April 2000
1986
MECHANISM OF THIAMIN UPTAKE IN THE HUMAN JEJUNAL
BRUSH·BORDER MEMBRANE VESICLES.
Sangeeta Tyagi, Reena J. Kavilaveetlil, Ravinder Gill, Hamid M. Said,
Pradeep K. Dudeja, Univ of Illinois, Chicago, IL; VAMC, Long Beach,
CA; VA: Westside, Chicago, IL; Univ of CA, Irvine, Long Beach, CA.
The water-soluble vitamin thiamin is essential for normal cellular function,
growth and development. Thiamin deficiency, even in developed countries
represents a significant problem in alcoholics, elderly, critically ill children
and in patients suffering from diabetes, coeliac disease and renal disease.
To date, however, very little is known about the mechanism of absorption
of dietary thiamin in the human small intestine. The current studies were,
therefore, undertaken to elucidate the mechanism of thiamin transport in
luminal brush-border membranes (BBM) of the human jejunum. BBMV
were isolated and purified from mucosal scrapings of organ donor jejunum
using a differential centrifugation and divalent cation (Mg+2) precipitation
technique. 3H thiamin uptake was measured by a rapid filtration technique.
Our results are summarized as follows: i) An outwardly directed H+-
gradient (pH 5.5in/7.Sout) significantly increased 3H thiamin uptake into
BBMV; ii) Amiloride significantly inhibited H+-gradient driven 3H thia-
min uptake in a dose dependent manner, Dixon plot analysis in the
presence of 0.25 I-tM or 10 I-tM 3H thiamin demonstrated that inhibition of
thiamin uptake by amiloride was competitive in nature with K; for amilo-
ride of -0.18 mM; iii) Thiamin uptake was sensitive to temperature and
osmolarity of the incubation medium; iv) 3Hthiamin (0.25I-tM) uptake was
markedly inhibited by 50 I-tM concentration of cold thiamin and structural
analogs amprolium, oxythiamine and pyrithiamine; v) 3H thiamin (0.25
I-tM) uptake into these vesicles was not significantly inhibited in the
presence of 50 or 100 I-tM concentrations of organic cations [tetraethyl-
ammonium (TEA), N-methylnicotinamide (NMN) or choline) in the incu-
bation medium; vi) 3H thiamin uptake was not affected by the presence of
ImM Mg+ 2-ATP in vesicles or incubation medium; vii?H thiamin uptake
was potential insensitive; viii) 3H thiamin uptake into these vesicles (after
correcting for diffusion component) demonstrated saturation kinetics with
an apparent K", for thiamin of 0.61 ± 0.081-tM(n=4) and a Vmax of 1.00 ±
0.47 pmol/mg prot./IO sec (n=4). Additional studies with the human ileal
BBMV also demonstrated the presence of a thiamin transporter in this
region of the small intestine with similar apparent affinity for thiamin.
Conclusion: These results demonstrate, for the first time, the existence of
a pH-dependent, amiloride-sensitive, electroneutral carrier-mediated mech-
anism for thiamin absorption in native human small intestinal BBMV.
1987
EVALUATION OF FECAL ELASTASE·1 CONCENTRATIONS IN
VARIOUS MALABSORPTION CONDITIONS.
Jaroslaw Walkowiak, Karl-Heinz Herzig, Karol Marcinkoswki Univ, In-
stitute of Pediatrics, Poznan, Poland; I. Dept of Internal Medicine, Chris-
tian-Albrechts-Univ, Kiel, Germany.
Fecal elastase- I (EI) is a very sensitive and reliable test in the assessment
of exocrine pancreatic function in chronic pancreatitis and cystic fibrosis.
However, levels of elastase-I in other malabsorption conditions are un-
known. Therefore, the aim of the present study was to evaluate EI spec-
ificity in the assessment of exocrine pancreatic insufficiency (EPI) in
malabsorption conditions other than CF. Methods: The study was carried
out in 56 patients referred for evaluation with borderline chloride concen-
trations in sweat test. 70 age-matched healthy children (He) and 102 cystic
fibrosis (CF) patients with EPI served as control groups. El concentrations
were measured in all subjects. Results: The referred patients were diag-
nosed as celiac disease (CD; n = 16), secondary malabsorption syndrome
(SMS; n = 18) and food allergy (FA; n = 20). In all patients, screening for
CFTR gene mutations was negative. El levels were 794.5+440.1 I-tglg in
HC, 73.7+ 132.3 I-tg/g in CF patients (p<O.OOI vs. HC), 249.6+ 188.2
I-tg/g in CD (p<O.OOI vs. HC. p<0.OO5 vs. CF), 212.2+ 120.6 I-tg/g in
SMS (p<O.OOI vs. HC, p<O.ool vs. CF) and 682.3+378.9 in FA (non
significant vs. HC, p<O.OOI vs. CF) patients, respectively. With a cut-off
level of 200 I-tglg (Walkowiak et al. 1999), abnormal concentrations were
found in 87.2% in the CF group, in 56.2% and 55.5% in CD and SMS
subgroups, respectively. In none of FA patients, El values were below the
normal range. In 18 out of 19 CD and SMS patients (with low El levels)
monitored for at least 12 months on a gluten-free diet, El levels increased
above the cut-off value to normal concentrations. Two out of the 54
referred patients were finally diagnosed as having CF. one with stable low
El levels and the second with finally normal values. Conclusion: Fecal
elastase-I is a very useful diagnostic test for the assessment of exocrine
pancreatic function but it is not specific for determination of the underlying
disease. Supported in part by the Scientific Research Committee, Poland.
AGAA375
1988
NUTRITIONAL INTAKE, PLASMA ANTIOXIDANT VITAMINS
AND OXIDATIVE STRESS IN CROHN'S DISEASE PATIENTS
COMPARED TO HEALTHY CONTROLS.
Barbara E. Wendland, Elaheh Aghdassi, Carolyn Tam, Julie Carrier, A.
Hillary Steinhart, Stephen L. Wolman, David Baron, Kursheed N. Jeejee-
bhoy, Alvin Newman, Johane P. Allard, Univ of Toronto, Toronto, ON,
Canada.
Background! Aims: During Crohn's disease (CD) activated neutrophils in
the inflamed intestinal mucosa produce reactive oxygen species (ROS).
This increased production of ROS may reduce plasma antioxidant vitamins
resulting in oxidative stress. The purpose of this study was to compare
dietary intake of macro- and micro-nutrients, plasma antioxidant vitamins,
and lipid peroxidation, a measure of ROS production, between CD patients
and healthy controls (He). Methods: Thirty-seven non-smoking CD pa-
tients were matched to an equal number of healthy controls (He), who
were matched for age and gender. In patients, CD activity was measured:
30% had a COAl;:" 150. Medication use included: 5-ASA, 40%; antibi-
otics, 22%; oral corticosteroids, 30%; azathioprine, 19%; no medications,
22%. Seven day food intake records were completed by both groups. Lipid
peroxidation was measured by breath pentane and ethane output, and
F2-isoprostane. Results: There were no significant differences in mean
macro-and micro-nutrient intakes, and no differences in body mass index
(BMI) between groups. However, plasma antioxidant vitamins (ascorbic
acid, alpha- and beta-carotene, lycopene, and beta-cryptoxanthin) were
found to be significantly lower in CD compared to He. Oxidative stress,
assessed by breath pentane output (CD:7.47±0.98 vs HC:4.97±0.48 pmol/
kg/min, pSO.025), breath ethane output (CD: I 1.24±I.I7 vs
HC:5.46±0.71 pmol/kg/min, pSO.OOO5), and F2-isoprostane
(CD:78.6±8.0 vs HC:60.6±3.7 pglml, psO.047) was significantly higher
in CD compared to HC. Conclusion: Although there was no difference in
macro- and micro-nutrient intake, plasma antioxidant vitamins were sig-
nificantly lower in these stable CD patients compared to He. This, along
with increased oxidative stress, suggests antioxidant vitamin consumption
during the free radical scavenging process. This work was supported by the
Crohn's and Colitis Foundation of Canada.
1989
THE SEVERITY OF INFECTIOUS GASTROENTERITIS IS NOT
RELATED TO PATIENT ANXIETY AND DEPRESSION.
Mark R. Borgaonkar, John K. Marshall, Elizabeth Churchill, Debra Mar-
sillo, Eric Matthews, David King, e. Bannon, A. Haley; Cathy Buffett,
Elizabeth Richardson, David Stronach, Stephen M. Collins, McMaster
Univ, Hamilton, ON, Canada; Hamilton-Wentworth Regional Dept of
Public Health, Hamilton, ON, Canada; Halton Regional Public Health
Dept, Hamilton, ON, Canada.
Rationale: There is evidence supporting a relationship between psychoso-
cial factors and the development of post-infectious irritable bowel syn-
drome (PI-IBS). The relationship between the symptoms of acute infec-
tious gastroenteritis (GE) and psychological factors has not been studied.
Purpose: To examine the relationship between reported symptoms of
bacterial GE and psychological distress. Methods: A cohort with docu-
mented bacterial GE within the preceeding month was prospectively re-
cruited. Questionnaires regarding patient demographics, details of the
symptoms of the GE, and the Hospital Anxiety and Depression Scale
(HADS) were completed. Linear regression was used to assess the rela-
tionship between HADS score and symptom severity. Results: From April
to November 1999, 252 cases of bacterial GE occurred within the region of
Hamilton-Wentworth. The majority of cases were due to Campylobaeter
(33.7%), Giardi (25.4%), and Salmonella (23%) species. 61 patients (24%)
were eligible and agreed to participate. 33 had Campylobaeter (54.1%), 15
Giardia (24.6%), and 8 Salmonella (13.1%). The mean age was 43.3 years
with 33 (54.1%) females. 98% of patients had diarrhea, 88% abdominal
pain, 54% fever, and 34% vomiting. The mean number of days with
diarrhea, pain, fever, and vomiting was 13.2,9.5,3.3, and 2.0, respectively.
68.3% of patients missed at least some of their usual activities. The mean
time to return to all regular activities was 14.8 days. The mean HADS
anxiety (0-12) and depression scores (0-10) were 6.0 (SO 3.7, range 12)
and 3.0 (SD 2.7, range 10) respectively. There was no relationship between
the presence or duration of any symptoms and the HADS anxiety or
depression scores. Symptoms did not differ among the specific bacterial
pathogens. Conclusions: These data suggest that psychological factors have
no impact on the reported severity of acute bacterial GE, which is in
contrast to the association of psychological distress and development of
PI-IBS. This ongoing study will explore chronic symptoms after acute GE
and the incidence of PI-IBS.