Jou nal o
Clinical Medicine
A icle
T ans e in Iso o ms, Old bu New Bioma ke s in He edi a y
F uc ose In ole ance
Aina a Cano 1, Ca los Alcalde 2, Amaya Belange -Quin ana 3, El i a Cañedo-Villa oya 4, Le icia Cebe io 5,
Sil ia Chumillas-Calzada 6, Pa icia Co eche 7, Ma ía Luz Couce 8, Dolo es Ga cía-A enas 9, Igo Gómez 10,
Tomás He nández 11, Elsa Izquie do-Ga cía12, Dáma is Ma ínez Chicano 9, Mon se a Mo ales 6,
Consuelo Ped ón-Gine 13, Es ella Pe ina Jáu egui 14, Luis Peña-Quin ana 15 , Paula Sánchez-Pin os 8,
Juliana Se ano-Nie o 16, Ma ía Unce a Sua ez 17, Isid o Vi o ia Miñana 7and Ja ie de las He as 1,18,19,*
Ci a ion: Cano, A.; Alcalde, C.;
Belange -Quin ana, A.;
Cañedo-Villa oya, E.; Cebe io, L.;
Chumillas-Calzada, S.; Co eche , P.;
Couce, M.L.; Ga cía-A enas, D.;
Gómez, I.; e al. T ans e in Iso o ms,
Old bu New Bioma ke s in He edi a y
F uc ose In ole ance. J. Clin. Med.
2021,10, 2932. h ps://doi.o g/
10.3390/jcm10132932
Academic Edi o : John G i i h Jones
Recei ed: 21 May 2021
Accep ed: 28 June 2021
Published: 30 June 2021
Publishe ’s No e: MDPI s ays neu al
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1Bioc uces Bizkaia Heal h Resea ch Ins i u e, 48093 Ba akaldo, Spain; [email p o ec ed]
2Paedia ics Uni , Río Ho ega Uni e si y Hospi al, 47012 Valladolid, Spain; [email p o ec ed]
3Me abolic Diseases Uni , Depa men o Paedia ics, Ramon y Cajal Hospi al, 28034 Mad id, Spain;
[email p o ec ed]
4Depa men o Me abolism Diseases and Nu i ion, Niño Jesús Uni e si y Child en’s Hospi al,
28009 Mad id, Spain; [email p o ec ed]
5In e nal Medicine Se ice, C uces Uni e si y Hospi al, 48903 Ba akaldo, Spain;
[email p o ec ed]
6
12 de Oc ub e Uni e si y Hospi al, CIBERER, 28041 Mad id, Spain; sil [email protected] id.o g (S.C.-C.);
[email p o ec ed] (M.M.)
7Nu i ion and Me abolic diseases Uni , La Fe Uni e si y Hospi al, 46026 Valencia, Spain;
[email p o ec ed] (P.C.); [email p o ec ed] (I.V.M.)
8Uni o Diagnosis and T ea men o Congeni al Me abolic Diseases, Depa men o Paedia ics, IDIS-Heal h
Resea ch Ins i u e o San iago de Compos ela, CIBERER, Me abERN, San iago de Compos ela Uni e si y
Clinical Hospi al, 15704 San iago de Compos ela, Spain; [email p o ec ed] (M.L.C.);
[email p o ec ed] (P.S.-P.)
9Depa men o Paedia ic Gas oen e ology, Hepa ology and Nu i ion, San Joan de Déu Hospi al,
08950 Ba celona, Spain; dga [email p o ec ed]g (D.G.-A.);
[email p o ec ed]g (D.M.C.)
10 A aba Uni e si y Hospi al, 01009 Gas eiz, Spain; igo [email p o ec ed]
11
Paedia ic Se ice, Albace e Uni e si y Hospi al, 02006 Cas illa-La Mancha, Spain; ohe be [email protected]
12 Pha macy Depa men , In an a Leono Uni e si y Hospi al, 28031 Mad id, Spain;
[email p o ec ed]g
13 Gas oen e ology and Nu i ion Sec ion, Niño Jesús Uni e si y Child en’s Hospi al, 28009 Mad id, Spain;
[email p o ec ed]g
14 Clinical Nu i ion Sec ion, Na a a Uni e si y Hospi al, 31008 Pamplona, Spain;
[email p o ec ed]
15 Pedia ic Gas oen e ology, Hepa ology and Nu i ion Uni , Mo he and Child Insula Uni e si y Hospi al
Complex, Asociación Cana ia pa a la In es igación Pediá ica (ACIP), CIBEROBN, Uni e si y Ins i u e o
Resea ch in Biomedical and Heal h Sciences, Uni e si y o Las Palmas de G an Cana ia,
35016 Las Palmas de G an Cana ia, Spain; [email p o ec ed]
16 Paedia ic Se ice, Málaga Regional Uni e si y Hospi al (HRU), 29010 Málaga, Spain;
[email p o ec ed]
17 Biochemis y Labo a o y, Me abolism A ea, C uces Uni e si y Hospi al, 48903 Ba akaldo, Spain;
[email p o ec ed]
18 Di ision o Paedia ic Me abolism, CIBERER, C uces Uni e si y Hospi al, 48093 Ba akaldo, Spain
19 Depa men o Paedia ics, Uni e si y o he Basque Coun y (UPV/EHU), 48940 Leioa, Spain
*Co espondence: ja ie [email p o ec ed]
Abs ac :
He edi a y F uc ose In ole ance (HFI) is an au osomal ecessi e inbo n e o o me abolism
cha ac e ised by he de iciency o he hepa ic enzyme aldolase B. I s ea men consis s in adop ing
a uc ose-, suc ose-, and so bi ol (FSS)- es ic i e die o li e. Un ea ed HFI pa ien s p esen
an abno mal ans e in (T ) glycosyla ion pa e n due o he inhibi ion o mannose-6-phospha e
isome ase by uc ose-1-phospha e. Hence, ele a ed se um ca bohyd a e-de icien T (CDT) may
allow he p omp de ec ion o HFI. The CDT alues imp o e when an FSS- es ic i e die is ollowed;
howe e , p e ious da a on CDT and uc ose in ake co ela ion a e inconsis en . The e o e, we
examined he comple e se um sialoT p o ile and co ela ed i wi h FSS die a y in ake and wi h
J. Clin. Med. 2021,10, 2932. h ps://doi.o g/10.3390/jcm10132932 h ps://www.mdpi.com/jou nal/jcm
J. Clin. Med. 2021,10, 2932 2 o 14
hepa ic pa ame e s in a coho o paedia ic and adul uc osemic pa ien s. To do so, he p o iles
o se um sialoT om gene ically diagnosed HFI pa ien s on an FSS- es ic ed die (n= 37) and
hei age-, sex- and body mass index-pai ed con ols (n= 32) we e analysed by capilla y zone
elec opho esis. We ound ha in HFI pa ien s, asialoT co ela ed wi h die a y in ake o suc ose
(R = 0.575,
p< 0.001
) and FSS (R = 0.475, p= 0.008), and ha pen asialoT +hexasialoT nega i ely
co ela ed wi h die a y in ake o uc ose (R =
−
0.386, p= 0.024) and FSS (R =
−
0.400, p= 0.019).
In addi ion, he e asialoT /disialoT a io u h ully di e en ia ed ea ed HFI pa ien s om heal hy
con ols, wi h an a ea unde he ROC cu e (AUROC) o 0.97, 92% sensi i i y, 94% speci ici y and
93% accu acy.
Keywo ds:
he edi a y uc ose in ole ance; uc ose; suc ose; so bi ol; die ; sialo ans e in p o ile;
bioma ke ; aldolase B
1. In oduc ion
He edi a y uc ose in ole ance (HFI; OMIM 229600) is an au osomal ecessi e inbo n
e o o me abolism [
1
]. HFI was i s epo ed in 1956 by Chambe s and P a [
2
]. I is
cha ac e ized by de iciency o he enzyme uc ose-1,6-bisphospha e aldolase (aldolase B;
E.C. 4.1.2.13), which is p edominan ly exp essed in he li e , kidney and small in es ine [
3
].
Aldolase B ca alyses di e en eac ions including clea age o uc ose-1-phospha e (F1P)
and e e sible clea age o uc ose-1,6-bisphospha e (FBP) in o glyce aldehyde phospha e
and dihyd oxyace one phospha e (DHAP) [
1
]. The e o e, his enzyme plays a key ole in he
con ol o uc ose and glucose me abolism, egula ing bo h glycolysis and gluconeogenesis.
HFI is caused by homozygous o compound he e ozygous mu a ions in he aldolase B
gene (ALDOB; 612724) on ch omosome 9q31 [
4
]. Based on he ca ie equency o he mos
common mu a ions in neona es, i has been es ima ed ha he p e alence o HFI is a ound
1 in 26,000 li e bi hs in Eu ope [
5
] and 1 in 20,000 bi hs in he US [
6
], ye many au ho s
ag ee ha i may be signi ican ly unde diagnosed. Symp oms in HFI pa ien s a e usually
ini ia ed i e o six mon hs a e bi h due o he in oduc ion o complemen a y eeding in
he in an , who eac s wi h a a ie y o clinical signs such as ailu e o h i e, accompanied
by omi ing, abdominal pain and acu e li e ailu e [
7
]. HFI is also cha ac e ised by a se o
me abolic al e a ions ha include hypoglycaemia, me abolic acidosis, hypophospha emia,
hype u icemia, hype magnesemia and hype alaninaemia a e uc ose loading [
1
]. Ea ly
diagnosis o HFI p e en s he con inued in ake o uc ose, which would o he wise lead o
enal and hepa ic seizu es, coma and e en dea h [
1
]. I allows die a y ea men , consis ing
mainly o a uc ose-, suc ose-, and so bi ol (FSS)- es ic ed die o li e [
1
]. Recu en
symp oms make he diagnosis possible in childhood; howe e , many pa ien s emain
undiagnosed un il adul hood [8].
E en hough uc osemic pa ien s a e belie ed o emain qui e heal hy when kep on
a uc ose- es ic ed die , compliance wi h his egime is o en di icul , especially when
aking in o conside a ion he o e load o uc ose in ou alimen a y ou ines obse ed in
ecen yea s. The e is a g owing use o uc ose, as i is widely used as a ood addi i e, and
small amoun s o his suga a e hidden in many oods. Thus, i is no su p ising ha HFI
pa ien s de elop p e iously un epo ed complica ions, such as li e s ea osis [
7
,
9
] o signs
o p oximal ubula dys unc ion [10,11].
Whe eas easy de ec ion o galac osemia, ano he e o o ca bohyd a e me abolism,
can be pe o med by means o me abolic analysis using d ied blood spo (DBS) es ing [
12
],
no simple me abolic es is a ailable o he apid iden i ica ion o HFI. The diagnosis
o HFI used o be based on he clinical ea u es o uc ose in ole ance, li e biopsy o
assess aldolase B ac i i y, and/o a uc ose challenge es [
13
]. Nowadays, he diagnosis
o HFI consis s in iden i ying ei he biallelic pa hogenic a ian s o ALDOB by molecula
gene ic es ing o de icien hepa ic aldolase B ac i i y om a li e biopsy [
13
]. Despi e li e
biopsy being e y speci ic, i is in asi e and cos ly. The e o e, he high sensi i i y and
J. Clin. Med. 2021,10, 2932 3 o 14
he non-in asi e na u e o ALDOB gene ic es ing make i he p e e ed me hod o HFI
diagnosis [13].
In he mid-1970s, a empo a y change in he ans e in (T ) glyco o m p o ile in he
se um and ce eb ospinal luid was demons a ed o be associa ed wi h sus ained hea y al-
cohol consump ion [
14
]. The abno mal se um T pa e n, ini ially p esen ing as an inc eased
amoun o T bands wi h an isoelec ic poin a o abo e 5.7 (co esponding o asialo-,
monosialo-, and disialo-T ) in isoelec ic ocusing (IEF), imp o ed on abs inence, wi h a
hal -li e o ~10 days [
15
]. This p o ein ac ion, la e named se um ca bohyd a e-de icien T
(CDT), was sugges ed as a speci ic bioma ke , and i s measu emen was p oposed o iden-
i y sus ained hea y alcohol consump ion and moni o abs inence du ing ea men [
16
].
In i o
s udies using cul u ed a li e hepa ocy es sugges ha ace aldehyde esul ing
om alcohol dehyd ogenase oxida ion o e hanol causes he dis up ion o endoplasmic
e iculum unc ion and hus in e e es wi h glycosyla ion [
17
]. A p esen , loss o sialyla-
ion on se um T is used as a sc eening es bo h o ch onic alcohol consump ion and o
congeni al diso de s o glycosyla ion (CDG) [15,16].
Se um T is a glycop o ein ha anspo s i on (Fe
3+
), which is mainly syn hesised
and me abolised in li e hepa ocy es. Two complex chains o oligosaccha ides, which
a y in hei deg ee o ami ica ion, o m he glycolic po ion o he glycop o ein; each o
hem can ha e wo o h ee ex e nal chains o an hems, wi h a sialic acid esidue in he
e minal posi ion (Figu e 1). Then, T has di e en iso o ms depending on he numbe
o sialic acid esidues p esen on i s oligosaccha ide chain: asialo-, monosialo-, disialo-,
isialo-, pen asialo- and hexasialo-T . As s a ed by Helande e al. [
15
], T shows na u al
mic ohe e ogenei y, owing o a ia ions in i on load, amino acid sequence (i.e., gene ic
a ian s) and he s uc u e o he wo N-linked oligosaccha ides (N-glycans). In human
se um, e asialoT is usually he mos abundan glyco o m (~80%), ollowed by pen asialo-
(~14%), isialo- (~4%), disialo- (~1%) and hexasialoT (~1%) [18,19].
P e ious s udies ha e shown ha un ea ed uc osemic pa ien s ha e an abno mal T
glycosyla ion pa e n as a consequence o F1P-media ed compe i i e inhibi ion o mannose-
6-phospha e isome ase (MPI) [
20
]. In pa icula , pa ien s wi h HFI p esen a T isoelec ic
ocussing (IEF) ype Ib pa e n, in which de ec s in he syn hesis and assembly o he glycans
occu , indica ing ha HFI is a seconda y CDG synd ome [
21
]. The e o e, as glycosyla ion
o T is a measu e o in ahepa ic F1P concen a ions, ele a ed se um CDT alues allow
he quick de ec ion o HFI [
22
]. This bioma ke is aluable o de ec ing he pe sis ence o
some abno mali ies caused by ace amoun s o uc ose inges ion and/o non-adhe ence
o he p esc ibed die in HFI pa ien s. Mo eo e , CDT alues imp o e a e 2–4 weeks
when a uc ose- es ic ed die is aken, and hen, CDT measu emen s a e ecommended
o HFI diagnosis and ea men moni o ing [22].
Howe e , he e is no ag eemen among au ho s ega ding he co ela ion o he sialoT
p o ile wi h die a y uc ose consump ion, no abou which iso o m o T is mo e aluable
o moni o ing uc osemic pa ien s [
21
–
25
]. Thus, he objec i e o he p esen s udy was
o de e mine he use ulness o he en i e sialoT p o ile o moni o ing an FSS- es ic ed
die in HFI pa ien s by assessing he co ela ion be ween he di e en sialoT iso o ms and
die a y uc ose, suc ose, and so bi ol in ake.
J. Clin. Med. 2021,10, 2932 4 o 14
J. Clin. Med. 2021, 10, x FOR PEER REVIEW 3 o 13
assess aldolase B ac i i y, and/o a uc ose challenge es [13]. Nowadays, he diagnosis
o HFI consis s in iden i ying ei he biallelic pa hogenic a ian s o ALDOB by molecula
gene ic es ing o de icien hepa ic aldolase B ac i i y om a li e biopsy [13]. Despi e
li e biopsy being e y speci ic, i is in asi e and cos ly. The e o e, he high sensi i i y
and he non-in asi e na u e o ALDOB gene ic es ing make i he p e e ed me hod o
HFI diagnosis [13].
Figu e 1. The link be ween uc ose me abolism and he p o ile o sialo ans e ins in he edi a y uc ose in ole ance.
Die a y so bi ol and suc ose, i.e., a disaccha ide o med by glucose and uc ose, a e p ecu so s o uc ose. The ca abolic
pa hway o die a y uc ose, suc ose and so bi ol is al e ed in pa ien s wi h he edi a y uc ose in ole ance (HFI). In al-
dolase B de iciency, he ca abolism o uc ose-1-P (F1P) is impai ed ( ed ba ), and his molecule is accumula ed la gely in
he li e o HFI pa ien s. Consequen ly, HFI pa ien s ha e an abno mal ans e in (T ) glycosyla ion pa e n because o
F1P-media ed compe i i e inhibi ion o mannose-6-phospha e isome ase (MPI). T exhibi s di e en iso o ms depending
on he numbe o sialic acid esidues p esen on i s oligosaccha ide chain; asialo-, monosialo-, disialo-, isialo-, pen asialo-
, and hexasialo-T . The sum o asialo-, monosialo-, and disialo-T is called ca bohyd a e-de icien T (CDT).
In he mid-1970s, a empo a y change in he ans e in (T ) glyco o m p o ile in he
se um and ce eb ospinal luid was demons a ed o be associa ed wi h sus ained hea y
alcohol consump ion [14]. The abno mal se um T pa e n, ini ially p esen ing as an in-
c eased amoun o T bands wi h an isoelec ic poin a o abo e 5.7 (co esponding o
asialo-, monosialo-, and disialo-T ) in isoelec ic ocusing (IEF), imp o ed on abs inence,
wi h a hal -li e o ~10 days [15]. This p o ein ac ion, la e named se um ca bohyd a e-
de icien T (CDT), was sugges ed as a speci ic bioma ke , and i s measu emen was p o-
posed o iden i y sus ained hea y alcohol consump ion and moni o abs inence du ing
ea men [16]. In i o s udies using cul u ed a li e hepa ocy es sugges ha ace alde-
hyde esul ing om alcohol dehyd ogenase oxida ion o e hanol causes he dis up ion o
endoplasmic e iculum unc ion and hus in e e es wi h glycosyla ion [17]. A p esen ,
Figu e 1.
The link be ween uc ose me abolism and he p o ile o sialo ans e ins in he edi a y
uc ose in ole ance. Die a y so bi ol and suc ose, i.e., a disaccha ide o med by glucose and uc ose,
a e p ecu so s o uc ose. The ca abolic pa hway o die a y uc ose, suc ose and so bi ol is al e ed in
pa ien s wi h he edi a y uc ose in ole ance (HFI). In aldolase B de iciency, he ca abolism o uc ose-
1-P (F1P) is impai ed ( ed ba ), and his molecule is accumula ed la gely in he li e o HFI pa ien s.
Consequen ly, HFI pa ien s ha e an abno mal ans e in (T ) glycosyla ion pa e n because o F1P-
media ed compe i i e inhibi ion o mannose-6-phospha e isome ase (MPI). T exhibi s di e en
iso o ms depending on he numbe o sialic acid esidues p esen on i s oligosaccha ide chain; asialo-,
monosialo-, disialo-, isialo-, pen asialo-, and hexasialo-T . The sum o asialo-, monosialo-, and
disialo-T is called ca bohyd a e-de icien T (CDT).
2. Expe imen al Sec ion
2.1. Pa icipan s
A c oss-sec ional s udy was conduc ed om Oc obe 2019 o No embe 2020. The
s udy popula ion comp ised 37 gene ically diagnosed HFI pa ien s om 31 un ela ed
amilies. The ec ui ed uc osemic pa ien s had been on die a y ea men wi h uc ose,
suc ose and so bi ol exclusion o a leas wo yea s. Thei age-, sex-, and BMI-ma ched
heal hy con ols (n= 32) we e also s udied.
Ele en Spanish hospi als pa icipa ed in he s udy: C uces Uni e si y Hospi al, Basque
Coun y [hos ]; A aba Uni e si y Hospi al, Basque Coun y; Na a a Uni e si y Hospi al;
Na a a, 12 de Oc ub e Uni e si y Hospi al, Mad id; Niño Jesús Uni e si y Child en’s
Hospi al, Mad id; Ramón y Cajal Uni e si y Hospi al, Mad id; La Fe Uni e si y Hospi al,
Valencian Communi y; Málaga Regional Uni e si y Hospi al, Andalusia; San iago de
Compos ela Uni e si y Clinical Hospi al, Galicia; Río Ho ega Uni e si y Hospi al, Cas ile
and Leon; Mo he and Child Insula Uni e si y Hospi al complex, Cana y Islands.
The s udy p o ocol was pe o med acco ding o he e hical guidelines o he e ised
1975 Decla a ion o Helsinki [
26
] and app o ed by he Resea ch E hics Commi ee o he
J. Clin. Med. 2021,10, 2932 5 o 14
Basque Coun y (CEIm-E), e hic app o al code: PI2019072. W i en in o med consen was
ob ained om pa en s o legal gua dians o child en (below 18 yea s o age) and adul
s udy pa icipan s.
2.2. S udy Visi s
S udy isi s we e pe o med a he C uces Uni e si y Hospi al, Spain. Blood samples
we e collec ed a he same ime, a 8:30 a.m., a e a leas 8 h o as ing. Pa ien s we e
weighed, and hei heigh and abdominal pe ime e s we e measu ed. S anding heigh was
measu ed wi h a wall-moun ed s adiome e , and pa ien s we e weighed ba e oo , o he
nea es 100 g, wi h digi al scales.
2.3. Geno ype Analysis
HFI pa ien s we e gene ically diagnosed in hei hospi als o o igin by Sange sequenc-
ing o he ALDOB gene [27] o by NGS gene panel sc eening.
2.4. Biochemical Analyses
Concen a ions o i on, T and e i in in he plasma we e de e mined by ou ine
clinical echniques.
2.5. T ans e in Quan i ica ion
The T p o ile was de e mined in se um by capilla y zone elec opho esis (CZE) using
a comme cially a ailable sys em (Sebia, Capilla ys 2TM, F ance) in 37 HFI pa ien s and in
32 heal hy olun ee s as desc ibed be o e [
18
]. B ie ly, he T iso o ms we e sepa a ed in o
i e ac ions acco ding o hei deg ee o sialyla ion, i.e., asialoT (non-sialyla ed o m),
disialo-, isialo-, e asialo- and he sum o pen asialo- and hexasialo-T , and exp essed as
a pe cen age o he o al sialoT . Da a analysis was pe o med wi h he so wa e package
Pho esis 8.6.3 (Sebia, F ance). The sum o he asialoT and disialoT ac ions was exp essed
as a CDT pe cen age. Addi ionally, he e asialoT - o-disialoT a io was calcula ed.
2.6. Assessmen o Die
Die a y in o ma ion was collec ed in a sel -adminis e ed nu i ional eco d o die a y
in ake on h ee days ( wo du ing he week and one on he weekend) in 34 o he 37 HFI
pa ien s, wi h a special emphasis on de e mining uc ose, suc ose and so bi ol die a y
in ake. Once comple ed, he di e en ca bohyd a e composi ion, exp essed in mg pe
day, was calcula ed using he Nu i ional Calcula ion DIAL P og am (Ve sion 3.10.5.0), o
ALCEGENI INERIA [
28
]. In addi ion, o comple e he in o ma ion o uc ose and so bi ol
die a y in ake, he ollowing da abases we e consul ed: Aus alian Food Composi ion
Da abase—Release 1.0 [
29
], Danish F ida Food Da abase [
30
], and Ge man Food Composi-
ion and Nu i ion Tables [
31
]. Da a ob ained on e men able mono-, di-, oligosaccha ides
and polyols (FODMAP) we e also used [32].
2.7. S a is ical Analysis
Con inuous a iables we e ep esen ed as mean
±
s anda d de ia ion and ange.
F uc osemic pa ien s on an FSS- es ic i e die (n= 37) we e compa ed wi h hei age-, sex-,
and BMI-ma ched con ols (n= 32) by unpai ed S uden ’s - es . The Mann–Whi ney U
es was used o compa e con inuous a iables be ween HFI pa ien s wi h and wi hou
asialoT exp ession. he Wallis es was used o compa e con inuous a iables among he
FFS-in ake e iles. p- alues we e based on wo- ailed compa isons as app op ia e, and
hose less han 0.05 we e conside ed o indica e a s a is ically signi ican di e ence.
Pea son co ela ion was used o assess bi a ia e ela ionships be ween a iables.
The diagnos ic accu acy o he model ha di e en ia es be ween HFI pa ien s and non-
HFI pa ien s was assessed by he a ea unde he ROC cu e (AUC) by expo ing he
ob ained da a o he Me aboAnalys so wa e package ( e sion 5.0). The op imal sco e
cu -o alue o he es ima ion g oup was selec ed based on sensi i i y and speci ici y,
J. Clin. Med. 2021,10, 2932 6 o 14
and posi i e and nega i e likelihood a ios as ha a which he sum o sensi i i y and
speci ici y was maximum.
S a is ical analyses we e pe o med using SPSS so wa e, e sion 23 o Windows
(IBM, Chicago, IL), R So wa e (R e sion 3.2.0; R De elopmen Co e Team, 2010;
h p://c an. -p ojec .o g) and Mic oso O ice Excel ( 19.0).
3. Resul s
3.1. S udy Visi s
The s udy popula ion comp ised 37 gene ically diagnosed HFI pa ien s om 31 un-
ela ed amilies and hei 32 heal hy con ols (Table 1). The HFI pa ien s we e 22 emales
and 15 males; 23 pa ien s we e below 18 yea s o age, and 14 we e adul s. The con ols
we e 19 emales and 13 males, and o hese olun ee s, 20 we e below 18 yea s o age, and
12 we e adul s.
Table 1.
P incipal ea u es in HFI pa ien s and in hei heal hy con ols. Con inuous a iables a e ep esen ed as
mean ±s anda d
de ia ion and ange. Di e ences in con inuous a iables be ween HFI pa ien s and con ols we e
calcula ed by using he S uden ’s - es . T ans e in (T ).
Con ols HFI Pa ien s p-Value
n32 37
Women/men, n/n19/13 22/15 0.594
Age, yea 21.5 ±15.1 (2.1–61.3) 19.7 ±14.8 (3.7–63.4) 0.629
Weigh , kg 50.5 ±15.3 (11.9–71.0) 45.3 ±16.4 (17–73) 0.178
Heigh , cm 155.5 ±19.5 (85.5–181.0) 151.0 ±21.1(104.2–190.7) 0.368
BMI, kg/m220.2 ±3.2 (14.6–27.1) 19.0 ±3.1 (14.0–27.4) 0.107
Wais ci cum e ence, cm 68.3 ±9.4 (48–88) 67.5 ±9.7 (51–87) 0.736
CDT, % (asialoT + disialoT ) 0.7 ±0.1 (0.4–1.0) 2.4 ±1.8 (0.6–8.4)
9<0.001
AsialoT , % 0.0 ±0.0 (0.0–0.0) 0.10 ±0.3 (0.0–0.1) 0.027
DisialoT , % 0.7 ±0.1 (0.4–1.0) 2.3 ±1.6 (0.6–7.8) <0.001
T isialoT , % 4.4 ±1.7 (2.5–6.7) 5.3 ±1.2 (3.2–7.7) <0.001
Te asialoT , % 84.4 ±1.6 (81–87) 79.0 ±2.7 (74–84) <0.001
Pen a- and hexa-sialoT , % 10.8 ±1.1 (9.0–14.1) 13.3 ±1.7 (10.3–17.1) <0.001
Te a-/di-sialoT 133.4 ±27.7 (81.3–204.8) 51.8 ±32.9 (9.4–137.5) <0.001
Se um i on (mg/dL) 98.7 ±39.7 (34–205) 85.3 ±30.6 (53–216) 0.133
T (mg/dL) 270.9 ±37.8 (210–397) 288.1 ±36.5 (222–378) 0.064
Fe i in (mg/dL) 50.4 ±37.9 (10–176) 73.2 ±73.4 (4–284) 0.124
Signi ican p- alues a e ma ked in bold.
The majo i y o he HFI pa ien s en olled we e diagnosed du ing hei childhood,
i.e., a 3.0
±
2.6 yea s, excep ou pa ien s, who we e diagnosed a 34, 43, 48,and 63 yea s
o age. The e we e no di e ences in weigh , heigh , BMI o wais ci cum e ence be ween
he HFI pa ien s and hei espec i e con ols (Table 1).
3.2. Biochemical Analyses
The ela i e pe cen ages o all sialoT iso o ms analysed we e al e ed in HFI pa ien s
when compa ed wi h hei con ols, as shown in Table 1and in Figu e 2. The concen a ion
o CDT, disialo-, isialo- and he sum o pen asialo- and hexasialo-T in he HFI pa ien s
was highe by 3.7- old, 3.5- old, 28%,and 23%, espec i ely, when compa ed wi h he
con ols. In con as , e asialoT concen a ion was lowe by 6% in he HFI pa ien s in
compa ison wi h hei heal hy con ols. AsialoT was p esen in 6 ou o he 37 HFI pa ien s
and was no de ec ed in any con ol pa icipan .
J. Clin. Med. 2021,10, 2932 7 o 14
J. Clin. Med. 2021, 10, x FOR PEER REVIEW 7 o 13
Figu e 2. Sialo ans e in (sialoT ) iso o m pe cen ages in HFI pa ien s (n = 37) and in hei espec i e heal hy con ols (n
= 32). Di e ences in con inuous a iables be ween HFI and hei con ols we e calcula ed by using he S uden ’s - es .
3.3. Se um T ans e in Iso o ms, Hepa ic Pa ame e s and F uc ose Consump ion in Pa ien s
wi h F uc osemia
Table 2 shows T iso o ms, hepa ic pa ame e s and FSS consump ion in HFI pa ien s,
all oge he and di ided by FSS-in ake e iles, and shows ha he e we e no signi ican
di e ences in sialoT o he s udied hepa ic pa ame e s among he h ee e iles o FSS-
in ake in HFI pa icipan s.
Table 2. Die a y in ake o uc ose, suc ose, so bi ol and hei sum (FSS), sialoT pe cen ages and li e pa ame e s in HFI
pa ien s, all oge he (n = 34) and di ided in o FSS-in ake e iles: 1s (n = 12), 2nd (n = 11) and 3 d (n = 11). Con inuous
a iables a e ep esen ed as mean ± s anda d de ia ion and ange. Di e ences in con inuous a iables among he h ee
e iles we e calcula ed using he K uskal–Wallis es .
HFI pa ien s all
oge he
FSS-in ake
1s e ile
FSS-in ake
2nd e ile
FSS-in ake
3 d e ile p
Figu e 2.
Sialo ans e in (sialoT ) iso o m pe cen ages in HFI pa ien s (n= 37) and in hei espec i e heal hy con ols
(n= 32). Di e ences in con inuous a iables be ween HFI and hei con ols we e calcula ed by using he S uden ’s - es .
3.3. Se um T ans e in Iso o ms, Hepa ic Pa ame e s and F uc ose Consump ion in Pa ien s
wi h F uc osemia
Table 2shows T iso o ms, hepa ic pa ame e s and FSS consump ion in HFI pa ien s,
all oge he and di ided by FSS-in ake e iles, and shows ha he e we e no signi ican
di e ences in sialoT o he s udied hepa ic pa ame e s among he h ee e iles o FSS-
in ake in HFI pa icipan s.
J. Clin. Med. 2021,10, 2932 8 o 14
Table 2.
Die a y in ake o uc ose, suc ose, so bi ol and hei sum (FSS), sialoT pe cen ages and li e pa ame e s in HFI
pa ien s, all oge he (n= 34) and di ided in o FSS-in ake e iles: 1s (n= 12), 2nd (n= 11) and 3 d (n= 11). Con inuous
a iables a e ep esen ed as mean
±
s anda d de ia ion and ange. Di e ences in con inuous a iables among he h ee
e iles we e calcula ed using he K uskal–Wallis es .
HFI Pa ien s all
Toge he
FSS-In ake
1s Te ile
FSS-In ake
2nd Te ile
FSS-In ake
3 d Te ile p
n34 12 11 11
F uc ose in ake
(mg/day) 322 ±304 (0–1323) 70 ±78 (0–220) 296 ±161 (98–604) 621 ±311
(238–1323) <0.001
Suc ose in ake
(mg/day)
503 ±651
(15–3747) 131 ±99 (15–308) 378 ±70 (255–530) 1032 ±944
(344–3747) <0.001
So bi ol in ake
(mg/day) 50 ±93 (0–443) 11 ±20 (0–57) 32 ±40 (0–90) 11 ±142 (0–443) 0.049
FSS in ake
(mg/day)
1764 ±938
(15–4366) 212 ±157 (15–446) 707 ±193
(457–936)
1764 ±937
(1020–4366) <0.001
AsialoT , % 0.08 ±0.25 (0–0.8) 0.08 ±0.21
(0.0–0.6)
0.09 ±0.22
(0.0–0.8)
0.14 ±0.32
(0.00–1.00) 0.934
DisialoT , % 2.3 ±1.6 (0.6–7.8) 2.2 ±2 (0.6–7.8) 2.4 ±1.5 (0.9–5.4) 2.3 ±1.3 (0.9–4.6) 0.612
T isialoT , % 5.3 ±1.2 (3.2–7.7) 5.1 ±1.1 (3.4–6.8) 5.2 ±1.2 (3.2–7.1) 5.7 ±1.1 (4.0–7.7) 0.395
Te asialoT , % 79.0 ±2.7
(73.7–84.3)
78.6 ±2.9
(73.7–82.5)
79.6 ±2.9
(74.4–84.3)
78.7 ±2.1
(76.7–82.7) 0.674
Pen a- and
hexa-sialoT , %
13.3 ±1.7
(10.3–17.1)
14.1 ±1.8
(10.8–17.1)
12.7 ±1.6
(10.3–15.9)
13.1 ±1.5
(10.37–14.6) 0.262
Te a-/di-sialoT 51.8 ±32.9
(9.4–137.5)
62.2 ±40.8
(9.4–137.5)
47.3 ±29.1
(13.8–93.7)
46.1 ±27.9
(16.7–91.9) 0.758
GGT (U/L) 15.3 ±5.9 (9–36) 17.4 ±8.2 (9–36) 14.4 ±4.9 (9–27) 14.2 ±3.5 (10–21) 0.221
GOT, AST (U/L) 26.5 ±9.5 (16–66) 45.8 ±30.4 (19–45) 28.5 ±4.8 (19–33) 27.9 ±4.7 (16–66) 0.322
GPT, ALT (U/L) 30.4 ±31.4 (9–198) 44.3 ±50.7
(16–198) 22.8 ±9.5 (10–47) 24.7 ±15.7 (9–68) 0.234
INR 1.00 ±0.08
(0.9–1.1)
0.96 ±0.08
(0.9–1.1)
1.00 ±0.67
(0.9–1.1)
1.00 ±0.77
(0.9–1.1) 0.151
Signi ican p- alues a e ma ked in bold.
The e we e six uc osemic pa icipan s wi h asialoT exp ession. The e we e no
signi ican di e ences in he s udied hepa ic pa ame e s o in he FSS-in ake be ween HFI
pa ien s who exp essed asialoT and hose who did no exp ess i (Table 3).
Table 3.
Die a y in ake o uc ose, suc ose, so bi ol, hei sum (FSS), li e pa ame e s and sialoT
iso o m pe cen ages in HFI pa ien s wi h (n= 6) o wi hou (n= 31) he asialoT iso o m. Con-
inuous a iables a e ep esen ed as mean
±
s anda d de ia ion and ange. Di e ences in con in-
uous a iables be ween HFI pa ien s wi h o wi hou asialoT iso o m we e calcula ed using he
Mann–Whi ney U es . T ans e in (T ).
No AsialoT
Exp ession AsialoT Exp ession p-Value
F uc ose in ake
(mg/day) 354 ±321 (0–1323) 434 ±327 (220–620) 0.254
Suc ose in ake
(mg/day) 434 ±327 (15–1334)
1096
±
1496 (117–3747)
0.232
So bi ol in ake
(mg/day) 64 ±104 (0–443) 19 ±41 (0–93) 0.192
FSS in ake (mg/day) 868 ±596 (15–2119)
1535
±
1618 (337–4366)
0.487
J. Clin. Med. 2021,10, 2932 9 o 14
Table 3. Con .
No AsialoT
Exp ession AsialoT Exp ession p-Value
GGT (U/L) 15.5 ±6.1 (9–36) 14.7 ±5.2 (9–21) 1
GOT, AST (U/L) 27.2 ±10.3 (13–66) 23 ±2.8 (19–27) 0.606
GPT, ALT (U/L) 31.5 ±34.2 (9–198) 24.7 ±6.2 (19–35) 0.223
INR 1.0 ±0.1 (0.9–1.1) 1.0 ±0.1 (1.0–1.1) 0.295
CDT, %
(asialoT +disialoT ) 1.8 ±1.0 (0.6–4.5) 5.5 ±1.7 (3.9–8.4) <0.001
AsialoT , % 0 0.6 ±0.3 (0.2–1.0) <0.001
DisialoT , % 1.8 ±1.0 (0.6–4.5) 4.9 ±1.5 (3.7–7.8) <0.001
T isialoT , % 5.3 ±1.2 (3.2–7.7) 5.6 ±1.2 (4.0–7.1) 0.575
Te asialoT , % 79.5 ±2.5 (76.5–84.3) 76.5 ±2.2 (73.5–79.8) 0.005
Pen a- and
hexa-sialoT , % 58.6 ±31.7 (10.3–17.1) 16.6 ±4.3 (10.3–14.4) 0.427
Te a-/di-sialoT 13.4 ±1.7 (17.0–137.5) 12.5 ±1.6 (9.4–20.9) <0.001
Signi ican p- alues a e ma ked in bold.
3.4. Co ela ions be ween T ans e in Iso o ms, Hepa ic Pa ame e s and F uc ose Consump ion in
Pa ien s wi h F uc osemia
The co ela ions be ween he die a y in ake o uc ose, suc ose, so bi ol and hei sum
(FSS) wi h he en i e sialoT p o ile we e assessed in HFI pa ien s (Table 4). The highes co -
ela ion alues we e ound o he asialoT pe cen age, which co ela ed wi h die a y in ake
o suc ose (R = 0.575, p< 0.001) and FSS (R = 0.475, p= 0.008). The pen asialoT +hexasialoT
ac ion co ela ed nega i ely wi h uc ose (R =
−
0.386, p= 0.024) and wi h FSS in ake
(
R = −0.400
,p= 0.019), and e asialoT co ela ed wi h die a y in ake o so bi ol (R = 0.361,
p= 0.036).
Table 4.
Co ela ion analysis o sialo ans e in (T ) ac ion le els wi h die a y in ake o uc ose, suc ose, so bi ol and hei
sum (FSS) and wi h li e pa ame e s in HFI pa ien s. Pea son co ela ion analysis was used o assess bi a ia e ela ionships
be ween a iables. Signi ican p- alues a e ma ked in bold.
Die a y
In ake CDT, % A-T , % Di-T , % T i-T , % Te a-T , % Pen a- and
Hexa-T , %
Te a-/di-T
Ra io
F uc ose R0.063 0.107 0.046 0.244 0.092 −0.386 −0.085
p0.725 0.573 0.794 0.169 0.606 0.024 0.631
Suc ose R0.242 0.575 0.18 0.238 −0.073 −0.303 −0.187
p0.168 <0.001 0.309 0.175 0.68 0.08 0.29
So bi ol R−0.197 −0.175 −0.197 −0.181 0.361 −0.253 0.209
p0.264 0.354 0.264 0.305 0.036 0.15 0.236
FSS R0.187 0.475 0.133 0.25 0.016 −0.4 −0.151
p0.29 0.008 0.453 0.154 0.929 0.019 0.392
Li e Pa-
ame e s
GGT R0.061 0.006 0.077 0.049 −0.12 0.091 −0.107
p0.719 0.974 0.653 0.774 0.48 0.592 0.528
GOT
(AST)
R−0.213 −0.135 −0.212 −0.004 0.184 −0.059 0.184
p0.212 0.453 0.214 0.982 0.282 0.733 0.281
GPT
(ALT)
R0.044 −0.01 0.061 0.076 −0.185 0.193 −0.092
p0.795 0.955 0.72 0.656 0.272 0.253 0.587
INR R−0.026 0.082 −0.033 −0.064 −0.055 0.162 0.103
p0.882 0.662 0.849 0.717 0.752 0.351 0.555
