Psychophysiology, 39 ~2002!, 409–413. Cambridge University Press. Printed in the USA.

Copyright © 2002 Society for Psychophysiological Research

DOI: 10.1017.S0048577202394010

Variability of sleep parameters across multiple

laboratory sessions in healthy young subjects:
The “very first night effect”

JOSÉ-LUIS LORENZO a and MANUEL-JOSÉ BARBANOJ a,b

a
Àrea d’ Investigació Farmacològica, Institut de Recerca de l’ Hospital de la Santa Creu i Sant Pau, Barcelona, Spain
b
Departament de Farmacologia i Terapèutica, Universitat Autónoma de Barcelona, Spain

Abstract
Many studies have been carried out to assess the variability of sleep parameters. The first night effect is one of the most
important factors in this variability and has been extensively studied. However, the readaptation phenomenon when
subjects returned to the sleep laboratory after spending a certain period of time at home has been not systematically
evaluated. To investigate this phenomenon across multiple sleep laboratory sessions, polysomnographic data from 12
healthy young subjects for 12 nights ~three periods each of 4 consecutive nights, with a minimum of 1 month between
them! were collected. The first night effect was present only in the first night of the first period ~“very first night”! and
was significant only for REM sleep-related variables. We conclude that the results from the first nights of consecutive
periods within a specific protocol with healthy young subjects need not be discarded in subsequent analyses.
Descriptors: First night effect, Readaptation effects, Polysomnography, Healthy subjects

Temporal stability is an important factor when measuring sleep
variables. Sleep measures used for clinical or research analysis
should demonstrate the fundamental property of stability over time
~Wohlgemuth, Edinger, Fins, & Sullivan, 1999!. Many studies
have been performed to assess the stability of sleep variables
~Botzin et al., 1995; Clausen, Sersen, & Lidsky, 1974; Greenberg
& Pearlman, 1976; Hall, Dahl, Al-shabbout, Trubnick, & Ryan,
1998; Hoch, Reynolds, Kupfer, & Berman, 1988; Moses, Lubin,
Naitoh, & Johnson, 1972; Schmidt & Kaelbling, 1971; Wohlge-
muth et al., 1999!, and other factors such as sleep cycles trends
~Feinberg & Floyd, 1979! or nocturnal motor activity ~Kronholm,
Alanen, & Hyyppä, 1987!. Most studies agree with the need to
average each variable over multiple nights to reduce the random,
unpredictable fluctuations from one night to the next. One impor-
tant factor in such variability is the so-called “first night effect”
The authors thank C. Martínez and F. Segarra ~Unitat Alteracions del
Son, Institut Dexeus, Barcelona! and O. Romero ~Servei Neurofisilogia,
Hospital Vall d’Hebrón, Barcelona! for their scoring assistance, L. Da
Graça for her technical assistance, I. Gich for his advice on statistical
procedures, and J. L. Aldirez and A. Funes for their help with computer
assistance. The latter four individuals are all from the Àrea Investigació
Farmacològica, Institut de Recerca de l’Hospital de la Santa Creu i Sant
Pau.
A portion of these results were presented at the 14th European Con-
gress of Sleep Research held in Madrid ~Spain!, September 9–12, 1998.
Address reprint requests to: M.-J. Barbanoj, Àrea Investigació Farma-
cològica, Institut de Recerca, Hospital de la Santa Creu i Sant Pau, Av Sant
Antoni M. Claret, 167, 08025 Barcelona, Spain. E-mail: mbarbano@
santpau.es.
~FNE!. The FNE is a well-known phenomenon mainly character-
ized by lower sleep efficiency, increased wakefulness, reduction in
the amount of rapid eye movement ~REM! sleep, and a longer
sleep and REM sleep latencies ~Agnew, Webb, & Williams, 1966!.
It seems that the major factors responsible for this effect are the
unfamiliar surroundings of the sleep laboratory and the subjects’
adaptation to the application of several instruments to their body,
mainly electrodes ~Agnew et al., 1966; Mendels & Hawkins,
1967!. Data obtained from the first night are thus discarded and not
included in assessing a subject’s sleep. These trials consequently
become long and expensive. This phenomenon has been replicated
in many studies with healthy participants ~Agnew et al., 1966;
Browman & Cartwright, 1980; Schmidt & Kaelbling, 1971; Tous-
saint et al., 1997! and psychiatric patients ~Mendels & Hawkins,
1967; Toussaint et al., 1995, 2000; Woodward, Bliwise, Friedman,
& Gusman, 1996!. Most of them are based on several consecutive
nights of recording for each subject. Age, type of participant, and
the setting where the recording is done are the most important
factors involved not only in this phenomenon but in the variability
of sleep parameters across multiple laboratory nights.
Whereas the FNE on sleep laboratory trials has been well
studied, the possibility of readaptation effects when studies are in
interrupted form has not been systematically evaluated. These
effects, if they exist, are crucial because many designs require
subjects to sleep in the laboratory on several nonconsecutive nights
~Wyatt et al., 1995!. To assess the presence of readaptation effects
when trials are designed in several discontinuous periods, poly-
somnographic recordings of 12 healthy young subjects were ob-
tained over three periods of 4 consecutive nights, with a minimum
of 1 month between them.

409
410
Methods
Participants
Twelve healthy participants ~7 men, 5 women! ranging from 19 to
27 years of age ~mean age 25! were recruited for the study. All of
them completed the entire protocol. They had an extensive phys-
ical and psychological evaluation prior to participation and had
regular sleep–wake habits ~mean total score of Pittsburgh Sleep
Quality Index: 2.3, range: 0– 4!. They were requested to adhere to
a regular sleep–wake pattern during the month prior to study entry
~bedtime within the 10:00 to 12:00 p.m. time range, wake time
within 07:00 to 09:00 a.m. time range!, not to take any medication
in the month before and during the study, to refrain from napping
during day time prior to each night of monitoring and to reduce
their caffeine ~no more than two cups of coffee per day! and
alcohol consumption ~mean 24 g0day!. In addition, caffeine and
alcohol were not allowed after lunch on the days when the record-
ings were performed. Moderate smoking was permitted during the
study ~maximum 10 cigarettes0day!. Written informed consent
was obtained and the study was performed in accordance with the
rules and regulations for the performance of clinical trials stated by
the World Medical Assembly of Helsinki and subsequent updates.
The study protocol was supervised by the ethical committee of our
institution. Participants were paid for their participation.
Design
Polysomnograms were recorded at all three periods. Each period
consisted of 4 consecutive nights, with a minimum of 1 month
between them ~range 29–35 days!. Each participant arrived at the
laboratory at 10 p.m. and slept in a comfortable individual sound-
attenuated bedroom, under the supervision of nursing staff. Lights
out was around 11 p.m. and lights on around 7 a.m. The recording
lasted approximately 8 hr.
Recordings and Sleep Stage Classification
Data were recorded by means of a 16-channel digital polygraph
~Jaeger Sleep Lab. 1000P! including two electroencephalographic
leads ~C3-A2, C4-A1, according to the 10020 International Sys-
tem!, two electrooculographic leads ~left eye-A2, right eye-A1!,
and one chin electromyographic channel. To monitor the respira-
tory function during the recording, three channels were included:
one for airflow signal ~by means of a thermistor placed in the path
of airflow from nose and mouth! and two channels to record the rib
cage and abdominal motion ~by calibrated transducers!. Finally,
two channels ~linked electrodes on the left and right anterior
tibialis! were used to register limb movements. EEG and EOG
channels were filtered to a bandwidth of 0.1–75 Hz with a sensi-
tivity of 10 mV0mm. EMG was filtered to a bandwidth of 10–
75 Hz with a sensitivity of 5 mV0mm. A 50-Hz notch filter was
used to attenuate electrical noise. The electrodes were gold plated.
Channels were calibrated prior to each recording and the electrode
impedance was kept below 10 kV.
Sleep stages were scored visually and independently by two
expert scorers in 30-s epochs according to Rechtschaffen and
Kales’ ~1968! criteria. Experts were blind in relation to position of
the recording in the time sequence. Interexpert variability was
calculated in the laboratory based on the present records and
indicated an average agreement rate superior to 80%. The final
quantification was thus chosen at random between the two scorers.
All of the sleep variables were derived from the visual scoring
of recordings using standard criteria and were divided into two
groups: sleep continuity indices and sleep architecture indices. In
J.-L. Lorenzo and M.-J. Barbanoj
addition, the exact total time in bed was measured. Sleep continu-
ity indices included total sleep time, sleep efficiency ~time spent
asleep divided by the time in bed!, sleep onset latency ~the time
from lights out to the first epoch in stage 2!, delta sleep latency ~the
time from the first epoch in stage 2 to the first epoch in stage 3!,
REM sleep latency ~the time from sleep onset to the first epoch in
stage REM!, number of arousals ~scored according to the rules of
Atlas Task Force of the American Sleep Disorders Association,
1992!, number of epochs classified as movement time, and number
of awakenings. Sleep architecture indices included percentage of
time spent asleep in the different stages: awake, stage 1, stage 2,
REM, and delta sleep ~the sum of sleep stages 3 and 4!; and also
the mean duration of REM sleep and the mean duration of NREM0
REM cycles.
Statistical Analysis
The following procedures were applied to sleep variables under
study: A multivariate, nonparametric approach to obtain a global
vision of results was applied ~Saletu, Grunberger, & Linzmayer,
1986!. Time-effect relations were evaluated by means of Fried-
man’s rank ANOVA and Wilcoxon tests of 15 combined polysom-
nographic variables that have been identified as principal variables
modified by the first night effect ~Agnew et al., 1966!, whereby a
higher Friedman’s rank means a higher first night effect, which
means worse sleep quality, considering: decreased total sleep time,
decreased sleep efficiency, increased sleep onset latency, increased
REM sleep latency, increased delta sleep latency, increased num-
ber of awakenings, increased number of arousals, increased num-
ber of epochs considered as movement time, increased percentage
of stage 1, increased percentage of stage 2, decreased percentage
of REM sleep, decreased percentage of delta sleep, decreased
mean duration of REM sleep and increased mean duration of
NREM0REM cycles.
To assess individually which variables were significantly af-
fected by the first night effect, two-way ANOVAs for repeated
measures ~month: three levels! and ~day: four levels! were applied.
Any p # .05 was considered significant.
Results
Mean values of sleep parameters obtained over three periods of 4
consecutive nights in the group of 12 healthy young subjects are
presented in Table 1. Sleep efficiency presented a mean ratio of
91.14%, with mean total sleep time of 431.62 min. Mean sleep
onset latency was 24.17 min and mean delta sleep latency, 32.09.
Mean REM sleep latency was 85.88, with mean duration of REM
sleep, 24.69 min. The average length of NREM0REM cycles was
108.07 min. The mean percentage of different sleep stages was:
wake 5.42%, stage 1 5.05%, stage 2 53.53%, delta sleep 13.95%,
and REM sleep 21.21%. Finally, subjects showed an average of
3.98 awakenings, 31.05 arousals, and 6.09 epochs scored as move-
ment time.
The relation between time and effect calculated by means of
Friedman’s rank of 15 combined sign-controlled polysomno-
graphic parameters showed a clearly significant difference be-
tween all nights evaluated, x 2 ~11! 5 52.19, p , .00001. The
subsequent application of Wilcoxon tests indicated that only the
first night of the first month showed significant differences, when
it was pairwise compared with all the other nights. There was thus
a highly significant difference between the 12 nights with the first
night showing the highest rank, that is, lowest sleep quality. No
significant effect was found when the remaining nights were pair-
The “very first night effect” 411

Table 1. Sleep Parameters (Mean 6 SEM) Obtained for Three Periods of 4 Consecutive Nights in a Group of 12
Healthy Young Volunteers

Day 1 Day 2 Day 3 Day 4

Month 1
Time in bed ~min! 469.33 6 3.25 461.33 6 5.62 477.17 6 2.32 484.50 6 7.98
Sleep continuity indices
Total sleep time ~min! 400.58 6 25.98 427.83 6 10.67 438.83 6 14.98 431.75 6 16.69
Sleep efficiency ~%! 85.56 6 5.61 92.67 6 1.67 91.93 6 3.02 89.22 6 3.45
Sleep onset latency ~min! 32.83 6 7.07 20.88 6 3.74 22.21 6 6.62 30.04 6 12.44
Delta sleep latency ~min! 44.63 6 9.57 24.42 6 5.00 26.50 6 5.88 27.50 6 9.18
REM sleep latency ~min! 106.75 6 14.72 90.38 6 10.24 100.63 6 12.86 68.42 6 8.08
Number of arousals 26.25 6 4.14 33.83 6 5.16 34.33 6 4.78 33.00 6 4.76
Number of movements 6.58 6 1.09 6.58 6 1.78 5.75 6 1.21 7.00 6 1.21
Number of awakenings 5.33 6 2.20 3.75 6 1.45 4.25 6 2.25 4.17 6 1.59
Sleep architecture indices
% Awake 10.69 6 5.13 4.23 6 1.64 4.96 6 2.10 5.18 6 2.32
% Stage 1 5.05 6 4.99 5.22 6 1.31 6.48 6 1.14 6.82 6 1.06
% Stage 2 55.33 6 4.35 53.86 6 2.51 50.48 6 1.45 50.28 6 3.08
% Delta sleep 12.80 6 2.22 15.18 6 2.34 16.67 6 1.66 15.71 6 1.53
% REM sleep 16.35 6 1.23 21.33 6 1.35 21.28 6 2.06 21.93 6 1.78
Mean duration REM sleep ~min! 22.64 6 1.74 25.34 6 1.52 26.80 6 2.70 23.68 6 2.05
Mean duration NREM0REM cycle ~min! 139.77 6 11.07 110.61 6 3.73 117.01 6 6.58 100.99 6 5.15
Month 2
Time in bed ~min! 466.50 6 4.27 469.50 6 5.55 473.50 6 3.89 459.00 6 12.79
Sleep continuity indices
Total sleep time ~min! 434.92 6 15.77 447.08 6 9.40 433.75 6 10.84 405.83 6 18.88
Sleep efficiency ~%! 93.05 6 2.89 95.17 6 1.37 91.61 6 2.15 88.74 6 3.80
Sleep onset latency ~min! 14.13 6 3.89 18.33 6 2.97 24.33 6 8.26 26.17 6 5.24
Delta sleep latency ~min! 23.83 6 4.16 30.58 6 6.29 30.29 6 7.33 52.38 614.10
REM sleep latency ~min! 85.58 6 8.39 76.08 6 8.13 89.50 610.96 87.58 6 9.53
Number of arousals 29.58 6 4.65 30.25 6 2.51 30.58 6 3.87 24.58 6 3.38
Number of movements 5.75 6 1.61 5.58 6 1.08 5.08 6 0.91 6.92 6 1.68
Number of awakenings 3.50 6 1.51 3.42 6 1.24 4.67 6 2.13 6.50 6 2.27
Sleep architecture indices
% Awake 4.62 6 2.07 2.79 6 1.19 5.44 6 1.86 8.40 6 3.58
% Stage 1 5.57 6 0.79 5.93 6 0.73 4.75 6 0.75 7.19 6 1.92
% Stage 2 52.87 6 1.83 53.99 6 2.08 54.80 6 2.41 52.33 6 2.53
% Delta sleep 14.90 6 1.96 13.91 6 1.65 13.01 6 1.86 11.00 6 1.43
% REM sleep 21.98 6 1.46 23.23 6 1.68 21.88 6 1.63 20.84 6 2.77
Mean duration REM sleep ~min! 25.30 6 1.77 26.08 6 2.00 25.01 6 1.74 22.73 6 2.25
Mean duration NREM0REM cycle ~min! 105.60 6 2.86 106.68 6 6.90 103.04 6 4.47 101.78 6 5.19
Month 3
Time in bed ~min! 481.75 6 8.56 480.67 6 3.62 476.42 6 9.52 476.50 6 4.31
Sleep continuity indices
Total sleep time ~min! 441.33 6 17.71 448.42 6 9.87 433.50 6 13.99 435.67 6 12.08
Sleep efficiency ~%! 91.61 6 3.26 93.35 6 2.13 90.92 6 2.26 91.43 6 2.40
Sleep onset latency ~min! 21.33 6 6.35 22.87 6 4.43 23.46 6 4.82 33.54 6 11.47
Delta sleep latency ~min! 37.50 612.61 20.87 6 1.97 35.71 6 9.01 30.88 6 8.68
REM sleep latency ~min! 80.00 6 7.26 71.54 6 8.24 104.83 6 11.22 69.79 6 8.99
Number of arousals 28.83 6 4.09 32.25 6 3.01 32.75 6 4.42 36.42 6 5.91
Number of movements 6.92 6 1.26 6.17 6 1.08 5.42 6 0.84 5.42 6 1.17
Number of awakenings 3.92 6 1.38 2.50 6 0.86 3.58 6 1.23 2.25 6 0.45
Sleep architecture indices
% Awake 6.62 6 3.06 3.45 6 2.09 5.10 6 2.18 3.63 6 1.02
% Stage 1 5.11 6 0.56 5.11 6 0.84 5.93 6 1.08 6.23 6 0.88
% Stage 2 57.53 6 2.89 54.21 6 2.11 52.14 6 2.79 54.65 6 2.68
% Delta sleep 12.73 6 2.03 13.07 6 6.95 14.87 6 1.51 12.99 6 1.73
% REM sleep 17.83 6 1.81 23.43 6 1.31 21.87 6 1.59 22.61 6 1.12
Mean duration REM sleep ~min! 22.84 6 2.89 25.79 6 1.57 24.37 6 1.77 25.81 6 1.97
Mean duration NREM0REM cycle ~min! 106.31 6 6.14 95.13 6 8.28 107.21 6 5.65 102.74 6 4.86

wise compared ~Figure 1!. However, it is interesting to note that
although not statistically significant, a tendency of the third and the
fourth nights on the second and the third month to display a
relative increase of Friedman’s rank, that it is lower sleep quality,
in comparison to the second night, was evidenced.
The univariate analysis showed a significant effect only in
REM sleep percentage, factor day: F~3,30! 5 8.40, p , .0001,
REM sleep latency, factor day: F~3,30! 5 3.66, p 5 .023, and
mean duration of REM sleep, month0day interaction: F~6,60! 5
3.10, p 5 .016. These findings are consistent with the assumption
412
’
Figure 1. Means of Friedman’s rank applied to 15 combined polysomno-
graphic variables that have been identified as modified by the first night
effect. A higher value of Friedman’s rank means a higher first night effect,
that is, decreased total sleep time, decreased sleep efficiency, increased
sleep onset latency, increased REM sleep latency, increased delta sleep
latency, increased number of awakenings, increased number of arousals,
increased number of epochs scored as movement time, increased stage 1
percentage, increased stage 2 percentage, decreased REM sleep percentage,
decreased delta sleep percentage, decreased mean duration of REM sleep
and increased mean duration of NREM0REM cycles. Significant differ-
ences were observed on the first night of the first month only ~comparison
of Night 1.1 with 1.2**; 1.1 vs. 1.3**; 1.1 vs. 1.4**; 1.1 vs. 2.1**; 1.1 vs.
2.2**; 1.1 vs. 2.3**; 1.1 vs. 2.4*; 1.1 vs. 3.1*; 1.1 vs. 3.2**; 1.1 vs. 3.3**;
1.1 vs. 3.4**, where ** means Z . 2.58, p , .01; * means Z . 1.96, p ,
.05; after Wilcoxon Test!.
that the very first night effect is mainly due to variations in REM
sleep, a higher REM sleep latency and a lower mean duration of
REM sleep as compared to the remaining nights evaluated.
Discussion
Mean values of sleep parameters obtained across the study agree
with the normative data of previous studies in young subjects
without pathology ~Carskadon & Dement, 1994! and confirm the
presence of a first night effect in laboratory sleep recordings in
healthy young subjects. These results are consistent with previ-
ously published data ~Agnew et al., 1996; Schmidt & Kaelbling,
1971; Toussaint et al., 1995; Wyatt et al., 1995!. Even though the
first night effect is mainly characterized by a lower sleep effi-
ciency, increased wakefulness, longer REM sleep latency and a
reduction of the amount of REM sleep ~Agnew et al., 1966!, the
REM sleep-related variables ~percentage, latency, and mean dura-
tion of REM sleep! appear as the most sensitive parameters im-
plicated in the adaptation process to the laboratory conditions
across nights in our study. Coble, McPartland, Silva, and Kupfer
~1974! did not find significant differences between the first and
second night and attributed this to the “super” normal subjects
~with an extremely low level of psychopathology! selected in their
study and to the maximal comfortable conditions of the laboratory
where the polysomnography was performed. However, they con-
cluded that the first night effect does not always exist and depends
mainly on the environmental conditions. Even so, REM sleep
latency was the only parameter with significant relevance across
nights. Browman and Cartwright ~1980! reached a similar conclu-
sion. When they compared the sleep measures on Night 1 with
J.-L. Lorenzo and M.-J. Barbanoj
those on subsequent nights, all differences between means were
nonsignificant in a comfortable environment. They concluded that
the subjects’ characteristics and familiar setting ~comfortable hotel-
type laboratory! can reduce but not fully eliminate the first night
effect on sleep studies, inasmuch as they found consistent changes
in percentage of REM sleep and sleep efficiency. Sharpley, Solo-
mon, and Cowen ~1988! demonstrated that the FNE is not dis-
played when healthy participants sleep at home. In a study performed
to evaluate the influence of some factors ~e.g., sex and age! on the
sleep variables’ adaptation across nights, Schmidt and Kaelbling
~1971! demonstrated that only REM sleep parameters revealed
statistical significance, not only with age as independent variable,
but also in the interaction of age and night. Our study, performed
on healthy, young subjects in a comfortable environment, agrees
with the above-mentioned results. Even though reduced, the first
night effect is still present, and it is represented by changes in the
REM sleep characteristics. This therefore emphasizes the great
fragility of REM sleep referred to by earlier investigators, which
seems to constitute the first line of defense in the attempt to
stabilize delta sleep and to maintain cortical homeostasis in the
face of changing external influences ~Ephron & Carrington, 1966;
Schmidt & Kaelbling, 1971!.
Although the FNE is a well-studied phenomenon, the possibil-
ity of readaptation effects when subjects return to the sleep labo-
ratory following several nights away has not been extensively
evaluated. These effects are critical in sleep laboratory trials. Kales
and Kales ~1970!, in their study to establish methodological rec-
ommendations for sleep laboratory drug evaluation studies, em-
phasized the importance of this phenomenon when the results are
evaluated. In a study specifically intended to evaluate the readap-
tion effects in insomniac subjects, Scharf, Kales, and Bixler ~1975!
demonstrated the presence of this phenomenon when the subjects
returned to the laboratory for a second series of consecutive nights
after spending 1 week at home. Wyatt et al. ~1995! evaluated these
effects in older adults across multiple testing nights. Their results
showed that after the first night in the laboratory, some parameters
normalized, whereas others ~i.e., percentage of stage 1 and REM
sleep! still presented a FNE during subsequent pairs of nights. Our
principal contribution to the study of this phenomenon is the
finding that the first night effect is present only on the first night
of the whole study ~the very first night! and it does not persist
during the remaining first nights of the subsequent periods, even
when the study is interrupted by a minimum of 1 month. Age and
neuropsychological disturbances appear to be the factors most
related with the sleep laboratory adaptation. Children and elderly
persons need a much more prolonged adaptation to the laboratory
than young adults ~Schmidt & Kaelbling, 1971!. Results obtained
from studies with depressed patients indicated a greatly reduced
first night effect with respect to normal subjects ~Kupfer, Weiss,
Detre, & Foser, 1974!. It has been suggested that the first night
effect is more than a mere laboratory artefact and rather represents
the constitutional age-related functional adaptability of the central
nervous system ~Toussaint et al., 1995!. It is therefore possible that
in healthy young adults, the adaptation phenomenon after the very
first night persists across nights, even if the study is in an inter-
rupted form.
In conclusion, our study corroborates previous findings that
the first night effect, even if reduced in size, is still present
when the study is performed with healthy, young subjects in an
environment with maximal comfort. The sleep REM-related pa-
rameters are those most affected by this effect. Also, the finding
that the first night effect was present only on the very first night
The “very first night effect”
of the study suggests that the data obtained on the consecutive
first nights of interrupted trials should not be discarded and
could be used in the final analyses. In addition, it would seem
reasonable to consider the presence of the FNE in the sub-
sequent periods of investigation in young subjects as a sign of
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~Received March 14, 2001; Accepted September 27, 2001!