re someone who tends to beat yourself up a lot when you make a mistake or experience stress, know that and watch out for self-undermining behavior.

Remember that above all else, hiring managers are looking for people who will do good work as part of a team. Being positive during the process will show that you’re not only qualified to do the job, but to create an uplifting work environment for the group.

More Interview Questions and Answers

More Interview Questions and Answers
Typical job interview questions and sample answers.

Interview Questions to Ask
Questions for candidates for employment to ask the interviewer.


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$700 Billion Defense Bill Flies Through the Senate

Amidst a massive focus on the repeal of Obamacare and healthcare being the primary topic of conversation in the Senate, the Senate took up a $696.5 billion defense bill on Monday. With a vote of 89 to 8, the bill easily passed through the Senate on its way to President Trump’s desk. After passing through the House in July, the legislation kind of stalled out and was lost. It wasn’t until Monday that it resurfaced, and now is the largest single defense authorization in American history.

This massive defense bill has received largely bipartisan support in Congress, and 89% of Democratic senators voted yes on the new budget. For now, the way the legislation is worde, it seems that the money will simply be allotted to a multitude of defense facets, and more specific appropriation will take place later this year. Among the largest issues the bill tackles, it issues a government-wide ban on using software produced by Russia’s Kaspersky Lab. This action is a building block on the order that Homeland Security Secretary Elaine Duke wrote up last week, and now includes government and military contractors as well. Other actions and allotments included in the bill have spawned from the issues with Russia as well, and deal with cybersecurity issues and keep ahead of Russia in the arms and space race.

The Department of Defense is the only governmental agency that cannot pass a financial audit, and therefore questions arise as to why this bill was passed in the midst of the healthcare issue, as well as what the DOD will do with the almost $700 billion coming into their reserves. Regardless, virtually all of the Senate, regardless of party or views, supports the infusion of capital into the Department. Leader of the Armed Forces Committee John McCain weighed in, acknowledging that this bill is only part of the solution but that, “For too long our nation has asked our men and women in uniform to do too much with far too little,” McCain said Monday, warning that financially, “we are gambling with the lives of the best among us, and we’re now seeing the costs.” Senator McCain raises a great point, and one that can be seen in so many locations within our military. Although the number may seem staggering, the infusion into the defense budget is needed. And that’s something that the Senate can agree on, regardless of party preferences.