Original Paper

Neuropsychobiology 2010;61:27– Received: August 29, 2008

32 Accepted after revision: July 20, 2009
DOI: 10.1159/000258640 Published online: November 13, 2009

Urinary Excretion of 5-Hydroxyindoleacetic

Acid, Serotonin and 6-Sulphatoxymelatonin
in Normoserotonemic and Hyperserotonemic
Autistic Individuals
Erik J. Muldera George M. Andersonc Ramses F.J. Kempermanb
Alida Oosterloo-Duinkerkenb Ruud B. Minderaaa Ido P. Kemab
Departments of a Child and Adolescent Psychiatry and b Pathology and Laboratory Medicine, University Medical
Center Groningen, Groningen, The Netherlands; c Child Study Center, Yale University School of Medicine,
New Haven, Conn., USA

Key Words = 0.061) and 5-HT (p = 0.071) and a significant decrease

Autism · Serotonin · 5-Hydroxyindoleacetic acid ·
Melatonin · Gut
Abstract
Objective: A substantial proportion of individuals with
au- tism have elevated levels of platelet serotonin (5-HT).
We ex- amined whether platelet hyperserotonemia is
associated with increased gut 5-HT synthesis, altered 5-
HT catabolism or altered melatonin production.
Tujuan: Sebagian besar orang dengan autisme memiliki
kadar serotonin (5-HT) yang tinggi. Kami memeriksa
apakah platelet hiperserotonemia dikaitkan dengan
peningkatan sintesis 5-HT usus, mengubah katabolisme 5-
HT atau mengubah produksi melatonin.
for 6- SM were found (p = 0.027). The urinary 5-HIAA:5-HT
ratio was similar in the normo- versus the
hyperserotonemic groups.
Hasil: Pada kelompok hiperserotonemia,
peningkatan yang signifikan pada tingkat tren
dalam ekskresi urin 5-HIAA (p = 0,061) dan 5-HT (p =
0,071) dan penurunan signifikan untuk 6- SM
ditemukan (p = 0,027) . Rasio 5-HIAA kemih: 5-HT
serupa pada kelompok normo dibandingkan
kelompok hiperserotonemia.
Conclusions: The catabolism of 5-HT does not differ in
the groups, but greater exposure of the platelet to 5-HT
cannot

Methods: Urinary excre- tion of 5-hydroxyindoleacetic

acid (5-HIAA) and 5-HT was compared in 10
normoserotonemic and 10 hyperserotone- mic age-
matched autistic individuals. The relationship of uri- nary 6-
sulfatoxymelatonin (6-SM) excretion to platelet 5-HT, and
to urinary 5-HT and 5-HIAA excretion, was also exam-
ined.
Metode: Ekskresi asam 5-hydroxyindoleacetic (5-HIAA)
dan 5-HT dalam urin dibandingkan dalam 10 individu
autistik yang disesuaikan dengan usia normoserotonemia
dan 10 hyperserotone-mic. Hubungan ekskresi 6-
sulfatoxymelatonin (6-SM) uri dengan platelet 5-HT, dan
dengan ekskresi 5-HT dan 5-HIAA, juga diperiksa.

Results: In the hyperserotonemic group, significant in-

creases at trend level in urinary excretion of 5-HIAA (p
 be ruled out as a cause of the platelet
hyperserotonemia of autism. Although only trend level
significant, the data point to a need for larger studies to
examine more thoroughly the relationships between
platelet hyperserotonemia, gut 5-HT synthesis and
melatonin production.
Kesimpulan: Katabolisme 5-HT tidak berbeda pada
kelompok, tetapi paparan platelet yang lebih besar
terhadap 5-HT tidak dapat dikesampingkan sebagai
penyebab hiperserotonemia trombosit autisme.
Meskipun hanya tingkat tren yang signifikan, data
menunjukkan perlunya penelitian yang lebih besar
untuk memeriksa lebih teliti hubungan antara
hiperserotonemia trombosit, sintesis usus 5-HT dan
produksi melatonin
Introduction
Elevated levels of serotonin (5-hydroxytryptamine,
5- HT) [1] in blood platelets of individuals with autism
is one of the most well-replicated findings in biological
in the control groups. The possible bimodality of
platelet 5-HT in autism may permit improved strategies
for elu- cidating the mechanisms underlying the
elevation.
Baru-baru ini, kami mengamati distribusi bimodal dari
plate-let 5-HT pada individu Belanda yang didiagnosis
dengan gangguan spektrum autisme [4]. Tampak bahwa
sekitar setengah dari individu dengan gangguan
spektrum autisme memiliki nilai trombosit 5-HT yang
terdistribusi dalam mode atas, sementara mode yang
lebih rendah sebagian besar tumpang tindih dengan
distribusi yang terlihat pada kelompok kontrol.
Kemungkinan bimodalitas trombosit 5-HT dalam
autisme dapat memungkinkan strategi yang lebih baik
untuk menjelaskan mekanisme yang mendasari
peningkatan.
Hyperserotonemia has been hypothesized to be due
to either increased exposure of the platelet to 5-HT or to
an alteration in the platelet’s handling of 5-HT [2, 3].
More than 90% of peripheral 5-HT is produced from the
essen- tial amino acid tryptophan in the
enterochromaffin cells of the gut wall [5]. Released 5-
HT is metabolized locally by uptake into enterocytes
and, after entering the gen- eral circulation, by efficient
clearance by the endotheli- um, lungs and liver [6]. In
both cases, 5-HT is predomi- nantly metabolized by
MAO-A (monoamine oxidase A) to 5-
hydroxyindoleacetic acid (5-HIAA), and nearly all
urinary 5-HIAA is derived from gut 5-HT [1].
Hiperserotonemia telah dihipotesiskan karena
peningkatan eksposur trombosit menjadi 5-HT atau
karena perubahan dalam penanganan trombosit 5-HT
psy- chiatry. Typically, the group mean has been
reported to be elevated by 25–50% in groups of
individuals with autism compared to normally
developing individuals [2, 3].
pengantar
Peningkatan kadar serotonin (5-hydroxytryptamine, 5-
HT) [1] dalam trombosit darah individu dengan autisme
adalah salah satu temuan yang paling baik direplikasi dalam
psikiatri biologis. Biasanya, rata-rata kelompok telah
dilaporkan meningkat 25-50% pada kelompok individu
dengan autisme dibandingkan dengan individu yang
berkembang secara normal [2, 3].
Recently, we observed a bimodal distribution of
plate- let 5-HT in Dutch individuals diagnosed with
autism spectrum disorders [4]. It appeared that about
half of the individuals with autism spectrum disorders
had platelet 5-HT values distributed in the upper mode,
while the lower mode largely overlapped with the
distribution seen
[2, 3]. Lebih dari 90% perifer 5-HT diproduksi dari asam
amino triptofan esensial dalam sel enterokromafin
dinding usus [5]. Dirilis 5-HT dimetabolisme secara
lokal dengan penyerapan ke dalam enterosit dan, setelah
memasuki sirkulasi umum, dengan pembersihan efisien
oleh endotelium, paru-paru dan hati [6]. Dalam kedua
kasus, 5-HT sebagian besar dimetabolisme oleh MAO-A
(monoamine oxidase A) menjadi asam 5-
hydroxyindoleacetic (5-HIAA), dan hampir semua urin
5-HIAA berasal dari usus 5-HT [1].
Plasma free 5-HT can also be taken up by circulating
platelets, and nearly all 5-HT in the blood (199%) is
found stored within the platelet. Increased gut production
of 5- HT, as in carcinoid syndrome, a cancerous
proliferation of the enterochromaffin cells, leads to
increased levels of platelet 5-HT and increased urinary
excretion of 5-HIAA [6, 7]. Much of the urine 5-HT can
also be assumed to be derived from the enterochromaffin
cell.
5-HT plasma gratis juga dapat diambil dengan
trombosit yang bersirkulasi, dan hampir semua 5-HT
dalam darah (199%) ditemukan disimpan dalam
trombosit. Peningkatan produksi usus 5-HT, seperti pada
sindrom karsinoid, proliferasi kanker sel-sel
enterochromaffin, menyebabkan peningkatan kadar
trombosit 5-HT dan peningkatan ekskresi 5-HIAA kemih
[6, 7]. Sebagian besar urin 5-HT juga dapat diasumsikan
berasal dari sel enterochromaffin.
Several studies have measured urinary excretion of 5-
HIAA in autism. Most studies have not found differences
in urinary 5-HIAA excretion between autistic individu-
als and normal or intellectually disabled controls [8–14].
However, one study reported increased levels of urinary
5-HIAA and urinary 5-HT [15], and another reported Most of the limited data regarding melatonin levels
borderline significantly higher levels of 5-HIAA excre- in autism suggest that group mean nighttime melatonin
tion in hyperserotonemic autistic individuals [10]. Uri- production is reduced in autism [20–23]. Recently non-
nary 5-HT excretion in autism has been reported to be conservative variations in the gene for the last enzyme
elevated [11–13] or unaltered [16]. Taken together, the in the melatonin pathway, acetylserotonin
pri- or studies tend to suggest that gut 5-HT synthesis is methyltransfer- ase (ASMT; EC 2.1.1.4), have been
not altered in autism. However, group sizes usually have reported to be associ- ated with lower levels of daytime
been small and, importantly, only a limited number of plasma melatonin levels in families with subjects with
subjects with elevated platelet 5-HT have been studied. autistic spectrum disorders [24]. These prior results, and
the fact that 5-HT is an ob- ligate precursor for
Beberapa penelitian telah mengukur ekskresi urin 5- melatonin in the pineal gland and the gut, make 6-SM
HIAA pada autisme. Sebagian besar penelitian belum excretion of relevance to studies of plate- let
menemukan perbedaan dalam ekskresi 5-HIAA kemih hyperserotonemia.
antara individu autis dan kontrol yang normal atau cacat
intelektual [8-14]. Namun, satu penelitian melaporkan Sebagian besar data terbatas mengenai kadar
peningkatan kadar 5-HIAA kemih dan 5-HT kemih melatonin pada autisme menunjukkan bahwa rata-rata
[15], dan yang lain melaporkan batas signifikansi 5- produksi melatonin malam hari kelompok berkurang
HIAA yang lebih tinggi pada individu autis pada autisme [20-23]. Baru-baru ini variasi non-
hiperserotonemia [10]. Ekskresi 5-HT urin dalam konservatif dalam gen untuk enzim terakhir di jalur
autisme telah dilaporkan meningkat [11-13] atau tidak melatonin, asetilserotonin metiltransferase (ASMT; EC
berubah [16]. Secara bersama-sama, penelitian utama 2.1.1.4), telah dilaporkan dikaitkan dengan tingkat yang
cenderung menunjukkan bahwa sintesis usus 5-HT lebih rendah dari tingkat melatonin plasma siang hari
tidak diubah dalam autisme. Namun, ukuran kelompok pada keluarga dengan subjek. dengan gangguan
biasanya kecil dan, yang penting, hanya sejumlah kecil spektrum autistik [24]. Hasil-hasil sebelumnya, dan
subjek dengan trombosit tinggi 5-HT telah dipelajari. fakta bahwa 5-HT adalah prekursor oblik untuk
melatonin di kelenjar pineal dan usus, membuat 6-SM
The pineal neurohormone melatonin is synthesized ekskresi relevansi dengan studi hiperserotonemia
from 5-HT, and has a major role in the regulation of cir- trombosit
cadian rhythms, sleep and mood [17]. Another source of
melatonin is the enterochromaffin cells of the gut wall. We hypothesized that 5-HT production in autism – as
Studies in animals indicate that melatonin derived from indexed by levels of urinary 5-HIAA and 5-HT – would
the GI tract accounts for most of the meager daytime be higher in hyperserotonemic compared to normosero-
blood concentration of melatonin (typically 5 –10 times tonemic individuals. Given the reported differences in
lower than peak nighttime levels), whereas nighttime melatonin production in autism and the close metabolic
cir- culating levels of melatonin are determined by relationship between 5-HT and melatonin, melatonin
pineal gland production [18]. Melatonin is metabolized sulfate excretion and the interrelationships between the
in the liver and kidney to 6-sulphatoxymelatonin (6- 3 urinary measures were also examined.
SM), and urinary excretion rates of 6-SM closely follow
the mela- tonin blood concentration [19]. Kami berhipotesis bahwa produksi 5-HT dalam
autisme - seperti yang diindeks oleh level urin 5-HIAA
The melatonin neurohormon pineal disintesis dari 5- dan 5-HT - akan lebih tinggi pada hiperserotonemia
HT, dan memiliki peran utama dalam pengaturan ritme
dibandingkan dengan individu normosero-tonemik.
sirosis, tidur dan suasana hati [17]. Sumber lain
melatonin adalah sel-sel enterochromaffin pada dinding Mengingat perbedaan yang dilaporkan dalam produksi
usus. Studi pada hewan menunjukkan bahwa melatonin melatonin dalam autisme dan hubungan metabolisme
yang berasal dari saluran GI menyumbang sebagian yang erat antara 5-HT dan melatonin, ekskresi
besar konsentrasi darah melatonin siang hari yang melatonin sulfat dan keterkaitan antara 3 ukuran urin
sedikit (biasanya 5-10 kali lebih rendah dari tingkat juga diperiksa.
puncak malam hari), sedangkan kadar melatonin pada
malam hari ditentukan oleh produksi kelenjar pineal Methods
[ 18]. Melatonin dimetabolisme di hati dan ginjal
menjadi 6-sulphatoxymelatonin (6-SM), dan tingkat Subjects
Subjects from our prior study [4] examining the
ekskresi urin 6-SM erat mengikuti konsentrasi darah distribution of platelet 5-HT values in autism spectrum
melatonin [19]. disorders were invited to participate in a follow-up study of
indole metabolism. All sub- jects (n = 20) were male,
unmedicated and had an ADI-R (Autism Interview Schedule,
Revised [25]) classification of autism, cor- roborated by an
ADOS (Autism Diagnostic Observation Sched- ule, Generic
[26]) and a DSM-IV-TR [27] clinical diagnosis of au- tism or
pervasive developmental disorder-not otherwise speci- fied.
Autistic subjects were assigned to hyperserotonemic and Petunjuk tentang asupan makanan [7] dikeluarkan untuk orang tua
normoserotonemic subgroups using a cutoff of 4.5 nmol/109 dan tidak ada konsumsi makanan yang dilarang dilaporkan.
platelets derived from the aforementioned study [4]. Hyper-
and normoserotonemic subjects were matched on age; there
were no significant differences between groups for age (15.3
8 4.4 vs. 15.3
8 4.0 years) and IQ (80.5 8 36.4 vs. 46.3 8 20.5).
Approval for the study was given by the Central Committee
for Research in Humans in The Hague, The Netherlands.
Informed consent/as- sent was obtained from all subjects and
their parents.

Metode

Subjek
Subjek dari penelitian kami sebelumnya [4] memeriksa
distribusi nilai 5-HT platelet dalam gangguan spektrum autisme
diundang untuk berpartisipasi dalam studi tindak lanjut dari
metabolisme indole. Semua subjek (n = 20) adalah laki-laki,
tidak di-medis dan memiliki ADI-R (Jadwal Wawancara
Autisme, Direvisi [25]) klasifikasi autisme, dikoordinasikan
oleh ADOS (Autism Diagnostic Observation Schedule, Generic
[26] ]) dan diagnosis klinis DSM-IV-TR [27] tentang autisme
atau gangguan perkembangan yang menyebar - tidak
dinyatakan secara spesifik. Subjek autistik ditugaskan untuk
subkelompok hiperserotonemia dan normoserotonemia
menggunakan cutoff 4,5 nmol / 109 trombosit yang berasal dari
penelitian tersebut [4]. Subjek hiper dan normoserotonemia
cocok pada usia; tidak ada perbedaan yang signifikan antara
kelompok untuk usia (15,3 8 4,4 vs 15,3
8 4.0 tahun) dan IQ (80.5 8 36.4 vs. 46.3 8 20.5). Persetujuan
untuk penelitian ini diberikan oleh Komite Pusat untuk
Penelitian Manusia di Den Haag, Belanda. Persetujuan /
persetujuan yang diinformasikan diperoleh dari semua subjek
dan orang tua mereka.

Laboratory Measures
Blood samples were collected in 10-ml Vacutainer tubes
(Bec- ton-Dickinson, Meylan, France) containing 0.12 ml (0.34
mol/l) dipotassium EDTA solution. Platelet-rich plasma was
prepared within 1 h of the venepuncture by centrifuging for 30
min at 120 g and 4 ° C, and a platelet count was obtained.
Platelet-rich plasma 5-HT and plasma tryptophan were
determined using a previous- ly described HPLC method with
fluorometric detection [28].

Tindakan Laboratorium
Sampel darah dikumpulkan dalam tabung Vacutainer 10-ml
(Becton-Dickinson, Meylan, Prancis) yang mengandung 0,12 ml
larutan dipotassium EDTA. Plasma yang kaya trombosit dibuat
dalam waktu 1 jam dari venepuncture dengan sentrifugasi selama
30 menit pada 120 g dan 4 ° C, dan jumlah trombosit diperoleh.
Plasma kaya 5-HT dan triptofan plasma ditentukan menggunakan
metode HPLC yang dijelaskan sebelumnya dengan deteksi
fluorometrik [28].

Twenty-four-hour urine samples were collected from the

sub- jects in brown propylene bottles (Sarstedt, Nümbrecht,
Germa- ny). Instructions concerning dietary intake [7] were
issued to the parents and no consumption of prohibited foods
was reported.

Sampel urin dua puluh empat jam dikumpulkan dari subyek

dalam botol propylene coklat (Sarstedt, Nümbrecht, Germain).
 Table 1. Twenty-four-hour urinary excretion of 5-HIAA, 5-HT and 6-SM in normoserotonemic (n = 10) and
hyperserotonemic (n = 9) autistic subjects

Normoserotonemic Hyperserotonemic Mann-Whitney U p

(<4.5 nmol/109 platelets) (≥4.5 nmol/109 platelets) Z(2)

Platelet rich plasma 5-HT

nmol/109 platelets 3.1480.70 (2.95) 6.0281.33 (5.60) 3.787 <0.0001
Urinary 5-HIAA excretion
µmol/24 h 14.086.27 (14.2) 19.2810.3 (16.8) 1.55 0.0611
mmol/mol creatinine 1.6980.86 (1.70) 1.7980.75 (1.60) 0.328 0.3901
Urinary 5-HT excretion
µmol/24 h 0.5280.20 (0.55) 0.6880.26 (0.65) 1.47 0.0711
µmol/mol creatinine 60.1821.4 (59.0) 63.2819.2 (56.0) 0.368 0.3601
Urinary 6-SM excretion
nmol/24 h 77.1835.1 (84.2) 53.0832.7 (57.2) 1.470 0.142
µmol/mol creatinine 9.0384.55 (9.07) 4.9082.84 (4.81) 2.205 0.027

Data presented as means 8 SD, with medians in parentheses.

1
One-tailed p value.

menggunakan uji Mann-Whitney U non-metrik.

One subject was excluded because of incompleteness of the 24-
hour urine collection. Urinary 5-HIAA and 5-HT were measured
Korelasi peringkat Spearman digunakan untuk
using previously described methods [29, 30]. 6-SM analysis was menguji hubungan antara variabel biokimia. Untuk
performed by ELISA, using a 6-SM ELISA Kit (Bühlmann, urin 5-HIAA dan urin 5-HT, analisisnya adalah satu
Schönenbuch, Switzerland). The urine samples were diluted arah, karena ada hipotesis a priori peningkatan
prior to assay. The within-day and between-day coefficients of
varia- tion were 14.4 and 15.5% (n = 35), respectively, for a 55.0 produksi usus senyawa pada individu
nmol/l control sample value. hiperserotonemia. Terlepas dari jumlah
perbandingan yang direncanakan, a ditetapkan 0,05
Satu subjek dikeluarkan karena ketidaklengkapan
untuk mengurangi potensi kesalahan tipe kedua.
pengumpulan urin 24 jam. 5-HIAA dan 5-HT kemih
diukur menggunakan metode yang dijelaskan Analisis korelatif harus dipertimbangkan terutama
sebelumnya [29, 30]. Analisis 6-SM dilakukan oleh tentatif, mengingat terbatasnya jumlah subjek dan
ELISA, menggunakan 6-SM ELISA Kit (Bühlmann, interval kepercayaan besar yang terkait.
Schönenbuch, Swiss). Sampel urin diencerkan sebelum
pengujian. Koefisien variasi dalam hari dan antara hari
adalah 14,4 dan 15,5% (n = 35), masing-masing, untuk Results
nilai sampel kontrol 55,0 nmol / l.
Group means, SDs and medians observed for platelet
5-HT levels and urinary 5-HIAA, 5-HT and 6-SM excre-
Statistical Analysis tion rates are given in table 1, and the individual data are
Demographic and biochemical variables were compared in plotted in figure 1. Comparison of urinary 5-HIAA and
the hyper- and normoserotonemic groups using the nonpara- 5-HT excretion in hyper- and normoserotonemic groups
metric Mann-Whitney U test. Spearman rank correlations were revealed trend-level significant differences between
used to examine relationships between biochemical variables.
For urinary 5-HIAA and urinary 5-HT, the analyses are one-
tailed, since there was an a priori hypothesis of increased gut
production of the compounds in hyperserotonemic individuals.
Despite the number of planned comparisons, a was set to 0.05
to reduce potential for errors of the second type. The correlative
analyses must be considered especially tentative, given the
limited number of subjects and the associated large confidence
intervals.

Analisis statistik
Variabel demografis dan biokimia dibandingkan
pada kelompok hiper dan normoserotonemik
groups, with one-tailed p-values just above the 0.05
level. Excretion of urinary 6-SM appeared to be
decreased in the hyperserotonemic group at a trend
level when ex- pressed per 24 h, and was
significantly lower when ex- pressed per mol of
creatinine (table 1). Plasma trypto- phan
concentrations (µmol/l) were not significantly dif-
ferent in the 2 groups [mean 8 SD (median): 52.3
8 15.2
(50.1) vs. 55.3 8 9.89 (53.6); Mann-Whitney U test,
Z(2) =
0.643, p = 0.523]. Although subjects No. 9 and 7 might
be considered outliers on 1 or more of the measures
(fig. 1), they were included in the analyses since no
distinguish- ing subject characteristics could be
identified. The use of rank-order statistics mitigates
against over-weighting of outlier subjects.
Hasil
Berarti kelompok, SD dan median diamati untuk
tingkat trombosit 5-HT dan tingkat ekskresi 5-HIAA
kemih, 5-HT dan 6-SM diberikan pada tabel 1, dan
data individu diplot pada gambar 1. Perbandingan urin
5 Ekskresi -HIAA dan 5-HT pada kelompok hiper dan
normoserotonemia mengungkapkan perbedaan tingkat
tren yang signifikan antara kelompok, dengan nilai-P
satu-ekor tepat di atas tingkat 0,05. Ekskresi 6-SM
kemih tampak menurun pada kelompok
hiperserotonemia pada tingkat tren ketika
diekspresikan per 24 jam, dan secara signifikan lebih
rendah ketika diekspresikan per mol kreatinin (tabel 1).
Konsentrasi triptofan plasma (µmol / l) tidak berbeda
secara signifikan dalam 2 kelompok [rata-rata 8 SD
(median): 52,3 8 15,2
(50.1) vs. 55.3 8 9.89 (53.6); Uji Mann-Whitney U, Z
(2) =
0,643, p = 0,523]. Meskipun subjek No. 9 dan 7 dapat
dianggap outlier pada 1 atau lebih tindakan (gbr. 1),
mereka dimasukkan dalam analisis karena tidak ada
karakteristik subjek yang dapat diidentifikasi.
Penggunaan statistik urutan-peringkat mengurangi
terhadap overlier dari subjek outlier.
Across all subjects, urinary 5-HIAA and 5-HT excre-
tion rates (per 24 h) were not significantly correlated
with platelet 5-HT (p = 0.128, p = 0.60 and p = 0.149, p
= 0.54, respectively); however, urinary 5-HIAA and
urinary 5- HT were highly correlated (p = 0.840, p !
0.0001). Uri- nary 6-SM was significantly negatively
correlated with platelet 5-HT at the trend level (p = –
0.365, p = 0.125) and positively correlated with urinary
5-HIAA p = 0.458, p = 0.049) and 5-HT (p = 0.433, p =
0.064) when expressed on a 24 h basis. Plasma
tryptophan was significantly corre- lated only with
urinary 5-HT (p = 0.633, p = 0.004). Sim- ilar
correlative relationships were observed between the
biochemical measures when excretion was expressed
per mol of creatinine (data not given). However, when 6-
SM excretion was expressed as per mole of creatinine,
all cor-
 10 50 1.5 150

9 7
9
7 8
125
8 40
7
7 1
8 1.0 100 57
7 4 2
6 30
3 64 5 4
2 4
5 1 1 75 5
3 1
1 594 10
4 9 10 20 41 3 85 3
8 6 3 89
3 2 8
7 28 5 0.5 50
31 9 6
3 4 65 6
3 2 67 6 7
10 82 10
1 2
2 10 5
25 6
9
1
9 24
9
0 0 0 0
aNormoHyperbNormoHypercNormoHyperdNormoHyper

Fig. 1. Individual platelet 5-HT concentrations (a), urinary excretion of 5-HIAA (b), urinary 5-HT (c) and uri-
nary 6-SM (d) of normoserotonemic (n = 10; X) and hyperserotonemic (n = 9; S) autistic subjects. Subjects’
ID numbers are given, and group means are indicated by a horizontal bar.

0,515, p = 0,024). Rasio produk / substrat urin (5-HIAA:

relations were significant (vs. platelet 5-HT, p = –
0.491, p = 0.033; vs. urinary 5-HIAA, p = 0.553, p =
0.014; vs. urinary 5-HT, p = 0.515, p = 0.024). The
urinary product/ substrate (5-HIAA:5-HT) [31] ratios
observed in the hy- per- and normoserotonemic
groups were not significant- ly different [mean 8 SD
(median); 270 8 70 (280) vs. 280
8 50 (260), respectively; Mann-Whitney U; Z(2) =
0.82,
p = 0.935].
Di semua mata pelajaran, tingkat ekskresi 5-HIAA dan
5-HT (per 24 jam) tidak berkorelasi secara signifikan
dengan platelet 5-HT (p = 0,128, p = 0,60 dan p =
0,149, p = 0,54, masing-masing); Namun, urin 5-HIAA
dan urin 5- HT berkorelasi tinggi (p = 0,840, p! 0,0001).
Uranium 6-SM secara signifikan berkorelasi negatif
dengan trombosit 5-HT pada tingkat tren (p = -0,365, p
= 0,125) dan berkorelasi positif dengan urin 5-HIAA p
= 0,458, p = 0,049) dan 5-HT ( p = 0,433, p = 0,064)
ketika diekspresikan pada basis 24 jam. Triptofan
plasma secara signifikan berkorelasi hanya dengan 5-
HT urin (p = 0,633, p = 0,004). Hubungan korelatif
yang sama diamati antara tindakan biokimia ketika
ekskresi diekspresikan per mol kreatinin (data tidak
diberikan). Namun, ketika ekskresi 6-SM diekspresikan
sebagai per kreatinin, semua korelasi signifikan (vs
trombosit 5-HT, p = -0,491, p = 0,033; vs 5-HIAA
kemih, p = 0,553, p = 0,014 ; vs. 5-HT kemih, p =
5-HT) [31] yang diamati pada kelompok hiper dan
normoserotonemia tidak berbeda secara signifikan [rata-
rata 8 SD (median); 270 8 70 (280) vs. 280
8 50 (260), masing-masing; Mann-Whitney U; Z (2) =
0,82,
p = 0,935].
The group means 8 SD (medians) for moles of
cre- atinine/24 h excreted in the hyper- and
normoserotone- mic groups were similar [11.3 8
3.83 (12.40) vs. 9.07 8
3.48 (7.91) mmol/24 h; Mann-Whitney U, Z(2) = 1.470,
p = 0.156]; the amount of urine collected per 24 h was
also similar in the 2 groups [1,327 8 552 (1,330) vs.
1,075 8 555 (975) ml/24 h; Mann-Whitney U, Z(2) =
1.184, p = 0.236, respectively].
Kelompok ini berarti 8 SD (median) untuk mol
kreatinin / 24 jam diekskresikan dalam kelompok
hiper dan normoserotoneik yang serupa [11,3 8 3,83
(12,40) vs 9,07 8
3,48 (7,91) mmol / 24 jam; Mann-Whitney U, Z (2) =
1,470, p = 0,156]; jumlah urin yang dikumpulkan per
24 jam juga serupa pada 2 kelompok [1.327 8 552
(1.330) vs 1.075 8 555 (975) ml / 24 jam; Mann-
Whitney U, Z (2) = 1,184, p = 0,236, masing-masing].
 0.0001 and p = –0.402, p = 0.087), apparently mainly
due to a positive correlation of creatinine with age (p =
0.62, p = 0.005).

The excretion of 5-HIAA, 5-HT and 6-SM expressed

on a 24-hour basis was uncorrelated with age (p = –
Ekskresi 5-HIAA, 5-HT dan 6-SM yang
0.194, p = 0.43, p = 0.091, p = 0.71 and p = –0.042, p = diekspresikan 24 jam tidak berkorelasi dengan
0.89, re- usia (p = -0.194, p = 0.43, p = 0.091, p = 0.71
spectively). The excretion rates of 5-HIAA, 5-HT dan p = –0.042, p = 0,89, ulang
and 6- SM expressed per mole of creatinine were
negatively cor- related with age (p = –0.82, p ! secara spektakular). Tingkat ekskresi 5-HIAA,
0.0001, p = –0.80, p ! 5-HT dan 6- SM yang diekspresikan per mol
kreatinin berhubungan negatif dengan usia (p =
-0,82, p! 0,0001, p = -0,80, p! 0,0001 dan p =
-0,402 , p = 0,087), tampaknya terutama
karena korelasi positif kreatinin dengan usia (p
= 0,62, p = 0,005).

Discussion

The present study revealed trend-level significant

dif- ferences in urinary excretion of 5-HIAA and 5-HT
be- tween hyperserotonemic and normoserotonemic
indi- viduals with autism. Group mean excretion of 6-
SM ap- peared to be lower in the hyperserotonemic
individuals. In all individuals with autism
(normoserotonemic as well as hyperserotonemic),
excretion rates of 6-SM were nega- tively correlated to
platelet 5-HT and positively correlat- ed to urinary
excretion rates of 5-HIAA and 5-HT. The group mean
excretion values reported here for 5-HIAA, 5-HT and 6-
SM were within normal ranges established at the
University Medical Center Groningen, and were in
excellent agreement with prior reports [10, 11, 16, 32].

Diskusi

Penelitian ini mengungkapkan perbedaan tingkat tren

signifikan dalam ekskresi urin 5-HIAA dan 5-HT antara
individu hiperserotonemia dan normoserotonemia dengan
autisme. Kelompok berarti ekskresi 6-SM tampaknya lebih
rendah pada individu hiperserotonemia. Pada semua
individu dengan autisme (normoserotonemia dan juga
hiperserotonemia), tingkat ekskresi 6-SM secara negatif
berkorelasi dengan platelet 5-HT dan secara positif
terkorelasi dengan tingkat ekskresi urin 5-HIAA dan 5-HT.
Nilai rata-rata ekskresi kelompok yang dilaporkan di sini
untuk 5-HIAA, 5-HT dan 6-SM berada dalam kisaran
normal yang ditetapkan di University Medical Center
Groningen, dan berada dalam perjanjian yang sangat baik
dengan laporan sebelumnya [10, 11, 16, 32].

This test of the relative rates of 5-HIAA and 5-HT
ex- cretion in well-defined hyperserotonemic and
normoserotonemic subgroups suggests that greater
exposure of the platelet to 5-HT due to greater gut
production of 5- HT can not be ruled out as a cause of
the platelet hyper- serotonemia of autism. The similar
urinary 5-HIAA:5- HT ratio observed across normo-
and hyperserotonemic groups is a fairly strong
indication that the catabolism of 5-HT does not differ
in the groups.
Tes tingkat relatif 5-HIAA dan ekskresi 5-HT pada
subkelompok hiperserotonemia dan normoserotonemia
yang terdefinisi dengan baik menunjukkan bahwa
paparan platelet yang lebih besar menjadi 5-HT karena
produksi usus yang lebih besar dari 5-HT tidak dapat
dikesampingkan. sebagai penyebab hiper serotonemia
trombosit autisme. Rasio 5-HIAA kemih yang serupa:
5- HT yang diamati pada kelompok normo- dan
hiperserotonemia adalah indikasi yang cukup kuat
bahwa katabolisme 5-HT tidak berbeda dalam
kelompok.
The decreased excretion of 6-SM in
hyperserotonemic individuals and the negative
correlation of urinary excre- tion of 6-SM with platelet
5-HT provide internally con- sistent data suggesting that
there may be a link between altered (elevated) platelet
5-HT and abnormal (reduced) melatonin production in
autism. It should be noted that the 24-hour urinary 6-
SM data reported here, in all like- lihood, reflect
decreases in pineal melatonin production, rather than
alterations in gut melatonin synthesis. In contrast, the
daytime plasma melatonin data reported by Melke et al.
[24] presumably reflect gut production. Al- though most
prior studies have found reduced nighttime melatonin
production in autism [21–23], the picture with respect to
daytime production is much less consistent. There are
observations of elevated [20, 21] and of lower
[22, 24 and Tordjman, S, pers. commun., 2008] daytime
melatonin production in autism, as well as an
observation of normal daytime plasma melatonin levels
[Anderson GM, unpublished data] in individuals with
autism.
Penurunan ekskresi 6-SM pada individu
hiperserotonemia dan korelasi negatif ekskresi urin 6-
SM dengan platelet 5-HT memberikan data yang
konsisten secara internal yang menunjukkan bahwa
mungkin ada hubungan antara platelet 5-HT yang
diubah (meningkat). dan produksi melatonin abnormal
(berkurang) pada autisme. Harus dicatat bahwa data 6-
SM urin 24 jam yang dilaporkan di sini, kemungkinan
besar, mencerminkan penurunan produksi melatonin
pineal, daripada perubahan dalam sintesis melatonin
usus. Sebaliknya, data melatonin plasma siang hari
dilaporkan oleh Melke et al. [24] mungkin
mencerminkan produksi usus. Meskipun sebagian besar
penelitian sebelumnya telah menemukan pengurangan
produksi melatonin pada malam hari pada autisme [21-
23], gambaran sehubungan dengan produksi siang hari
jauh lebih tidak konsisten. Ada pengamatan
peningkatan [20, 21] dan lebih rendah [22, 24 dan
Tordjman, S, pers. commun., 2008] produksi melatonin
siang hari pada autisme, serta pengamatan kadar
melatonin plasma siang hari normal [Anderson GM,
data tidak dipublikasikan] pada individu dengan
autisme.
In summary, studies in large well-characterized
autistic and contrast groups and appropriate subgroups
are needed to sort out the underlying cause(s) of the
platelet hyperserotonemia. The observations reported
here are not inconsistent with theories seeking to
explain the hyperserotonemia on the basis of altered
regulation of gut 5-HT [33]. Additional studies are also
needed to charac- terize daytime and nighttime
melatonin production in autism, to understand the
possible causes and consequences of the alterations in
melatonin, and to elucidate their possible relationship to
the platelet hyperserotonemia.
Singkatnya, penelitian dalam kelompok besar autistik dan
kontras yang ditandai dengan baik dan subkelompok yang
tepat diperlukan untuk memilah penyebab yang mendasari
hiperserotonemia trombosit. Pengamatan yang dilaporkan
di sini tidak konsisten dengan teori yang berusaha
menjelaskan hiperserotonemia berdasarkan regulasi
regulasi usus 5-HT [33]. Studi tambahan juga diperlukan
untuk mengkarakterisasi produksi melatonin pada siang
dan malam hari dalam autisme, untuk memahami
kemungkinan penyebab dan konsekuensi dari perubahan
melatonin, dan untuk menjelaskan kemungkinan
hubungan mereka dengan hiperserotonemia trombosit.
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