Journal of Behavioral and Brain Science, 2011, 1, 134-139
doi:10.4236/jbbs.2011.13018 Published Online August 2011 (http://www.SciRP.org/journal/jbbs)
Examination of Sexually Dimorphic Behavior on the
Novel-Image Novel-Location Recognition Memory Test
Brian J. Piper1,2*#, Alia L. Yasen2#, Jeremy K. Miller2
1Department of Behavioral Neuroscience, Oregon Health & Science University, Portland, USA
2Department of Psycholo gy , Willamette University, Salem, USA
Received June 16, 2011; revised July 1, 2011; accepted July 9, 2011
Abstract
Objectives: Sex differences in object location memory favoring females appear to be a replicable phenome-
non but may also depend on the task demands. This investigation evaluated if females outperformed males at
both a short (immediate) and long (half-hour) interval between the learn and test condition using a recently
developed version of the Novel-Image Novel-Location (NINL) test (Piper et al. 2011, Physiology & Behav-
ior, 103, 513 - 522). Methods: Young-adults (N = 184) completed a standardized handedness inventory and
the NINL. Results: Participants assigned to the Immediate and Delayed conditions did not differ in age, sex,
or handedness. The NINL total score was higher among females at the Immediate, but not Delayed, interval.
However, within the Delayed condition, females excelled at correctly identifying the unchanged items with a
similar pattern for the Novel-Location (NL) scale. Conclusions: These findings are consistent with the view
that sexually dimorphic performance favoring females in neurocognitive function can also extend to tasks
that have a spatial component.
Keywords: Apolipoprotein E, Female, Learning, Memory
1. Introduction
Our understanding of the domains where there are sex
differences in neurobehavioral function continues to be
expanded and refined [1]. The standard view that women
excel at verbal tasks and men at spatial tasks is likely an
oversimplification as a female advantage has also been
identified on selected measures that also have a spatial
component [2]. One area that has repeatedly been shown
as sensitive to sex differences is the Object Location
Memory (OLM) in which participants are first instructed
to learn a large set of common objects (e.g. an umbrella)
and, on subsequent trials, identify which pairs of objects
have switched positions [3,4]. The OLM has also been
expanded to include a condition in which an object is
moved to a formerly empty space. In contrast to the loca-
tion-exchange, the location-shift condition did not show
a female advantage [5], although see [6].
The Novel-Image Novel-Location (NINL) test con-
tains some conceptual similarities to the OLM but is
procedurally different as it is based on the rodent object-
recognition paradigm [7,8]. Participants view sets of
three unfamiliar pictures (learn images) and are subse-
quently asked to identify whether one of the three pic-
tures has been replaced (the Novel-Image or NI condi-
tion) or relocated (the Novel-Location or NL condition
[9]. In a broadly aged sample (6 to 86), we recently iden-
tified a significant female advantage on both NI and NL
when the test was conducted immediately after learning
the images [10]. Interestingly, the retention interval may
be a key element in the detection of sex differences [11].
Therefore, the primary objective of the present report
was to attempt to replicate and extend upon the sex dif-
ference observed in NINL using a more homogenous
aged sample (young-adults) and at different intervals. As
the NINL instrument [10] has undergone some modifica-
tions relative to an earlier version [9], a secondary goal is
to complete a more refined analysis of each item and the
correspondence with the total NINL scores.
2. Methods
2.1. Procedures
The Institutional Review Board at Willamette University
#Both authors contributed equally.

B. J. PIPER ET AL.
135
approved all procedures. Subjects consisted of college
students from a small private school receiving credit for
experimental participation. Participants completed a ques-
tionnaire that queried about their age and sex and were
asked which hand they used to complete activities. A
Laterality Index was computed by adding up the number
of activities (e.g. writing, drawing), completed with each
hand and using the formula [(R – L)/(R + L)]* 100 [12].
A saliva sample was obtained for apolipoprotein E geno-
typing according to procedures outlined in [13].
The NINL, version 0.21, was an extension of the pre-
vious NINL test [9]. The differences compared to version
0.1 were: 1) stimuli were imported into a slide show in
Microsoft Power Point to precisely regulate the display
time; 2) more detailed instructions and a practice trial so
that the test could be more readily completed in a group
setting; 3) the number of pictures was doubled to eighty
with high resolution neutral images (e.g. a dustpan) from
the International Affective Picture System [14]; and 4)
the location of the changed item balanced across the four
potential quadrants. Each slide was shown for 8.0 sec
during the learn phase. Either directly or approximately
one-half hour after viewing these slides (henceforth re-
ferred to as Immediate or Delayed), testing commenced
(eight NI, eight NL, and eight No-Change or NC). Par-
ticipants in the Delayed condition completed other dis-
tracter tasks during the interval. The quadrant of the NI
or NL was dispersed across the four quadrants with item
type (NI, NL, or NC) staggered within the Learn and
Test sets (see Table 1 for further details). Scoring con-
sisted of 0 - 3 points per NI/NL item with zero points
awarded if the subject could not identify if a change had
occurred, one point for correctly identifying that a
change had happened, two points for also recognizing the
change type (NI or NL), and three points for the previous
plus accurately identifying the quadrant of the change.
Correct identification of a NC item resulted in three
points.
2.2. Data-Analysis
All statistics were performed using SPSS, version 16.0
(SPSS Inc., Chicago, IL) with an alpha level of 0.05.
Pearson correlations were completed for each item score
with the corresponding scale (NI, NL, or NC) and with
the NINL total (NI + NL + NC). Mean data is presented
with the SEM. Cohen’s d was determined for group dif-
ferences with 0.20 interpreted as small, 0.50 as medium,
and 0.80 as a large effect size.
3. Results
The Immediate and Delayed groups did not differ based
on age, sex, or handedness (Table 2). Table 3 shows the
item to scale and item to total correlations for the 24-
item NINL instrument were all positive and significant.
The item to scale associations were similar for the NC
(Min = 0.35 to Max = 0.63), NI (0.32 to 0.61), and NL
(0.30 to 0.59) scales. The item to total NINL score cor-
relations were generally homogenous for NC (0.21 to
0.52) and NI (0.26 to 0.51). Interestingly, the first NINL
item also had the lowest correlation (r = 0.17) but values
for the remaining seven NL items were higher (r = 0.29
to 0.43). Further, the NC scale was moderately correlated
with NI (r(167) = 0.39, p < 0.0005) and NL (r(169) =
0.39, p < 0.0005) and NI and NL showed a similar asso-
ciation (r(163) = 0.37, p < 0.0005).
Figure 1(a) shows that total NINL performance dif-
fered by both sex and retention interval. Females had
higher scores than males in the Immediate (d = 0.59) but
not Delayed condition. Females scored lower at the long
interval relative to the Immediate (d = 0.47). Further
analyses for each scale (Figure 1(b)) shows that sex,
NINL difficulty, and task demands each determined rec-
ognition memory. Within the Immediate, males were
A
. Total
Immediate Delayed
40
45
50
55 Male
Female
s
i
Point s
(a)
B. Sc a le
NC NI NLNC NI NL
10.0
12.5
15.0
17.5
20.0 Male
Female
Immediate Delayed
ils
s
n
n
ln l
n
n
l
Points
(b)
Figure 1. Novel-Image Novel-Location performance total (a)
and for each scale (b) by sex and retention interval. NC:
No-Change; NI: Novel-Image; NL: Novel-Location; sp <
0.05 sex difference within an interval; ip < 0.05 interval
differences versus delayed; np < 0.05 scale difference versus
NC; lp < 0.05 scale difference versus NL.

B. J. PIPER ET AL.
136
significantly lower on NI (d = 0.59) but did not differ
significantly on NC (t(104) = 1.94, p = 0.055, d = 0.38)
from females. In the Delayed, females outperformed
males on NC (d = 0.50) and showed a similar tendency
for NL (t(72) = 1.99, p = 0.051, d = 0.46). The NL was
lower than both NC and NI independent of sex and in-
terval.
4. Discussion
This report contributes to a very large and evolving lit-
erature on sex differences in neurocognitive function [1].
Clearly, the identification and direction of group differ-
ences based on sex depends on the sample size and na-
ture of the task employed including how it is scored [2].
Males outperform females on the mental rotation test
[4,15] and exhibit almost equally large group differences
on spatial navigation measures [9,10,16]. In contrast, less
is definitely known about other domains. Although the
NINL total was elevated among females only at the im-
mediate interval, examination of each scale revealed a
moderate effect size (d ≈ 0.50) for NI at the shorter pe-
riod and on both NL and identifying the unchanged items
at the longer period. This finding is broadly concordant
with our previous study with a lifespan sample [10] and
in line with a prior investigation with the OLM in which
college aged females scored higher than males on iden-
tifying drawings of new objects (d = 0.46) and location
exchanges (d = 0.44, [3]. However, in the location shift
of the OLM (the condition most similar to the NL), prior
results have been contradictory with some [6], but not all
[5], investigations identifying better performance among
women.
In the OLM, participants view the original learn array
and then, during repeated testing, may view elements of
this same array again which makes these conditions dif-
ficult to compare directly. As Levy et al. [6] noted, this
design could also introduce proactive interference. An
advantage of the NINL is that this instrument has differ-
ent learn sets that are subsequently tested in the NI, NL,
or NC. Rodents find the location test much more chal-
lenging than the novel-object test [7]. Similarly, human
participants did significantly less well on NL compared
to NI at both intervals indicating that this is a robust
phenomenon across species. The neural substrates that
mediate these task specific processes have not yet been
characterized on NINL but investigations with rats have
shown that lesions of the perirhinal cortex impact object
recognition memory [8]. It is tempting to speculate that
location recognition would be more dependent on the
dorsal stream (i.e. where) and image recognition on the
dorsal ventral stream (i.e. what) structures although the
integration of both occipto-parietal and occipito-temporal
regions may also be essential [17].
The interval between the learn and test phases is a key
element in task difficulty [18]. Adult female rats showed
object-recognition at intervals (1 - 3 hours) that males
could not. Conversely, males were capable of identifying
when an object had been moved to a new position at
several intervals that were too difficult for females [11].
A potential limitation is that only two intervals were
examined in the present study. A slightly longer (1 hour)
delay was evaluated during pilot testing and found to be
unfeasible for this participant population. A comparison
between an immediate and a 24-hour retention period
produced the anticipated reduction at the longer-interval
in a separate all-female sample [Thornburg & Murphy,
unpublished observations] but a multi-interval exami-
nation of sex differences should be a topic for further re-
search.
A fundamental characteristic of any cognitive test is
the type of the material to be learned. In addition to other
caveats, this factor may also limit the generalizability be-
tween rodent and human recognition memory tests. In-
vestigators utilizing laboratory animals can quite easily
use objects that their subjects have never before experi-
enced [18]. This frequently takes the form of children’s
toys [7] or commonplace household objects [19]. The
NINL learn set (Table 1) consists of pictures from a da-
tabase maintained for research purposes [14]. However,
the images are of objects or scenes that can readily be
labeled. Although the magnitude of sex differences in
generalized verbal abilities is a matter of some conten-
tion [20], we suspect that females were more likely to
employ verbal strategies to facilitate a deeper encoding of
the image sets. Therefore, it is quite interesting that when
abstract line drawings (which would limit the potential to
label) are used as stimuli, there is conflicting data whether
a female advantage is still detected [21-23]. Further, the
Design Memory subtest of the Weschsler Memory Scale-
IV includes a topographically more complicated NL type
element with geometric shapes and abstract stimuli and
sex differences are very minimal (personal communica-
tion from JM Laurer).
There are several future directions that may be worthy
of some consideration for investigators interested in in-
dividual differences in NINL performance. A fascinating
report determined that homosexual males substantially
outperformed heterosexual males on Object-Location
Memory [24]. Additional study with a more ethnically
and socioeconomically diverse sample in which other in-
formation (e.g. sexual orientation, IQ, EEG) is obtained
may prove fruitful as these factors could contribute to the
present neurobehavioral profile.
Overall, there are clear sex differences on NINL which,
when analyzed at the level of each scale, are independent

B. J. PIPER ET AL.
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Table 1. Images for the Novel-Image, Novel-Location test, Learn (L) and Test. Image numbers (in parentheses) are from the
International Affective Picture System ([14]). Quadrants are north-west (NW), north-east (NE), south-east (SE) and
south-west (SW) with empty quadrants depicted by a dash (-). Type is Novel Image (NI), Novel Location (NL) or No Change
(NC).
Learn Test
Slide Quadrant Slide
# NW NE SW SE # Basis Type Description
L01 - outlet (6150) mushroom (5531) train (7039) T01L07 NL car (8531) to NE
L02 dust pan (7040) pocket watch (7190) - towel (7002) T02L16 NC
L03 coffee cup (7057) dark cloud (5594) cube (7185) - T03L04 NI clock (7211) at SE
L04 hammer (7034) book (7090) - abstract painting (7830) T04L11 NI crimps (7056) at NW
L05 yard (5130) - shoes (7031) woven basket (7010) T05L10 NC
L06 set table (5849) canyon (5661) - row boats (5390) T06L24 NL pins (7052) to NW
L07 hair dryer (7050) - convertible car (8531)fan (7020) T07L05 NC
L08 - leaves (5750) fork (7080) parking lot (7595) T08L17 NI stool (7025) at SW
L09 fireworks (5480) - unlit light bulb (7055)waste can (7060) T09L18 NC
L10 - satellites (5471) blue cup (7009) headlight (7095) T10L02 NL pocket (7190) to SW
L11 rolling pin (7000) bus (7140) tiger lilly (5030) - T11L08 NI clothes (7242) at SW
L12 drill (7043) latch (7059) - Native Amer pattern (7179)T12L03 NL cube (7185) to SE
L13 blue door (5731) bridge (7547) pink flower(1604) - T13L06 NC
L14 - power lines (9080) spoon (7004) abstract painting (7161) T14L15 NL bulb (7170) to SE
L15 lit bulb (7170) ship (5395) large baskets (7041)- T15L19 NC
L16 orchid (5010) - leaves (5740) empty pool (9360) T16L09 NI rack (7217) at NW
L17 earth (5890) mountain top (5660) airplane (7620) - T17L23 NI field (5250) at NW
L18 - white bowl (7006) tissue (7950) sports car (8510) T18L22 NC
L19 clear glass (7035) hydrant (7100) gold bars (8500) - T19L13 NC
L20 - ferris wheel (7508) orange flower (5020)file cabinet (7705) T20L20 NL wheel (7508) to NW
L21 dumbells (7042) - lamp (7175) yellow sail-boat (8210) T21L21 NI universe (5300) at SE
L22 shoes (7038) scarf (7205) building (7491) - T22L12 NL Native (7179) to SW
L23 flowers (5000) semi-truck (7130) - freeway (7560) T23L14 NI plate (7233) at NE
L24 - clothes pins (7052) umbrella (7150) snow day (5635) T24L01 NL train (7039) at NE
of the retention interval. The cognitive (e.g. attention or
encoding) or biological (e.g. distinct neural substrates)
mechanisms responsible for the female advantage on this
and other similar tasks will be the subject of additional
study. A strength of the NINL is that this procedure is
based on the rodent object recognition test which may
aid in translating the substantial knowledge base from
rodents to humans.
5. Acknowledgements
Special thanks to Jacob Raber, PhD and Mark Stewart,
PhD for their support and to Matthew B. Herson, Donna
M. Nolan, Hannah M. Gandsey, Caitlin St. John, Casey

B. J. PIPER ET AL.
138
Table 2. Participant characteristics by Novel-Image Novel-
Location retention interval. apoE4: Apoliprotein E4 allele.
Immediate (N = 106) Delay (N = 78)
Age 19.0 (0.1) 18.7 (0.1)
Sex (% Female) 61.3% 53.8%
apoE4+ 21.7% 17.9%
Laterality Index 0.23 (0.06) 0.33 (0.07)
Table 3. Pearson correlations for each item with the scale
and total score (in parentheses) for the Novel-Image Novel-
Location test. The p value was ≤ 0.001 for all correlations
except ap < 0.05.
Item No Change Novel-Image Novel-Location
1 0.51 (0.41) 0.61 (0.46) 0.30 (0.17a)
2 0.50 (0.40) 0.32 (0.33) 0.46 (0.43)
3 0.45 (0.21) 0.39 (0.25) 0.59 (0.41)
4 0.48 (0.36) 0.50 (0.31) 0.53 (0.35)
5 0.57 (0.39) 0.42 (0.26) 0.51 (0.38)
6 0.35 (0.28) 0.58 (0.51) 0.51 (0.37)
7 0.63 (0.52) 0.42 (0.32) 0.50 (0.43)
8 0.41 (0.35) 0.60 (0.47) 0.47 (0.29)
Conzatti, and Alan Curtis for assistance in data collection.
Some of this data is also contained in the honor’s thesis
of ALY. This work was supported by the National Insti-
tute of Environmental Health Sciences (T32 ES007060-
31A1), and the National Institute of Drug Abuse (T32DA
07262 & L30 DA027582-01).
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