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A Polymorphism and Serum Levels in Patients with Acute Pancreatitis
S2093
as severe based on early organ failure, while in obese patients 24 of 65 (37%) were severe (OR 3.8; CI 1.8, 7.8; p=0.0002). Serum IL-6 levels were significantly higher in severe AP (n=5) than mild AP (n=20) at 48 hours (5,037 vs. 64 pg/ml, p=0.01). There was no association between IL-6 -174 genotype and serum levels in lean or obese subjects with or without AP. The distribution of IL-6 -174 C/C, C/G, G/G, genotypes were as follows: controls 59, 175, 133; lean AP patients 21, 52, 46 and obese AP patients 5, 23, 37, respectively. Susceptibility to AP was associated with the G/G genotype (OR 1.4; CI 1.0, 2.1; p=0.04) with dose effect in obese subjects (OR 2.3; CI 1.2, 4.3; p=0.01). Conclusions: We confirm that obese subjects are at risk for severe AP and that serum IL-6 levels correlate with AP severity. The IL-6 -174 polymorphism does not influence AP severity, suggesting that IL-6 production during the acute inflammatory response is not dependent on the -174 locus. The IL-6 -174 G/G genotype appears to increase susceptibility to AP in obese, but not lean subjects. The mechanism of increased susceptibility to AP in obese subjects with the IL-6 -174 G allele is unknown.
AGA Abstracts
Epidermal Growth Factor +61 G/A Polymorphism and Serum Levels in Patients with Acute Pancreatitis Georgios I. Papachristou, Venkata Muddana, Kim Corrado, Michael R. O'Connell, Adam Slivka, David C. Whitcomb Background: Epidermal growth factor (EGF) binds to pancreatic acinar cells and facilitates recovery from acute pancreatitis (AP). In animal models, EGF protects against cerulein induced pancreatic injury and prevents septic complications. Recently, a single nucleotide polymorphism (SNP) has been described at position +61 (G/A) with the G allele increasing EGF production. Aim: To assess any association of the EGF +61 G/A SNP with susceptibility and severity in AP. To assess EGF serum level response in subjects with mild and severe forms of AP. Methods: 179 AP patients were prospectively enrolled and compared with 189 unrelated controls. The +61G/A polymorphism was genotyped by restriction fragment length polymorphism (RFLP) with selected confirmation by DNA sequencing. EGF serum levels were measured using a standard Luminex assay in 58 controls and 25 subjects with AP between 24 and 48 hours from onset of pain. Comparisons were made using chi-square and trend test. Results: 38 patients developed organ failure and classified as severe AP (SAP; 21%). The genotypic frequencies were similar among controls, mild AP (MAP) and SAP patients (Table 1). Serum EGF in AP patients reached trough levels on day 2 and was significantly lower when compared to controls (13.0 vs. 25.2 pg/ml; p=0.03). There was a trend towards lower EGF levels in SAP patients when compared to MAP patients (8.0 vs. 14.2 pg/ml; p=0.08). Conclusions: EGF serum levels are suppressed in AP patients when compared to controls. They are even lower in patients who develop severe disease trending towards statistical significance. Thus, EGF suppression might delay recovery and increase the risk for severe disease and complications in humans with AP. EGF +61 genotype appears to have no effect on either susceptibility or severity in AP. Table 1
S2095a Leptin As a Substitute for Insulin Therapy in Insulin-Deficient Diabetes: Correction of Hyperglycemia and Hyperphgia and Surviving Without Diabetic Complications Shinya Kojima, Haruka Amitani, Marie Sameshima, Akihiro Asakawa, Akio Inui, Pushpa S. Kalra, Satya P. Kalra Objective: The objective of this study was to evaluate the effects of central leptin gene therapy on insulin-deficient diabetes. Methods: Twenty-five animals were divided into 3 groups: A streptozotocin (STZ)-induced diabetic group that received rAAV encoding leptin (rAAV-LEP) (STZ-LEP), an STZ-induced diabetic group that received an injection of rAAV encoding green fluorescent protein (rAAV-GFP) (STZ-GFP), and a control nondiabetic group (control). The nonimmunogenic, nonpathogenic and replicative-deficient rAAV-LEP or rAAVGFP vectors were packaged, purified, concentrated, and titered in the Vector Core Laboratory at the University of Florida. These vectors were used in the experiment. Diabetes was induced by a single intraperitoneal infusion of STZ (200 mg/kg). The control nondiabetic group received an injection of citrate buffer only. At the time of vector injection, the STZ mice whose blood glucose concentration was >350 mg/dl were defined as diabetic. Results: The mice in the STZ-GFP group died by week 6; however, the STZ-LEP mice and the control mice survived for more than 1 year. The STZ diabetic mice demonstrated hyperphagia; however, leptin treatment blocked hyperphagia. Food intake in the STZ-LEP mice was normalized after 14 weeks, and food intake was lower in the STZ-LEP mice than in the control group after 20 weeks. Blood glucose concentrations were increased after STZ treatment; however, the blood glucose concentration decreased after 8 weeks of administration of rAAV-LEP vectors and reached the near normal range. The STZ-LEP mice did not develop diabetic complications within 1 year; however, these mice showed regeneration of pancreatic beta cells. Conclusions: Central leptin gene therapy suppressed food intake over a long period of time and successfully treated insulin-deficient diabetes. This study suggests that leptin administration can be used as an alternative to insulin therapy in insulin-deficient diabetes, thereby providing an insight into the therapeutic implications of leptin as a substitute for insulin therapy.
S2094 Visfatin Polymorphisms and Elevated BMI Interact to Increase the Susceptibility to Acute Pancreatitis Arun Sharma, Venkata Muddana, Janette Lamb, Adam Slivka, David C. Whitcomb, Georgios I. Papachristou Background: Acute pancreatitis (AP) is an acute inflammatory event that originates within the pancreas. Most patients develop mild AP (MAP); however, in some patients, there is a systemic inflammatory response that results in severe AP (SAP). Obesity is a known risk factor for developing SAP. Adipokines released from adipose tissue play a role in the inflammatory response. We hypothesize that adipokines mediate the effect of obesity in AP. Visfatin is a pro-inflammatory adipokine released from visceral adipose tissue. Polymorphisms in the visfatin gene have been described but have not yet been studied in AP. Methods: 134 AP patients and 94 controls were prospectively enrolled. SAP was defined as the presence of remote organ failure. The polymorphisms of interest (-1543 C/T, -1001 T/G, -948 G/T, -423 A/G, +21181 C/T and +21427 G/A) were evaluated by polymerase chain reaction amplification and DNA sequencing. Statistical analyses were performed using chi-square and Fisher's exact tests. Results: There were 69 females and 65 males with AP; their mean age was 51 years. The most common etiologies of AP were biliary (35%), idiopathic (20%), post-endoscopic retrograde cholangio-pancreatography (19%), and alcohol (14%). Of the 134 AP patients, 26 had SAP (19%) and 108 had MAP (81%). Although there were trends in the genotypes of the visfatin polymorphisms when comparing AP patients and controls, they were not statistically significant. However, in patients with elevated BMI (≥25 kg/m2), the -1543 C/C and +21181 T/T genotypes resulted in an increased risk of developing AP (p=0.010; OR 2.45 and p=0.013; OR 3.32, respectively). Also among patients with elevated BMI, the -1001 T/T and -948 G/G genotypes were found more frequently in SAP than MAP patients, but these differences did not achieve statistical significance. Conclusions: In the overall population, polymorphisms in the visfatin gene have a small or no effect on either susceptibility or severity in AP. However, in overweight and obese patients, the -1543 C/C and +21181 T/T genotypes increase the susceptibility to AP.
S2095b Gastric Phase 3 Is a Hunger Signal in the Interdigestive State in Man Daphne C. Ang, Heleen Nicolai, Rita Vos, Pieter Vanden Berghe, Daniel Sifrim, Inge Depoortere, Theo L. Peeters, Jozef Janssens, Jan F. Tack Background: Appetite is controlled by the brain based on input from the GI tract (GIT). In the fasting state, hunger is expected to rise over time, but the GIT displays a cyclical secretomotor pattern, the migrating motor complex (MMC). Aims: To elucidate the relationship between appetite and MMC phases (ph), we studied hunger scores during spontaneous and induced ph3. Materials and Methods: Twelve healthy subjects (6 males; 30.9±2.0 years) underwent interdigestive gastroduodenal manometry on 2 separate occasions. After a full MMC cycle, erythromycin(EM) i.v.40mg or octreotide(OCT) s.c.100µg was administered. Hunger scores (100mm visual analogue scales, VAS) and GI hormones were measured. Analysis used visual recognition of MMC phases and computerized motility index (MI). We compared hunger scores in different MMC phases, and linear correlation with MI. Results: Over 268±10 mins, 47 spontaneous phase3 occurred with gastric origin in 18. Compared to phase2, gastric ph3 was associated with higher hunger scores (35.9±5.4vs.62.5±7.5,p<0.005) while ph1 scores were lower (17.0±2.8,p=0.04)(Fig.). In contrast, hunger scores during ph3 with small bowel origin (38.2±4.9) did not differ significantly from ph1 and ph2 scores (34.6±7.4 and 41.0±3.9). Hunger scores were significantly higher during spontaneous gastric compared to small bowel ph3(p=0.01). A significant correlation was found between gastric MI and hunger scores before and after spontaneous gastric (r=0.6021,p<0.001), but not small bowel (r=0.0941, NS) origin ph3. EM-induced gastric ph3, but not OC-induced intestinal ph3, had a hunger peak (61.7±8.0 vs. 30.4± 7.6,p=0.01). Conclusions: In the fasting state, hunger scores are closely correlated with gastric motility. Peak hunger scores are associated with the spontaneous or pharmacologically induced gastric phase3, demonstrating that gastric phase 3 is a hunger signal from the GIT in the fasting state.
S2095 The IL-6 -174 G/G Genotype Increases Susceptibility to Acute Pancreatitis (AP) in Obese Subjects Venkata Muddana, Arun Sharma, Janette Lamb, Adam Slivka, David C. Whitcomb, Georgios I. Papachristou Introduction: Interleukin 6 (IL-6) is a multifunctional cytokine with complex regulatory controls that increases in the serum of subjects with AP in parallel to severity. The IL-6 -174 C/G polymorphism alters IL-6 protein levels in response to LPS, IL-1, obesity, weight loss and diabetes mellitus. Up to 50% of serum IL-6 is derived from visceral adipose tissue and visceral obesity is a risk factor for development of AP and AP severity. Aim: To determine whether IL-6 -174 genotype is associated with AP susceptibility or severity in lean or obese subjects. Methods: 184 prospectively ascertained patients with AP and 367 control subjects were genotyped from the SAPS and NAPS2 studies respectively. IL-6 -174 genotyping was performed by restriction fragment length polymorphism (RFLP) with selected confirmation by DNA sequencing. IL-6 serum levels were measured using a standard Luminex assay in 58 controls and 25 subjects with AP between 24 and 48 hours from onset of pain. Comparisons were made using chi-square and trend test. Results: 65 patients (35%) and 101 controls (28%) had a BMI≥30 (p=0.06). In lean patients with AP, 16 of 119 (13%) were classified
AGA Abstracts
A-314
as severe based on early organ failure, while in obese patients 24 of 65 (37%) were severe (OR 3.8; CI 1.8, 7.8; p=0.0002). Serum IL-6 levels were significantly higher in severe AP (n=5) than mild AP (n=20) at 48 hours (5,037 vs. 64 pg/ml, p=0.01). There was no association between IL-6 -174 genotype and serum levels in lean or obese subjects with or without AP. The distribution of IL-6 -174 C/C, C/G, G/G, genotypes were as follows: controls 59, 175, 133; lean AP patients 21, 52, 46 and obese AP patients 5, 23, 37, respectively. Susceptibility to AP was associated with the G/G genotype (OR 1.4; CI 1.0, 2.1; p=0.04) with dose effect in obese subjects (OR 2.3; CI 1.2, 4.3; p=0.01). Conclusions: We confirm that obese subjects are at risk for severe AP and that serum IL-6 levels correlate with AP severity. The IL-6 -174 polymorphism does not influence AP severity, suggesting that IL-6 production during the acute inflammatory response is not dependent on the -174 locus. The IL-6 -174 G/G genotype appears to increase susceptibility to AP in obese, but not lean subjects. The mechanism of increased susceptibility to AP in obese subjects with the IL-6 -174 G allele is unknown.
AGA Abstracts
Epidermal Growth Factor +61 G/A Polymorphism and Serum Levels in Patients with Acute Pancreatitis Georgios I. Papachristou, Venkata Muddana, Kim Corrado, Michael R. O'Connell, Adam Slivka, David C. Whitcomb Background: Epidermal growth factor (EGF) binds to pancreatic acinar cells and facilitates recovery from acute pancreatitis (AP). In animal models, EGF protects against cerulein induced pancreatic injury and prevents septic complications. Recently, a single nucleotide polymorphism (SNP) has been described at position +61 (G/A) with the G allele increasing EGF production. Aim: To assess any association of the EGF +61 G/A SNP with susceptibility and severity in AP. To assess EGF serum level response in subjects with mild and severe forms of AP. Methods: 179 AP patients were prospectively enrolled and compared with 189 unrelated controls. The +61G/A polymorphism was genotyped by restriction fragment length polymorphism (RFLP) with selected confirmation by DNA sequencing. EGF serum levels were measured using a standard Luminex assay in 58 controls and 25 subjects with AP between 24 and 48 hours from onset of pain. Comparisons were made using chi-square and trend test. Results: 38 patients developed organ failure and classified as severe AP (SAP; 21%). The genotypic frequencies were similar among controls, mild AP (MAP) and SAP patients (Table 1). Serum EGF in AP patients reached trough levels on day 2 and was significantly lower when compared to controls (13.0 vs. 25.2 pg/ml; p=0.03). There was a trend towards lower EGF levels in SAP patients when compared to MAP patients (8.0 vs. 14.2 pg/ml; p=0.08). Conclusions: EGF serum levels are suppressed in AP patients when compared to controls. They are even lower in patients who develop severe disease trending towards statistical significance. Thus, EGF suppression might delay recovery and increase the risk for severe disease and complications in humans with AP. EGF +61 genotype appears to have no effect on either susceptibility or severity in AP. Table 1
S2095a Leptin As a Substitute for Insulin Therapy in Insulin-Deficient Diabetes: Correction of Hyperglycemia and Hyperphgia and Surviving Without Diabetic Complications Shinya Kojima, Haruka Amitani, Marie Sameshima, Akihiro Asakawa, Akio Inui, Pushpa S. Kalra, Satya P. Kalra Objective: The objective of this study was to evaluate the effects of central leptin gene therapy on insulin-deficient diabetes. Methods: Twenty-five animals were divided into 3 groups: A streptozotocin (STZ)-induced diabetic group that received rAAV encoding leptin (rAAV-LEP) (STZ-LEP), an STZ-induced diabetic group that received an injection of rAAV encoding green fluorescent protein (rAAV-GFP) (STZ-GFP), and a control nondiabetic group (control). The nonimmunogenic, nonpathogenic and replicative-deficient rAAV-LEP or rAAVGFP vectors were packaged, purified, concentrated, and titered in the Vector Core Laboratory at the University of Florida. These vectors were used in the experiment. Diabetes was induced by a single intraperitoneal infusion of STZ (200 mg/kg). The control nondiabetic group received an injection of citrate buffer only. At the time of vector injection, the STZ mice whose blood glucose concentration was >350 mg/dl were defined as diabetic. Results: The mice in the STZ-GFP group died by week 6; however, the STZ-LEP mice and the control mice survived for more than 1 year. The STZ diabetic mice demonstrated hyperphagia; however, leptin treatment blocked hyperphagia. Food intake in the STZ-LEP mice was normalized after 14 weeks, and food intake was lower in the STZ-LEP mice than in the control group after 20 weeks. Blood glucose concentrations were increased after STZ treatment; however, the blood glucose concentration decreased after 8 weeks of administration of rAAV-LEP vectors and reached the near normal range. The STZ-LEP mice did not develop diabetic complications within 1 year; however, these mice showed regeneration of pancreatic beta cells. Conclusions: Central leptin gene therapy suppressed food intake over a long period of time and successfully treated insulin-deficient diabetes. This study suggests that leptin administration can be used as an alternative to insulin therapy in insulin-deficient diabetes, thereby providing an insight into the therapeutic implications of leptin as a substitute for insulin therapy.
S2094 Visfatin Polymorphisms and Elevated BMI Interact to Increase the Susceptibility to Acute Pancreatitis Arun Sharma, Venkata Muddana, Janette Lamb, Adam Slivka, David C. Whitcomb, Georgios I. Papachristou Background: Acute pancreatitis (AP) is an acute inflammatory event that originates within the pancreas. Most patients develop mild AP (MAP); however, in some patients, there is a systemic inflammatory response that results in severe AP (SAP). Obesity is a known risk factor for developing SAP. Adipokines released from adipose tissue play a role in the inflammatory response. We hypothesize that adipokines mediate the effect of obesity in AP. Visfatin is a pro-inflammatory adipokine released from visceral adipose tissue. Polymorphisms in the visfatin gene have been described but have not yet been studied in AP. Methods: 134 AP patients and 94 controls were prospectively enrolled. SAP was defined as the presence of remote organ failure. The polymorphisms of interest (-1543 C/T, -1001 T/G, -948 G/T, -423 A/G, +21181 C/T and +21427 G/A) were evaluated by polymerase chain reaction amplification and DNA sequencing. Statistical analyses were performed using chi-square and Fisher's exact tests. Results: There were 69 females and 65 males with AP; their mean age was 51 years. The most common etiologies of AP were biliary (35%), idiopathic (20%), post-endoscopic retrograde cholangio-pancreatography (19%), and alcohol (14%). Of the 134 AP patients, 26 had SAP (19%) and 108 had MAP (81%). Although there were trends in the genotypes of the visfatin polymorphisms when comparing AP patients and controls, they were not statistically significant. However, in patients with elevated BMI (≥25 kg/m2), the -1543 C/C and +21181 T/T genotypes resulted in an increased risk of developing AP (p=0.010; OR 2.45 and p=0.013; OR 3.32, respectively). Also among patients with elevated BMI, the -1001 T/T and -948 G/G genotypes were found more frequently in SAP than MAP patients, but these differences did not achieve statistical significance. Conclusions: In the overall population, polymorphisms in the visfatin gene have a small or no effect on either susceptibility or severity in AP. However, in overweight and obese patients, the -1543 C/C and +21181 T/T genotypes increase the susceptibility to AP.
S2095b Gastric Phase 3 Is a Hunger Signal in the Interdigestive State in Man Daphne C. Ang, Heleen Nicolai, Rita Vos, Pieter Vanden Berghe, Daniel Sifrim, Inge Depoortere, Theo L. Peeters, Jozef Janssens, Jan F. Tack Background: Appetite is controlled by the brain based on input from the GI tract (GIT). In the fasting state, hunger is expected to rise over time, but the GIT displays a cyclical secretomotor pattern, the migrating motor complex (MMC). Aims: To elucidate the relationship between appetite and MMC phases (ph), we studied hunger scores during spontaneous and induced ph3. Materials and Methods: Twelve healthy subjects (6 males; 30.9±2.0 years) underwent interdigestive gastroduodenal manometry on 2 separate occasions. After a full MMC cycle, erythromycin(EM) i.v.40mg or octreotide(OCT) s.c.100µg was administered. Hunger scores (100mm visual analogue scales, VAS) and GI hormones were measured. Analysis used visual recognition of MMC phases and computerized motility index (MI). We compared hunger scores in different MMC phases, and linear correlation with MI. Results: Over 268±10 mins, 47 spontaneous phase3 occurred with gastric origin in 18. Compared to phase2, gastric ph3 was associated with higher hunger scores (35.9±5.4vs.62.5±7.5,p<0.005) while ph1 scores were lower (17.0±2.8,p=0.04)(Fig.). In contrast, hunger scores during ph3 with small bowel origin (38.2±4.9) did not differ significantly from ph1 and ph2 scores (34.6±7.4 and 41.0±3.9). Hunger scores were significantly higher during spontaneous gastric compared to small bowel ph3(p=0.01). A significant correlation was found between gastric MI and hunger scores before and after spontaneous gastric (r=0.6021,p<0.001), but not small bowel (r=0.0941, NS) origin ph3. EM-induced gastric ph3, but not OC-induced intestinal ph3, had a hunger peak (61.7±8.0 vs. 30.4± 7.6,p=0.01). Conclusions: In the fasting state, hunger scores are closely correlated with gastric motility. Peak hunger scores are associated with the spontaneous or pharmacologically induced gastric phase3, demonstrating that gastric phase 3 is a hunger signal from the GIT in the fasting state.
S2095 The IL-6 -174 G/G Genotype Increases Susceptibility to Acute Pancreatitis (AP) in Obese Subjects Venkata Muddana, Arun Sharma, Janette Lamb, Adam Slivka, David C. Whitcomb, Georgios I. Papachristou Introduction: Interleukin 6 (IL-6) is a multifunctional cytokine with complex regulatory controls that increases in the serum of subjects with AP in parallel to severity. The IL-6 -174 C/G polymorphism alters IL-6 protein levels in response to LPS, IL-1, obesity, weight loss and diabetes mellitus. Up to 50% of serum IL-6 is derived from visceral adipose tissue and visceral obesity is a risk factor for development of AP and AP severity. Aim: To determine whether IL-6 -174 genotype is associated with AP susceptibility or severity in lean or obese subjects. Methods: 184 prospectively ascertained patients with AP and 367 control subjects were genotyped from the SAPS and NAPS2 studies respectively. IL-6 -174 genotyping was performed by restriction fragment length polymorphism (RFLP) with selected confirmation by DNA sequencing. IL-6 serum levels were measured using a standard Luminex assay in 58 controls and 25 subjects with AP between 24 and 48 hours from onset of pain. Comparisons were made using chi-square and trend test. Results: 65 patients (35%) and 101 controls (28%) had a BMI≥30 (p=0.06). In lean patients with AP, 16 of 119 (13%) were classified
AGA Abstracts
A-314