- Email: [email protected]
Compensatery lacreasses in glutamine transport in bovine cryptosporidiosis
California. In an effort to develop a strategy to further lower the incidence of diarrhea, our AIM in this analysis was to identify the pattern of diarrhea peaks important to the spread of the outbreakfrom one county to the next in Southern California.METHODS:Diarrheaspecimens were submitted to one central laboratory (So. California PermanenteRegional Lab in North Hollywood, CA) from the hospitals and clinics of the Southern California PermanenteMedical Group. Fecal analysis data for bacterial identification, site of origin and month of the year were acquired for 1997-99. Examinationof the relationship between seasonaldiarrhea incidence peaks (> = mean+ 1standarddeviation) and one month lag correlation coefficient (cc) in surmueding five counties (Los Angeles (LA), San Semardino (SB), Riverside(RS), Orange (OR) and San Diego (SD)) was done, RESULTS:For Campyiobacterjejuni, the 1 lag cc between counties showed SB having the most influence on the subsequent month in the other four counties (cc 0.531). For Shigella sonnei RS and OR countieswere the drivers for this infection with a strong tendency for the infection peaks to move towards SO County after a month. For Shioella flexneri SD was an important driver with a strong 1 month lag cc to LA, 0.709. Salmonella group D produced a strong simultaneous cc between LA and SD coonbes, 0.604, and then JnfectJonpeak shifted fo RS County after a one-month lag. CONCLUSIONS:Specific outbreaks from specific counties spreading to other counties were identified in spite of enhanced state and local health regulations. Evidence for a lead lag phenomenon were discernible which can potentially identify infection before it spreads suggesting important implications for developing preventive strategies. organism
Driver county
C. jujuni
SB
S.Sonnei
RS, OR SD LA, SO
S.flexned Salmonellagp.D
production in response to known allergens. Histamine and TIfF production was each clearly lower when pancreaticenzymeswere given, suggestungthat pancreatic enzymesmay activate allergen degradation, histamine catabolism or medlate other not yet identified immunologic effects.
Methiyhlsta~ne A÷P: I ~ H I ~ A,P: I~TNF A÷P:
Pro,.chatlengeday
Provocation
Post-challenge day
5.8÷1 5.6+1
7.3+5 3.7±T
16.3+22 5.4±3
0.2~0.~ 0.4~0.2
0.9±1.3 0.1±0.05
0.2±0.1 0.1±0.2
6.0±2 6.2~2
14±15 7.5±3
7.5±3 5.7±1.6
"p<0.03
1136
I month lag cc to other counties
CeiqNmmly llu=wses In Glutamioe Transport in 6ovine CWptesporidiosis Elaine L Hunt, Anthony T. Blikslager, North Carolina State Univ, Raleigh, NC; Richard L. Guermnt, Univ of Virginia, Charlottesville,VA; J Marc Rhoeds, Univ of North Carolina, Chapel Hill, NC; Robert A. Argenzio, North Carolina State Univ, Raleigh, NC
LA 0.410, RS 0.395, OR 0.316, SD 0.287 $8 0.522, SO 0.484, SD 0.362 RS 0.336 RS 0.336 RS 0.354
BACKGROUND:Cryptosporidium parvum infection is a significant zoonofic diarrheal disease. Cryptospoddiosis in the human mimics that of the calf and the calf is a common reservoir of human infection. The parasite is resistant to antimicrobial treatment, and intravenous or oral rehydratlon with glucose-based solutions remains the single-most effective therapy. However,the severeVillousatrophy accompanyingthe infection would be expectedto destroy the mature eflterocyteswhich are principally responsiblefor the glucose-coupledNaabsorption upon which all oral rehydratlon solutions are based. Previous studies in infected piglet have shown that glntamioe is more effective than glucose in stimulating Na absorption despite villous atrophy and despite the presence of prostanoids, which inhibit electroneutral NaCI absorption. We studied the effects of luminally-applied glutamine and indomethacin on NaCI absorption from infected calf ileum. METHODS:Colostrum fed day-old Holstein calves were rn~nt~ned free of Cparvum infection until day 7 of life, at which time sham or oral infection was performed with 10~ oocysts. Four days later, ileal mucose was stripped and mounted in Ussing chambers. Morphomctric, immunohistochemical, and Western analyses were performed on select tissues. RESULTS: Infected ileum displayed a decrease in both mucocal surface area and NaCIabsorption as comparedto uninfected control. Indomethacin increased oct NaCIabsorption in infected tissue to control levels. Conversely,addition of fpJ~ PGEeto control tissue mimicked the altered transport of the infection, Glutamine and the stable dipelrddeaianyf-glutamineincreasednet Na absorption and Isc by lp.Ecl/cm2.hin both control and infected ileum. The addition of indomethacinto glutamine-treatedinfected tissue further stimulated net Na and CI absorption from that seen with glutamine alone and resulted in a significant net change from the untreated tissue of +3.5+-0.09 pEq/cm2.h (P < 0.001). Immunohlatochemical and Western blot studies showed that the Na-dependentASC neutral amino acid transport system was strongly expressedon the apical border of infected crypts. CONCLUSIONS:These results indicate that endogenous prostaglandins mediate the altered salt and water absorption in this infection. The combined use of indomethacinand glutamine can stimulate maximal NaCI absorption despite severe villous atrophy, an effect associated with increased expression of the Na-dependentglutamine transporter in crypt epithelium.
1134 Clinical Significance of Unusual Inflammntop/Bowel Diesese-Ubo PMbololli¢ Features in Lymphocytic and Collaoeoous Colitis Sarathchandra I. Reddy, Brigham and Women's Hasp, Boston, MA; Francis A. Farraye, Boston Medical Ctr, Boston, MA; Heidi Sapp, Tom Brien, Brigham and Women's Hosp, Boston, MA; Helen Wang, Beth Israel DeaconessMedical Ctr, Boston, MA; Robert D. Odze, Brigham and Women's Hosp, Boston, MA Background/Objective:The clinical course and natural history of lymphocytic colitis (LC) and coliagenouscolitis (CC) are quite variableand poorly understood.We have noted, anecdutaJly, the presence of unusual inflammatory bowel disease (IBD)-Iike histologic features, such as neutrophilic cryptitis and crypt abscesses, paneth cell metapiasia and crypt architectural disarray in patients with these disorders. The aim of this study was to evaluatethe clinical significance of a variety of typical and unusual histolngic features in LC and CC. Methods: Routinely processed H & E stained mucosal biopsies from 84 patients with clinically and pathologically confirmed LC (39 patients, F/M ratio: 31/8, mean age: 56 years) or CC (45 patients, F/M ratio: 36/9, mean age: 60 years) were evaluated for a number of histolngic features including neutrophilic cryptifis or crypt abscesses, paneth cell metsplasia, crypt architectural disarrayand thicknessof the subepithelia} collagen layer (CCon/y). The pathologic features were correlated with a variety of symptoms such as frequency and consistency of diarrhea, abdominal pain, tenesmus, NSAID or prednisone use, duration of disease, etc. as determined by a retrospective review of the patients medical records. Categoricalvariables were analyzedby Chi square test or Fischer exact test dependent on sample size while the continuous variables were analyzed by the Kroskal-Waifis test. Results: Cryptflis or crypt abscesses, paneth cell metaplasiaand crypt architectural disarray were seen in 28%, 8.6%, 5.1% of the LC patients, respectivelyand 16%, 35%, 4.4% of the CC patients, respectively. Paneth cell metaplasiawas significantly more common in CC (P=0.01). LC and CC patients had similar clinical features except that watery stools were more common in CC than LC, whereas stools with mucous were more common in LC than CC (P=0.01). The mean subepithelial collagen layer thickness in CC was 18.4 microns (range: 3-56). An association was observed between the severity of diarrhea, or presence of tsnesmus, and increasing collagen layer thickness (P = 0.06). Other histologic features were not associatedwith any of the clinical features, including drug use or outcome. Conclusions: A significant percentage of patients with LC or CC contain unusual histologic features, which may cause diagnostic confusion with IBD if not recognized. Increasingthickness of the sobepithellalcollagen layer in CC may indicate more severe disease.
1137 Tissue Traesglntamlnase Antibodies in Patients With Confirmed Food Allergy Michael Weidenhiller, Sandra Winterkamp, Dieter Schwab, Eckart Georg Hahn, Detlef Schuppan, Martin Raithel, Dept of Medicine I, Friedrich-Alexander-Universitaet,Erlangen Germany Backgmund~llergic enteropathy (gastrointestinal or food allergy ) is a rare condition and thought to involve toE, mast cells and eosinophils of the small or large bowel. In severecases of allergic entempathy the mucosa of duodenum and small intestine sometimes resembles celiac disease, with different degrees of partial or total villus atrophy. In addition, allergic individuals may also present with diarrhea or other gastrointestinal complaints, when food hypersensitivityprimarily involvesthe gastrointestinaltract. Gluten entempathy(celiacdisease) is a diseaseof the small bowel mediated by cytotoxic T-cells. Tissue transolutaminase (FrG) has been characterizedas an auto-antigenand antibodies can usually be found in high titers. Antibodies to serum-transglutaminase can also be found in Crohn s disease and indicate acltvfly of the disease. We lovestigeted, whether antibodies to TTG also exist in patients with proven allergic enteropathy (proven by double blind placebo controlled food challenge), Mcthods:Sera of 47 consecutive patients with allergic eoteropathywere investigated for the presence of "n'G antibodies by a commercially availableELISA.The cut-off for normal values has previously been set up at 5 mg/mL Resuits:0nly 1 of 47 patients (2 %) had borderline elavated TTG-antibndies.Her course differed from those of the other allergic patients in that she had been previously diagnosedas suffering from ulcerativecolitis. However,after specific allergen elimination diet, she was free of symptoms. No signs of celiac diseasewere obvious in this patient. Conclusion:Antibodiesto TTG can usually not be found in patients with allergic enteropathy. Low titer antibodies are not specific for celiac disease and seem to be a paraallergic phenomenon in the one positive patient. l-rG antibodies do not seem to have a diagnostic or pathogenetic role in the managementof allergic enteropethy-patients,but they can help to differentiate from celiac disease in unclear cases of allergic enteropathy with severe changes on upper endoscopy and histology.
1135 Application of Pancreatic Enzymes Significantly Reduces the Expression of Allergic Inflammation in Patients with Food Allergy (FA| Martin Raithel, Michael Weidenhiller, Eckhart Georg Hahn, Thomas Schneider, Dept Medicine I, Erlangen Germany FA involves 5% of the general population with increasing rates of incidence. Apart from allergen avoidancemany patients with FA try severaltherapeuticsduring their diseasecourse like mast cell stabilisers or antihistamines.Some patientsreport clinical benefitfrom pancreatic enzymesin terms of relief of gastrointestinalsymptoms. Sincethere are no dataon immunological effects of pancreaticenzymes,7 patients with known FA were repeatedlychallengedeither with the allergen alone (A) or in combination with pancreatic enzymes (A+P; 1200001E Panzytrat/challenge). All food challenge tests (n=22) were performed in a double-blind, placebo-controlled manner with fresh homogenizedfoods using a nasogestrictube. Clinical symptoms [points], urine methylhistamine [p.g/mmol crea x m2BSA], plasma TNF and histamine [both ng/ml x m2BSA]wereprospectivelyevaluatedbefore,dudng and affer the challenge. Results see table: Similarly to the decrease of symptom scores (A 7 +- 4, A+P 4.8 ± 3), application of pancreatic enzymes in FA strikingly reduced the amount of allergy mediator
A-216
Driver county
C. jujuni
SB
S.Sonnei
RS, OR SD LA, SO
S.flexned Salmonellagp.D
production in response to known allergens. Histamine and TIfF production was each clearly lower when pancreaticenzymeswere given, suggestungthat pancreatic enzymesmay activate allergen degradation, histamine catabolism or medlate other not yet identified immunologic effects.
Methiyhlsta~ne A÷P: I ~ H I ~ A,P: I~TNF A÷P:
Pro,.chatlengeday
Provocation
Post-challenge day
5.8÷1 5.6+1
7.3+5 3.7±T
16.3+22 5.4±3
0.2~0.~ 0.4~0.2
0.9±1.3 0.1±0.05
0.2±0.1 0.1±0.2
6.0±2 6.2~2
14±15 7.5±3
7.5±3 5.7±1.6
"p<0.03
1136
I month lag cc to other counties
CeiqNmmly llu=wses In Glutamioe Transport in 6ovine CWptesporidiosis Elaine L Hunt, Anthony T. Blikslager, North Carolina State Univ, Raleigh, NC; Richard L. Guermnt, Univ of Virginia, Charlottesville,VA; J Marc Rhoeds, Univ of North Carolina, Chapel Hill, NC; Robert A. Argenzio, North Carolina State Univ, Raleigh, NC
LA 0.410, RS 0.395, OR 0.316, SD 0.287 $8 0.522, SO 0.484, SD 0.362 RS 0.336 RS 0.336 RS 0.354
BACKGROUND:Cryptosporidium parvum infection is a significant zoonofic diarrheal disease. Cryptospoddiosis in the human mimics that of the calf and the calf is a common reservoir of human infection. The parasite is resistant to antimicrobial treatment, and intravenous or oral rehydratlon with glucose-based solutions remains the single-most effective therapy. However,the severeVillousatrophy accompanyingthe infection would be expectedto destroy the mature eflterocyteswhich are principally responsiblefor the glucose-coupledNaabsorption upon which all oral rehydratlon solutions are based. Previous studies in infected piglet have shown that glntamioe is more effective than glucose in stimulating Na absorption despite villous atrophy and despite the presence of prostanoids, which inhibit electroneutral NaCI absorption. We studied the effects of luminally-applied glutamine and indomethacin on NaCI absorption from infected calf ileum. METHODS:Colostrum fed day-old Holstein calves were rn~nt~ned free of Cparvum infection until day 7 of life, at which time sham or oral infection was performed with 10~ oocysts. Four days later, ileal mucose was stripped and mounted in Ussing chambers. Morphomctric, immunohistochemical, and Western analyses were performed on select tissues. RESULTS: Infected ileum displayed a decrease in both mucocal surface area and NaCIabsorption as comparedto uninfected control. Indomethacin increased oct NaCIabsorption in infected tissue to control levels. Conversely,addition of fpJ~ PGEeto control tissue mimicked the altered transport of the infection, Glutamine and the stable dipelrddeaianyf-glutamineincreasednet Na absorption and Isc by lp.Ecl/cm2.hin both control and infected ileum. The addition of indomethacinto glutamine-treatedinfected tissue further stimulated net Na and CI absorption from that seen with glutamine alone and resulted in a significant net change from the untreated tissue of +3.5+-0.09 pEq/cm2.h (P < 0.001). Immunohlatochemical and Western blot studies showed that the Na-dependentASC neutral amino acid transport system was strongly expressedon the apical border of infected crypts. CONCLUSIONS:These results indicate that endogenous prostaglandins mediate the altered salt and water absorption in this infection. The combined use of indomethacinand glutamine can stimulate maximal NaCI absorption despite severe villous atrophy, an effect associated with increased expression of the Na-dependentglutamine transporter in crypt epithelium.
1134 Clinical Significance of Unusual Inflammntop/Bowel Diesese-Ubo PMbololli¢ Features in Lymphocytic and Collaoeoous Colitis Sarathchandra I. Reddy, Brigham and Women's Hasp, Boston, MA; Francis A. Farraye, Boston Medical Ctr, Boston, MA; Heidi Sapp, Tom Brien, Brigham and Women's Hosp, Boston, MA; Helen Wang, Beth Israel DeaconessMedical Ctr, Boston, MA; Robert D. Odze, Brigham and Women's Hosp, Boston, MA Background/Objective:The clinical course and natural history of lymphocytic colitis (LC) and coliagenouscolitis (CC) are quite variableand poorly understood.We have noted, anecdutaJly, the presence of unusual inflammatory bowel disease (IBD)-Iike histologic features, such as neutrophilic cryptitis and crypt abscesses, paneth cell metapiasia and crypt architectural disarray in patients with these disorders. The aim of this study was to evaluatethe clinical significance of a variety of typical and unusual histolngic features in LC and CC. Methods: Routinely processed H & E stained mucosal biopsies from 84 patients with clinically and pathologically confirmed LC (39 patients, F/M ratio: 31/8, mean age: 56 years) or CC (45 patients, F/M ratio: 36/9, mean age: 60 years) were evaluated for a number of histolngic features including neutrophilic cryptifis or crypt abscesses, paneth cell metsplasia, crypt architectural disarrayand thicknessof the subepithelia} collagen layer (CCon/y). The pathologic features were correlated with a variety of symptoms such as frequency and consistency of diarrhea, abdominal pain, tenesmus, NSAID or prednisone use, duration of disease, etc. as determined by a retrospective review of the patients medical records. Categoricalvariables were analyzedby Chi square test or Fischer exact test dependent on sample size while the continuous variables were analyzed by the Kroskal-Waifis test. Results: Cryptflis or crypt abscesses, paneth cell metaplasiaand crypt architectural disarray were seen in 28%, 8.6%, 5.1% of the LC patients, respectivelyand 16%, 35%, 4.4% of the CC patients, respectively. Paneth cell metaplasiawas significantly more common in CC (P=0.01). LC and CC patients had similar clinical features except that watery stools were more common in CC than LC, whereas stools with mucous were more common in LC than CC (P=0.01). The mean subepithelial collagen layer thickness in CC was 18.4 microns (range: 3-56). An association was observed between the severity of diarrhea, or presence of tsnesmus, and increasing collagen layer thickness (P = 0.06). Other histologic features were not associatedwith any of the clinical features, including drug use or outcome. Conclusions: A significant percentage of patients with LC or CC contain unusual histologic features, which may cause diagnostic confusion with IBD if not recognized. Increasingthickness of the sobepithellalcollagen layer in CC may indicate more severe disease.
1137 Tissue Traesglntamlnase Antibodies in Patients With Confirmed Food Allergy Michael Weidenhiller, Sandra Winterkamp, Dieter Schwab, Eckart Georg Hahn, Detlef Schuppan, Martin Raithel, Dept of Medicine I, Friedrich-Alexander-Universitaet,Erlangen Germany Backgmund~llergic enteropathy (gastrointestinal or food allergy ) is a rare condition and thought to involve toE, mast cells and eosinophils of the small or large bowel. In severecases of allergic entempathy the mucosa of duodenum and small intestine sometimes resembles celiac disease, with different degrees of partial or total villus atrophy. In addition, allergic individuals may also present with diarrhea or other gastrointestinal complaints, when food hypersensitivityprimarily involvesthe gastrointestinaltract. Gluten entempathy(celiacdisease) is a diseaseof the small bowel mediated by cytotoxic T-cells. Tissue transolutaminase (FrG) has been characterizedas an auto-antigenand antibodies can usually be found in high titers. Antibodies to serum-transglutaminase can also be found in Crohn s disease and indicate acltvfly of the disease. We lovestigeted, whether antibodies to TTG also exist in patients with proven allergic enteropathy (proven by double blind placebo controlled food challenge), Mcthods:Sera of 47 consecutive patients with allergic eoteropathywere investigated for the presence of "n'G antibodies by a commercially availableELISA.The cut-off for normal values has previously been set up at 5 mg/mL Resuits:0nly 1 of 47 patients (2 %) had borderline elavated TTG-antibndies.Her course differed from those of the other allergic patients in that she had been previously diagnosedas suffering from ulcerativecolitis. However,after specific allergen elimination diet, she was free of symptoms. No signs of celiac diseasewere obvious in this patient. Conclusion:Antibodiesto TTG can usually not be found in patients with allergic enteropathy. Low titer antibodies are not specific for celiac disease and seem to be a paraallergic phenomenon in the one positive patient. l-rG antibodies do not seem to have a diagnostic or pathogenetic role in the managementof allergic enteropethy-patients,but they can help to differentiate from celiac disease in unclear cases of allergic enteropathy with severe changes on upper endoscopy and histology.
1135 Application of Pancreatic Enzymes Significantly Reduces the Expression of Allergic Inflammation in Patients with Food Allergy (FA| Martin Raithel, Michael Weidenhiller, Eckhart Georg Hahn, Thomas Schneider, Dept Medicine I, Erlangen Germany FA involves 5% of the general population with increasing rates of incidence. Apart from allergen avoidancemany patients with FA try severaltherapeuticsduring their diseasecourse like mast cell stabilisers or antihistamines.Some patientsreport clinical benefitfrom pancreatic enzymesin terms of relief of gastrointestinalsymptoms. Sincethere are no dataon immunological effects of pancreaticenzymes,7 patients with known FA were repeatedlychallengedeither with the allergen alone (A) or in combination with pancreatic enzymes (A+P; 1200001E Panzytrat/challenge). All food challenge tests (n=22) were performed in a double-blind, placebo-controlled manner with fresh homogenizedfoods using a nasogestrictube. Clinical symptoms [points], urine methylhistamine [p.g/mmol crea x m2BSA], plasma TNF and histamine [both ng/ml x m2BSA]wereprospectivelyevaluatedbefore,dudng and affer the challenge. Results see table: Similarly to the decrease of symptom scores (A 7 +- 4, A+P 4.8 ± 3), application of pancreatic enzymes in FA strikingly reduced the amount of allergy mediator
A-216