- Email: [email protected]
Tu2019 Role of Vitamin D in the Inflammatory Response of Morbidly Obese Patients With Non-Alcoholic Fatty Liver Disease (NAFLD)
Tu2016
assay and the human TGFB1 assay. RNA was extracted from the VAT samples using BioRad's Aurum Total RNA Fatty and Fibrous Tissue Kit and was converted to cDNA using SABiosciences' RT2 First Strand Kit. This cDNA was subjected to qPCR using custom designed primers for FOXP3, COL1A1, UCP1 and TGFB1. Clinical and demographic variables were also available. Spearman's rank-sum correlation analysis was performed to determine the co-expression patterns.RESULTS: 241 patients (58.6% insufficient for Vitamin D, 28.6% type 2 diabetes, 39.1% NASH, age 43.4±11.7, BMI 46.4±10.9) were included. Vitamin D serum levels were found to positively correlate with the apoptosis marker M30 in serum (r = 0.17, p ≤ 1.1e-04), ALT (r = 0.35, p-value ≤ 2.1e-05), and AST (r = 0.34, p-value ≤ 4.2e-05) and to negatively correlate with BMI (r = -0.25, p-value ≤ 0.002) and LDL (r = -0.3, p-value ≤ 0.002). Serum levels of M30 positively correlate with ALT (r = 0.32, p-value ≤ 1.7e-04) and AST (r = 0.36, p-value ≤ 1.4e-05). Correlation analysis also showed that patients with higher overall Vitamin D showed a decrease in the expression levels of TGFB1 (r = -0.31, p-value ≤ 0.05), FOXP3 (r = -0.34, p-value ≤ 0.03), COL1A1 (r = -0.42, p-value ≤ 0.01), and UCP1 (r = -0.41, p-value ≤ 0.01) inVAT. In addition there was a substantial positive correlation with serum levels of IL-17 (r = 0.38, p-value ≤ 4e-06), IL-4 (r = 0.28, p-value ≤ 7e-04), and IL-5 (r = 0.32, p-value ≤ 1e-04) and a negative correlation with serum levels of TGFB1 (r = -0.21, p-value ≤ 0.014). CONCLUSIONS: These results suggest that Vitamin D may have an impact TGFB1 expression in VAT thus affecting the levels of inflammatory cytokines systemically in the body. This possible alteration of the inflammatory pathways may have an impact on the pathogenesis of liver disease in morbidly obese patients. Further research is necessary to determine the full impact of Vitamin D on these inflammatory pathways and the subsequent effect on liver disease.
AGA Abstracts
Establishment of a Novel Automated Method to Assess the Food Intake Microstructure in Rats Lixin Wang, Pauline Teuffel, Miriam Goebel-Stengel, Peter Kobelt, Matthias Rose, Burghard F. Klapp, Joseph R. Reeve, Andreas Stengel Background: Food intake is a parameter frequently assessed in rodent models. The classical approach is to measure food intake manually after defined time intervals. Although this technique is easy to learn and use, it holds several disadvantages: the animals are disturbed by the investigator, a real assessment of dark phase feeding is not possible, the measurement is rather rough as short-lasting effects may be missed and lastly, there is no information on the underlying feeding microstructure. Therefore, several approaches have been undertaken in order to assess this feeding microstructure including the use of liquid, powder or micropelleted food, which, however, does not represent the physiological solid food of rats. Aim: To evaluate an automated method to assess the physiological feeding microstructure in rats. Methods: Male Sprague Dawley rats were used and food intake was monitored manually and in an automated fashion using an automated episodic feeding monitoring system recently established for the use in mice (BioDAQ, Research Diets). In a separate experiment, intracerebroventricularly (ICV) cannulated rats were compared to naive rats (n=8/group). Results: Rats showed a linear daily body weight gain when housed in groups (3.1±1.5g) which did not significantly change directly after separation into single housing cages with feeding from the cage top (3.6±1.3g) or from the hopper of the automated monitoring device (2.7±0.1g, P=0.81). Also, the nighttime food intake, the period of the maximum food intake in rodents, was similar when assessed by the automated system on day 1 and 5 after starting the measurement (17.7±0.8 vs. 19.1±0.7g, P=0.24), indicating a rapid accommodation to this feeding device. In addition, daily food intake did not differ when assessed manually or in an automated fashion (21.9±0.4 vs. 21.1±0.2g, P=0.17), indicating a good concordance between both methods. The automated assessment allowed the measurement of parameters of the food intake microstructure, namely latency to first meal, inter-meal interval, meal size, duration and frequency, eating rate and the satiety ratio. The investigators observed the satiety sequence of rats in the automated feeding cages with a decrease in dark phase feeding behavior and an increase in grooming, exploration and resting indicating a physiological behavior. Lastly, in cages with an enlarged opening of the cage mount, ICV cannulated rats showed a similar dark phase food intake as naive rats (16.6±0.6 vs. 16.4±0.5g, P=0.86), indicating that the system can also be used in head device-bearing rats. Conclusions: The automated feeding monitoring device allows a detailed assessment of the food intake microstructure in undisturbed rats housed in regular cages and fed physiological rat chow. Therefore, this device will be a valuable tool in behavioral studies investigating food intake.
Tu2020 Do Serum 25-Hydroxyvitamin D Levels Affect the Prevalence of Dysplasia in Barrett's Esophagus? Prashanthi N. Thota, Prabhdeep Singh, Linda C. Cummings, Rocio Lopez Introduction: Vitamin D has several anti-carcinogenic properties such as suppressing cell proliferation, promoting cell differentiation and regulating apoptosis. There is growing interest in the protective role of Vitamin D against development of many cancers such as colon cancer and breast cancer. The relationship between Vitamin D levels and esophageal cancer seems to be inconsistent. In fact, lower levels of vitamin D were reported to be associated with statistically decreased risk of upper gastrointestinal cancers. To date, there are no studies that have investigated the association between Vitamin D levels and Barrett's esophagus, the precursor lesion for esophageal adenocarcinoma. Our aim was to see if low vitamin D levels were associated with increased prevalence of dysplasia or cancer in patients with Barrett's esophagus. Methods: Patients with Barrett's esophagus (BE) seen our department of gastroenterology from Jan 2000 t o Dec 2012 who had serum 25-hydroxyvitamin D (25(OH)D) levels checked were included. Demographic data such as age, gender, race, BMI, Vitamin D levels and endoscopic findings such as BE length, hiatal hernia size, and biopsy findings were recorded. Data are presented as mean ± standard deviation, median [25th, 75th percentiles] or N (%).The prevalence of dysplasia was estimated as the % of subjects out of each Vitamin D group that had each biopsy reading. Jonckheere-Terpstra tests were used to assess differences between the vitamin D groups. Results: There were 2370 patients with Barrett's esophagus seen during that time period. 1896 patients did not have 25(OH)D levels checked and 45 patients did not have biopsy results. A total of 429 subjects met the inclusion criteria. Average age was 61 ± 12 years, 65.5% (281/429) were male and 94.5% were Caucasian. Average BE length was 2.5± 3 cm and hiatal hernia size was 2.2± 3.9 cm. The mean 25(OH)D levels were 28.8±12.5 ng/mL. Vitamin D levels were < 25ng/mL in 175(40.8%), between 25-37.4ng/ml in 177(41.3%) and ≥ 37.5 ng/mL in 77 (17.9%). The histological findings were no dysplasia in 340 (79.3%), low-grade dysplasia (LGD) in 52(12.1%), high grade dysplasia (HGD) in 15(3.5%) and adenocarcinoma (EAC) in 22 (5.1%). The histological findings BE in different groups of serum 25(OH)D levels are presented in Table 1. There was no evidence to suggest an association between serum 25(OH)D levels and dysplasia (p = 0.90). Conclusions: The mean serum Vitamin D levels in our Barrett's population were low. There was no significant association between degree of dysplasia and Vitamin D levels in patients with Barrett's esophagus. Larger population studies are needed in this area of research.
Tu2017 Non-Transferrin Bound Iron Transporter ZIP 14 in the Liver of Pediatric NonAlcoholic Steatohepatitis Patients Maan Khatib, Susan S. Baker, Wensheng Liu, Diana Moya, Ricardo A. Arbizu, Robert D. Baker, Michael D. Garrick, Lixin Zhu Background: Non-alcoholic steatohepatitis (NASH), the severe form of non-alcoholic fatty liver disease (NAFLD), is characterized by steatosis and inflammation. Iron may be an important factor in the pathogenesis of NASH as it catalyzes the production of potent reactive oxygen species, which could lead to liver inflammation. Non-transferrin bound iron (NTBI) uptake by ZRT/IRT-like protein (ZIP14, or SLC39A14) is an important mechanism for scavenging plasma iron during iron overload in rodents. In iron overloaded rodents, liver ZIP14 is elevated. The purpose of this study was to examine iron status and hepatic expression of iron related genes in pediatric NASH. Methods: Serum iron concentration, ferritin, and soluble transferrin receptor 1 of biopsy proven NASH patients were compared to those in the National Health and Nutrition Examination Survey database. Iron related gene expression was examined in NASH livers and normal livers, using microarray and quantitative realtime PCR. Results: Serum iron concentration and soluble transferrin receptor 1 were decreased while serum ferritin was increased in NASH patients, compared to controls. No detectable iron was observed in NASH liver by Perls' Prussian blue staining. Our microarray analysis and qRT-PCR analysis consistently demonstrated that, in NASH livers, transferrin mRNA and transferrin receptor 2 mRNA were elevated, while ZIP14 mRNA was decreased (realtime PCR, 83% decrease in NASH p=0.016). Conclusion: Pediatric NASH patients tend to be iron deficient. Increased liver gene expression of transferrin and transferrin receptor 2 supports a status of iron deficiency. Our observation of decreased ZIP14 expression in pediatric NASH patients who are iron deficient is in harmony with previous observation that ZIP14 expression in rodent livers is positively correlated to liver iron load. Our study points to a role for ZIP14 in human liver iron homeostasis. Tu2019 Role of Vitamin D in the Inflammatory Response of Morbidly Obese Patients With Non-Alcoholic Fatty Liver Disease (NAFLD) Katherine Doyle, Maria Keaton, Lei Wang, Rohini Mehta, Massih Abawi, Zahra Younoszai, Thomas Jeffers, Hazem A. Elarainy, Amir H. Moazez, Aybike Birerdinc, Ancha Baranova, Zobair M. Younossi
Values presented as N (column%) p-value corresponds to Joncheere-Terpstra test Tu2027
INTRODUCTION: Vitamin D regulates many genes related to cell proliferation, differentiation, and apoptosis and recent studies have suggested a role for Vitamin D deficiency in liver disease as well as obesity. Vitamin D receptor is expressed by macrophages and dendritic cells suggesting that this highly lipidophilic hormone also plays a role in the regulation of the body's inflammatory response which is particularly important in patients who are morbidly obese.AIM: To determine the anti-inflammatory roles of vitamin D in a cohort of morbidly obese NAFLD patients.METHODS: Serum samples obtained in conjunction with visceral adipose tissue from patients with biopsy-proven NAFLD during Bariatric surgery. Serum was assayed for Vitamin D levels using 25(OH)-Vitamin D3 assays (Alpco, Salem, NH). These samples were also run on Biorad's Bioplex pro using the 17-plex human cytokine
AGA Abstracts
Treatment with the CSF-1 Receptor kinase (CSF-1R) Inhibitor JNJ40346527 Suppresses Murine Colitis Beverley Moore, Hao Liu, Matthew Loza, Holly A. Raymond, Stanley M. Belkowski, Carl Manthey Background: The role of macrophages (Mø) in inflammatory bowel disease is as yet poorly understood. It is theorized that Mø recruited to the intestinal lamina propria differentiate into an inflammatory phenotype with myeloid DC properties (Mø/mDC), which then express high levels of TNFα and IL-23 upon activation by enteric bacteria. IL-23 expands innate lymphocyte cell populations, and bacterial antigen presentation by Mø/mDC induces proliferation of Th1 T cells, while IFN-g produced by NK and Th1 cells perpetuates the inflammatory Mø phenotype. Colony stimulating factor (CSF-1) is prominent among the several factors
S-900
assay and the human TGFB1 assay. RNA was extracted from the VAT samples using BioRad's Aurum Total RNA Fatty and Fibrous Tissue Kit and was converted to cDNA using SABiosciences' RT2 First Strand Kit. This cDNA was subjected to qPCR using custom designed primers for FOXP3, COL1A1, UCP1 and TGFB1. Clinical and demographic variables were also available. Spearman's rank-sum correlation analysis was performed to determine the co-expression patterns.RESULTS: 241 patients (58.6% insufficient for Vitamin D, 28.6% type 2 diabetes, 39.1% NASH, age 43.4±11.7, BMI 46.4±10.9) were included. Vitamin D serum levels were found to positively correlate with the apoptosis marker M30 in serum (r = 0.17, p ≤ 1.1e-04), ALT (r = 0.35, p-value ≤ 2.1e-05), and AST (r = 0.34, p-value ≤ 4.2e-05) and to negatively correlate with BMI (r = -0.25, p-value ≤ 0.002) and LDL (r = -0.3, p-value ≤ 0.002). Serum levels of M30 positively correlate with ALT (r = 0.32, p-value ≤ 1.7e-04) and AST (r = 0.36, p-value ≤ 1.4e-05). Correlation analysis also showed that patients with higher overall Vitamin D showed a decrease in the expression levels of TGFB1 (r = -0.31, p-value ≤ 0.05), FOXP3 (r = -0.34, p-value ≤ 0.03), COL1A1 (r = -0.42, p-value ≤ 0.01), and UCP1 (r = -0.41, p-value ≤ 0.01) inVAT. In addition there was a substantial positive correlation with serum levels of IL-17 (r = 0.38, p-value ≤ 4e-06), IL-4 (r = 0.28, p-value ≤ 7e-04), and IL-5 (r = 0.32, p-value ≤ 1e-04) and a negative correlation with serum levels of TGFB1 (r = -0.21, p-value ≤ 0.014). CONCLUSIONS: These results suggest that Vitamin D may have an impact TGFB1 expression in VAT thus affecting the levels of inflammatory cytokines systemically in the body. This possible alteration of the inflammatory pathways may have an impact on the pathogenesis of liver disease in morbidly obese patients. Further research is necessary to determine the full impact of Vitamin D on these inflammatory pathways and the subsequent effect on liver disease.
AGA Abstracts
Establishment of a Novel Automated Method to Assess the Food Intake Microstructure in Rats Lixin Wang, Pauline Teuffel, Miriam Goebel-Stengel, Peter Kobelt, Matthias Rose, Burghard F. Klapp, Joseph R. Reeve, Andreas Stengel Background: Food intake is a parameter frequently assessed in rodent models. The classical approach is to measure food intake manually after defined time intervals. Although this technique is easy to learn and use, it holds several disadvantages: the animals are disturbed by the investigator, a real assessment of dark phase feeding is not possible, the measurement is rather rough as short-lasting effects may be missed and lastly, there is no information on the underlying feeding microstructure. Therefore, several approaches have been undertaken in order to assess this feeding microstructure including the use of liquid, powder or micropelleted food, which, however, does not represent the physiological solid food of rats. Aim: To evaluate an automated method to assess the physiological feeding microstructure in rats. Methods: Male Sprague Dawley rats were used and food intake was monitored manually and in an automated fashion using an automated episodic feeding monitoring system recently established for the use in mice (BioDAQ, Research Diets). In a separate experiment, intracerebroventricularly (ICV) cannulated rats were compared to naive rats (n=8/group). Results: Rats showed a linear daily body weight gain when housed in groups (3.1±1.5g) which did not significantly change directly after separation into single housing cages with feeding from the cage top (3.6±1.3g) or from the hopper of the automated monitoring device (2.7±0.1g, P=0.81). Also, the nighttime food intake, the period of the maximum food intake in rodents, was similar when assessed by the automated system on day 1 and 5 after starting the measurement (17.7±0.8 vs. 19.1±0.7g, P=0.24), indicating a rapid accommodation to this feeding device. In addition, daily food intake did not differ when assessed manually or in an automated fashion (21.9±0.4 vs. 21.1±0.2g, P=0.17), indicating a good concordance between both methods. The automated assessment allowed the measurement of parameters of the food intake microstructure, namely latency to first meal, inter-meal interval, meal size, duration and frequency, eating rate and the satiety ratio. The investigators observed the satiety sequence of rats in the automated feeding cages with a decrease in dark phase feeding behavior and an increase in grooming, exploration and resting indicating a physiological behavior. Lastly, in cages with an enlarged opening of the cage mount, ICV cannulated rats showed a similar dark phase food intake as naive rats (16.6±0.6 vs. 16.4±0.5g, P=0.86), indicating that the system can also be used in head device-bearing rats. Conclusions: The automated feeding monitoring device allows a detailed assessment of the food intake microstructure in undisturbed rats housed in regular cages and fed physiological rat chow. Therefore, this device will be a valuable tool in behavioral studies investigating food intake.
Tu2020 Do Serum 25-Hydroxyvitamin D Levels Affect the Prevalence of Dysplasia in Barrett's Esophagus? Prashanthi N. Thota, Prabhdeep Singh, Linda C. Cummings, Rocio Lopez Introduction: Vitamin D has several anti-carcinogenic properties such as suppressing cell proliferation, promoting cell differentiation and regulating apoptosis. There is growing interest in the protective role of Vitamin D against development of many cancers such as colon cancer and breast cancer. The relationship between Vitamin D levels and esophageal cancer seems to be inconsistent. In fact, lower levels of vitamin D were reported to be associated with statistically decreased risk of upper gastrointestinal cancers. To date, there are no studies that have investigated the association between Vitamin D levels and Barrett's esophagus, the precursor lesion for esophageal adenocarcinoma. Our aim was to see if low vitamin D levels were associated with increased prevalence of dysplasia or cancer in patients with Barrett's esophagus. Methods: Patients with Barrett's esophagus (BE) seen our department of gastroenterology from Jan 2000 t o Dec 2012 who had serum 25-hydroxyvitamin D (25(OH)D) levels checked were included. Demographic data such as age, gender, race, BMI, Vitamin D levels and endoscopic findings such as BE length, hiatal hernia size, and biopsy findings were recorded. Data are presented as mean ± standard deviation, median [25th, 75th percentiles] or N (%).The prevalence of dysplasia was estimated as the % of subjects out of each Vitamin D group that had each biopsy reading. Jonckheere-Terpstra tests were used to assess differences between the vitamin D groups. Results: There were 2370 patients with Barrett's esophagus seen during that time period. 1896 patients did not have 25(OH)D levels checked and 45 patients did not have biopsy results. A total of 429 subjects met the inclusion criteria. Average age was 61 ± 12 years, 65.5% (281/429) were male and 94.5% were Caucasian. Average BE length was 2.5± 3 cm and hiatal hernia size was 2.2± 3.9 cm. The mean 25(OH)D levels were 28.8±12.5 ng/mL. Vitamin D levels were < 25ng/mL in 175(40.8%), between 25-37.4ng/ml in 177(41.3%) and ≥ 37.5 ng/mL in 77 (17.9%). The histological findings were no dysplasia in 340 (79.3%), low-grade dysplasia (LGD) in 52(12.1%), high grade dysplasia (HGD) in 15(3.5%) and adenocarcinoma (EAC) in 22 (5.1%). The histological findings BE in different groups of serum 25(OH)D levels are presented in Table 1. There was no evidence to suggest an association between serum 25(OH)D levels and dysplasia (p = 0.90). Conclusions: The mean serum Vitamin D levels in our Barrett's population were low. There was no significant association between degree of dysplasia and Vitamin D levels in patients with Barrett's esophagus. Larger population studies are needed in this area of research.
Tu2017 Non-Transferrin Bound Iron Transporter ZIP 14 in the Liver of Pediatric NonAlcoholic Steatohepatitis Patients Maan Khatib, Susan S. Baker, Wensheng Liu, Diana Moya, Ricardo A. Arbizu, Robert D. Baker, Michael D. Garrick, Lixin Zhu Background: Non-alcoholic steatohepatitis (NASH), the severe form of non-alcoholic fatty liver disease (NAFLD), is characterized by steatosis and inflammation. Iron may be an important factor in the pathogenesis of NASH as it catalyzes the production of potent reactive oxygen species, which could lead to liver inflammation. Non-transferrin bound iron (NTBI) uptake by ZRT/IRT-like protein (ZIP14, or SLC39A14) is an important mechanism for scavenging plasma iron during iron overload in rodents. In iron overloaded rodents, liver ZIP14 is elevated. The purpose of this study was to examine iron status and hepatic expression of iron related genes in pediatric NASH. Methods: Serum iron concentration, ferritin, and soluble transferrin receptor 1 of biopsy proven NASH patients were compared to those in the National Health and Nutrition Examination Survey database. Iron related gene expression was examined in NASH livers and normal livers, using microarray and quantitative realtime PCR. Results: Serum iron concentration and soluble transferrin receptor 1 were decreased while serum ferritin was increased in NASH patients, compared to controls. No detectable iron was observed in NASH liver by Perls' Prussian blue staining. Our microarray analysis and qRT-PCR analysis consistently demonstrated that, in NASH livers, transferrin mRNA and transferrin receptor 2 mRNA were elevated, while ZIP14 mRNA was decreased (realtime PCR, 83% decrease in NASH p=0.016). Conclusion: Pediatric NASH patients tend to be iron deficient. Increased liver gene expression of transferrin and transferrin receptor 2 supports a status of iron deficiency. Our observation of decreased ZIP14 expression in pediatric NASH patients who are iron deficient is in harmony with previous observation that ZIP14 expression in rodent livers is positively correlated to liver iron load. Our study points to a role for ZIP14 in human liver iron homeostasis. Tu2019 Role of Vitamin D in the Inflammatory Response of Morbidly Obese Patients With Non-Alcoholic Fatty Liver Disease (NAFLD) Katherine Doyle, Maria Keaton, Lei Wang, Rohini Mehta, Massih Abawi, Zahra Younoszai, Thomas Jeffers, Hazem A. Elarainy, Amir H. Moazez, Aybike Birerdinc, Ancha Baranova, Zobair M. Younossi
Values presented as N (column%) p-value corresponds to Joncheere-Terpstra test Tu2027
INTRODUCTION: Vitamin D regulates many genes related to cell proliferation, differentiation, and apoptosis and recent studies have suggested a role for Vitamin D deficiency in liver disease as well as obesity. Vitamin D receptor is expressed by macrophages and dendritic cells suggesting that this highly lipidophilic hormone also plays a role in the regulation of the body's inflammatory response which is particularly important in patients who are morbidly obese.AIM: To determine the anti-inflammatory roles of vitamin D in a cohort of morbidly obese NAFLD patients.METHODS: Serum samples obtained in conjunction with visceral adipose tissue from patients with biopsy-proven NAFLD during Bariatric surgery. Serum was assayed for Vitamin D levels using 25(OH)-Vitamin D3 assays (Alpco, Salem, NH). These samples were also run on Biorad's Bioplex pro using the 17-plex human cytokine
AGA Abstracts
Treatment with the CSF-1 Receptor kinase (CSF-1R) Inhibitor JNJ40346527 Suppresses Murine Colitis Beverley Moore, Hao Liu, Matthew Loza, Holly A. Raymond, Stanley M. Belkowski, Carl Manthey Background: The role of macrophages (Mø) in inflammatory bowel disease is as yet poorly understood. It is theorized that Mø recruited to the intestinal lamina propria differentiate into an inflammatory phenotype with myeloid DC properties (Mø/mDC), which then express high levels of TNFα and IL-23 upon activation by enteric bacteria. IL-23 expands innate lymphocyte cell populations, and bacterial antigen presentation by Mø/mDC induces proliferation of Th1 T cells, while IFN-g produced by NK and Th1 cells perpetuates the inflammatory Mø phenotype. Colony stimulating factor (CSF-1) is prominent among the several factors
S-900