Western Abstracts 54-1 01/05/2006 8:12 AM Page S126
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Days in Force/Gel n Number of Cells per
Culture (dynes) Std Error Cross-Section Std Error n
1 31 6 8 992 8 4
4 41 7 14 1,024 3 4
8 57 9 10 2,688 18 4
12 89 9 12 3,520 23 4
Conclusion: We have for the first time directly measured the force generated by HSCs after
a range of days in culture. Both the force of contraction and the number of cells within a
cross-section of a gel increased with the number of days in culture. This suggests that the
time-dependent increase in force generation may be due to an increase in HSC number
rather than an increase in force generated by each cell.
268
PREDNISOLONE BLOCKS NUCLEAR FACTOR KAPPA B TRANSCRIPTIONAL
ACTIVATION THROUGH THE GLUCOCORTICOID RESPONSE ELEMENT. M.A. Boivin,
J. Kennedy, D. Ye, T.Y. Ma, Department of Medicine, University of New Mexico,
Albuquerque, NM.
Altered intestinal permeability is an important potential cause of Crohn’s disease.
Cytokines, an important therapeutic target in Crohn’s disease, can cause disruption of the
intestinal tight junction barrier, leading to increased permeability. Tumor necrosis factor
alpha (TNF-) activates nuclear factor kappa B (NF-
B), causing an increase in expression respect to smell by 22 (53.7%) of subjects and taste by 33 (78.6%). CM was rated as the most
of the enzyme myosin light chain kinase (MLCK) that modulates the tight junction barrier
defect. We have shown this increase in expression is due to NF-
B activating the MLCK pro- rated the most offensive taste by 12 (28.6%) of subjects. The VAS score for F was statistically
moter through specific NF-
B binding sites. Furthermore, prednisolone (an important lower than all other beverages with respect to both smell and taste (p < .005). Conclusion:
treatment of Crohn’s disease) protects the intestinal epithelial tight junction barrier by
blocking NF-
B–mediated MLCK promoter activation. In examining the MLCK promoter, considered as an option when administering oral NAC to adults.
we determined that it contained a sequence that matched the consensus for a glucocorti-
coid response element (GRE). In this abstract, we determine the mechanism through which
glucocorticoids prevent NF-
B activation of the MLCK promoter. Methods: Filter-grown 272
epithelial Caco-2 cells were used as an in vitro model of the intestinal barrier. The 2 kb
upstream of the MLCK translational start site was cloned into pGL3 vector and used for pro-
moter activity studies. Results: (1) Prednisolone blocked the TNF-–induced increase in
intestinal epithelial permeability. (2) Prednisolone inhibited the TNF-–induced increase
in MLCK expression. (3) Prednisolone did not inhibit NF-
B translocation to the nucleus. CA; Targeted Diagnostics & Therapeutics Inc., West Chester, PA; National Cancer Institute,
(4) Prednisolone caused translocation of glucocorticoid receptor (GR) to the nucleus. (5)
Prednisolone increased binding of GR to its transcriptional factor binding site, the GRE. (6)
Prednisolone blocked the TNF-–induced up-regulation of MLCK promoter activity. (7)
Prednisolone did not block the TNF- up-regulation of MLCK promoter if (a) the GRE was
rendered inactive through site-directed mutagenesis or (b) RU-486 (GRE binding antago-
nist) was present. (8) Mutation of the NF-
B site 30 bp downstream of the MLCK GRE site detection of colorectal carcinoma. Recent studies have identified detectable GCC in the
blocked TNF-–induced promoter up-regulation. Conclusions: For the first time, we
demonstrate that glucocorticoids can block NF-
B–mediated promoter activation through the significance of GCC expression in lymph nodes from CRC patients with clinical out-
a GRE site. Further studies will identify the interaction between the GRE site on the MLCK
promoter and the nearby NF-
B binding site using mutagenesis, binding studies, and inser- sis under an institutional review board–approved protocol from 103 CRC patients (stage I:
tions.
269
CRITICAL ROLE OF INTERLEUKIN-17 AND INTERLEUKIN-17 RECEPTOR SIGNALING
IN TNBS-INDUCED COLITIS. Z. Zhang,1 M. Zheng,2 J. Bindas,2 P. Schwarzenberger,3 J.K.
Kolls,2 1Oregon Health & Science University, Portland, OR; 2Children’s Hospital of
Pittsburgh, Pittsburgh, PA; 3Louisiana State University Health Sciences Center, New
Orleans, LA.
Inflammatory bowel diseases (IBD) are characterized by recurrent inflammation in the gas-
trointestinal tract. Infiltration of CD4+ lymphocytes and neutrophils is one of the predom-
inant features in IBD. Recently, interleukin (IL)-23 and the downstream T-cell derived
cytokine IL-17 have been found to be elevated in IBD patients. However, the role of IL-17
and IL-17 receptor (IL-17R) signaling in gastrointestinal inflammation is unknown. To
examine their role, we investigated colonic inflammation in wild-type or IL-17R KO mice.
Using a model of TNBS-induced colitis, we found that IL-17 was produced in colon tissue
at 24 and 48 hours. IL-17R KO mice were significantly protected against TNBS-induced
weight loss, IL-6 production, local MIP-2 induction, as well as colonic inflammation. This
protection occurred in the presence of equivalent induction of local IL-23 and higher lev-
els of IL-12p70 and IFN-gamma in IL-17R KO mice compared to wild-type mice. Moreover,
IL-17R KO mice exhibited a reduced tissue neutrophil infiltration as assessed by myeloper-
oxidase activity. Furthermore, overexpression of an IL-17R IgG1 fusion protein significantly
attenuated colonic inflammation after TNBS challenge. These results demonstrate that IL-
17 and IL-17R signaling play a critical role in the development of TNBS-induced colitis and
may represent a target for therapeutic intervention for IBD.
270
REDUCE THE RISK OF COLON CANCER WITH COLON CANCER SCREENING. K. Kempe,
University of Washington School of Medicine, Seattle, WA.
Purpose: There are very low rates of colon cancer screening in White Sulphur Springs, Mon-
tana, population 1,010. An educational project was designed to increase community aware-
ness of the risk of colon cancer and the benefits from colon cancer screening. Four modal-
ities for colon cancer screening are available: colonoscopy, barium enema, sigmoidoscopy,
and fecal occult. Mountainview Medical Center is the only medical facility within a 100-
mile radius. A gastroenterologist travels from Helena, MT, to perform screening exams in
White Sulphur Springs on a quarterly basis, but the service is currently underutilized. Meth-
ods: A literature search using MEDLINE and UW Healthlinks was completed looking at the
epidemiology of colon cancer, risk factors, and screening efficacy. In addition, insurance
coverage and costs of each procedure were evaluated. Interviews were conducted with local
health care providers to identify available opportunities for colon cancer screening as well
as possible barriers to screening in the community. Results: Colorectal cancer ranks second
to lung cancer as a cause of cancer death. Colon cancer screening, including colonoscopy,
can significantly reduce the risk of colon cancer and prevent colon cancer. An article was
written, including information about colon cancer risks, prevention, and costs, and was
S126
distributed to every household in the county by the local community hospital. Conclusion:
Education, increased awareness of the risks of colon cancer, screening modalities, and
Medicare’s willingness to pay for colon cancer screening may increase dialogue between
patient and practitioner regarding colon cancer as well as increase colon cancer screenings
in White Sulphur Springs, Montana.
271
MASKING THE SMELL AND TASTE OF ACETYLCYSTEINE: WHAT IS THE BEST OPTION?
C.E. Dandoy, B.I. Crouch, E.M. Caravati, Utah Poison Control Center, University of Utah,
Salt Lake City, UT.
Objective: To determine what beverage best masks the smell and taste of acetylcysteine
(NAC). Methods: A 5% solution of NAC was prepared using the following five diluents: water
(W), FrescaTM (F), Coca ColaTM (CC), cranberry juice (CJ), chocolate milk (CM). Subjects
used a 10 cm visual analog scale (VAS) to rate the smell and taste for each 5% solution with
the left margin representing a non-offensive odor or taste and the right margin represent-
ing a very offensive odor or taste. For the taste portion, approximately 10 mL of each 5%
solution was placed in a cup with a lid and straw. Each subject tasted each solution in a dif-
ferent order than for smell. The subjects ate soup crackers between each taste test to limit
carryover between solutions. The difference in VAS for taste and smell was measured using
the Kruskal-Wallis ranked sum test. Multiple comparisons were performed using the Mann
Whitney U test with the Bonferoni correction with significance at p < .005. Results: A total
of 42 subjects participated in the study. The mean VAS score (cm) for smell and taste,
respectively, of each dilutent was (F) 1.1738, 4.2833; (CC) 2.6405, 7.2714; (CJ) 3.9762, 8.1786;
(W) 4.1071, 7.7810; and (CM) 5.1286, 8.076. F was rated as the least offensive diluent with
offensive beverage with respect to smell by 22 (53.7%) and to taste by 11 (26.2%). CJ was
NAC diluted with F to a 5% solution had the least offensive odor and taste and should be
DETECTION, ANALYSIS AND SIGNIFICANCE OF GUANYLYL CYCLASE C IN THE LYMPH
NODES FROM PATIENTS WITH COLORECTAL CANCER. C. Tang, A. Wu, E. Leong, T.
Fears, R. Azzi, D. O’Shannessy, A. Lin, University of California at Los Angeles, Los Angeles,
NIH, Bethesda, MD; Santa Clara Valley Medical Center, San Jose, CA.
Purpose: Colorectal cancer (CRC) is one of the top leading causes of cancer-related mor-
tality in the United States and worldwide. Guanylyl cyclase C (GCC) is a receptor that medi-
ates fluid and electrolyte secretion and is expressed only in mucosal cells lining the intes-
tine but not by other extraintestinal tissues. Expression of GCC has been used for the
peripheral blood from patients diagnosed with CRC. This study was designed to correlate
comes. Methods Used: Paraffin-embedded lymph nodes (PELN) were obtained for analy-
11%; stage II: 31%; stage III: 34%; stage IV: 23%; median follow-up 3.4 years, range 0.1–12.1
years). The male to female ratio was 57:46. Follow-up information was obtained from hos-
pital tumor registry and RT-PCR assays were performed in 273 PELN blocks. Kaplan-Meier
methods were used to estimate overall survival. Cox proportional hazard models were used
for univariate as well as multivariate analyses. Results: The Spearman coefficient of corre-
lation between the expression of GCC and clinical stage was 0.67 (p < .01). Survival was
associated with clinical stage (p < .001) and GCC expression (p < .01) but not chemother-
apy (p = .44) or tumor grade (p = .11). Conclusion: GCC expression by RT-PCR assay of the
lymph nodes significantly correlates with clinical stage. In addition to clinical stage, its
expression in the lymph nodes is associated with survival in patients with CRC. Further
studies with a larger sample size are needed to confirm these findings.
General Neonatology II
Concurrent Session
1:30 PM
Friday, February 3, 2006
273
THE EFFECTS OF LACTOFERRIN ON THE ANTIMICROBIAL ACTIVITY OF
PASTEURIZED HUMAN MILK. G.M. Chan, University of Utah Health Science Center, Salt
Lake City, UT.
Human milk provides both nutritional and immunologic benefits for the infant. Pasteur-
ized human milk (PHM) is available commercially, but pasteurization has been reported to
decrease the milk’s antimicrobial properties. Lactoferrin (Lf ) is a major human milk protein
with antimicrobial activity that is dependent on its iron binding capabilities. This protein
has been isolated and purified but never added to PHM. The major aim of this study was to
evaluate the addition of Lf in PHM and study its in vitro effects against 3 major organisms
in the NBICU, E. coli, E. sakazakii, and group B streptococcus. Human milk samples were
collected from mothers with infants in NBICU between the 7th and 112th postpartum day.
All lactating mothers were healthy and were on no medications except vitamins with iron.
All samples were frozen and used within 4 weeks. Pasteurization was by the Holder method
of heating the milk to 62.5 C for 30 minutes. Lf was obtained from recombinant human Lf
(Ventria Bioscience, Sacramento, CA). Two Lfs were tested: one with no iron and another
that was > 50% saturated with iron (Lf/Fe). 1.1 mg/mL of each Lf were added to the PHM.
Bacterial growth of the 3 organisms was analyzed by inoculating 1 mL of 107 bacteria into
the milk samples and counting the number of colony-forming units after 3.5 hours of incu-
bation at 37 C. Analyses of variance with the Bonferroni post test were used to analyze the
data.
JOURNAL OF INVESTIGATIVE MEDICINE
 Downloaded from http://jim.bmj.com/ on February 16, 2018 - Published by group.bmj.com

271 MASKING THE SMELL AND TASTE OF

ACETYLCYSTEINE: WHAT IS THE BEST
OPTION?
C. E. Dandoy, B. I. Crouch and E. M. Caravati

J Investig Med2006 54: S126

doi: 10.2310/6650.2005.X0004.270

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