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Expression of monosaccharide transporters in the intestine of human diabetics
April 2000
AGAA885
4893
4895
EXPRESSION OF MONOSACCHARIDE TRANSPORTERS IN THE INTESTINE OF HUMAN DIABETICS.
BILE ACID MALABSORPTION (BAM) IN PATIENTS WITH FUNCTIONAL CHRONIC DIARRHEA: RESPONSE TO CHOLESTYRAMINE.
Jane Dyer, Altaf Palejwala, Anthony Ellis, Soraya P. Shirazi-Beechey, The Univ of Liverpool, Liverpool, United Kingdom. D-glucose and Dvgalactose are transported across the brush-border membrane, into enterocytes by the Na+Iglucose co-transporter (SGLTl). Fructose is transported from the intestinal lumen into the enterocytes by the fructose transporter (GLUTS). These monosaccharides exit the cell across the basolateral membrane by the monosaccharide transporter, GLUT2. It has been shown in rats with experimentally induced diabetes that the capacity of the small intestine to absorb D-glucose increases. This increase is mainly due to enhanced activity and abundance of SGLTl and GLUT2. No studies have been carried out to investigate if a similar situation occurs in the intestine of diabetic humans. We have assessed the expression of SGLTl, GLUTS, disaccharidases, and the structural proteins villin and l3-actin in the duodenum of healthy control individuals and patients with non-insulin dependent diabetes mellitus (NIDDM). Biopsy samples were removed, during endoscopy, from the second part of the duodenum of both control and diabetic subjects. Purified brush-border membrane vesicles (BBMV) were prepared from human biopsies using an established technique. A 4-fold increase in the abundance of SGLTI in the BBMV isolated from diabetic patients (85.2 ± 13.4 pmollmg protein, n=6) compared with controls (20.0 ± 5.6 pmollmg protein, n=6), was shown by quantitative western blotting. A similar increase in the initial rate of Na+-dependent Dsglucose transport in BBMV isolated from the intestine of diabetic patients compared with controls was also shown. The abundance of GLUTS, disaccharidases, villin and l3-actin were determined in the same BBMV by western blotting with appropriate antibodies. Our results indicate that levels of the fructose transporter, GLUTS, are also increased significantly in diabetics compared to controls. In contrast the abundance of lactase, sucrase, and the structural protein villin were only slightly elevated in diabetic patients. Total protein recoveries were equal in biopsies taken from both control and diabetic patients. We report here for the first time an increase in the capacity of the human intestine to absorb monosaccharides in NIDDM. This increase appears to be due to a combination of villus hypertrophy and a specific increase in the expression of SGLTl and GLUTS.
4894 ULTRASONOGRAPHY AND ENDOSCOPY FOR THE DIAGNOSIS OF PENICILLIN-INDUCED HEMORRHAGIC SEGMENTAL COLITIS - COMPARISON WITH PSEUDOMEMBRANOUS COLITIS. Christoph F. Dietrich, Barbara Braden, Hans Seifert, Bernhard Lembcke, Wolfgang F. Caspary, Till Wehrmann, Univ Frankfurt, Frankfurt, Germany. Penicillin-induced segmental hemorrhagic colitis (she) is a characteristic and striking but rare diagnosed clinical entity. Bloody diarrhea and abdominal cramps start few days after the intake of oral penicillin-derivates. We report the ultrasonographic, endoscopic and clinical findings of 9 she-patients and compare the results with the findings in 10 patients with antibiotic-related pseudomembranous colitis (pmc). Methods: 9 consecutive she-patients (age: 32 ± 10 years) in whom pmc was ruled out by endoscopy findings, stool cultures and clostridium difficile toxin-tests and 10 pmc-patients (age: 50 ± 18 years) with positive endoscopy and clostridium difficile toxin findings were examined by high resolution ultrasonography. The sonographic and endoscopic findings as well as the clinical course of the diseases were documented. Results: In all 9 shepatients the wall of the ascending colon was asymmetrically thickened with loss of the layer structure. Neither the small bowel nor the coecum were involved in she-patients. In all patients a distinct border between involved and uninvolved colonic wall was found, which was endoscopically confirmed. During follow-up all patients recovered soon after stopping antibiotic treatment and symptomatic care. In 7 pmc-patients pancolitis was found whereas in the remaining 3 patients left-sided colitis could be revealed at ultrasonography. In all 10 pmc-patients bowel wall was symmetrically thickened with the layers remaining distinct. The clinical course was more protracted. Conclusion: Clinical course and sonographic findings are helpful in differentiating patients with she and pmc. The knowledge of the clinical characteristics and sonographic findings of the penicillininduced segmental hemorrhagic colitis may reduce the need for invasive endoscopic and radiological investigations in diagnosis and follow-up.
Fernando Femandez-Banares, Maria Esteve, Jorge C. Espinos, Montserrat Forne, Josep Martin-Comin, Josep M. Viver, Hosp Univ Mutua de Terrassa, Terrassa, Spain; CSUB Bellvitge, L'Hospitalet, Spain. Frequency of BAM in patients with chronic diarrhea of functional characteristics is variable, ranging from 25 to 80% in the different studies. It has been stated that contribution of BAM to diarrhea in a given patient can only be determined by a therapeutic trial of cholestyramine. Aim: To study the frequency of BAM in patients with functional chronic diarrhea in our geographical area. To assess the response to cholestyramine. Methods: 32 patients (11 m, 21 w; age: 53±2 yrs) with functional chronic diarrhea of > 3 months of duration were included. Nine had previous cholecystectomy. Previous time with diarrhea was a median of 48 months (interquartile range -IQ-, 6-72) and daily stool number was a median of 5 (lQ, 4-7). In all cases routine blood biochemistry and hematology, parasitological exam of feces, serum T4-TSH, C reactive protein, IgA-antigliadin/ antiendomysium antibodies, small bowel barium meal and full colonoscopy with multiple biopsies were normal. BAM was assessed by means of the 75SeHCAT abdominal retention test (N) 11% on day 7). Patients with BAM received cholestyramine (sachets) at a starting dose of 4 g1day, which was modified depending on clinical response (2 to 12 g1day). Treatment success was considered as the passage of two or less formedlsemiformed stools per day. Results: BAM was detected in 24 of the 32 patients (75%; 95% CI, 57-88.5%), and in 15 of the 23 without cholecystectomy (65%). BAM was severe (value<5%) in 12. Diarrhea spontaneously resolved in 2 patients and with loperamide in I. In the other 21 patients cholestyramine was administered at a median dose of 8 g1day (lQ, 4-12). Response was not valuable by associated medical problems in 1 of them. In the other 20 diarrhea stopped (response: 100%; 95% CI, 83-100%); median daily stool number passed from 5 (IQ, 4-8) to I (IQ, 1-2) (p=0.OOO2), and feces increased their consistency. Conclusions: Frequency of BAM in patients with chronic diarrhea of functional characteristics is high in our geographical area. The excellent response to cholestyramine suggests that BAM is the cause of diarrhea in such patients.
4896 HIGH ANTI·TOXIN A ANTIBODY LEVELS ARE ASSOCIATED WITH PROTECTION AGAINST RECURRENT CLOSTRIDIUM DIFFICILE DIARRHEA. Lorraine Kyne, Michel Wamy, Amir Qamar, Ciaran P. Kelly, Beth Israel Deaconess Med Ctr & Harvard Med Sch, Boston, MA. Background & Aim: We reported recently that asymptomatic carriers of toxigenic C. difficile have high serum IgG anti-toxin A antibody levels. The aim of this study was to determine whether antibody responses to toxin A during an episode of C. difficile diarrhea influence the risk for recurrent disease. Methods: We studied all patients with antibiotic-associated C. difficile diarrhea in our hospital during a 5 month period. We prospectively determined the clinical severity and duration of diarrhea and followed patients for 60 days after discharge to identify relapses. Serial serum IgA, IgG, and IgM antibody levels against C. difficile toxin A, toxin B and non-toxin antigens were measured and results expressed as ELISA units (EU). Results: We identified 63 patients with C. difficile diarrhea all of whom agreed to participate in the study. Patients with severe C. difficile diarrhea (n=20) had lower serum IgG anti-toxin A levels than those with mild disease (median levels: 0.66 versus 1.60 EU, p=O.04). Patients with prolonged (> 14 days) C. difficile diarrhea (n= 10) had a later peak in serum IgG anti-toxin A levels compared to those with a shorter duration of symptoms (median: day 9 versus day 3, p=0.03). Patients with recurrent C. difficile diarrhea (n=22) had lower serum IgM anti-toxin A levels on day 3 of their first episode of diarrhea (0.85 versus 1.72 EU, p=O.OO4) and lower serum IgG anti-toxin A levels on day 12 (0.66 versus 3.87, p=0.009) compared to those who did not suffer a relapse. Multivariable analysis indicated that patients with a low serum IgM anti-toxin A level on day 3 had a much greater risk of recurrence compared to patients with high antitoxin levels (adjusted odds ratio 21.9, 95% CI: 1.8 to 274.1, p=0.02). Conclusions: An antibody response against toxin A acquired during an initial episode of C. difficile diarrhea is associated with a 22-fold reduced risk of recurrence. Our findings indicate that the host immune response to toxin A plays a substantial role in determining the clinical course of C. difficile diarrhea. This study also provides further evidence that acquired immunity against toxin A may protect against C. difficile diarrhea in humans.
AGAA885
4893
4895
EXPRESSION OF MONOSACCHARIDE TRANSPORTERS IN THE INTESTINE OF HUMAN DIABETICS.
BILE ACID MALABSORPTION (BAM) IN PATIENTS WITH FUNCTIONAL CHRONIC DIARRHEA: RESPONSE TO CHOLESTYRAMINE.
Jane Dyer, Altaf Palejwala, Anthony Ellis, Soraya P. Shirazi-Beechey, The Univ of Liverpool, Liverpool, United Kingdom. D-glucose and Dvgalactose are transported across the brush-border membrane, into enterocytes by the Na+Iglucose co-transporter (SGLTl). Fructose is transported from the intestinal lumen into the enterocytes by the fructose transporter (GLUTS). These monosaccharides exit the cell across the basolateral membrane by the monosaccharide transporter, GLUT2. It has been shown in rats with experimentally induced diabetes that the capacity of the small intestine to absorb D-glucose increases. This increase is mainly due to enhanced activity and abundance of SGLTl and GLUT2. No studies have been carried out to investigate if a similar situation occurs in the intestine of diabetic humans. We have assessed the expression of SGLTl, GLUTS, disaccharidases, and the structural proteins villin and l3-actin in the duodenum of healthy control individuals and patients with non-insulin dependent diabetes mellitus (NIDDM). Biopsy samples were removed, during endoscopy, from the second part of the duodenum of both control and diabetic subjects. Purified brush-border membrane vesicles (BBMV) were prepared from human biopsies using an established technique. A 4-fold increase in the abundance of SGLTI in the BBMV isolated from diabetic patients (85.2 ± 13.4 pmollmg protein, n=6) compared with controls (20.0 ± 5.6 pmollmg protein, n=6), was shown by quantitative western blotting. A similar increase in the initial rate of Na+-dependent Dsglucose transport in BBMV isolated from the intestine of diabetic patients compared with controls was also shown. The abundance of GLUTS, disaccharidases, villin and l3-actin were determined in the same BBMV by western blotting with appropriate antibodies. Our results indicate that levels of the fructose transporter, GLUTS, are also increased significantly in diabetics compared to controls. In contrast the abundance of lactase, sucrase, and the structural protein villin were only slightly elevated in diabetic patients. Total protein recoveries were equal in biopsies taken from both control and diabetic patients. We report here for the first time an increase in the capacity of the human intestine to absorb monosaccharides in NIDDM. This increase appears to be due to a combination of villus hypertrophy and a specific increase in the expression of SGLTl and GLUTS.
4894 ULTRASONOGRAPHY AND ENDOSCOPY FOR THE DIAGNOSIS OF PENICILLIN-INDUCED HEMORRHAGIC SEGMENTAL COLITIS - COMPARISON WITH PSEUDOMEMBRANOUS COLITIS. Christoph F. Dietrich, Barbara Braden, Hans Seifert, Bernhard Lembcke, Wolfgang F. Caspary, Till Wehrmann, Univ Frankfurt, Frankfurt, Germany. Penicillin-induced segmental hemorrhagic colitis (she) is a characteristic and striking but rare diagnosed clinical entity. Bloody diarrhea and abdominal cramps start few days after the intake of oral penicillin-derivates. We report the ultrasonographic, endoscopic and clinical findings of 9 she-patients and compare the results with the findings in 10 patients with antibiotic-related pseudomembranous colitis (pmc). Methods: 9 consecutive she-patients (age: 32 ± 10 years) in whom pmc was ruled out by endoscopy findings, stool cultures and clostridium difficile toxin-tests and 10 pmc-patients (age: 50 ± 18 years) with positive endoscopy and clostridium difficile toxin findings were examined by high resolution ultrasonography. The sonographic and endoscopic findings as well as the clinical course of the diseases were documented. Results: In all 9 shepatients the wall of the ascending colon was asymmetrically thickened with loss of the layer structure. Neither the small bowel nor the coecum were involved in she-patients. In all patients a distinct border between involved and uninvolved colonic wall was found, which was endoscopically confirmed. During follow-up all patients recovered soon after stopping antibiotic treatment and symptomatic care. In 7 pmc-patients pancolitis was found whereas in the remaining 3 patients left-sided colitis could be revealed at ultrasonography. In all 10 pmc-patients bowel wall was symmetrically thickened with the layers remaining distinct. The clinical course was more protracted. Conclusion: Clinical course and sonographic findings are helpful in differentiating patients with she and pmc. The knowledge of the clinical characteristics and sonographic findings of the penicillininduced segmental hemorrhagic colitis may reduce the need for invasive endoscopic and radiological investigations in diagnosis and follow-up.
Fernando Femandez-Banares, Maria Esteve, Jorge C. Espinos, Montserrat Forne, Josep Martin-Comin, Josep M. Viver, Hosp Univ Mutua de Terrassa, Terrassa, Spain; CSUB Bellvitge, L'Hospitalet, Spain. Frequency of BAM in patients with chronic diarrhea of functional characteristics is variable, ranging from 25 to 80% in the different studies. It has been stated that contribution of BAM to diarrhea in a given patient can only be determined by a therapeutic trial of cholestyramine. Aim: To study the frequency of BAM in patients with functional chronic diarrhea in our geographical area. To assess the response to cholestyramine. Methods: 32 patients (11 m, 21 w; age: 53±2 yrs) with functional chronic diarrhea of > 3 months of duration were included. Nine had previous cholecystectomy. Previous time with diarrhea was a median of 48 months (interquartile range -IQ-, 6-72) and daily stool number was a median of 5 (lQ, 4-7). In all cases routine blood biochemistry and hematology, parasitological exam of feces, serum T4-TSH, C reactive protein, IgA-antigliadin/ antiendomysium antibodies, small bowel barium meal and full colonoscopy with multiple biopsies were normal. BAM was assessed by means of the 75SeHCAT abdominal retention test (N) 11% on day 7). Patients with BAM received cholestyramine (sachets) at a starting dose of 4 g1day, which was modified depending on clinical response (2 to 12 g1day). Treatment success was considered as the passage of two or less formedlsemiformed stools per day. Results: BAM was detected in 24 of the 32 patients (75%; 95% CI, 57-88.5%), and in 15 of the 23 without cholecystectomy (65%). BAM was severe (value<5%) in 12. Diarrhea spontaneously resolved in 2 patients and with loperamide in I. In the other 21 patients cholestyramine was administered at a median dose of 8 g1day (lQ, 4-12). Response was not valuable by associated medical problems in 1 of them. In the other 20 diarrhea stopped (response: 100%; 95% CI, 83-100%); median daily stool number passed from 5 (IQ, 4-8) to I (IQ, 1-2) (p=0.OOO2), and feces increased their consistency. Conclusions: Frequency of BAM in patients with chronic diarrhea of functional characteristics is high in our geographical area. The excellent response to cholestyramine suggests that BAM is the cause of diarrhea in such patients.
4896 HIGH ANTI·TOXIN A ANTIBODY LEVELS ARE ASSOCIATED WITH PROTECTION AGAINST RECURRENT CLOSTRIDIUM DIFFICILE DIARRHEA. Lorraine Kyne, Michel Wamy, Amir Qamar, Ciaran P. Kelly, Beth Israel Deaconess Med Ctr & Harvard Med Sch, Boston, MA. Background & Aim: We reported recently that asymptomatic carriers of toxigenic C. difficile have high serum IgG anti-toxin A antibody levels. The aim of this study was to determine whether antibody responses to toxin A during an episode of C. difficile diarrhea influence the risk for recurrent disease. Methods: We studied all patients with antibiotic-associated C. difficile diarrhea in our hospital during a 5 month period. We prospectively determined the clinical severity and duration of diarrhea and followed patients for 60 days after discharge to identify relapses. Serial serum IgA, IgG, and IgM antibody levels against C. difficile toxin A, toxin B and non-toxin antigens were measured and results expressed as ELISA units (EU). Results: We identified 63 patients with C. difficile diarrhea all of whom agreed to participate in the study. Patients with severe C. difficile diarrhea (n=20) had lower serum IgG anti-toxin A levels than those with mild disease (median levels: 0.66 versus 1.60 EU, p=O.04). Patients with prolonged (> 14 days) C. difficile diarrhea (n= 10) had a later peak in serum IgG anti-toxin A levels compared to those with a shorter duration of symptoms (median: day 9 versus day 3, p=0.03). Patients with recurrent C. difficile diarrhea (n=22) had lower serum IgM anti-toxin A levels on day 3 of their first episode of diarrhea (0.85 versus 1.72 EU, p=O.OO4) and lower serum IgG anti-toxin A levels on day 12 (0.66 versus 3.87, p=0.009) compared to those who did not suffer a relapse. Multivariable analysis indicated that patients with a low serum IgM anti-toxin A level on day 3 had a much greater risk of recurrence compared to patients with high antitoxin levels (adjusted odds ratio 21.9, 95% CI: 1.8 to 274.1, p=0.02). Conclusions: An antibody response against toxin A acquired during an initial episode of C. difficile diarrhea is associated with a 22-fold reduced risk of recurrence. Our findings indicate that the host immune response to toxin A plays a substantial role in determining the clinical course of C. difficile diarrhea. This study also provides further evidence that acquired immunity against toxin A may protect against C. difficile diarrhea in humans.