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Interlukin-6 and Stem Cell Factor as Prognostic Indicators in Acute Liver Failure
between sham-operation and PHX but increased 11-fold after IR+PHX, indicating poor liver function. Daidzein increased BrdU incorporation to ~30% and prevented hyperbilirubinemia in mice subjected to IR+PHX. While all mice survived after PHX, survival decreased to ~13% after IR+PHX. Daidzein increased survival after IR+PHX to ~88%. Conclusion. Suppression of MB after IR+PHX plays an important role in inhibition of liver regeneration and function. MB is a novel therapeutic target to improve the outcome of MLR (NIDDK).
to analysis. Results: Proteomic analysis of the major insoluble liver proteins after FasL exposure for 4-5 hours identified fibrinogen-γ (FIB-γ) dimers and FIB-γ-containing high molecular mass complexes among the major insoluble proteins that are visible by Coomassie blue staining. Presence of the FIB-γ-containing products was confirmed using FIB-γ-specific antibodies. The FIB-γ-containing products partition selectively and quantitatively into the liver parenchyma after inducing apoptosis, and are also readily detected after acetaminopheninduced liver injury. The observed intrahepatic intravascular coagulation raised the possibility that heparin therapy may ameliorate the FasL-mediated liver injury. Notably, heparin subcutaneous administration into mice 4 hours before or up to 2 hours after FasL injection had a dramatic reduction of the liver injury including liver hemorrhage, serum alanine aminotransferase, caspase activation and liver apoptosis as compared to heparin-untreated mice. Heparin-treated mice survived the FasL induced liver injury longer compared to heparinuntreated mice. Time course analysis suggests that the liver injury first involves endothelial cell apoptosis followed by epithelial cell apoptosis and IC, followed by increased apoptosis possibly due to the intra-hepatic IC. Conclusions: Formation of FIB-γ dimers and their high molecular mass products are readily detectable within the liver upon IC during mouse apoptotic liver injury. Heparin provides a potential therapeutic modality since it not only prevents the extensive FasL-related liver injury but also limits the extent of injury if given at early stages of injury exposure.
Mo1001 Outcomes in Pregnancy Related Acute Liver Failure: Report of the Acute Liver Failure Study Group (Alfsg) Lisa C. Casey, Robert J. Fontana, Corron M. Sanders, William M. Lee Background: Acute Liver failure is a rare complication of pregnancy that can lead to substantial maternal and fetal morbidity and mortality. However, the presenting laboratory and clinical features associated with a favorable outcome are not well described. Aim: To determine the etiologies and presenting features associated with mortality and with spontaneous maternal survival in 30 consecutive women with ALF during pregnancy enrolled in the ALFSG registry. Methods: The US ALFSG is a 24-center network investigating the etiologies and outcomes of adult patients with ALF since 1998. Admission demographic, laboratory and clinical features as well as outcomes at 3-weeks were prospectively collected at all sites. Pregnancy related ALF was classified as HELLP, acute fatty liver of pregnancy (AFLP) and other miscellaneous etiologies. The baseline and dynamic features of spontaneous survivors were compared to those who died or underwent liver transplant (OLT) using descriptive statistics. Results: The mean age was 29.1 years, 18 were white (60%), 9 black (30%), 1 Egyptian, 1 Asian and 1 Hispanic. Identified etiologies of ALF included 25 with presumed HELLP/ AFLOP, 1 autoimmune hepatitis, 1 lymphoma, 1 drug induced liver injury, and 3 acetaminophen overdose cases. All were seen post-partum: 11 patients (37%) died or were transplanted by 3 weeks while 19 survived without grafting. The median time to OLT was 8 days (range 0-48 days) and median time to death was 6 days (range 0-10 days). Taking into account recommended weight gain with pregnancy, 72% of patients with available data (18/25) were still noted to meet BMI criteria for obesity . Although none of the baseline features were significantly associated with spontaneous survival, ethnicity, intubation and infection status trended towards significance. The need for hemodialysis during the hospital course was marginally associated with adverse outcomes (6/11 vs X/19, P=.29). Infections were common with 67% of patients overall having documented bacterial or fungal infections. Of those that died or were transplanted, 90% (10/11) had documented infections versus only 53% of those that survived without OLT (10/19). Of those that died, 71% (5/7) had documented fungal infection. Conclusions: Pregnancy-related ALF is associated with only fair outcomes, given the young population. All cases are not HELLP and AFLOP although these are the most commonly identified causes. High BMI during pregnancy may be a risk factor for ALF and/or poor outcome. Infectious complications were common and may relate to prolonged post-partum state with unresolved liver failure.
Mo1004
Background: A recent study of acetaminophen induced acute liver failure (ALF) in mice found that serum liver-specific micro RNA (miR-122) level might be a sensitive bio-marker for drug-induced liver injury. Increased levels of human genomic DNA (hgDNA) have been found in the sera of patients with liver diseases but their significance is unknown. This preliminary study was performed to quanitfy serum miR-122 and/or hgDNA in patients with acetaminophen-induced ALF (APAP-ALF), other forms of ALF and patients with chronic hepatitis C (CHC) to determine whether changes in values of either under different circumstances reflected severity of disease or prognosis. Method: Sera from 35 ALF (20 APAPALF, 15 other etiologies ALF), 37 CHC patients and 2 healthy controls were tested. The serum miR-122 levels were measured with a Taqman MicroRNA assay. The single copy gene β-actin was chosen to represent hgDNA. Serum β-actin DNA levels were measured with a Taqman Real-time PCR assay. Results: Serum miR-122 and β-actin DNA levels significantly correlated with serum ALT levels (Spearman's correlation for miR-122: r=0.45, P<0.001; for β-actin DNA: r=0.63, P<0.001). The median serum miR-122 level in ALF and in CHC patients were significantly higher than those in controls (P=0.019 and 0.018 respectively). There was no significant difference in serum miR-122 levels between ALF and CHC patients (P=0.29), despite a big disparity in ALT levels. ALF patients had significantly higher serum β-actin DNA levels compared to CHC patients (P<0.001) and controls (P= 0.027). In ALF patients, those with acetaminophen-induced ALF had significantly higher serum miR-122 and beta-actin DNA levels as compared to other etiologies (P=0.004 and 0.036, respectively); levels in non-APAP cases were still several logs higher than controls. In CHC patients, those with abnormal ALTs had higher serum miR-122 levels than those with normal ALTs (<40U/L, P=0.008). However, 14 patients with normal ALT had values of serum miR-122 comparable to non-APAP-ALF, and thus did not reflect simply necrosis but specific miR-122 secretion in response to hepatitis C; β-actin DNA levels were similar to controls. Conclusions: In several liver injury settings, miR-122 levels were at least 3 logs higher than β-actin-DNA levels, suggesting that serum miR-122 is more sensitive for detecting liver injury. MiR-122 is also highly expressed in hepatitis C patients and does not reflect the level hepatocyte injury whereas hg-DNA does. Serum miR-122 and hg-DNA levels may be useful bio-markers for liver injury but will require further study. Supported by NIH U01 DK 58369
Mo1002 Interlukin-6 and Stem Cell Factor as Prognostic Indicators in Acute Liver Failure Virendra Singh, Ashish Bhalla, Saujatya Chakraborty, Arun Sharma, Ck Nain Background: The prognosis in acute liver failure (ALF) is grave and the resource allocation at centers with liver transplant is a challenge. Various prognostic scores have been used in the past to address this problem. Many cytokines have been identified for stimulating stem cells. Interleukin 6(IL-6) and stem cell factor (SCF) are strongly associated with the liver regenerative process. We studied the prognostic role of IL-6 and SCF in patients with acute liver failure. Methods: We prospectively evaluated patients with ALF, admitted to medical high dependency unit of a tertiary care centre between July 2008 to December 2009. Twenty patients with acute viral hepatitis attending the outpatient department served as controls. Demographic data and baseline biochemical and hematological parameters were noted. Serum SCF and IL-6 levels were measured at baseline and at 48 hours. These patients were followed up till the time of death or discharge. All the patients were managed conservatively. For analysis of data the patients with ALF were grouped as survivors, non-survivors and controls. The statistical analysis was carried out using Statistical Package for Social Sciences (SPSS Inc., Chicago, IL, version 15.0 for Windows). Results: A total of 36 patients had ALF. Out of these 20 died and 16 were alive. The demographic parameters were comparable in all the three groups. There was a statistically significant difference in baseline serum levels of interleukin-6 among survivors, non-survivors and controls, however, the levels of stem cell factors did not differ. IL-6 serum level of 20.52 pg/ml, was found to have sensitivity of 90%, specificity 87.5%, positive predictive value of 90%, and a negative predictive value of 87.8% for a poorer prognosis. Conclusions: High baseline serum IL-6 value is a bad prognostic indicator in ALF patients, while SCF level is not useful in this respect. Mo1003 Fibrinogen-γ Proteolysis and Solubility Dynamics During Apoptotic Mouse Liver Injury: Heparin Prevents and Treats Liver Damage Sujith Weerasinghe, David Moons, Peter J. Altshuler, Yatrik Shah, Bishr Omary Background: Fas ligand (FasL) mediated hepatocyte apoptosis occurs in the context of acute liver injury that can also be accompanied by intravascular coagulation (IC). We tested the hypothesis that analysis of selected protein fractions from livers undergoing apoptosis will shed light on mechanisms that are involved in liver injury that may be amenable to intervention. Methods: FasL was injected intraperitoneally into age and sex-matched mice to induce apoptosis in the liver. Heparin was administered by subcutaneous injection. Mice were sacrificed by CO2 inhalation at the indicated time points, or survival time was recorded for the lethality experiments. Livers were isolated and were either stored in liquid nitrogen for biochemical analysis, or fixed in formalin for histological analysis. Blood samples were collected from the sacrificed mice by intracardiac puncture and stored overnight (4 C) prior
1=All ALF combined 2=acetaminophen-ALF 3=other etiologies-ALF 4=All CHC combined 5=CHC, ALT<40 6=CHC, ALT>40 7=healthy controls
S-951
AASLD Abstracts
AASLD Abstracts
Serum miR-122 and Human Genomic DNA in Patients With Drug-Induced Acute Liver Failure or With Chronic Hepatitis C R Kenton Devine, Hejun Yuan, Mamta K. Jain, Corron M. Sanders, Nahid Attar, William M. Lee
to analysis. Results: Proteomic analysis of the major insoluble liver proteins after FasL exposure for 4-5 hours identified fibrinogen-γ (FIB-γ) dimers and FIB-γ-containing high molecular mass complexes among the major insoluble proteins that are visible by Coomassie blue staining. Presence of the FIB-γ-containing products was confirmed using FIB-γ-specific antibodies. The FIB-γ-containing products partition selectively and quantitatively into the liver parenchyma after inducing apoptosis, and are also readily detected after acetaminopheninduced liver injury. The observed intrahepatic intravascular coagulation raised the possibility that heparin therapy may ameliorate the FasL-mediated liver injury. Notably, heparin subcutaneous administration into mice 4 hours before or up to 2 hours after FasL injection had a dramatic reduction of the liver injury including liver hemorrhage, serum alanine aminotransferase, caspase activation and liver apoptosis as compared to heparin-untreated mice. Heparin-treated mice survived the FasL induced liver injury longer compared to heparinuntreated mice. Time course analysis suggests that the liver injury first involves endothelial cell apoptosis followed by epithelial cell apoptosis and IC, followed by increased apoptosis possibly due to the intra-hepatic IC. Conclusions: Formation of FIB-γ dimers and their high molecular mass products are readily detectable within the liver upon IC during mouse apoptotic liver injury. Heparin provides a potential therapeutic modality since it not only prevents the extensive FasL-related liver injury but also limits the extent of injury if given at early stages of injury exposure.
Mo1001 Outcomes in Pregnancy Related Acute Liver Failure: Report of the Acute Liver Failure Study Group (Alfsg) Lisa C. Casey, Robert J. Fontana, Corron M. Sanders, William M. Lee Background: Acute Liver failure is a rare complication of pregnancy that can lead to substantial maternal and fetal morbidity and mortality. However, the presenting laboratory and clinical features associated with a favorable outcome are not well described. Aim: To determine the etiologies and presenting features associated with mortality and with spontaneous maternal survival in 30 consecutive women with ALF during pregnancy enrolled in the ALFSG registry. Methods: The US ALFSG is a 24-center network investigating the etiologies and outcomes of adult patients with ALF since 1998. Admission demographic, laboratory and clinical features as well as outcomes at 3-weeks were prospectively collected at all sites. Pregnancy related ALF was classified as HELLP, acute fatty liver of pregnancy (AFLP) and other miscellaneous etiologies. The baseline and dynamic features of spontaneous survivors were compared to those who died or underwent liver transplant (OLT) using descriptive statistics. Results: The mean age was 29.1 years, 18 were white (60%), 9 black (30%), 1 Egyptian, 1 Asian and 1 Hispanic. Identified etiologies of ALF included 25 with presumed HELLP/ AFLOP, 1 autoimmune hepatitis, 1 lymphoma, 1 drug induced liver injury, and 3 acetaminophen overdose cases. All were seen post-partum: 11 patients (37%) died or were transplanted by 3 weeks while 19 survived without grafting. The median time to OLT was 8 days (range 0-48 days) and median time to death was 6 days (range 0-10 days). Taking into account recommended weight gain with pregnancy, 72% of patients with available data (18/25) were still noted to meet BMI criteria for obesity . Although none of the baseline features were significantly associated with spontaneous survival, ethnicity, intubation and infection status trended towards significance. The need for hemodialysis during the hospital course was marginally associated with adverse outcomes (6/11 vs X/19, P=.29). Infections were common with 67% of patients overall having documented bacterial or fungal infections. Of those that died or were transplanted, 90% (10/11) had documented infections versus only 53% of those that survived without OLT (10/19). Of those that died, 71% (5/7) had documented fungal infection. Conclusions: Pregnancy-related ALF is associated with only fair outcomes, given the young population. All cases are not HELLP and AFLOP although these are the most commonly identified causes. High BMI during pregnancy may be a risk factor for ALF and/or poor outcome. Infectious complications were common and may relate to prolonged post-partum state with unresolved liver failure.
Mo1004
Background: A recent study of acetaminophen induced acute liver failure (ALF) in mice found that serum liver-specific micro RNA (miR-122) level might be a sensitive bio-marker for drug-induced liver injury. Increased levels of human genomic DNA (hgDNA) have been found in the sera of patients with liver diseases but their significance is unknown. This preliminary study was performed to quanitfy serum miR-122 and/or hgDNA in patients with acetaminophen-induced ALF (APAP-ALF), other forms of ALF and patients with chronic hepatitis C (CHC) to determine whether changes in values of either under different circumstances reflected severity of disease or prognosis. Method: Sera from 35 ALF (20 APAPALF, 15 other etiologies ALF), 37 CHC patients and 2 healthy controls were tested. The serum miR-122 levels were measured with a Taqman MicroRNA assay. The single copy gene β-actin was chosen to represent hgDNA. Serum β-actin DNA levels were measured with a Taqman Real-time PCR assay. Results: Serum miR-122 and β-actin DNA levels significantly correlated with serum ALT levels (Spearman's correlation for miR-122: r=0.45, P<0.001; for β-actin DNA: r=0.63, P<0.001). The median serum miR-122 level in ALF and in CHC patients were significantly higher than those in controls (P=0.019 and 0.018 respectively). There was no significant difference in serum miR-122 levels between ALF and CHC patients (P=0.29), despite a big disparity in ALT levels. ALF patients had significantly higher serum β-actin DNA levels compared to CHC patients (P<0.001) and controls (P= 0.027). In ALF patients, those with acetaminophen-induced ALF had significantly higher serum miR-122 and beta-actin DNA levels as compared to other etiologies (P=0.004 and 0.036, respectively); levels in non-APAP cases were still several logs higher than controls. In CHC patients, those with abnormal ALTs had higher serum miR-122 levels than those with normal ALTs (<40U/L, P=0.008). However, 14 patients with normal ALT had values of serum miR-122 comparable to non-APAP-ALF, and thus did not reflect simply necrosis but specific miR-122 secretion in response to hepatitis C; β-actin DNA levels were similar to controls. Conclusions: In several liver injury settings, miR-122 levels were at least 3 logs higher than β-actin-DNA levels, suggesting that serum miR-122 is more sensitive for detecting liver injury. MiR-122 is also highly expressed in hepatitis C patients and does not reflect the level hepatocyte injury whereas hg-DNA does. Serum miR-122 and hg-DNA levels may be useful bio-markers for liver injury but will require further study. Supported by NIH U01 DK 58369
Mo1002 Interlukin-6 and Stem Cell Factor as Prognostic Indicators in Acute Liver Failure Virendra Singh, Ashish Bhalla, Saujatya Chakraborty, Arun Sharma, Ck Nain Background: The prognosis in acute liver failure (ALF) is grave and the resource allocation at centers with liver transplant is a challenge. Various prognostic scores have been used in the past to address this problem. Many cytokines have been identified for stimulating stem cells. Interleukin 6(IL-6) and stem cell factor (SCF) are strongly associated with the liver regenerative process. We studied the prognostic role of IL-6 and SCF in patients with acute liver failure. Methods: We prospectively evaluated patients with ALF, admitted to medical high dependency unit of a tertiary care centre between July 2008 to December 2009. Twenty patients with acute viral hepatitis attending the outpatient department served as controls. Demographic data and baseline biochemical and hematological parameters were noted. Serum SCF and IL-6 levels were measured at baseline and at 48 hours. These patients were followed up till the time of death or discharge. All the patients were managed conservatively. For analysis of data the patients with ALF were grouped as survivors, non-survivors and controls. The statistical analysis was carried out using Statistical Package for Social Sciences (SPSS Inc., Chicago, IL, version 15.0 for Windows). Results: A total of 36 patients had ALF. Out of these 20 died and 16 were alive. The demographic parameters were comparable in all the three groups. There was a statistically significant difference in baseline serum levels of interleukin-6 among survivors, non-survivors and controls, however, the levels of stem cell factors did not differ. IL-6 serum level of 20.52 pg/ml, was found to have sensitivity of 90%, specificity 87.5%, positive predictive value of 90%, and a negative predictive value of 87.8% for a poorer prognosis. Conclusions: High baseline serum IL-6 value is a bad prognostic indicator in ALF patients, while SCF level is not useful in this respect. Mo1003 Fibrinogen-γ Proteolysis and Solubility Dynamics During Apoptotic Mouse Liver Injury: Heparin Prevents and Treats Liver Damage Sujith Weerasinghe, David Moons, Peter J. Altshuler, Yatrik Shah, Bishr Omary Background: Fas ligand (FasL) mediated hepatocyte apoptosis occurs in the context of acute liver injury that can also be accompanied by intravascular coagulation (IC). We tested the hypothesis that analysis of selected protein fractions from livers undergoing apoptosis will shed light on mechanisms that are involved in liver injury that may be amenable to intervention. Methods: FasL was injected intraperitoneally into age and sex-matched mice to induce apoptosis in the liver. Heparin was administered by subcutaneous injection. Mice were sacrificed by CO2 inhalation at the indicated time points, or survival time was recorded for the lethality experiments. Livers were isolated and were either stored in liquid nitrogen for biochemical analysis, or fixed in formalin for histological analysis. Blood samples were collected from the sacrificed mice by intracardiac puncture and stored overnight (4 C) prior
1=All ALF combined 2=acetaminophen-ALF 3=other etiologies-ALF 4=All CHC combined 5=CHC, ALT<40 6=CHC, ALT>40 7=healthy controls
S-951
AASLD Abstracts
AASLD Abstracts
Serum miR-122 and Human Genomic DNA in Patients With Drug-Induced Acute Liver Failure or With Chronic Hepatitis C R Kenton Devine, Hejun Yuan, Mamta K. Jain, Corron M. Sanders, Nahid Attar, William M. Lee